Novel compounds containing biguanidyl radicals and uses thereof

Abstract

To provide novel compounds comprising biguanidyl radicals which are more potent than metformin in the treatment of cancer.SOLUTION: The present invention relates to novel compounds of formula (I), and their uses.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] Funding The project leading to this application has received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (grant agreement no. [647973]).

[0002] The present invention relates to the field of medicine, in particular to the field of oncology.The present invention relates to novel compounds containing biguanidyl radicals. [Background technology]

[0003] Various derivatives of biguanides are used as medicines. Most of them are used as antihyperglycemic agents, especially for the treatment of diabetes mellitus and prediabetes. The most widely used drug is metformin.

[0004] [ka]

[0005] Biguanide derivatives are also used as antimalarials, such as proguanil and chlorproguanil, and as disinfectants, such as chlorhexidine, polyaminopropyl biguanine, polyhaxanide, and alexidine.

[0006] More recently, metformin and its derivatives have been described for their use in the treatment of cancer (Safe et al., 2018, Biol Chem, 399, pp. 321-335; WO2017 / 192602). More specifically, patent applications disclose derivatives of metformin (EP2522653, EP3222614, WO2013 / 022279, WO2014 / 123364, WO2015 / 160220, WO2016 / 025725, and WO2016 / 155679). [Prior art documents]

Patent Documents

[0007]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Patent Document 5

Patent Document 6

Patent Document 7

Patent Document 8

Non-Patent Documents

[0008]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Non-Patent Document 5

Summary of the Invention

[0009] However, new drugs are constantly needed, especially for the treatment of cancer. [Means for solving the problem]

[0010] The present inventors provide a novel compound that contains biguanidyl radical, which is more potent than metformin in the treatment of cancer.In addition, the present inventors have confirmed that the compound of the present invention targets mitochondrial copper and can selectively target a population of cancer cells with mesenchymal characteristics.

[0011] Thus, the present invention relates to a compound of formula (I)

[0012] [ka]

[0013] {In the formula, R1 is C1-C6 alkyl, -(CH2) a -C≡CH, -(CH2) b -P + Ph3, C3~C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12 heteroaryl, wherein "a" is an integer selected from 1 to 6, preferably 2 to 4, and "b" is an integer selected from 5 to 11, preferably 6 to 10, wherein said alkyl, cycloalkyl, cycloheteroalkyl, aryl, or heteroaryl is optionally substituted with an R or R' group; R2 is - -(CH2) d -C≡CH, where "d" is an integer selected from 1 to 6; and - -(CH2) e -CH=CH2 (wherein "e" is an integer selected from 2 to 6) selected from the group consisting of; Or, R1 is H, C1-C6 alkyl, -(CH2) a -C≡CH, -(CH2) b -P + Ph3, C3~C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12 heteroaryl, wherein "a" is an integer selected from 1 to 6, preferably 2 to 4, and "b" is an integer selected from 5 to 11, preferably 6 to 10, wherein said alkyl, cycloalkyl, cycloheteroalkyl, aryl, or heteroaryl is optionally substituted with an R or R' group; R2 is - -(CH2) f -CR4=CH-CH=CR5-(CH2) g -R3, where R4 and R5 are H, an R group, or together form a 6-membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3 to 6 members, and "f" and "g" are integers independently selected from 1 to 3; - -(CH2) f -C≡CC≡C-(CH2) g -R3, where "f" and "g" are integers independently selected from 1 to 3; - -(CH2) i -R3, where "i" is an integer selected from 7 to 9, preferably 8; - -(CH2) j -CH[(CH2) k -P + Ph3]-(CH2) l -R3, where "j" and "l" are integers independently selected from 1 to 6, and "k" is an integer selected from 1 to 6; - -(CH2) m -Cyclobutanyl-(CH2)n -R3, where "m" and "n" are integers independently selected from 1 to 3; and - -(CH2) p -CHR6-CH=CH-CHR7-(CH2) q -R3, where R6 and R7 are H, R groups, or together form a 4-6 membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3-6 members, and "p" and "q" are integers independently selected from 1 to 3. selected from the group consisting of; R3 is

[0014] [ka]

[0015] [Wherein, R8 is H, C1-C6 alkyl, -(CH2) a -C≡CH, -(CH2) b -P + Ph3, C3~C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12 heteroaryl, wherein "a" is an integer selected from 1 to 6, preferably 2 to 4, and "b" is an integer selected from 5 to 11, preferably 6 to 10, and wherein said alkyl, cycloalkyl, cycloheteroalkyl, aryl, or heteroaryl is optionally substituted with an R or R' group. and; R is C1-C6 alkyl, C1-C6 alkyloxy, C1-C6 halogenoalkyl, C3-C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12heteroaryl, wherein the group is optionally substituted with an R′ group; R' is C1-C6 alkyl, C1-C6 alkyloxy, C1-C6 thioalkyl, halogen, C1-C6 halogenoalkyl, hydroxyl (-OH), cyano (-CN), nitro, amino (-NH2), carboxyl (-COOH), phosphate (PO4 - ), amide (-CONH2), -COOR'', -NHR'', -NR''R''', -COR'', -CONHR'', -NHCOR'', -NHSO2R'', -SOR'', -SO2R'', -SONR''R'''', -SO2NR''R''', C3~C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12 and heteroaryl, wherein said cycloalkyl, cycloheteroalkyl, aryl or heteroaryl is selected from the group consisting of halogen, hydroxyl, cyano, nitro, amino, or C 1~ and optionally substituted with C3 alkoxy, R'' and R'' are H or C1-C6 alkyl. or a pharma- ceutically acceptable salt thereof.

[0016] Preferably, R3 is present in the compound.

[0017] Optionally, R2 is - -(CH2) d -C≡CH, where "d" is an integer selected from 2 to 4; - -(CH2) e -CH=CH2 (wherein "e" is an integer selected from 2 to 4); - -(CH2) f -CR4=CH-CH=CR5-(CH2) g-R3, where R4 and R5 are H, an R group, or together form a 6-membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3 to 6 members, and "f" and "g" are integers independently selected from 1 to 3, and "f" + "g" is 2 to 4; - -(CH2) f -C≡CC≡C-(CH2) g -R3 (wherein "f" and "g" are integers independently selected from 1 to 3, and "f" + "g" is 2 to 4); - -(CH2) i - R3, where "i" is an integer selected from 7 to 9; - -(CH2) j -CH[(CH2) k -P + Ph3]-(CH2) l -R3, where "j" and "l" are integers independently selected from 1 to 6, "j" + "l" is 5 to 7, and "k" is an integer selected from 2 to 4; - -(CH2) m -Cyclobutanyl-(CH2) n -R3, where "m" and "n" are integers independently selected from 1 to 3, and "m" + "n" is 2 to 4; and - -(CH2) p -CHR6-CH=CH-CHR7-(CH2) q -R3, where R6 and R7 are H, R groups, or together form a 4-6 membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3-6 members, and "p" and "q" are integers independently selected from 1 to 3, and "p" + "q" is 2 to 4. is selected from the group consisting of:

[0018] Optionally, R1 is methyl, -(CH2) a -C≡CH and -(CH2)b -P + Ph3 (wherein "a" is an integer selected from 2 to 4 and "b" is an integer selected from 6 to 10); R2 is - -(CH2) d -C≡CH, where "d" is an integer selected from 2 to 4; and - -(CH2) e -CH=CH2 (wherein "e" is an integer selected from 2 to 6) is selected from the group consisting of:

[0019] Optionally, R1 is H, methyl, -(CH2) a -C≡CH and -(CH2) b -P + Ph3 (wherein "a" is an integer selected from 2 to 4 and "b" is an integer selected from 6 to 10); R2 is - -(CH2) f -CR4=CH-CH=CR5-(CH2) g -R3 (wherein R4 and R5 are H, "f" and "g" are integers independently selected from 1 to 3, and "f" + "g" is 2 to 4); - -(CH2) f -C≡CC≡C-(CH2) g -R3 (wherein "f" and "g" are integers independently selected from 1 to 3, and "f" + "g" is 2 to 4); - -(CH2) i - R3, where "i" is an integer selected from 7 to 9; - -(CH2) j -CH[(CH2) k -P + Ph3]-(CH2) l -R3, where "j" and "l" are integers independently selected from 1 to 6, "j" + "l" is 5 to 7, and "k" is an integer selected from 1 to 6; - -(CH2) m -Cyclobutanyl-(CH2)n -R3, where "m" and "n" are integers independently selected from 1 to 3, and "m" + "n" is 2 to 4; and - -(CH2) p -CHR6-CH=CH-CHR7-(CH2) q -R3, wherein R6 and R7 are H or together form a 4-membered ring, "p" and "q" are integers independently selected from 1 to 3, and "p" + "q" is 2 to 4. selected from the group consisting of; R3 is

[0020] [ka]

[0021] [Wherein, R8 is H, methyl, -(CH2) a -C≡CH and -(CH2) b -P + Ph3 (wherein "a" is an integer selected from 1 to 6, preferably 2 to 4, and "b" is an integer selected from 5 to 11, preferably 6 to 10). It is.

[0022] Optionally, at least one of R1, R2, and R8 is -(CH2) a -C≡CH, where "a" is an integer from 1 to 6.

[0023] Optionally, the compound is

[0024] [ka]

[0025] (wherein n is an integer selected from 3 to 9);

[0026] [ka]

[0027] (wherein n is an integer independently selected from 0 to 5);

[0028] [ka]

[0029] (wherein n is an integer independently selected from 0 to 5); [In the formula, A dotted line may be present or absent, and represents one or two atoms with a covalent bond; R is C1-C6 alkyl, C1-C6 alkyloxy, C1-C6 halogenoalkyl, C3-C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12 heteroaryl, the group being optionally substituted with an R' group; R' being C1-C6 alkyl, C1-C6 alkyloxy, C1-C6 thioalkyl, halogen, C1-C6 halogenoalkyl, hydroxyl (-OH), cyano (-CN), nitro, amino (-NH2), carboxyl (-COOH), phosphate (PO4 - ), amide (-CONH2), -COOR'', -NHR'', -NR''R''', -COR'', -CONHR'', -NHCOR'', -NHSO2R'', -SOR'', -SO2R'', -SONR''R'''', -SO2NR''R''', C3~C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12 and heteroaryl, wherein said cycloalkyl, cycloheteroalkyl, aryl or heteroaryl is selected from the group consisting of halogen, hydroxyl, cyano, nitro, amino, or C 1~ and optionally substituted with C3 alkoxy, where R'' and R'' are H or C1-C6 alkyl. or a pharma- ceutically acceptable salt thereof.

[0030] Optionally, the compound is

[0031] [ka]

[0032] (wherein n is an integer selected from 3 to 9); and

[0033] [ka]

[0034] (wherein n is an integer independently selected from 0 to 5); or a pharma- ceutically acceptable salt thereof.

[0035] The present invention also relates to a conjugate comprising a compound disclosed herein covalently bound to a drug, toxin, or label. More specifically, the present invention relates to a conjugate obtained by reacting a compound according to the present disclosure having an alkyne group with a drug, toxin, or label covalently bound to an azide (N3) group.

[0036] The present invention further relates to a pharmaceutical or veterinary composition comprising a compound or conjugate disclosed herein. Optionally, the pharmaceutical or veterinary composition further comprises another drug, preferably an anti-tumor drug, more preferably a drug selected from the group consisting of a chemotherapeutic drug, an anti-cancer antibody, a hormonal therapy drug, an immunotherapy drug, a kinase inhibitor drug, and combinations thereof.

[0037] The present invention relates to a compound, conjugate or pharmaceutical composition as disclosed herein for use as a medicament for treating cancer, in particular cancer selected from the group consisting of rectal cancer, colon cancer, gastric cancer, head and neck cancer, thyroid cancer, cervical cancer, uterine cancer, breast cancer, in particular triple negative breast cancer, ovarian cancer, brain tumors, in particular glioblastoma and neuroblastoma, lung cancer, in particular small cell lung cancer and non-small cell lung cancer, skin cancer, bladder cancer, blood cancer, kidney cancer, liver cancer, prostate cancer, multiple myeloma, and endometrial cancer. Optionally, the compound, conjugate or pharmaceutical composition is used in combination with radiation therapy and / or another drug, preferably an antitumor drug, more preferably a drug selected from the group consisting of chemotherapy drugs, hormonal therapy drugs, and immunotherapy drugs.

[0038] Finally, the present invention relates to the use of compounds containing an alkyne group, as disclosed herein, for medical imaging or diagnosis. [Brief description of the drawings]

[0039] [Figure 1] FIG. 1A: Metformin and the more potent compound metforminyn preferentially alter the proliferation of cancer cells in the mesenchymal state. (FIG. 1A) Molecular structures of metformin, phenformin and metformin synthesis. (FIG. 1B) Dose-response viability curves of cancer cells treated for 72 hours as indicated. (FIG. 1C) Dose-response viability curves of epithelial and mesenchymal state cells of MDA-MB-468 cells treated for 72 hours as indicated. (FIG. 1D) Dose-response viability curves of epithelial and mesenchymal state HMLER cells treated for 72 hours as indicated. Data points and error bars, mean and standard deviation of three independent biological replicates. [Diagram 2]Figure 2 shows that biguanides directly target mitochondria and promote copper oxidation. (Figure 2A) Schematic of the labeling of metformin in cells using click chemistry. (Figure 2B) Fluorescence microscopy image of labeled metformin. Cells were treated with metformin and click labeling was performed as described in Methods. Mitochondria were detected using cytochrome c immunostaining and nuclear DNA stained with 4',6-diamidino-2-phenylindole (DAPI). Scale bar is 10 μm. (Figure 2C) Fluorescence microscopy detection of copper(II) in cancer cells treated for 48 h as indicated. Scale bar is 10 μm. (Figure 2D) Flow cytometry analysis of copper(II) in cancer cells treated for 48 h as indicated. (Figure 2E) Cyclic voltammetry measurements towards the oxidation potential (arrow) of copper(I) solution. Data recorded in the absence and presence of metformin (2 mol equiv). Redox peak potential is indicated by dashed line. (FIG. 2F) Flow cytometry analysis of iron(II) in cancer cells treated for 48 h as indicated. MDA-MB-468 cells were used in FIG. 2C, D, and D(F) and treated as described in Methods. [Figure 3A] Figure 1 shows fluorescence microscopy imaging of labeled Met. The indicated cell lines were treated with metformin and click labeling was performed. Metformin was labeled intracellularly as described with the addition of a copper catalyst. Mitochondria were detected using cytochrome c immunostaining or MitoTracker and nuclear DNA stained with DAPI. Scale bar is 10 μm. The term no Cu indicates that no copper catalyst was added to the cells to facilitate labeling. [Figure 3B] Figure 1 shows fluorescence microscopy imaging of labeled Met. The indicated cell lines were treated with metformin and click labeling was performed. Metformin was labeled intracellularly as described, without the addition of a copper catalyst. Mitochondria were detected using cytochrome c immunostaining or MitoTracker, and nuclear DNA was stained with DAPI. Scale bar is 10 μm. The term no Cu indicates that no copper catalyst was added to the cells to facilitate labeling. [Figure 4A] Western blot analysis of Ctr1 levels.MDA-MB-468 cells were treated as indicated for 72 hours. [Figure 4B] Flow cytometry analysis of Ctr1 levels.MDA-MB-468 cells were treated as indicated for 72 hours. [Diagram 5] Figure 5: Copper is required to maintain the mesenchymal state of cancer cells. (Figure 5A) Western blot analysis of mesenchymal marker (vimentin), Ctr1, and copper-containing mitochondrial proteins (SOD1, Cox4) levels in cells treated with EGF for 72 h. (Figure 5B) Flow cytometry analysis of Ctr1 protein levels in cells treated with EGF for 72 h. (Figure 5C) Flow cytometry analysis of copper(II) in cells treated with EGF for 72 h. (Figure 5D) Western blot analysis of mesenchymal markers (vimentin, fibronectin) in cells treated as indicated for 72 h. (Figure 5E) Fluorescence microscopy images of E-cadherin and phalloidin in cells treated as indicated for 72 h. DAPI stains nuclear DNA. Scale bar is 10 μm. (Figure 5F) Western blot analysis of mesenchymal markers and EMT-TFs in cancer cells treated as indicated for 72 h. (FIG. 5G) Flow cytometry analysis and corresponding quantification of cell surface markers of cancer cells treated for 72 h as indicated. Bars and error bars, mean and standard deviation of three independent biological replicates. MDA-MB-468 cells were used throughout the figure. [Figure 6] Figure 1 Flow cytometry analysis of CD44 high / CD24 low cells. MDA-MB-468 cells were treated with EGF and CuCl2 for 72 hours as indicated. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0040] definition For example, when the term C1-C3 is used, it means that the corresponding hydrocarbon chain may contain 1 to 3 carbon atoms, particularly 1, 2 or 3 carbon atoms. For example, when the term C1-C6 is used, it means that the corresponding hydrocarbon chain may contain 1 to 6 carbon atoms, particularly 1, 2, 3, 4, 5 or 6 carbon atoms.

[0041] The term "alkyl" refers to a saturated, straight or branched chain aliphatic group. The term "(C1-C3)alkyl" more specifically means methyl, ethyl, propyl, or isopropyl. The term "(C1-C6)alkyl" more specifically means methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, or hexyl. In a preferred embodiment, "alkyl" is methyl, ethyl, propyl, isopropyl, or tert-butyl, more preferably methyl.

[0042] The term "alkoxy" or "alkyloxy" corresponds to an alkyl group as defined above attached to the molecule by an -O- (ether) bond. (C1-C3)alkoxy includes methoxy, ethoxy, propyloxy, and isopropyloxy. (C1-C6)alkoxy includes methoxy, ethoxy, propyloxy, isopropyloxy, butyloxy, isobutyloxy, tert-butyloxy, pentyloxy, and hexyloxy. In a preferred embodiment, the "alkoxy" or "alkyloxy" is methoxy.

[0043] The term "thioalkyl" corresponds to an alkyl group as defined above bonded to the molecule by an -S- (thioether) bond. Thio-(C1-C6)alkyl groups include thio-methyl, thio-ethyl, thio-propyl, thio-butyl, thio-pentyl and thio-hexyl.

[0044] The term "thioalkyl" corresponds to an alkyl group as defined above bonded to the molecule by an -S- (thioether) bond. Thio-(C1-C6)alkyl groups include thio-methyl, thio-ethyl, thio-propyl, thio-butyl, thio-pentyl and thio-hexyl.

[0045] The term "heterocycloalkyl" refers to saturated or unsaturated cycloalkyl groups as defined above further containing at least one heteroatom, such as a nitrogen, oxygen, or sulfur atom. Heterocycloalkyl also includes fused, bridged, or spiro-linked heterocycloalkyl groups. Representative heterocycloalkyl groups include, but are not limited to, 3-dioxolane, benzo[1,3]dioxolyl, pyrazolinyl, pyranyl, thiomorpholinyl, pyrazolidinyl, piperidyl, piperazinyl, 1,4-dioxanyl, imidazolinyl, pyrrolinyl, pyrrolidinyl, piperidinyl, imidazolidinyl, morpholinyl, 1,4-dithianyl, pyrrolidinyl, oxozolinyl, oxazolidinyl, isoxazolinyl, isoxazolidinyl, thiazolinyl, thiazolidinyl, isothiazolinyl, isothiazolidinyl, dihydropyranyl, tetrahydro-2H-pyranyl, tetrahydrofuranyl, and tetrahydrothiophenyl. The term "heterocycloalkyl" may also refer to 5-10 membered bridged heterocyclyls, such as 7-oxabicyclo[2,2,1]heptanyl. In preferred embodiments, the heterocycloalkyl group is tetrahydro-2H-pyranyl, tetrahydro-2H-pyranyl, tetrahydrothiophenyl, morpholinyl, or piperazinyl.

[0046] The term "aryl" refers to a monocyclic or bicyclic aromatic hydrocarbon having 6 to 12 carbon atoms. For example, the term "aryl" includes phenyl, biphenyl, or naphthyl. In a preferred embodiment, aryl is phenyl.

[0047] The term "heteroaryl" as used herein refers to an aromatic, monocyclic or polycyclic group containing between 5 and 14 atoms and containing at least one heteroatom such as a nitrogen, oxygen or sulfur atom. Examples of such monocyclic and polycyclic heteroaryl groups are pyridinyl, thiazolyl, thiophenyl, furan, pyrrolyl, pyrazole, imidazolyl, triazolyl, tetrazolyl, benzofuran, pyridinyl, thiazolyl, thiophenyl, furanyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, benzofuranyl, thianaphthalenyl, indolyl, indolinyl, quinolinyl, isoquinolinyl, benzimidazolyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, triazinyl, thianthrenyl, isobenzofuranyl, chromenyl, xanthenyl, phenoxanthinyl, isothiazolyl, isoxazolyl, pyrazinyl, pyridazinyl, indolizinyl, isoindoline, cyclohexane ... The aryl group may be aryl, indazolyl, purinyl, quinolidinyl, phthalazinyl, naphthyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl, pteridinyl, carbazolyl, β-carbolinyl, phenanthridinyl, acridinyl, pyrimidinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, furazanyl, phenoxazinyl, isochromanyl, chromanyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, indolinyl, isoindolinyl, oxazolidinyl, benzotriazolyl, benzisoxazolyl, oxindolyl, benzoxazolinyl, benzothienyl, benzothiazolyl, isathinyl, dihydropyridyl, pyrimidinyl, s-triazinyl, oxazolyl, or thiofuranyl. In preferred embodiments, the heteroaryl group is thiophenyl, pyridinyl, pyrazinyl, or thiazolyl.

[0048] The term "halogen" corresponds to a fluorine, chlorine, bromine or iodine atom, preferably fluorine, chlorine or bromine, more preferably chlorine or fluorine.

[0049] In the context of the present invention, the term treatment means curative, symptomatic and preventive treatment. The pharmaceutical composition, kit, product and combined preparation of the present invention can be used in humans with existing cancer or tumor, including early or late stage of cancer progression. The pharmaceutical composition, kit, product and combined preparation of the present invention do not necessarily cure the patient with cancer, but delay or slow the progression or prevent further progression of the disease, thereby improving the patient's condition. In particular, the pharmaceutical composition, kit, product and combined preparation of the present invention reduce the occurrence of tumors, reduce tumor burden, cause tumor regression and / or prevent the occurrence of metastasis and recurrence of cancer in a mammalian host. In the treatment of cancer, the pharmaceutical composition of the present invention is administered in a therapeutically effective amount.

[0050] The term "cancer" or "tumor" as used herein refers to the presence of cells that have characteristics typical of cancer-causing cells, such as uncontrolled proliferation, and / or immortality, and / or metastatic potential, and / or rapid growth, and / or proliferation rate, and / or certain characteristic morphological traits. The term refers to any type of malignant tumor (primary or metastatic) in any type of subject. In particular, the term encompasses prostate cancer at any stage of progression.

[0051] Throughout this specification, whenever "treatment of cancer" or the like is referred to in relation to the pharmaceutical composition of the invention, it means: a) a method for treating cancer, which method comprises administering the pharmaceutical composition of the invention to a subject in need of such treatment; b) the use of the pharmaceutical composition of the invention for the treatment of cancer; c) the use of the pharmaceutical composition of the invention for the manufacture of a medicament for the treatment of cancer; and / or d) the pharmaceutical composition of the invention for use in the treatment of cancer.

[0052] "Therapeutically effective amount" means an amount of the pharmaceutical composition of the present invention that prevents, eliminates or reduces the harmful effects of cancer in a mammal, including a human, either alone or in combination with other active ingredients of a pharmaceutical composition, kit, product or combination preparation. It is understood that the dosage may be less than the "therapeutically effective amount" defined for each compound in the composition, either alone or in combination with other treatments other than the combination described herein. The "therapeutically effective amount" of the composition will be adapted by those skilled in the art depending on the patient, the condition, the mode of administration, etc.

[0053] As used herein, the term "antitumor chemotherapy" or "chemotherapy" refers to a therapeutic treatment of cancer using chemical or biochemical agents, in particular using one or more anti-cancer drugs. The term "hormonal therapy" refers to a cancer treatment aimed at blocking, adding, or removing hormones. For example, in breast cancer, the female hormones estrogen and progesterone may promote the growth of some breast cancer cells. The term "immunotherapy" refers to a therapeutic treatment of cancer that uses the immune system to reject cancer. The therapeutic treatment stimulates the patient's immune system to attack malignant tumor cells.

[0054] compound The present application relates to a compound of formula (I)

[0055] [ka]

[0056] {In the formula, R1 is H, C1-C6 alkyl, preferably C1-C3 alkyl, -(CH2) a -C≡CH, -(CH2) b -P + Ph3, C3~C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12heteroaryl, wherein "a" is an integer selected from 1 to 6, preferably 2 to 4, and "b" is an integer selected from 5 to 11, preferably 6 to 10, wherein said alkyl, cycloalkyl, cycloheteroalkyl, aryl, or heteroaryl is optionally substituted with an R or R' group; R2 is - -(CH2) d -C≡CH, where "d" is an integer selected from 1 to 6; - -(CH2) e -CH=CH2, where "e" is an integer selected from 2 to 6; - -(CH2) f -CR4=CH-CH=CR5-(CH2) g -R3, where R4 and R5 are H, an R group, or together form a 6-membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3 to 6 members, and "f" and "g" are integers independently selected from 1 to 3; - -(CH2) f -C≡CC≡C-(CH2) g -R3, where "f" and "g" are integers independently selected from 1 to 3; - -(CH2) i -R3, where "i" is an integer selected from 7 to 9, preferably 8; - -(CH2) j -CH[(CH2) k -P + Ph3]-(CH2) l -R3, where "j" and "l" are integers independently selected from 1 to 6, and "k" is an integer selected from 1 to 6; - -(CH2) m -Cyclobutanyl-(CH2) n -R3, where "m" and "n" are integers independently selected from 1 to 3; and - -(CH2) p -CHR6-CH=CH-CHR7-(CH2)q -R3, where R6 and R7 are H, R groups, or together form a 4-6 membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3-6 members, and "p" and "q" are integers independently selected from 1 to 3. selected from the group consisting of; R3 is

[0057] [ka]

[0058] [In the formula, R8 is H, C1 to C6 alkyl, preferably C1 to C3 alkyl, -(CH2) a -C≡CH, -(CH2) b -P + Ph3, C3~C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12 heteroaryl, wherein "a" is an integer selected from 1 to 6, preferably 2 to 4, and "b" is an integer selected from 5 to 11, preferably 6 to 10, and wherein said alkyl, cycloalkyl, cycloheteroalkyl, aryl, or heteroaryl is optionally substituted with an R or R' group. and; R is C1-C6 alkyl, C1-C6 alkyloxy, C1-C6 halogenoalkyl, C3-C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12 heteroaryl, wherein the group is optionally substituted with an R′ group; R' is C1-C6 alkyl, C1-C6 alkyloxy, C1-C6 thioalkyl, halogen, C1-C6 halogenoalkyl, hydroxyl (-OH), cyano (-CN), nitro, amino (-NH2), carboxyl (-COOH), phosphate (PO4 - ), amide (-CONH2), -COOR'', -NHR'', -NR''R''', -COR'', -CONHR'', -NHCOR'', -NHSO2R'', -SOR'', -SO2R'', -SONR''R'''', -SO2NR''R''', C3~C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12 and heteroaryl, wherein said cycloalkyl, cycloheteroalkyl, aryl or heteroaryl is selected from the group consisting of halogen, hydroxyl, cyano, nitro, amino, or C 1~ and optionally substituted with C3 alkoxy, R'' and R'' are H or C1-C6 alkyl. or a pharma- ceutically acceptable salt thereof.

[0059] More specifically, the present invention relates to a compound of formula (I):

[0060] [ka]

[0061] {In the formula, R1 is C1-C6 alkyl, -(CH2) a -C≡CH, -(CH2) b -P + Ph3, C3~C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12heteroaryl, wherein "a" is an integer selected from 1 to 6, preferably 2 to 4, and "b" is an integer selected from 5 to 11, preferably 6 to 10, wherein said alkyl, cycloalkyl, cycloheteroalkyl, aryl, or heteroaryl is optionally substituted with an R or R' group; R2 is - -(CH2) d -C≡CH, where "d" is an integer selected from 1 to 6; and - -(CH2) e -CH=CH2 (wherein "e" is an integer selected from 2 to 6) selected from the group consisting of; Or, R1 is H, C1-C6 alkyl, -(CH2) a -C≡CH, -(CH2) b -P + Ph3, C3~C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12 heteroaryl, wherein "a" is an integer selected from 1 to 6, preferably 2 to 4, and "b" is an integer selected from 5 to 11, preferably 6 to 10, wherein said alkyl, cycloalkyl, cycloheteroalkyl, aryl, or heteroaryl is optionally substituted with an R or R' group; R2 is - -(CH2) f -CR4=CH-CH=CR5-(CH2) g -R3, where R4 and R5 are H, an R group, or together form a 6-membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3 to 6 members, and "f" and "g" are integers independently selected from 1 to 3; - -(CH2) f -C≡CC≡C-(CH2) g-R3, where "f" and "g" are integers independently selected from 1 to 3; - -(CH2) i -R3, where "i" is an integer selected from 7 to 9, preferably 8; - -(CH2) j -CH[(CH2) k -P + Ph3]-(CH2) l -R3, where "j" and "l" are integers independently selected from 1 to 6, and "k" is an integer selected from 1 to 6; - -(CH2) m -Cyclobutanyl-(CH2) n -R3, where "m" and "n" are integers independently selected from 1 to 3; and - -(CH2) p -CHR6-CH=CH-CHR7-(CH2) q -R3, where R6 and R7 are H, R groups, or together form a 4-6 membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3-6 members, and "p" and "q" are integers independently selected from 1 to 3. selected from the group consisting of; R3 is

[0062] [ka]

[0063] [Wherein, R8 is H, C1-C6 alkyl, -(CH2) a -C≡CH, -(CH2) b -P + Ph3, C3~C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12heteroaryl, wherein "a" is an integer selected from 1 to 6, preferably 2 to 4, and "b" is an integer selected from 5 to 11, preferably 6 to 10, and wherein said alkyl, cycloalkyl, cycloheteroalkyl, aryl, or heteroaryl is optionally substituted with an R or R' group. and; R is C1-C6 alkyl, C1-C6 alkyloxy, C1-C6 halogenoalkyl, C3-C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12 heteroaryl, wherein the group is optionally substituted with an R′ group; R' is C1-C6 alkyl, C1-C6 alkyloxy, C1-C6 thioalkyl, halogen, C1-C6 halogenoalkyl, hydroxyl (-OH), cyano (-CN), nitro, amino (-NH2), carboxyl (-COOH), phosphate (PO4 - ), amide (-CONH2), -COOR'', -NHR'', -NR''R''', -COR'', -CONHR'', -NHCOR'', -NHSO2R'', -SOR'', -SO2R'', -SONR''R'''', -SO2NR''R''', C3~C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12 and heteroaryl, wherein said cycloalkyl, cycloheteroalkyl, aryl or heteroaryl is selected from the group consisting of halogen, hydroxyl, cyano, nitro, amino, or C 1~ and optionally substituted with C3 alkoxy, R'' and R'' are H or C1-C6 alkyl. or a pharma- ceutically acceptable salt thereof.

[0064] In a first particular embodiment, R1 is methyl, -(CH2)a -C≡CH and -(CH2) b -P + Ph3 (wherein "a" is an integer selected from 2 to 4, and "b" is an integer selected from 6 to 10); selected from the group consisting of; R2 is - -(CH2) d -C≡CH, where "d" is an integer selected from 2 to 4; and - -(CH2) e -CH=CH2 (wherein "e" is an integer selected from 2 to 6) is selected from the group consisting of:

[0065] In a second particular embodiment, R1 is H, methyl, -(CH2) a -C≡CH and -(CH2) b -P + Ph3, wherein "a" is an integer selected from 2 to 4 and "b" is an integer selected from 6 to 10; R2 is selected from the group consisting of - -(CH2) f -CR4=CH-CH=CR5-(CH2) g -R3 (wherein R4 and R5 are H, "f" and "g" are integers independently selected from 1 to 3, and "f" + "g" is 2 to 4); - -(CH2) f -C≡CC≡C-(CH2) g -R3 (wherein "f" and "g" are integers independently selected from 1 to 3, and "f" + "g" is 2 to 4); - -(CH2) i - R3, where "i" is an integer selected from 7 to 9; - -(CH2) j -CH[(CH2) k -P + Ph3]-(CH2) l -R3, where "j" and "l" are integers independently selected from 1 to 6, "j" + "l" is 5 to 7, and "k" is an integer selected from 1 to 6; - -(CH2) m -Cyclobutanyl-(CH2) n -R3, where "m" and "n" are integers independently selected from 1 to 3, and "m" + "n" is 2 to 4; and - -(CH2) p -CHR6-CH=CH-CHR7-(CH2) q -R3, wherein R6 and R7 are H or together form a 4-membered ring, "p" and "q" are integers independently selected from 1 to 3, and "p" + "q" is 2 to 4. selected from the group consisting of; R3 is

[0066] [ka]

[0067] [Wherein, R8 is H, methyl, -(CH2) a -C≡CH and -(CH2) b -P + Ph3, (wherein "a" is an integer selected from 1 to 6, preferably 2 to 4, and "b" is an integer selected from 5 to 11, preferably 6 to 10). It is.

[0068] A particular aspect of the present disclosure relates to compounds that have at least one alkyne group (-C≡CH).One advantage of these compounds is that terminal alkyne is suitable for labeling by click chemistry.Another advantage is based on the ability of alkyne group to chelate copper, especially mitochondrial copper.

[0069] Another specific embodiment of the present disclosure relates to a compound having two biguanidyl radicals.The advantage of increasing the number of biguanidyl radicals in a compound is that it increases the copper chelation, especially of mitochondrial copper, and then increases the effect of the compound on cancer cells, especially cancer mesenchymal cells.

[0070] Finally, one aspect of the present disclosure relates to compounds that have both at least one alkyne group (-C≡CH) and two biguanidyl radicals, both attributes that are optimized for copper chelation and subsequently the effect of the compound on cancer cells, particularly cancer mesenchymal cells.

[0071] Compounds containing at least one alkyne group In one embodiment, the compound has at least one alkyne group (-C≡CH). Optionally, the compound may have one, two, three, or four alkyne groups. In this regard, at least one of R1, R2, and R8 is -(CH2) a -C≡CH, where "a" is an integer from 1 to 6. "a" can be an integer selected from the group consisting of 1, 2, 3, 4, 5, and 6, or an integer selected from 2 to 6, 3 to 6, 4 to 6, 2 to 5, 3 to 5, 4 to 5, 2 to 4, or 2 to 3.

[0072] Optionally, both R1 and R2 are alkyne groups. Alternatively, one of R1 or R2 is an alkyne group. Thus, the compound has the formula (I): a When -C≡CH, R1 and R2 are H, C1-C6 alkyl, -(CH2) a -C≡CH, -(CH2) b -P + Ph3, C3~C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12 wherein "a" is an integer selected from 1 to 6, preferably 2 to 4, and "b" is an integer selected from 5 to 11, preferably 6 to 10; said alkyl, cycloalkyl, cycloheteroalkyl, aryl, or heteroaryl being selected from the group consisting of R or R' groups, and -(CH2) eOptionally, at least one of R1 and R2 is -(CH2) a When -C≡CH, R1 and R2 are H, methyl, -(CH2) a -C≡CH and -(CH2) b -P + Ph3.

[0073] Optionally, one of R1 or R2 is an alkyne group and R8 is an alkyne group, in particular R1 and R8 are alkyne groups.

[0074] Thus, the compound has the formula (I) [In the formula, R1 is -(CH2) a -C≡CH; R2 is - -(CH2) f -CR4=CH-CH=CR5-(CH2) g -R3, where R4 and R5 are H, an R group, or together form a 6-membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3 to 6 members, and "f" and "g" are integers independently selected from 1 to 3; - -(CH2) f -C≡CC≡C-(CH2) g -R3, where "f" and "g" are integers independently selected from 1 to 3; - -(CH2) i - R3, where "i" is an integer selected from 7 to 9; - -(CH2) j -CH[(CH2) k -P + Ph3]-(CH2) l -R3, where "j" and "l" are integers independently selected from 1 to 6, and "k" is an integer selected from 1 to 6; - -(CH2) m-Cyclobutanyl-(CH2) n -R3, where "m" and "n" are integers independently selected from 1 to 3; and - -(CH2) p -CHR6-CH=CH-CHR7-(CH2) q -R3, where R6 and R7 are H, R groups, or together form a 4-6 membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3-6 members, and "p" and "q" are integers independently selected from 1 to 3. selected from the group consisting of; R3 is

[0075] [ka]

[0076] (Wherein, R8 is -(CH2) a -C≡CH is] It is of the following.

[0077] Optionally, R8 is an alkyne group and R1 and R2 are not alkyne groups.

[0078] Thus, the compound has the formula (I) [In the formula, R1 is H, C1-C6 alkyl, -(CH2) b -P + Ph3, C3~C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12 heteroaryl, wherein "b" is an integer selected from 5 to 11, preferably 6 to 10, wherein said alkyl, cycloalkyl, cycloheteroalkyl, aryl, or heteroaryl is optionally substituted with an R or R' group; R2 is - -(CH2) f -CR4=CH-CH=CR5-(CH2) g -R3, where R4 and R5 are H, an R group, or together form a 6-membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3 to 6 members, and "f" and "g" are integers independently selected from 1 to 3; - -(CH2) f -C≡CC≡C-(CH2) g -R3, where "f" and "g" are integers independently selected from 1 to 3; - -(CH2) i - R3, where "i" is an integer selected from 7 to 9; - -(CH2) j -CH[(CH2) k -P + Ph3]-(CH2) l -R3, where "j" and "l" are integers independently selected from 1 to 6, and "k" is an integer selected from 1 to 6; - -(CH2) m -Cyclobutanyl-(CH2) n -R3, where "m" and "n" are integers independently selected from 1 to 3; and - -(CH2) p -CHR6-CH=CH-CHR7-(CH2) q -R3, where R6 and R7 are H, R groups, or together form a 4-6 membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3-6 members, and "p" and "q" are integers independently selected from 1 to 3. selected from the group consisting of; R3 is

[0079] [ka]

[0080] (Wherein, R8 is -(CH2) a -C≡CH is] It is of the following.

[0081] Optionally, R1 is H, methyl, -(CH2) a -C≡CH and -(CH2) b -P + Ph3.

[0082] Optionally, R2 is - -(CH2) f -CR4=CH-CH=CR5-(CH2) g -R3, where R4 and R5 are H, an R group, or together form a 6-membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3 to 6 members, and "f" and "g" are integers independently selected from 1 to 3, and "f" + "g" is 2 to 4; - -(CH2) f -C≡CC≡C-(CH2) g -R3 (wherein "f" and "g" are integers independently selected from 1 to 3, and "f" + "g" is 2 to 4); - -(CH2) i - R3, where "i" is an integer selected from 7 to 9; - -(CH2) j -CH[(CH2) k -P + Ph3]-(CH2) l -R3, where "j" and "l" are integers independently selected from 1 to 6, "j" + "l" is 5 to 7, and "k" is an integer selected from 2 to 4; - -(CH2) m -Cyclobutanyl-(CH2) n -R3, where "m" and "n" are integers independently selected from 1 to 3, and "m" + "n" is 2 to 4; and - -(CH2)p -CHR6-CH=CH-CHR7-(CH2) q -R3, where R6 and R7 are H, R groups, or together form a 4-6 membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3-6 members, and "p" and "q" are integers independently selected from 1 to 3, and "p" + "q" is 2 to 4. is selected from the group consisting of:

[0083] For example, such compounds are in the following groups:

[0084] [ka]

[0085] (wherein n is an integer selected from 3 to 9, preferably 4 to 8); and

[0086] [ka]

[0087] (wherein n is an integer independently selected from 0 to 5, preferably 1 to 3); and It may be selected from its pharma- ceutically acceptable salts.

[0088] Compounds with two biguanidyl radicals In a second embodiment, which can be optionally combined with the first embodiment, the compound has two biguanidyl radicals, in other words, R3 is present in the compound.

[0089] Thus, the compound of formula (I) R1 is H, C1-C6 alkyl, -(CH2) a -C≡CH, -(CH2) b -P + Ph3, C3~C10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12 heteroaryl, wherein "a" is an integer selected from 1 to 6, preferably 2 to 4, and "b" is an integer selected from 5 to 11, preferably 6 to 10, wherein said alkyl, cycloalkyl, cycloheteroalkyl, aryl, or heteroaryl is optionally substituted with an R or R' group; R2, - -(CH2) f -CR4=CH-CH=CR5-(CH2) g -R3, where R4 and R5 are H, an R group, or together form a 6-membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3 to 6 members, and "f" and "g" are integers independently selected from 1 to 3; - -(CH2) f -C≡CC≡C-(CH2) g -R3, where "f" and "g" are integers independently selected from 1 to 3; - -(CH2) i -R3, where "i" is an integer selected from 7 to 9, preferably 8; - -(CH2) j -CH[(CH2) k -P + Ph3]-(CH2) l -R3, where "j" and "l" are integers independently selected from 1 to 6, and "k" is an integer selected from 1 to 6; - -(CH2) m -Cyclobutanyl-(CH2) n -R3, where "m" and "n" are integers independently selected from 1 to 3; and - -(CH2) p -CHR6-CH=CH-CHR7-(CH2) q-R3, where R6 and R7 are H, R groups, or together form a 4-6 membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3-6 members, and "p" and "q" are integers independently selected from 1 to 3. selected from the group consisting of; R3,

[0090] [ka]

[0091] [Wherein, R8 is H, C1-C6 alkyl, -(CH2) a -C≡CH, -(CH2) b -P + Ph3, C3~C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12 heteroaryl, wherein "a" is an integer selected from 1 to 6, preferably 2 to 4, and "b" is an integer selected from 5 to 11, preferably 6 to 10, and wherein said alkyl, cycloalkyl, cycloheteroalkyl, aryl, or heteroaryl is optionally substituted with an R or R' group. That is, It's like that.

[0092] Optionally, R1 and R8 are H, methyl, -(CH2) a -C≡CH and -(CH2) b -P + Ph3.

[0093] Optionally, R2 is - -(CH2) f -CR4=CH-CH=CR5-(CH2) g-R3, where R4 and R5 are H, an R group, or together form a 6-membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3 to 6 members, and "f" and "g" are integers independently selected from 1 to 3, and "f" + "g" is 2 to 4; - -(CH2) f -C≡CC≡C-(CH2) g -R3 (wherein "f" and "g" are integers independently selected from 1 to 3, and "f" + "g" is 2 to 4); - -(CH2) i - R3, where "i" is an integer selected from 7 to 9; - -(CH2) j -CH[(CH2) k -P + Ph3]-(CH2) l -R3, where "j" and "l" are integers independently selected from 1 to 6, "j" + "l" is 5 to 7, and "k" is an integer selected from 2 to 4; - -(CH2) m -Cyclobutanyl-(CH2) n -R3, where "m" and "n" are integers independently selected from 1 to 3, and "m" + "n" is 2 to 4; and - -(CH2) p -CHR6-CH=CH-CHR7-(CH2) q -R3, where R6 and R7 are H, R groups, or together form a 4-6 membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3-6 members, and "p" and "q" are integers independently selected from 1 to 3, and "p" + "q" is 2 to 4. is selected from the group consisting of:

[0094] For example, such compounds are in the following groups:

[0095] [ka]

[0096] (wherein n is an integer independently selected from 0 to 5, preferably 1 to 3);

[0097] [ka]

[0098] (wherein n is an integer independently selected from 0 to 5, preferably 1 to 3);

[0099] [ka]

[0100] [In the formula, A dotted line may be present or absent, and represents one or two atoms with a covalent bond; R is C1-C6 alkyl, C1-C6 alkyloxy, C1-C6 halogenoalkyl, C3-C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12 heteroaryl, the group being optionally substituted with an R' group; R' being C1-C6 alkyl, C1-C6 alkyloxy, C1-C6 thioalkyl, halogen, C1-C6 halogenoalkyl, hydroxyl (-OH), cyano (-CN), nitro, amino (-NH2), carboxyl (-COOH), phosphate (PO4 - ), amide (-CONH2), -COOR'', -NHR'', -NR''R''', -COR'', -CONHR'', -NHCOR'', -NHSO2R'', -SOR'', -SO2R'', -SONR''R'''', -SO2NR''R''', C3~C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12Aryl and C5-C 12 and heteroaryl, wherein said cycloalkyl, cycloheteroalkyl, aryl or heteroaryl is selected from the group consisting of halogen, hydroxyl, cyano, nitro, amino, or C 1~ optionally substituted with C3 alkoxy, where R'' and R'' are H or C1-C6 alkyl; or a pharma- ceutically acceptable salt thereof.

[0101] Compounds with an alkyne group and two biguanidyl radicals In certain embodiments, the compound has at least one alkyne group, but also has two biguanidyl radicals.

[0102] Thus, the compound of formula (I) is a compound in which at least one of R1 and R8 is -(CH2) a When -C≡CH, R1 is H, C1-C6 alkyl, -(CH2) a -C≡CH, -(CH2) b -P + Ph3, C3~C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12 heteroaryl, wherein "a" is an integer selected from 1 to 6, preferably 2 to 4, and "b" is an integer selected from 5 to 11, preferably 6 to 10, wherein said alkyl, cycloalkyl, cycloheteroalkyl, aryl, or heteroaryl is optionally substituted with an R or R' group; R2, - -(CH2) f -CR4=CH-CH=CR5-(CH2) g-R3, where R4 and R5 are H, an R group, or together form a 6-membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3 to 6 members, and "f" and "g" are integers independently selected from 1 to 3; - -(CH2) f -C≡CC≡C-(CH2) g -R3, where "f" and "g" are integers independently selected from 1 to 3; - -(CH2) i -R3, where "i" is an integer selected from 7 to 9, preferably 8; - -(CH2) j -CH[(CH2) k -P + Ph3]-(CH2) l -R3, where "j" and "l" are integers independently selected from 1 to 6, and "k" is an integer selected from 1 to 6; - -(CH2) m -Cyclobutanyl-(CH2) n -R3, where "m" and "n" are integers independently selected from 1 to 3; and - -(CH2) p -CHR6-CH=CH-CHR7-(CH2) q -R3, where R6 and R7 are H, R groups, or together form a 4-6 membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3-6 members, and "p" and "q" are integers independently selected from 1 to 3. selected from the group consisting of; R3,

[0103] [ka]

[0104] [Wherein, R8 is H, C1-C6 alkyl, -(CH2)a -C≡CH, -(CH2) b -P + Ph3, C3~C 10 Cycloalkyl, C 3~ C 10 Cycloheteroalkyl, C6-C 12 Aryl and C5-C 12 heteroaryl, wherein "a" is an integer selected from 1 to 6, preferably 2 to 4, and "b" is an integer selected from 5 to 11, preferably 6 to 10, and wherein said alkyl, cycloalkyl, cycloheteroalkyl, aryl, or heteroaryl is optionally substituted with an R or R' group. That is, It's like that.

[0105] Optionally, R1 and R8 are H, methyl, -(CH2) a -C≡CH and -(CH2) b -P + Ph3.

[0106] Optionally, R2 is - -(CH2) f -CR4=CH-CH=CR5-(CH2) g -R3, where R4 and R5 are H, an R group, or together form a 6-membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3 to 6 members, and "f" and "g" are integers independently selected from 1 to 3, and "f" + "g" is 2 to 4. - -(CH2) f -C≡CC≡C-(CH2) g -R3 (wherein "f" and "g" are integers independently selected from 1 to 3, and "f" + "g" is 2 to 4); - -(CH2) i - R3, where "i" is an integer selected from 7 to 9; - -(CH2) j -CH[(CH2)k -P + Ph3]-(CH2) l -R3, where "j" and "l" are integers independently selected from 1 to 6, "j" + "l" is 5 to 7, and "k" is an integer selected from 2 to 4; - -(CH2) m -Cyclobutanyl-(CH2) n -R3, where "m" and "n" are integers independently selected from 1 to 3, and "m" + "n" is 2 to 4; and - -(CH2) p -CHR6-CH=CH-CHR7-(CH2) q -R3, where R6 and R7 are H, R groups, or together form a 4-6 membered ring, optionally substituted with one or two R' groups, or fused to a cycloalkyl, aryl, cycloheteroalkyl, or heteroaryl having 3-6 members, and "p" and "q" are integers independently selected from 1 to 3, and "p" + "q" is 2 to 4. is selected from the group consisting of:

[0107] For example, such compounds are

[0108] [ka]

[0109] (wherein n is an integer independently selected from 0 to 5, preferably 1 to 3). or a pharma- ceutically acceptable salt thereof.

[0110] A pharma- ceutically acceptable salt of a compound of the present invention may be, for example, a salt with an acid selected from the group consisting of formic acid, acetic acid, propionic acid, lactic acid, butyric acid, isobutyric acid, trifluoroacetic acid, malic acid, malonic acid, fumaric acid, succinic acid, glutamic acid, tartaric acid, oxalic acid, citric acid, glycolic acid, glucuronic acid, ascorbic acid, benzoic acid, phthalic acid, salicylic acid, anthranilic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, dichloroacetic acid, aminooxyacetic acid, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, and boric acid.

[0111] Conjugates The present invention relates to a conjugate comprising a compound of formula (I) disclosed herein covalently bound to a toxin, drug, or label. More specifically, the present invention relates to a conjugate obtainable or obtained by reacting a compound according to the present disclosure having an alkyne group with a drug, toxin, or label covalently bound to an azide (N3) group.

[0112] Since the compound of formula (I) has the ability to target mitochondria, the toxin, drug or label is selected from among the toxins or drugs that are specifically related to targeting mitochondria. The drug can be an antidiabetic drug, an anticancer drug, an antiparasitic drug, an antiviral drug, an antiepileptic drug, an anesthetic drug, an antiinflammatory drug, and an antiarrhythmic drug. The drug can be an inhibitor of mitochondrial respiratory chain complexes, an uncoupler and an ionophore, a regulator of the permeability transition pore, a regulator of the mitochondrial potassium channel, an antioxidant, an inhibitor of the mitochondrial ATPase, an apoptosis regulator, and the like. For example, a non-exhaustive list of drugs includes rotenone, antimycin A, myxothiazol, cyanide, dinitrophenol, valinomycin, nigericin, cyclosporine A, bongkrekic acid, betulinic acid, CD437, sangliferin, Ro68-3400, atractyloside, protoporphyrin IX, diazoxide, pinacidil, nicorandil, cromakalim, glibenclamide, 5-HD, NS1619, NS004, CGS7184, paxillline, IbTx, ChTx, ophthalmic solution ... Ligomycin C, apoptolysin A, resveratrol, Bz-423, diindolylmethane, aurovertin, PK11195, R207910, NSAIDs, minocycline, KB-R7943, dimebon, CoQ10, idebenone, lipoic acid, melatonin, vitamin E, nicotinamide, carnitine, L-carnitine, acetyl-L-carnitine, sirtuits, FK506, deferoxamine, 3-bromopyruvate, 2-deoxyglucose, methyl jasmonate, mannoheptulose, gossypol.ABT-737, A-385358, ABT-263, HA14-1, AT-101, obatoclax, isothiocyanates, arsenic trioxide, lonidamine, arsenite, GSAO, clodronate, Ro5-4684, a-TOS, tamoxifen, piceatannol, rhodamine-123, dichloroacetic acid, CAP-232 / TLN-232, MJE3, vitamin K3, fialuridine, 2-methoxyestradiol, β-lapachone, menadione, STA-4783, sanglifehrin, NIM811, Debio025, SSR180575, Ro5-4684; MitoSNO1, nitrites, mangafodipir, edaravone, statins These include, but are not limited to, UQ, cariporide, propofol, MCI-186, pyruvate, minoxidil, alpha-lipoic acid, Alda-1, etomoxir, perhexiline, ranolazine, trimetazidine, NAO, diazoxide, BMS-191095, ascofuranone, nafrezin, atovaquone, flutolanil, atopenine A5, nafrezin-c (c-lactone derivative), thiazolidine-diones, fibrates, thizolidinediones, beta-aminoisobutyric acid, SRT1720, trolox, ursodeoxycholic acid, doxorubicin, zidovudine, valproic acid, indomethacin, and carvedinol. In addition, those skilled in the art have sufficient knowledge of the drug of interest, as shown in Olszewska et al. (IUBMB Life. 2013, Mar;65(3):273-81).

[0113] In the context of cancer therapy, conjugates of interest are more particularly those compounds of formula (I) and 3-bromopyruvate, 2-deoxyglucose, methyl jasmonate, mannoheptulose, gossypol, ABT-737, A-385358, ABT-263, HA14-1, AT-101, obatoclax, isothiocyanates, arsenic trioxide, lonidamine, arsenite, GSAO, clodronate, It may be a conjugate with a drug selected from Ro5-4684, a-TOS, tamoxifen, piceatannol, rhodamine-123, dichloroacetic acid, CAP-232 / TLN-232, MJE3, vitamin K3, fialuridine, 2-methoxyestradiol, β-lapachone, menadione, STA-4783, sanglifehrin, NIM811, Debio025, SSR180575, Ro5-4684; MitoSNO1, nitrite, and mangafodipir.

[0114] Optionally, the drug is an anti-tumor drug, particularly as disclosed below.

[0115] In a particular embodiment, the conjugates can be prepared by click chemistry using an alkyne group for covalent attachment to a toxin, drug or label. The toxin, drug or label can be covalently attached to the azide (N3) group, in particular by copper-catalyzed azide-alkyne cycloaddition (CuAAC), which reacts with a compound of formula (I) bearing an alkyne group. For a review on bioorthogonal chemistry, including click chemistry, see Sletten and Bertozzi (Angew. Chem. Int. Ed. Engl. 2009, 48(38):6974-6998, the disclosure of which is incorporated herein by reference). The reaction of an alkyne with an azide is a [3+2] cycloaddition reaction, resulting in a triazole ring.

[0116] The label may be a directly or indirectly detectable moiety. The label may be selected from among dyes, radioactive labels, affinity tags. In particular, the dye may be selected from the group consisting of fluorescent, luminescent, or phosphorescent dyes, preferably dansyl dyes, fluorescein dyes, acridine dyes, rhodamine dyes, coumarin dyes, BODIPY dyes, and cyanine dyes. More particularly, the fluorescent dye may be selected from among dyes sold by Thermo Fisher, such as Alexa Fluor dyes, Pacific dyes, Texas Red, or dyes sold by other suppliers for cyanines 3, 5, 7. In particular, azide-containing dyes for CuAAC are commercially available in the following classes: Alexa Fluor® 488, 55, 594, 647, and TAMRA (tetramethylrhodamine). In a second embodiment, the label may be an affinity tag. Such affinity tags can be selected from the group consisting of, for example, biotin, His-tag, Flag-tag, strep-tag, sugar, lipid, sterol, PEG linker, and cofactor. In a particular embodiment, the label is a biotinylated label. Biotin bound to azide is commercially available (biotin azide). Finally, the label can be a radioactive label. The radioactive label can be selected from the group consisting of radioactive forms of hydrogen, carbon, phosphorus, sulfur, and iodine, including tritium, carbon-11, carbon-14, phosphorus-32, phosphorus-33, sulfur-33, iodine-123, and iodine-125.

[0117] Pharmaceutical composition or veterinary composition The present invention relates to pharmaceutical or veterinary compositions comprising a compound of formula (I) as disclosed herein, or a conjugate comprising a compound of formula (I) as disclosed herein and a drug or toxin.

[0118] The pharmaceutical compositions contemplated herein may contain a pharma- ceutically acceptable carrier in addition to the active ingredient. The term "pharma-ceutically acceptable carrier" is meant to encompass any carrier (e.g., a support, a substance, a solvent, etc.) that does not interfere with the effectiveness of the biological activity of the active ingredient and is not toxic to the host to which it is administered. For example, for parenteral administration, the active compound may be formulated in a unit dosage form for injection in a vehicle such as saline, dextrose solution, serum albumin, and Ringer's solution.

[0119] Pharmaceutical compositions can be formulated in a manner known in the art as solutions in pharma- ceutically compatible solvents, or as emulsions, suspensions or dispersions in suitable pharmaceutical solvents or vehicles, or as pills, tablets or capsules containing a solid vehicle. Formulations of the invention suitable for oral administration can be in the form of discrete units as capsules, sachets, tablets or lozenges, each containing a predetermined amount of the active ingredient in powder or granule form; in the form of a solution or suspension in an aqueous or non-aqueous liquid; or in the form of an oil-in-water or water-in-oil emulsion. Formulations suitable for parental administration conveniently comprise a sterile oily or aqueous preparation of the active ingredient, preferably isotonic with the blood of the recipient. All such formulations may also contain other pharma-ceutically compatible and non-toxic auxiliary substances, such as, for example, stabilizers, antioxidants, binders, dyes, emulsifiers or flavorings. Thus, the formulations of the invention comprise the active ingredient in association with a pharma- ceutically acceptable carrier, and optionally other therapeutic ingredients. The carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not harmful to its recipient. The pharmaceutical composition is advantageously applied by injection or intravenous infusion of a suitable sterile solution, or by the digestive tract, such as by oral administration. Methods for safely and effectively administering most of these chemotherapeutic agents are known to those skilled in the art. In addition, their administration is described in the standard literature.

[0120] The present invention relates to a compound of formula (I) as disclosed herein, or a conjugate comprising a compound of formula (I) as disclosed herein and a drug or toxin, for use as a medicament, in particular for the treatment of a disease or disorder as defined below.The present invention relates to the use of a compound of formula (I) as disclosed herein, or a conjugate comprising a compound of formula (I) as disclosed herein and a drug or toxin, for the manufacture of a medicament, in particular for the treatment of a disease or disorder as defined below.

[0121] The compounds of formula (I) disclosed herein or the conjugates comprising the compounds of formula (I) disclosed herein and drugs or toxins can be used as drugs.In particular, the compounds of formula (I) disclosed herein or the conjugates comprising the compounds of formula (I) disclosed herein and drugs or toxins can be used to treat diseases or disorders selected from a non-exhaustive list including cancer, diabetes, especially diabetes mellitus, obesity, hyperlipidemia, hypercholesterolemia, preeclampsia, liver and kidney diseases, especially fatty liver, cardiovascular diseases, especially coronary artery disease, osteoporosis, metabolic syndrome, multiple sclerosis, polycystic ovarian disease, muscle pain, muscle cell damage, rhabdomyolysis, erectile dysfunction, cognitive dysfunction, regulation of gut microbiota, neurodegenerative disorders and diseases, such as Alzheimer's disease and Parkinson's disease, and as antiparasitic drugs, antiviral drugs, antiepileptic drugs, anesthetic drugs, antiinflammatory drugs, and antiarrhythmic drugs.

[0122] The pharmaceutical or veterinary compositions disclosed herein may further comprise additional active ingredients or drugs.

[0123] cancer In certain aspects of the present invention, a compound of formula (I) disclosed herein, or a conjugate comprising a compound of formula (I) disclosed herein and a drug or toxin, can be used to treat cancer.

[0124] The terms "cancer," "cancerous," or "malignant" refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth. Examples of cancer include, for example, leukemia, lymphoma, blastoma, carcinoma, and sarcoma.

[0125] For example, the cancer may be selected from a non-exhaustive list including rectal cancer, colon cancer, gastric cancer, head and neck cancer, thyroid cancer, cervical cancer, uterine cancer, breast cancer, particularly triple-negative breast cancer, ovarian cancer, brain tumors, particularly glioblastoma and neuroblastoma, lung cancer, particularly small cell lung cancer and non-small cell lung cancer, skin cancer, bladder cancer, blood cancer, kidney cancer, liver cancer, prostate cancer, multiple myeloma, and endometrial cancer. In a particular embodiment, the cancer may be selected from a list including pancreatic cancer, prostate cancer, liver cancer, cervical cancer, endometrial cancer, ovarian cancer, breast cancer, colon cancer, glioblastoma, and neuroblastoma. In a very particular embodiment, the cancer may be selected from a list including ovarian cancer, breast cancer, colon cancer, glioblastoma, and neuroblastoma. The cancer may be mesenchymal ovarian cancer, particularly high-grade ovarian cancer of the serous type, or invasive breast cancer, and / or metastases thereof.

[0126] In a preferred embodiment, cancer has mesenchymal cells.Indeed, the compounds of formula (I) disclosed herein can selectively target a population of cancer cells that have mesenchymal characteristics.In certain aspects, the compounds of formula (I) disclosed herein present the advantage of acting against circulating tumor cells, particularly cancer stem cells.

[0127] Thus, the present invention relates to compounds of formula (I) disclosed herein for use in inhibiting cancer recurrence or cancer metastasis, and methods for inhibiting cancer recurrence or cancer metastasis by administering the compounds disclosed herein.

[0128] In certain embodiments, the subject has metastatic cancer. In another particular embodiment, the subject has cancer that is resistant to conventional anti-tumor drugs. In further particular embodiments, the subject has a secondary or recurrent tumor.

[0129] The compounds of formula (I) disclosed herein can be used in combination with another drug. Thus, the present invention relates to - a pharmaceutical composition comprising a compound of formula (I) as disclosed herein and another drug as defined herein, and optionally a pharma- ceutically acceptable carrier, in particular for use in the treatment of cancer; - an article of manufacture or a kit comprising a compound of formula (I) as disclosed herein and another drug as defined herein as a combined preparation for simultaneous, separate or sequential use, in particular in the treatment of cancer; - a combined preparation comprising a compound of formula (I) disclosed herein and another drug as defined herein for simultaneous, separate or sequential use, especially in the treatment of cancer; - a pharmaceutical composition comprising a compound of formula (I) as disclosed herein for use in the treatment of cancer in combination with another drug treatment and / or with radiation therapy as defined herein; - the use of a pharmaceutical composition comprising a compound of formula (I) as disclosed herein for the manufacture of a medicament for the treatment of cancer in combination with another drug treatment and / or with radiation therapy as defined herein; - use of a pharmaceutical composition comprising a compound of formula (I) disclosed herein, another drug as defined herein, and optionally a pharma- ceutically acceptable carrier, for the manufacture of a medicament for the treatment of cancer; - a method for treating cancer in a subject in need thereof, comprising administering an effective amount of a pharmaceutical composition comprising a) a compound of formula (I) as disclosed herein, and b) another drug as defined herein, and a pharma- ceutically acceptable carrier; - a method for treating cancer in a subject in need thereof, comprising administering an effective amount of a pharmaceutical composition comprising a compound of formula (I) as disclosed herein and an effective amount of a pharmaceutical composition comprising another drug as defined herein; - a method for treating cancer in a subject in need thereof, comprising administering an effective amount of a pharmaceutical composition comprising a compound of formula (I) disclosed herein and another drug as defined herein. Regarding.

[0130] The terms "kit", "product" or "combination preparation" as used herein specifically define a "kit of parts" in the sense that the combination partners defined above can be administered independently or by using different fixed combinations with distinct amounts of the combination partners, i.e. simultaneously or at different times. In this case, the parts of the kit of parts can be administered, for example, simultaneously or staggered, i.e. at different times and with equal or different time intervals for any part of the kit of parts. The ratio of the total amounts of the administered combination partners in the combination preparation can be varied. The combination partners can be administered by the same route or by different routes.

[0131] Radiation therapy includes, but is not limited to, gamma rays, X-rays, and / or direct delivery of radioisotopes to tumor cells. Other radiation therapies include microwave and UV irradiation. Other approaches to radiation therapy are also contemplated in the present invention.

[0132] The other drugs may be selected from a non-exhaustive list including chemotherapeutic drugs, immune checkpoint inhibitors, hormonal therapy drugs, and immunotherapy drugs.

[0133] Chemotherapeutic agents may include inhibitors of topoisomerase I or II, DNA cross-linking agents, DNA alkylating agents, antimetabolites, and / or inhibitors of mitotic spindles.

[0134] Inhibitors of topoisomerase I and / or II include, but are not limited to, etoposide, topotecan, camptothecin, irinotecan, amsacrine, intoplecin, anthracyclines such as doxorubicin, epirubicin, daunorubicin, idarubicin, and mitoxantrone. Inhibitors of topoisomerase I and II include, but are not limited to, intoplecin.

[0135] DNA crosslinking agents include, but are not limited to, cisplatin, carboplatin, and oxaliplatin.

[0136] Antimetabolites block enzymes involved in nucleic acid synthesis or are incorporated into DNA, which results in the generation of erroneous genetic code and apoptosis. Non-exhaustive examples include, but are not limited to, folate antagonists, pyrimidine analogs, purine analogs and adenosine deaminase inhibitors, more specifically, methotrexate, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, pentostatin, 5-fluorouracil, gemcitabine and capecitabine.

[0137] Alkylating agents include, but are not limited to, nitrogen mustards, ethylenimine derivatives, alkylsulfonates, nitrosoureas, metal salts, and triazenes, non-exhaustive examples of which include uracil mustard, chlormethine, cyclophosphamide (CYTOXAN®), ifosfamide, melphalan, chlorambucil, pipobroman, triethylenemelamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, fotemustine, cisplatin, carboplatin, thiotepa, streptozocin, dacarbazine, oxaliplatin, and temozolomide.

[0138] Mitotic spindle inhibitors include paclitaxel, docetaxel, vinorelbine, larotaxel (also known as XRP9881; Sanofi-Aventis), XRP6258 (Sanofi-Aventis), BMS-184476 (Bristol-Meyer-Squibb), BMS-188797 (Bristol-Meyer-Squibb), BMS-275183 (Bristol-Meyer-Squibb), ortataxel (also known as IDN 5109, BAY 59-8862, or SB-T-101131; Bristol-Meyer-Squibb), RPR 109881A (Bristol-Meyer-Squibb), RPR Examples of suitable anti-inflammatory drugs include, but are not limited to, 116258 (Bristol-Meyer-Squibb), NBT-728 (TAPESTRY), PG-paclitaxel (also known as CT-2103, PPX, paclitaxel poliglumex, polyglutamated paclitaxel, or Xyotax™), ABRAXANE® (also known as Nab-paclitaxel, ABRAXIS BIOSCIENCE), tesetaxel (also known as DJ-927), IDN 5390 (INDENA), Taxoplexin (also known as docosahexaenoic acid-paclitaxel, PROTARGA), DHA-paclitaxel (also known as Taxoplexin®), and MAC-321 (WYETH). See also the review by Hennenfent & Govindan (2006, Annals of Oncology, 17, 735-749).

[0139] The immune checkpoint inhibitor may be selected from the group consisting of anti-CTLA-4 (cytotoxic T-lymphocyte associated protein 4) therapy, such as ipilimumab, PD-1 (programmed cell death protein 1) inhibitors, such as nivolumab, pembrolizumab, or BGB-A317, PDL1 (programmed cell death ligand) inhibitors, such as atezolizumab, avelumab, or durvalumab, LAG-3 (lymphocyte activation gene 3) inhibitors, such as BMS-986016, TIM-3 (T-cell immunoglobulin and mucin domain containing-3) inhibitors, TIGIT (T-cell immunoreceptor with Ig and ITIM domains) inhibitors, BLTA (B-lymphocyte and T-lymphocyte attenuator) inhibitors, IDO1 inhibitors, such as epacadostat, or combinations thereof.

[0140] Hormonal therapy drugs include, for example, Tamoxifen, Fareston, Arimidex, Aromasin, Femara, Zoladex / Lupron, Megas, and Halotestin.

[0141] Imaging and Diagnostics The compounds of formula (I) disclosed herein having an alkyne group are suitable for forming detectable entities in situ, and therefore for labeling mitochondria. In fact, by carrying out click chemistry, in particular copper-catalyzed azide-alkyne cycloaddition (CuAAC), a label having an azide (N3) group can be covalently bound to the compounds of formula (I) disclosed herein having an alkyne group.

[0142] Copper-catalyzed azide-alkyne cycloaddition (CuAAC) generally requires the presence of a copper (I) catalyst. However, as shown in the examples, the addition of copper is not essential. In fact, the compounds of formula (I) disclosed herein are specifically targeted and localized in mitochondria, so that the natural mitochondrial abundance of endogenous copper is sufficient to facilitate the labeling of the compounds of the present invention in native conditions.

[0143] The present invention relates to a kit comprising a compound of formula (I) disclosed herein bearing an alkyne group and a label bearing an azide group, which may preferably be a fluorescent label or a biotinylated label as defined in the section "Conjugates" above.

[0144] The present invention also relates to the in vitro, ex vivo or in vivo use of a compound of formula (I) as disclosed herein bearing an alkyne group or of the above kit, e.g. as a research tool for medical imaging or diagnosis, in particular for visualizing mitochondria.

[0145] The present invention provides an in vitro method for visualizing mitochondria in cells, comprising: - contacting the cells with a compound of formula (I) disclosed herein having an alkyne group; - contacting said cells with a label carrying an azide group, preferably a fluorescent label; - detecting a label, preferably a fluorescent label, within said cells; The present invention relates to a method comprising the steps of:

[0146] Preferably, the cells are permeabilized and then fixed prior to the step of contacting the cells with a label carrying an alkyne group.

[0147] Further aspects and advantages of the present invention are described in the following examples, which should be considered as illustrative and not limiting. EXAMPLES

[0148] The clinically approved drug metformin has been shown to selectively kill viable cancer cells by a mechanism that is not fully understood. To provide further putative mechanisms, the inventors developed novel compounds that are more potent than metformin against a panel of cancer cells and can be labeled in situ by click chemistry. With this technique, the inventors were able to provide the first evidence of mitochondrial targeting by this class of drugs. A combination of fluorescence microscopy and cyclic voltammetry showed that metformin and the novel compounds target mitochondrial copper and induce reactive oxygen species generation, mitochondrial dysfunction, and apoptosis within this organelle. Importantly, this study revealed that mitochondrial copper is required for the maintenance of the mesenchymal state in human cancer cells, and that metformin and the novel compounds can block epithelial-mesenchymal transition, a biological process that usually accounts for the development of viable cancer cells, by directly targeting copper.

[0149] result Development of novel compounds that are potent clickable metformin derivatives. To provide further putative mechanisms, we set out to develop derivatives of metformin that would allow direct visualization of the intracellular site of action of biguanide drugs. To this end, we synthesized an alkyne-containing analogue, named metforminin (Met) (3) (Figure 1A). This analogue was designed to mimic metformin and phenformin, with the distal methyl / benzyl substituent replaced by a terminal alkyne suitable for intracellular labelling via click chemistry.

[0150] Comparative evaluation of metformin and metformin against a series of triple-negative human breast cancer cells and circulating tumor cells (CTCs) from colorectal cancer patients showed that the synthetic analog metformin exhibited higher potency compared to metformin (Figure 1B: IC of metformin). 50 is the IC of metformin 502.5, 4.7, 2.8, and 3.5 mM, compared with 12.1, 15.1, 7.7, and 10.9 mM, respectively.

[0151] Cancer cells can undergo phenotypic changes reminiscent of those observed during normal embryonic development, acquiring physical properties that allow these cells to detach from the primary site and give rise to distant secondary tumors. This process of epithelial-mesenchymal transition (EMT) can be artificially induced in normal epithelial breast cells or triple-negative breast cancer cells to generate cells with a pronounced mesenchymal phenotype. It is noteworthy that cells capable of adopting such a metastable state have been shown to be resistant to conventional chemotherapy. These surviving cancer cells are therefore associated with cancer recurrence.

[0152] Importantly, in these models metformin was more potent compared to metformin and exhibited higher toxicity against MDA-MB-468 triple-negative breast cancer cells that were transiently induced into a mesenchymal state using epidermal growth factor (EGF) (Figure 1C: IC of metformin 50 is the IC of metformin 50 (12.6, 5.2, 0.8, and 0.9 mM, compared with 24.3, 12.1, 5.5, and 2.3 mM, respectively). 高 / CD24 低 Compared with corresponding epithelial cells, HMLER CD44 低 / CD24 高 This was observed in mesenchymal cells (Figure 1D). These data indicated that metformin is suitable for cell imaging studies.

[0153] Metformin targets mitochondrial copper. Metformin-treated cancer cells were labeled intracellularly by click chemistry to identify the putative mechanistic site of action of the biguanides (Figure 2A). In breast cancer cells, labeled metformin colocalized with cytochrome c, a cellular component that uniquely characterizes mitochondria, with weak nuclear staining and large foci similar to those observed in nucleolar proteins (Figures 2B and 3A). In contrast, labeled metformin could not be detected in the nuclei of CTCs in response to treatment, consistent with the idea that mitochondria are the functional target organelles of biguanides (Figure 2B). Notably, intracellular metformin labeling, a chemical reaction normally catalyzed by copper(I), could be performed in situ without the need to incubate cells with additional copper catalyst (Figure 3B). Thus, the natural mitochondrial abundance of endogenous copper was sufficient to promote metformin labeling in native conditions. Furthermore, we demonstrated that metformin chemically interacts with copper within this organelle.

[0154] This finding prompted the inventors to investigate the effects of metformin and metforminin on mitochondrial copper(II) levels using a selective turn-off fluorescent probe. Metformin and metformin induced a decrease in the fluorescence of this turn-off probe in MDA-MB-468 cells, indicating an increase in the levels of copper(II) (Figure 2C and Figure 2D). In addition, metformin altered the redox properties of copper, defined by the occurrence of a cyclic voltammetric peak characteristic of a quasi-reversible couple (Figure 2E), and metformin treatment reduced the expression of copper uptake protein 1 (Ctr1) in these cells (Figure 4). These data indicated that metformin increased mitochondrial copper(II) levels not by increasing the cellular uptake of copper, but rather by directly interacting with copper(I) in this organelle and promoting its oxidation, resulting in an increase in the levels of mitochondrial copper(II). Interestingly, biguanides also increased levels of mitochondrial iron(II) as defined by flow cytometry using a selective turn-off probe (Figure 2F), and cyclic voltammetry showed that metformin altered the redox properties of this metal, suggesting a direct interaction. These data supported a model in which metformin directly inhibits complex I through iron binding and can hijack iron(III) from the Fe-S cluster. Alternatively, metformin treatment also consumes iron(III), acting as an electron scavenger for copper(I) oxidation. Consistent with metformin acting as a pro-oxidant of copper in mitochondria and altering the ion balance between copper and iron, treatment of MDA-MB-468 cells with metformin or metforminin increased levels of reactive oxygen species (ROS) in this organelle, accompanied by a decrease in mitochondrial membrane potential and changes in mitochondrial morphology. Furthermore, metformin- or metforminin-induced cell death was rescued by the caspase-3 inhibitor Z-VAD-FMK, but not by the ferroptosis and necroptosis inhibitors ferrostatin-1 and necrostatin-1, respectively, consistent with apoptotic cell death occurring as a result of mitochondrial dysfunction.

[0155] Copper is an essential component for epithelial-mesenchymal transition. The striking effect of metformin on mesenchymal cancer cells, the accumulation of this biguanide in mitochondria, and the effect of metformin on copper homeostasis in this organelle led us to investigate the putative dependency of mesenchymal cancer cells on mitochondrial copper. Western blot analysis revealed that EGF-induced EMT significantly increased mitochondrial copper-containing proteins, including superoxide dismutase 1 (SOD1) and cytochrome c oxidase subunit 4 (Cox4), and increased levels of Ctr1 and mitochondrial copper (Figures 5A-C). In agreement with previous findings showing that mitochondrial load is higher in surviving cancer cells, these data suggested that in this cellular state there is a higher need for energy production, a biological process that is highly dependent on metalloproteins and metal cofactors essential to the mitochondrial ETC. Importantly, the addition of copper synergized with EGF treatment, shifting cancer cells towards a mesenchymal phenotype, as indicated by increased levels of mesenchymal markers (e.g., vimentin, fibronectin) and CD44. 高 / CD24 低 as defined by the percentage of cells containing the cell surface marker pattern (Figure 5D and Figure 6).

[0156] Finally, the induction of EMT in MDA-MB-468 cells using EGF was prevented by metformin and metforminin, as demonstrated by the subcellular localization and phalloidin staining of E-cadherin, the levels of the mesenchymal markers fibronectin, vimentin, the EMT transcription factors (TFs) Zeb1, Snail, and Slug, and the CD44 高 / CD24 低 As defined by the percentage of cells containing the cell surface marker pattern (Figure 5E-G), these data reveal the dependency of mesenchymal cancer cells on mitochondrial copper and provide a rationale for the heightened sensitivity of these cells to biguanides.

[0157] method Reagents. Copper chloride (CuCl2, 459097, Sigma Aldrich), ferrostatin-1 (Fer-1, SML0583, Sigma Aldrich, 10 μM for 72 hours), human epidermal growth factor (EGF, 130-093-825, Miltenyi Biotech, 100 ng / mL for 96 hours (additional 72 hours for cell viability measurements), carbonyl cyanide m-chlorophenylhydrazone (CCCP, 50 μM), metformin (1,1-dimethylbiguanide hydrochloride, J63361, Alfa Aesar, 12 mM unless otherwise stated), metforminin (Met, in-house discovery, 2 mM unless otherwise stated), necrostatin-1 (N9037, Sigma Aldrich, 20 μM for 72 h), turn-off Cu(II) probe (10 μM for 1 h), turn-off Fe(II) probe (10 μM for 1 h), and Z-VAD-FMK (550377, BD Biosciences, 50 μM for 72 h).

[0158] antibody. Caspase 3 (9665S, Cell Signaling Technology, WB 1:500), CD24-APC (2155590, Sony Biotechnology Inc., FC 1:100), CD44-PE (FAB4948P, R&D Systems, FC 1:100), copper uptake protein 1 (Ctr1, ab129067, Abcam, WB 1:1000 or FC 1:100), cytochrome c (12963, Cell Signaling Technology, IF 1:500), cytochrome c oxidase subunit 4 (Cox4, ab16056, Abcam, WB 1:2000), E-cadherin (20023195, Cell Signaling Technology, WB 1:1000), E-cadherin (610181, BD Antibodies against IF-1:200, Fibronectin (F0791, Sigma Aldrich, WB 1:1000), DyLight™ 488 phalloidin (12935S, Cell Signaling Technology, IF 1:40), Slug (9585S, Cell Signaling Technology, WB 1:500), Snail (3895, Cell Signaling Technology, WB 1:500), Superoxide dismutase 1 (SOD1, ab13498, Abcam, WB 1:1000), γ-tubulin (T5326, Sigma Aldrich, WB 1:2000), Vimentin (3932, Cell Signaling Technology, WB 1:500), Zeb1 (sc-81428, Santa Cruz Biotechnology, WB Secondary antibodies for WB: HRP anti-mouse (A90-116P, Bethyl Laboratories, WB 1:30000) and HRP anti-rabbit (A120-108P, Bethyl Laboratories, 1:30000).Secondary antibodies for IF and FC: Alexa Fluor 488 conjugate (A-11017 mouse, A-11008 rabbit, Life Technologies, IF 1:500), Alexa Fluor 594 conjugate (A-11032 mouse, A-11072 rabbit, Life Technologies, IF 1:500), Alexa Fluor 647 conjugate (A-20991 rabbit, A-20990 mouse, Life Technologies, IF 1:500). All antibodies were diluted in blocking solution (2 or 5% BSA, 0.1% Tween-20 / TBS).

[0159] Cell culture: Dulbecco's Phosphate Buffered Saline (14190-094, 500 mL, Gibco), Dulbecco's Modified Eagle Medium (DMEM) / F12 (31331-028, 500 mL, Gibco), DMEM GlutaMAX™ (61965059, Thermo Fisher Scientific), Fetal Bovine Serum (FBS, 10270-106, Gibco), Hydrocortisone (H0888, Sigma Aldrich), Insulin (I0516 or I9278, Sigma Aldrich), PEN-STREP (DE17-602E, BioWhittaker, Lonza), Puromycin dihydrochloride (A11138-02, Life Technologies). MCF-7 (ATCC®, HTB-22™), MDA-MB-231 (ATCC®, HTB-26™), MDA-MB-468 (ATCC®, HTB-132™), and circulating tumor cells were grown in DMEM supplemented with 10% FBS and 1% penicillin (100 U / mL), streptomycin (100 μg / mL), and incubated at 37° C. with 5% CO2. To induce EMT, cells were treated with epidermal growth factor EGF (100 ng / mL) (Miltenyi Biotec) for 72 hours. Human mammary epithelial cell lines, designated HMLER cells, infected with retroviruses carrying hTERT, SV40, and oncogenic alleles of HrasV12, were a generous gift from A. Puisieux. HMLER cells were cultured in DMEM / F12 supplemented with 10% FBS, 10 μg / mL insulin, 0.5 μg / mL hydrocortisone, 10 ng / mL hEGF, and 0.5 μg / mL puromycin. Mycoplasma testing was performed using a PCR mycoplasma detection kit (G238, Applied Biological Materials).

[0160] Cell viability. Cell viability was performed by plating 1000 cells / well in 96-well plates using the CellTiter-Blue® viability assay according to the manufacturer's protocol. Cells were treated for 72 hours as indicated. CellTiter-Blue® reagent (G8081, Promega) was added after 72 hours of treatment and cells were incubated for 1-2 hours before recording fluorescence intensity (e.g., 560 / 20 nm; em. 590 / 10 nm) using a Perkin Elmer Wallac 1420 Victor2 microplate reader.

[0161] Cell imaging. For immunofluorescence, cells were blocked with 2% BSA, 0.2% Tween-20 / PBS (blocking buffer) for 20 min at room temperature. Coverslips were incubated with 50-100 μL of diluted primary antibody in blocking buffer for 1 h at room temperature. Coverslips were then washed 3 times with blocking buffer and incubated with the appropriate secondary antibody for 1 h as described above. Coverslips were washed 3 times with PBS and mounted using Vectashield Mounting Medium with DAPI (H-1200, VECTOR Laboratories). Fluorescent images were acquired using a Deltavision real-time microscope (Applied Precision). 2D and 3D images were acquired using 60× / 1.4NA and 100× / 1.4NA objectives, which were deconvolved with SoftWorx (Ratio conservative-15 replicates, Applied Precision) and processed with ImageJ®.

[0162] Chemical labeling of Met in cells. Cells were cultured at approximately 80% confluence and treated with 0.3 mM Met for 3 h. Cells were fixed with formaldehyde (2% in PBS, 12 min), then permeabilized (Triton X-100, 0.1% in PBS, 5–10 min) and washed three times with 1% BSA / PBS. Click reaction cocktail was prepared from Click-iT® EdU Imaging Kit (C10337, Life Technologies) according to the manufacturer's protocol. Briefly, 430 μL of 1× Click-iT® reaction buffer was mixed with 20 μL of CuSO4 solution, 1.2 μL of AlexaFluor® azide, and 50 μL of reaction buffer additive (sodium ascorbate) to a final volume of approximately 500 μL. Coverslips were incubated with the click reaction cocktail in the dark at room temperature for 30 min and then washed three times with PBS. Immunofluorescence was then performed as indicated.

[0163] Western blotting. Cells were treated as indicated and then washed with PBS. Proteins were solubilized in Laemmli buffer supplemented with benzonase (70664-3, VWR, 1:100). Extracts were incubated for 1 h at 37°C. Proteins were separated using precast protein gels (Biorad) and the Trans-Blot® Turbo™ transfer system (Biorad). Membranes were incubated overnight with primary antibodies in 5% FBS in PBS, 0.1% Tween-20. Primary antibodies were detected using horseradish peroxidase (HRP)-conjugated secondary antibodies (Jackson Laboratories) and the Super Signal enhanced chemiluminescence detection kit (Thermo Scientific). Signals were revealed using a Fusion Solo S Imaging System (Vilber).

[0164] Flow cytometry. Metal-specific probes were synthesized as described above. Cells were treated as indicated. For mitochondrial copper and iron probes, cells were incubated with the relevant probe before analysis by flow cytometry. Cells were trypsinized and washed twice with ice-cold PBS. For antibodies, cells were suspended in ice-cold PBS containing 2% FBS and 1 mM EDTA (incubation buffer) and incubated with the relevant antibody for 20 min at 4° C. Cells were then washed twice with ice-cold PBS and suspended in incubation buffer before analysis by flow cytometry. At least 100,000 cells were counted for each condition. Data were recorded on a BD Accuri™ C6 (BD Biosciences) and processed using Cell Quest (BD Biosciences) and FlowJo (FlowJo, LLC) software.

[0165] Copper and iron levels. Cells were incubated with the relevant probes as described in the Reagents section. Cells were then analyzed by either fluorescence microscopy or flow cytometry as described in the Cell Imaging and Flow Cytometry section.

[0166] Cyclic voltammetry. Cyclic voltammetry experiments were performed using a three-electrode cell under argon. A saturated calomel electrode (SCE) was used as the reference, a stationary glassy carbon (GC) electrode with a diameter of 3 mm was selected as the working electrode, and a platinum wire was selected as the counter electrode. All cyclic voltammograms were recorded at room temperature using a μ-autolab III from Metrohm at a scan rate of 0.5 V / s using Nova software. MeCN was used as degassed HPLC grade from Carlo Erba. Water was mQ H2O. A copper-containing solution was prepared with 0.3 M nBu4NBF4 (180 mg) in MeCN (1.8 mL) and 200 μL of a 20 mM Cu(MeCN)4PF6 stock solution in MeCN (7.4 mg / mL). Then, 20 μL of a 400 mM metformin stock solution in mQ H2O (or PBS) (66.2 mg / mL) was added. Other MeCN solutions were prepared with 0.3 M nBu4NBF4 (200 mg) in MeCN (2 mL) and 20 μL of a 200 mM Fe(NO3)3 stock solution in mQ HO (48.4 mg / mL) and / or 20 μL of a 400 mM metformin stock solution in mQ HO (66.2 mg / 1 mL). All aqueous solutions were prepared with 0.3 M Na2SO4 (85.2 mg) in mQ HO (2 mL) and 20 μL of a 200 mM Fe(NO3)3 stock solution in mQ HO (48.4 mg / mL) and / or 20 μL of a 400 mM metformin stock solution in mQ HO (66.2 mg / mL).

[0167] ROS levels. Mitochondrial ROS were measured using MitoSOX™ Red Mitochondrial Superoxide Indicator (M36008, Thermo Fisher Scientific) according to the manufacturer's protocol. Briefly, cells were treated with red fluorescent dye for the indicated times, trypsinized, washed, and resuspended in ice-cold PBS containing 2% FBS and 1 mM EDTA, and then analyzed by flow cytometry.

[0168] Mitochondrial membrane potential. Measurements were performed using the MitoProbe™ JC-1 Assay Kit (M34152, Thermo Fischer Scientific) according to the manufacturer's protocol. Briefly, cells were treated with 2 μM JC-1 or CCCP (m-chlorophenylhydrazone) control for the indicated times. Cells were then trypsinized, washed, and resuspended in ice-cold PBS containing 2% FBS and 1 mM EDTA, and then analyzed by flow cytometry.

[0169] Cell death. Cells were treated as indicated in the figures. After treatment, cell death was quantified using the Annexin V-FITC(A) / Propidium Iodide (PI) assay according to the manufacturer's protocol (Annexin V-FITC Apoptosis Detection Kit II, 556570, BD Pharmingen™). Data were analyzed by an LSRFortessa™ flow cytometer (BD Biosciences, San Jose, CA) and processed using Cell Quest (BD Biosciences) and FlowJo (FLOWJO, LLC) software.

[0170] chemical synthesis General information: All starting materials were purchased from commercial sources and were either used without further purification or purified according to Purification of Laboratory Chemicals (Armarego, WLF, Chai, CLL 5th edition). Reactions were monitored by thin layer chromatography (TLC) using TLC silica gel coated aluminum plates 60F-254 (Merck). Metformin was purified using a Combiflash RF+Teledyne Isco system. 1 H NMR spectra were recorded at 300 MHz. 13C NMR was recorded at 75 MHz, 298 K. Chemical shifts are reported as δ values ​​in ppm, residual non-deuterated solvent was used as internal standard, and coupling constants (J) are expressed in Hz. The following abbreviations are used: s, singlet; t, triplet; td, double triplet. UPLC traces and electrospray mass spectra were obtained on a Waters Acquity SQD2 UPLC-MS system. HRMS was measured using a Bruker maXis LC-QTOF.

[0171] Synthesis of metformin (Met). In a sealed tube, commercially available dicyandiamide (169 mg, 2.0 mmol) and N-methylbut-3-yn-1-amine (200 mg, 2.4 mmol) were suspended in 20 mL of xylene, then 2.5 mL of 1 M aqueous HCl was added. The tube was closed and heated at 140° C. for 5 h. After this time the reaction was cooled to room temperature, MeOH was added, and the solvent was then removed under reduced pressure. The crude residue was purified by chromatography (DCM / MeOH, 10-20%) using a Combiflash instrument to give metformin as a white solid (97 mg, 24%). 1 H NMR (300 MHz, MeOD) δ3.60 (t, J = 7.0 Hz, 1H), 3.09 (s, 1H), 2.50 (td, J = 7.0, 3.0 Hz, 1H), 2.36 (t, J = 3.0 Hz, 1H). 13 C NMR (75 MHz, MeOD) δ160.8, 160.5, 81.9, 71.4, 50.3, 36.7, 18.3. C7H 14 N5[M+H] + HRMS (ESI-TOF) calculated for 168.1244, found 168.1246.

Claims

1. Formula (I) 【Chemistry 1】 {In the formula, ・R 1 is C 1 ~C 6 Alkyl, -(CH 2 ) a -C≡CH, -(CH 2 ) b -P + Ph 3 , C 3 ~C 10 Cycloalkyl, C 3～ C 10 Cycloheteroalkyl, C 6 ~C 12 Aryl and C 5 ~C 12 heteroaryl, wherein "a" is an integer selected from 1 to 6, preferably 2 to 4, and "b" is an integer selected from 5 to 11, preferably 6 to 10, said alkyl, cycloalkyl, cycloheteroalkyl, aryl, or heteroaryl being optionally substituted with an R or R' group; R 2 teeth, - -(CH 2 ) d -C≡CH, where "d" is an integer selected from 1 to 6; and - -(CH 2 ) e -CH=CH 2 where "e" is an integer selected from 2 to 6. selected from the group consisting of; ・R is C 1 ~C 6 Alkyl, C 1 ~C 6 Alkyloxy, C 1 ~C 6 Halogenoalkyl, C 3 ~C 10 Cycloalkyl, C 3～ C 10 Cycloheteroalkyl, C 6 ~C 12 Aryl and C 5 ~C 12 heteroaryl, wherein the group is optionally substituted with an R′ group; ・R' is C 1 ~C 6 Alkyl, C 1 ~C 6 Alkyloxy, C 1 ~C 6 Thioalkyl, halogen, C 1 ~C 6 Halogenoalkyl, hydroxyl (-OH), cyano (-CN), nitro, amino (-NH 2 ), carboxyl (-COOH), phosphate (PO 4 - ), amide (-CONH 2 ), -COOR'', -NHR'', -NR''R''', -COR'', -CONHR'', -NHCOR'', -NHSO 2 R'', -SOR'', -SO 2 R'', -SONR''R'''', -SO 2 N.R.'R.', C 3 ~C 10 Cycloalkyl, C 3～ C 10 Cycloheteroalkyl, C 6 ~C 12 Aryl and C 5 ~C 12 and heteroaryl, wherein said cycloalkyl, cycloheteroalkyl, aryl or heteroaryl is selected from the group consisting of halogen, hydroxyl, cyano, nitro, amino, or C 1～ C 3 Optionally substituted with alkoxy, R'' and R'' are H or C 1 ~C 6 alkyl} or a pharma- ceutically acceptable salt thereof.

2. R 2 but, - -(CH 2 ) d -C≡CH, where "d" is an integer selected from 2 to 4; and - -(CH 2 ) e -CH=CH 2 wherein "e" is an integer selected from 2 to 4; 2. The compound of claim 1, selected from the group consisting of:

3. ・R 1 Methyl, -(CH 2 ) a -C≡CH and -(CH 2 ) b -P + Ph 3 wherein "a" is an integer selected from 2 to 4 and "b" is an integer selected from 6 to 10; ・R 2 but, - -(CH 2 ) d -C≡CH, where "d" is an integer selected from 2 to 4; and - -(CH 2 ) e -CH=CH 2 where "e" is an integer selected from 2 to 6.

2. The compound of claim 1, selected from the group consisting of:

4. R 1 and R 2 At least one of the groups is -(CH 2 ) a 4. The compound of claim 3, wherein "a" is an integer from 1 to 6. 【Request 5】 【Chemical 2】 wherein n is an integer selected from 3 to 9; 2. The compound of claim 1, which is selected from the group consisting of: 【Request 6】 【Chemical 3】 wherein n is an integer selected from 3 to 9; 2. The compound of claim 1, which is selected from the group consisting of:

7. 10. A conjugate comprising a compound of any one of claims 1 to 6 covalently attached to a drug, toxin, or label.

8. 8. A pharmaceutical or veterinary composition comprising a compound of formula (I) as defined in any one of claims 1 to 6 or a pharma- ceutically acceptable salt thereof, or a conjugate as defined in claim 7.

9. 9. A compound according to any one of claims 1 to 6, a conjugate according to claim 7, or a pharmaceutical composition according to claim 8, for use as a medicament.

10. 9. A compound according to any one of claims 1 to 6, a conjugate according to claim 7, or a pharmaceutical composition according to claim 8, for use in the treatment of cancer.

11. 11. The compound, conjugate or pharmaceutical composition for use according to claim 10, wherein the cancer is selected from the group consisting of rectal cancer, colon cancer, gastric cancer, head and neck cancer, thyroid cancer, cervical cancer, uterine cancer, breast cancer, ovarian cancer, brain cancer, lung cancer, skin cancer, bladder cancer, blood cancer, kidney cancer, liver cancer, prostate cancer, multiple myeloma, and endometrial cancer.

12. 12. The compound, conjugate or pharmaceutical composition for use according to claim 11, wherein the breast cancer is triple negative breast cancer, and / or the brain cancer is glioblastoma or neuroblastoma, and / or the lung cancer is small cell lung cancer or non-small cell lung cancer.

13. 13. The compound, conjugate or pharmaceutical composition for use according to claim 11 or 12 in combination with radiation therapy and / or another drug.

14. 14. The compound, conjugate or pharmaceutical composition for use according to claim 13, wherein said further drug is an anti-tumor drug.

15. 10. A composition comprising a compound as defined in claim 4 or 7 for use in medical imaging or diagnosis.

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