Novel therapeutic vectors and prodrugs for cancer treatment

Abstract

The present invention relates to compounds having formula (I), as well as corresponding prodrugs, pharmaceutical compositions containing said compounds, particularly for treating cancer. TIFF2022531612000041.tif40146

Description

[Technical field]

[0001] The present invention relates to novel therapeutic vectors that allow the immobilization of anti-cancer drugs and their corresponding prodrugs. The present invention also relates to said vectors and prodrugs for use in cancer therapy. [Background technology]

[0002] Cancer is currently among the most common etiologies. In particular, it is today one of the main causes of death in France as well as in most industrialized countries. Death usually results from the occurrence of metastases and the lack of therapeutic approaches, at least for the most frequent tumor sites: breast, colorectal, prostate, pancreatic and lung cancer. In later cancer stages, cancer chemotherapy remains the primary and often the only available treatment method. However, chemotherapy does not show complete success against many common solid tumor types. Most anticancer drugs lack inherent antitumor selectivity. Secondly, chemotherapy is often accompanied by severe side effects due to the destruction of normal tissues. As a result, the amount of drug that can be administered is usually insufficient to deliver a lethal concentration of anticancer drugs to the tumor site. Moreover, the lack of selectivity of cytotoxic drugs significantly increases the risk of the development of cellular resistance by tumor cells. Thus, in view of the non-specific toxicity of most anticancer drugs, the development of more selective chemotherapy approaches is of major interest to fight cancer.

[0003] Within the framework of this concept, intensive research has been carried out on the development of self-responsive chemical systems programmed to selectively deliver potent cytotoxic drugs to tumor sites. Such systems are usually complex molecular assemblies assembled into structures consisting of (1) targeting units that allow the recognition of tumor-associated specificities and (2) either enzymatic or chemical triggers that can be activated only in cancer tissues to induce the release of the drugs in a strictly controlled manner.

[0004] Most of the drug delivery systems developed to date have been designed to target the surface specificities (e.g., membrane receptors or antigens) of cancer cells. In this approach, the molecular assembly includes either monoclonal antibodies or low molecular weight ligands that display high affinity for the corresponding tumor-associated cell surface markers. Upon detection by the targeting unit, the entire system is internalized into the cell by receptor-mediated endocytosis. Once inside the cell, a trigger is activated to induce the release of the active drug selectively in the intracellular medium. Several drug delivery systems of this type are currently undergoing clinical evaluation for a wide variety of applications in oncology. Summary of the Invention [Problem to be solved by the invention]

[0005] However, the Achilles heel of drug delivery systems designed to target cell surface specificity lies in the heterogeneity of cancer tissue. In fact, not all cells in a tumor mass are identical and display different concentrations of a given cell surface marker. Thus, only cells expressing sufficient levels of the selected tumor-associated marker will be directly acted upon by this type of targeting system. In this regard, the use of enzyme-reactive prodrugs, which can be selectively activated by the corresponding enzymes naturally overexpressed in the tumor microenvironment, offers a valid alternative to this targeting approach. In this case, the anticancer drug is released into the extracellular medium and can passively penetrate inside various types of surrounding malignant cells, whatever their membrane properties. [Means for solving the problem]

[0006] It is an object of the present invention to provide a system for the selective release of anticancer drugs in the tumor microenvironment.

[0007] Thus, the present invention provides a compound of formula (I):

[0008] [ka]

[0009] [During the ceremony: R a may be optionally interrupted by H or one or more oxygen atoms, (C 1 ~C 12 ) represents an alkyl group, R a is preferably H; L represents a group having the formula (II):

[0010] [ka]

[0011] where: A is an anticancer drug; Y is an electron-withdrawing group; X is -O-; G is a glucuronyl group or a derivative thereof; L 1 is a linker represented by formula (III): -A 1 -A 2 -A 3 -A 4 -A 5 -A 6 -A 7 - (III) where: A 1 (C 1 ~C 6 ) an alkylene group; A 2 is a group obtained by click chemistry; A 3 (C 1 ~C 6 ) an alkylene group; A 4 is -C(=O)-NR b -, -C(=S)-NR b -, -NR b -C(=O)-, -NRb -C(=S)- and NR b R b is H or (C 1 ~C 12 ) represents an alkyl group; A 4 is preferably -C(=O)-NR b - group, more preferably -C(=O)-NH-; A 5 is interrupted by at least one oxygen atom (C 1 ~C 32 ) alkylene group, preferably interrupted by at least one oxygen atom (C 1 ~C 6 ) alkylene groups, and more preferably polyoxyalkylenated groups; A 6 is a group obtained by click chemistry; and A 7 (C 1 ~C 6 ) an alkylene group;

[0012] L' represents a group having the formula (IV): -A 8 -A 9 -L”(IV) where A 8 is interrupted by at least one oxygen atom (C 1 ~C 6 ) an alkylene group, preferably a polyoxyalkylenated group; A 9 -NR c -, -O-, and S-, where R c is H or (C 1 ~C 12 ) represents an alkyl group; and L" is a group capable of reacting with an amino, hydroxyl or thiol function. or a pharma- ceutically acceptable salt, or a racemate, diastereomer, or enantiomer thereof.

[0013] The compound of the present invention is a compound having a gallic acid structure capable of fixing three molecules of an anticancer drug (A) via a linker L.

[0014] The compounds described herein may have asymmetric centers. Compounds of the present invention containing asymmetrically substituted atoms may be isolated in optically active or racemic forms. Methods for preparing optically active forms are well known in the art, for example, by resolution of racemic forms or by synthesis from optically active starting materials. All chiral, diastereomeric, racemic, and geometric isomeric forms of the compounds are intended, unless the stereochemistry or isotopic form is specifically indicated.

[0015] The term "pharmaceutical acceptable salts" refers to salts that retain the biological effectiveness and properties of the compounds of the present invention and are not biologically or otherwise undesirable. In many cases, the compounds of the present invention can form salts with acids and / or bases due to the presence of amino and / or carboxyl groups or groups similar thereto. On the one hand, pharmaceutical acceptable acid addition salts can be prepared from inorganic and organic acids, while pharmaceutical acceptable base addition salts can be prepared from inorganic and organic bases. For a review of pharmaceutical acceptable salts, see Berge et al. (1977) J. Pharm. Sd. Vol. 66, 1. The expression "non-toxic pharmaceutical acceptable salts" refers to non-toxic salts formed with non-toxic pharmaceutical acceptable inorganic or organic acids or inorganic or organic bases. For example, such salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and the like, as well as those prepared from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, fumaric acid, methanesulfonic acid, toluenesulfonic acid, and the like.

[0016] As mentioned above, A is an anticancer drug.

[0017] According to one embodiment, the anticancer agent is selected from a cytostatic agent, antimetabolite, a DNA intercalating agent, a topoisomerase I and II inhibitor, a tubulin inhibitor, an alkylating agent, neocarzinostatin, calicheamicin, dynemicin or esperamicin A, a liposome inhibitor, a tyrosine phosphokinase inhibitor, a compound that induces cell differentiation, a histone deacetylase inhibitor, a small molecule immunomodulator or a small molecule that targets cancer stem cells.

[0018] Even more particularly, the anticancer agent of the present invention is selected from the group consisting of cytostatics and antimetabolites, such as 5-fluorouracil, 5-fluorocytidine, 5-fluorouridine, cytosine arabinoside or methotrexate, DNA intercalators, such as doxorubicin, daunomycin, idarubicin, epirubicin or mitoxantrone, topoisomerase I and II inhibitors, such as camptothecin, etoposide or m-AMSA, tubulin inhibitors, such as vincristine, vinblastine, vindesine, taxol, nocodazole or colchicine, alkylating agents, such as cyclophosphamide, mitomycin C, rachelmycin, cisplatin, mustard gas phosphoramide, melphalan, bleomycin, N-biphenylsulfonyl esterase, cyclophosphamide ... from (2-chloroethyl)-4-hydroxyaniline, or from neocarzinostatin, calicheamicin, dynemicin or esperamicin A, or from ribosomal inhibitors such as verrucarin A, from tyrosine phosphokinase inhibitors such as quercetin, genistein, erbstatin, tyrphostin or rohitukin and derivatives thereof, from compounds inducing cell differentiation such as retinoic acid, butyric acid, phorbol esters or aclacinomycin, from histone deacetylase inhibitors such as CI-994 or MS275, from immunomodulators such as Imiquimod, and from small molecules targeting cancer stem cells such as hedgehog (pathway) inhibitors such as cyclopamine derivatives.

[0019] According to a preferred embodiment, A is an anti-cancer drug that contains at least one primary or secondary amine function.

[0020] Preferably, according to the present invention, the anti-cancer drug A is modified so that it can be attached to the -C(=O)-O- group of the L group of formula (II) above.

[0021] According to one embodiment, A is a group of the dolastatin family or a derivative thereof. In the context of the present invention, a "derivative" of the dolastatin family refers to a compound that is structurally closely related and possesses comparable biological properties, in particular the ability to inhibit tubulin polymerization, ultimately to inhibit cell mitosis. This may be an issue especially in the case of substitution or deletion derivatives.

[0022] The dolastatin family represents a class of compounds having a structure of at least four amino acids, at least three of which are unique thereto, i.e., different from the 20 amino acids commonly found in nature.

[0023] Reference is made in particular to document WO2004 / 010957, which describes compounds which correspond to those suitable for the present invention.

[0024] In one particularly preferred embodiment of the invention, A represents a group derived from dolastatin 10, auristatin PE, auristatin E, monomethylauristatin E, and derivatives thereof, preferably a group derived from monomethylauristatin E or a derivative thereof.

[0025] The structural difference between dolastatin 10 and the synthetic compounds of the auristatin subfamily resides in the replacement of the aminothiazole phenethyl group, particularly at the C-terminal position of dolastatin 10, with a norephedrine unit in the case of auristatin PE, auristatin E or monomethylauristatin.

[0026] For the purposes of the present invention, a derivative of dolastatin 10, auristatin PE, auristatin E or monomethylauristatin E has a chemical structure closely related to at least one of the active agents and has the antimitotic activity attributed to the dolastatin family of compounds. The structural difference can in particular be, for example, that at least one of the four amino acids that compose it is substituted on at least one side chain. This substitution is made so as to include or represent a linear, cyclic and / or branched alkyl group, an aryl group, a heterocycle or a carbocycle. This structural difference can also consist of a modification of the dolastatin 10, auristatin PE or auristatin E molecule, for example at the level of its tertiary amine at the N-terminal position, in order to make the tertiary amine function compatible with the establishment of a covalent bond with a linker arm.

[0027] According to a preferred embodiment, the anticancer agent A is monomethyl auristatin E (MMAE) or a derivative thereof.

[0028] Preferably, A is represented by the following formula (A-1):

[0029] [ka]

[0030] In the context of the present invention, an "electron-withdrawing group" refers to the property of an atom or group of atoms to withdraw electrons.

[0031] Preferably, in formula (II), Y is halogen, NO 2 and CF 3 More preferably, Y is an electron withdrawing group selected from the group consisting of NO 2 It is.

[0032] As described above, in formula (II), G is a glucuronyl group or a derivative thereof.

[0033] In the context of the present invention, G is configured to be enzymatically removed to provide for intramolecular rearrangement of the linker arm connecting it to a molecule of the dolastatin family, resulting in release of the active molecule (the A group).

[0034] Additionally, the enzymatically hydrolyzable glucuronyl groups of the present invention can impart tissue and / or cell specificity to the conjugates and prodrugs of the present invention.

[0035] β-glucuronidase is known to be an enzyme naturally occurring in high concentrations in the vicinity of many tumors. The conjugates and prodrugs of the present invention containing a glucuronyl group can therefore be advantageously activated at extracellular levels during prodrug monotherapy (or PMT). In the context of the present invention, the term "activation" refers to the release at the tumor site, for example the release of groups of the dolastatin family, so that they can exert their antimitotic biological activity.

[0036] In the context of the present invention, a "derivative" of the glucuronyl group refers to a compound that is structurally closely related and possesses comparable biological properties, in particular the ability to be an enzymatic substrate for β-glucuronidase, which can be problematic especially in the case of derivatives with substitution or deletion of one or more hydroxyl (-OH) or carboxylic acid (-COOH) groups.

[0037] Derivatives of the glucuronyl group include glucuronide ester groups.

[0038] As described above in formula (II), the linker L may be a divalent linker L bonded directly to an oxygen atom of the gallic acid structure of the compound of the invention. 1 Also includes.

[0039] In other words, the linker L 1 Each A 7 corresponds to the terminal group of each L group linked to each oxygen atom of the gallic acid structure. 7is linked to the oxygen atom carrying the L group.

[0040] Within the scope of this application, the term "alkyl", unless otherwise specified, means a saturated or unsaturated aliphatic hydrocarbon group which may be straight or branched having 1 to 12 carbon atoms in the chain. Preferred alkyl groups have 1 to 6 carbon atoms in the chain. "Branched" means that one or a lower alkyl group, such as a methyl, ethyl or propyl group, is attached to a linear alkyl chain. "Lower alkyl" means 1 to 4 carbon atoms in the chain which may be straight or branched.

[0041] As used herein, unless otherwise specified, the term "alkylene" refers to a divalent group containing 1 to 6 carbon atoms. The group has the formula (CH 2 ) n where n is an integer ranging from 1 to 6.

[0042] According to the present invention, in formula (III), A 2 and A 6 are groups obtainable by click chemistry. These groups are obtainable by click chemistry reactions.

[0043] The above-mentioned click chemistry reactions include, in particular, cycloadditions of unsaturated compounds, in particular the Diels-Alder reaction between dienophiles and dienes, and in particular the azide-alkyne 1,3-dipolar cycloaddition, preferably the copper-catalyzed azide-alkyne cycloaddition (CuAAC).

[0044] Other click chemistry reactions include those involving thiol functions, such as the formation of thioethers from alkenes and mixed disulfides, and reactions involving non-aldol electrophilic carbonyl groups, such as the formation of oxime ethers from oxyamines, hydrazones from hydrazines, or even thiosemicarbazones from thiosemicarbazines.

[0045] Click chemistry reactions can include reactions involving thiocarboxylic acids or thioesters, such as those which result in the formation of thioesters and amides, as well as reactions between azides and phosphines (e.g., the Staudinger ligation reaction).

[0046] Preferably, the group A 2 and A 6 is obtained by a reaction between two reactive functional groups, said reaction being selected from the group consisting of: Reaction between azides and alkynes, The reaction between an aldehyde or ketone and a hydrazide, Reaction between aldehydes or ketones and oxyamines, The reaction between azides and phosphines, Reaction between alkenes and tetrazines, The reaction between isonitriles and tetrazines, and Reaction between thiols and alkenes (thiol-ene reaction).

[0047] According to a preferred embodiment, A 2 and A 6 is a triazole group.

[0048] Preferably, A in formula (III) 2 is a triazole group, preferably a group having the formula (V):

[0049] [ka]

[0050] Preferably, A in formula (III) 6 is a triazole group, preferably a group having the formula (VI):

[0051] [ka]

[0052] According to one embodiment, L has the following formula (VII):

[0053] [ka]

[0054] A, Y and L 1 is as defined above. Preferably, in formula (VII), Y is a nitro group.

[0055] Preferably, in formula (VII), A is monomethylauristatin E (MMAE) or a derivative thereof, more preferably having the formula (A-1) defined above.

[0056] According to one embodiment, L has the following formula (VIII):

[0057] [ka]

[0058] Here, A, A 1 , A 3 , A 4 , A 5 and A 7 is as defined above.

[0059] Preferably, in formula (VII), A is monomethylauristatin E (MMAE) or a derivative thereof, more preferably having the formula (A-1) defined above.

[0060] According to one embodiment, in formula (VIII), A 4 is -C(=O)-NH-.

[0061] According to one embodiment, in formula (VIII), A 5 is the formula -CH 2 -(CH 2 -O-CH 2 ) n -CH 2-group, where n is an integer from 1 to 12, inclusive.

[0062] The present invention relates to compounds having formula (I) as defined above, wherein L has the following formula (IX):

[0063] [ka]

[0064] i is an integer from 1 to 6, inclusive; j is an integer from 1 to 10, inclusive; n is an integer from 1 to 12, inclusive; and k is an integer from 1 to 6 inclusive.

[0065] Preferably, in formula (IX), i=1. Preferably, in formula (IX), j=4. Preferably, in formula (IX), n=10. Preferably, in formula (IX), k=1.

[0066] In formula (I), as described above, L″ (which is attached to the gallic acid structure via the linker L′) is a group capable of reacting with an amino, hydroxyl or thiol function.

[0067] In the context of the present invention, "groups capable of reacting with amino, hydroxyl or thiol functions" refers to groups, usually hydrocarbon-based groups or units, that have a chemical function that is capable of interacting with a free secondary amino, hydroxyl or thiol function, thereby establishing a covalent bond between the conjugate molecule and a separate chemical entity carrying this function, and that is compatible with generating this covalent bond function. In the context of the present invention, this separate chemical entity is more particularly a macromolecule naturally occurring in a living organism, and preferably an endogenous albumin molecule, such as human serum albumin.

[0068] According to a preferred embodiment, L'' comprises a maleimide group. According to a preferred embodiment, L″ has the formula:

[0069] [ka]

[0070] where L''' is optionally substituted with an electron withdrawing group, particularly halo(C 1 ~C 6 ) alkyl groups, e.g., CF 3 It may be replaced by (C 1 ~C 12 ) an alkylene group or a phenylene group which may be optionally substituted with electron-withdrawing groups, particularly halogens.

[0071] According to a preferred embodiment, L'' is a maleimidocaproyl group.

[0072] According to a preferred embodiment, L″ is a group having one of the following formulae:

[0073] [ka]

[0074] According to one embodiment, in formula (IV), A 8 is the formula -CH 2 -(CH 2 -O-CH 2 ) m -CH 2 -, where m is an integer from 1 to 12 inclusive, and in particular is equal to 3.

[0075] The present invention relates to compounds having formula (I) as defined above, wherein L' has the following formula (X):

[0076] [ka]

[0077] m is an integer from 1 to 6, inclusive, preferably 3; and p is an integer from 1 to 6 inclusive, and is preferably 5.

[0078] According to one embodiment, the compound of the invention, which may also be referred to as a conjugate, has formula (I) as defined above, wherein the linker L has formula (VII), preferably formula (VIII), more preferably formula (IX) as defined above, and the linker L' has formula (X) as defined above.

[0079] Preferred compounds of the invention have the following formula (XI):

[0080] [ka]

[0081] Another preferred compound of the present invention has the following formula (XI'):

[0082] [ka]

[0083] Another preferred compound of the present invention has the following formula (XI″):

[0084] [ka]

[0085] The present invention also relates to a prodrug comprising a compound of formula (I) as defined above, linked via a covalent bond to albumin or a derivative or fragment thereof.

[0086] According to the present invention, the term "prodrug" refers to a molecule capable of transporting an anticancer drug (A) in an inactive form in an organism and releasing said compound in an organ, tissue or cell, where it can be specifically targeted under the action of β-glucuronidase.

[0087] More specifically, such prodrugs advantageously correspond to the following general formula (XII):

[0088] [ka]

[0089] Here, L and R a is as defined above.

[0090] The L' units result from the reaction between L' groups whose constituent parts contain units capable of reacting on the one hand with free amino, hydroxyl or thiol functions, in particular with free thiol functions carried by macromolecules, preferably albumin molecules, more preferably serum albumin.

[0091] Within the scope of this application, prodrugs can be formed in vivo or in vitro with macromolecules, preferably albumin molecules.

[0092] Thus, endogenous or exogenous albumin may be envisaged, in particular human serum albumin, recombinant albumin or even fragments of albumin.

[0093] According to one embodiment, the formation of a covalent bond between a molecule of the complex (conjugate) described according to the invention and a molecule of endogenous albumin, in particular a molecule of human serum albumin, or a derivative thereof, is carried out in vivo.

[0094] In one embodiment, the prodrug of the present invention comprises at least one molecule of the conjugate of the present invention linked via a thioether bond to the sulfur atom of cysteine ​​at position 34 of a molecule of endogenous albumin.

[0095] Indeed, covalent bonds have been shown to be spontaneously established in vivo between compounds, one of which can react with the thiol function of cysteine ​​34 of human serum albumin (Kratz et al., 2002, J. Med. Chem.).

[0096] According to one embodiment, the present invention also relates to a prodrug of formula (XII) as defined above, wherein L' 1 Albumin has the following formula (XIII):

[0097] [ka]

[0098] m and p are as defined above.

[0099] According to another embodiment, the prodrugs of the present invention may be formed in vitro by at least one conjugate molecule of formula (I) linked via a covalent bond to an albumin molecule, a recombinant albumin molecule or a fragment of an albumin molecule or a derivative thereof.

[0100] For the purposes of the present invention, "fragment of an albumin molecule" means a fragment of an albumin molecule having a size sufficient to ensure sufficient bioavailability, permeability with respect to tumor tissue and impermeability with respect to the endothelial barrier of healthy tissue of the produced prodrug.

[0101] In this particular embodiment, in vitro coupling between the conjugate of general formula (I) and the albumin molecule, recombinant albumin molecule or fragment of an albumin molecule via its L' group can be achieved using free complementary reactive functional groups present on the albumin molecule, recombinant albumin molecule or fragment of an albumin molecule.

[0102] In one embodiment, the fragment of the albumin molecule may contain a cysteine ​​corresponding to cysteine ​​at position 34 of the endogenous albumin sequence. Surprisingly, coupling of the conjugate of general formula (I) with an albumin molecule results in the formation of a prodrug with the ability to be specifically delivered and targeted into the microenvironment of the tissue to be treated; the ability to be cleaved by β-glucuronidase in the microenvironment of the tissue to be treated, and Ability to undergo rearrangement of the linker arms to release the anticancer drug after cleavage of the glucuronyl group has no effect on

[0103] Furthermore, the coupling of the conjugate of general formula (I) through its L' group to the amino, hydroxyl or thiol functions of an albumin molecule, in particular an endogenous albumin molecule, does not have any effect on the ability of the anti-cancer drug to be consequently released and perform its biological activity.

[0104] Finally, the coupling of the conjugate of general formula (I) via its L' group with the amino, hydroxyl or thiol functions of albumin molecules, in particular endogenous albumin molecules, limits the excretion of the prodrugs by the kidney. The blood half-life of the prodrugs according to the invention is increased compared to that of prodrugs represented by anticancer drugs functionalized with glucuronyl groups. [Brief description of the drawings]

[0105] [Figure 1] FIG. 1: MIA PaCa2 tumor growth inhibition under the treatment with vehicle and RC-Alb (a compound of the invention of formula (XI)) (4.32 and 6.48 mg / kg). Injections were administered on days 37, 44, 50 and 64. The solid curve with black circles (-●-) corresponds to vehicle, the dotted curve with white circles (--○--) corresponds to RC-Alb at 4.32 mg / kg, and the dotted curve with black circles (--●--) corresponds to RC-Alb at 6.48 mg / kg. [Diagram 2] Figure 2. MIA PaCa2 tumor growth inhibition under therapy with Br-Alb (2 mg / kg, 1.1 x 10-6 mol / kg), RC-Alb (4.32 mg / kg, 0.7 x 10-6 mol / kg) and RC-Alb (6.48 mg / kg, 1.1 x 10-6 mol / kg). Injections were performed on days 37, 44, 50 and 64. The solid curve with black squares (-■-) corresponds to vehicle, the dotted curve with white circles (--○--) corresponds to RC-Alb at 4.32 mg / kg, and the dotted curve with black circles (--●--) corresponds to RC-Alb at 6.48 mg / kg. [Diagram 3] Body weights of mice treated with a single iv injection of RC-Alb, YG-Alb and AB-Alb at 9.72 mg / kg or 12.96 mg / kg. [Figure 4] Survival curves showing survival as a function of time in days. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0106] In another embodiment of the invention, the albumin molecule or albumin fragment of the prodrug may also be modified, particularly by glycosylation or by pegylation.

[0107] The present invention also relates to a compound or a conjugate of the invention having formula (I) as defined above, or a prodrug as defined above, for its use as a medicament.

[0108] The present invention also relates to a pharmaceutical composition comprising a compound having formula (I) as defined above, or a prodrug as defined above, or a pharma- ceutically acceptable salt thereof, and also at least one pharma- ceutically acceptable excipient.

[0109] While the compounds of the invention having formula (I) can be administered alone, it is preferred to present them as pharmaceutical compositions. Pharmaceutical compositions useful according to the invention, both for vertebrate and human use, comprise at least one compound having formula (I) as defined above, one or more pharma- ceutical acceptable carriers, and optionally, another therapeutic ingredient.

[0110] In certain preferred embodiments, the active ingredients required in a combination therapy may be combined in a single pharmaceutical composition for simultaneous administration.

[0111] As used herein, the terms "pharmaceutical acceptable" and grammatical variations thereof, when they refer to compositions, carriers, diluents, and reagents, that materials can be administered to a mammal without undesirable physiological effects, such as nausea, dizziness, stomach upset, and the like.

[0112] The preparation of pharmacological compositions containing active ingredients dissolved or dispersed therein is well known in the art and need not be limited based on formulation. Typically, such compositions are prepared as injectables, either as liquid solutions or suspensions. However, solid forms suitable for liquid or suspension, which are brought to a liquid state before use, can also be prepared. Such preparations may be emulsified. In particular, pharmaceutical compositions can be formulated in solid dosage forms, such as capsules, tablets, pills, powders, dragees or granules.

[0113] The choice of vehicle and the content of active substance in the vehicle are usually determined according to the solubility and chemical properties of the active compound, the specific method of administration, and the prescriptions to be observed in medical practice. For example, excipients such as lactose, sodium citrate, calcium carbonate, dicalcium phosphate, starch, alginic acid, and certain complex silicates in combination with lubricants such as magnesium stearate, sodium lauryl sulfate, and talc can be used to prepare tablets. For preparing capsules, lactose and high molecular weight polyethylene glycols are advantageously used. When aqueous suspensions are used, they can contain emulsifiers and agents that promote suspension. Sucrose, ethanol, polyethylene glycol, propylene glycol, glycerol, and chloroform, or mixtures thereof, may be used.

[0114] The compounds of formula (I) or conjugates, prodrugs or pharmaceutical compositions of the present invention may be administered orally, parenterally (subcutaneously, intravenously or intramuscularly) or locally by topical application to the skin or mucosa.

[0115] The conjugates, prodrugs or pharmaceutical compositions of the invention can in particular be administered alone or in combination with chemotherapy or radiotherapy, or in combination with other therapeutic agents, for example, in particular anti-cancer or anti-mitotic agents, as well as in combination with anti-inflammatory agents.

[0116] The dosages suitable for the present invention can be determined according to routine procedures commonly used in the art, and adjustment of said dosages is clearly part of the normal competence of a person skilled in the art.

[0117] In fact, it depends especially on the weight, age and sex of the individual to be treated, as well as on the advanced stage of the disease to be treated.

[0118] The present invention also relates to the compounds or prodrugs of formula (I) as defined above for use in the treatment and / or prevention of cancer.

[0119] In the context of the present invention, the terms "treat" or "treatment" as used herein means to reverse, alleviate, or inhibit the progression of the disease or condition or one or more symptoms of the disease or condition for which such term is used.

[0120] The present invention also relates to a method for treating cancer, comprising administering a conjugate or a prodrug of formula (I) as defined above or a pharmaceutical composition according to the present invention in combination with another treatment selected from the group comprising chemotherapy, radiation therapy, treatment with at least one anti-inflammatory drug, and combinations thereof.

[0121] According to one embodiment, the cancer is selected from solid cancers.

[0122] Solid tumors may include: neuroblastoma, glioma, osteosarcoma, retinoblastoma, soft tissue sarcoma, cancer of the central nervous system, nephroblastoma, lung cancer, breast cancer, prostate cancer, colorectal cancer, thyroid cancer, cervical cancer, endometrial cancer, ovarian cancer, kidney cancer, liver cancer, brain tumors, testicular cancer, pancreatic cancer, osteosarcoma, skin cancer, small intestine cancer, stomach cancer, esophageal cancer, pharyngeal cancer, and bladder cancer.

[0123] In one embodiment, the solid cancer is selected from the group comprising pancreatic cancer, lung cancer and breast cancer, preferably pancreatic cancer. EXAMPLES

[0124] chemistry General Experimental Method All reactions were carried out under an argon atmosphere. Solvents used were of HPLC quality unless otherwise stated. Chemicals were of analytical grade from commercial sources and were used without further purification. Reaction progress was monitored on precoated silica gel TLC plates MACHEREY-NAGEL ALUGRAM® SIL G / UV254 (0.2 mm silica gel 60). Spots were visualized by placing the TLC plate under 254 nm UV light and / or by immersing the TLC plate in a solution of phosphomolybdic acid (3 g) in ethanol (100 mL) followed by heating with a heat gun.

[0125] Automated chromatography was carried out using a COMBIFLASH® RF 200I TELEDYNE ISCO instrument equipped with UV and ESLD detectors and using flash cartridges Interchim® Silica 15 or 50 μm for normal phase chromatography and HP C18 RediSep® GOLD 4 g or 15.5 g for reversed phase chromatography.

[0126] 1 H, 19 F and 13 C NMR spectra were recorded on a Bruker 400 Avance III instrument equipped with an ultrashield magnet and a BBFO 5 mm broadband probe at 400 MHz, 376 MHz, and 100 MHz, respectively. Chemical shifts (δ) are reported in parts per million (ppm) from downfield to upfield and were referenced to residual solvent. Coupling constants (J) are reported in Hertz (Hz).

[0127] Exact masses were determined for all derivatives through their injection on a high-resolution ESI mass spectrometer at the CBM / ICOA FR2708 of the University of Orléans and at the Organic Analysis Center of the IC2MP of the University of Poitiers.

[0128] Analytical RP-HPLC was performed on a Dionex Ultimate 3000 system equipped with a UV / Visible variable wavelength detector and reverse phase column chromatography was performed on a MACHEREY-NAGEL NUCLEOSHELL® (150 / 4.6, RP18, 5 μm) at 30° C. and 1 mL / min.

[0129] Method 1 consisted of A (0.2% TFA in water) and B (CH 3 A linear gradient consisting of 1000 µL ...

[0130] Synthetic strategy to access RC-A1b

[0131] [ka]

[0132] 1. Synthesis of Compound 5

[0133] [ka]

[0134] Azide reduction PPh 3 (730 mg, 2.78 mmol, 1.5 equiv.) in THF (8 mL) 3 (590 mg, 1.85 mmol, 1 equiv.). The mixture was stirred at room temperature for 20 h and then at 50° C. for 5 h. After completion of the reaction, the mixture was cooled to room temperature, then water (2.5 mL) was added and the mixture was stirred for 18 h. The solvent was removed under reduced pressure to give the crude amine 4 was used in the next step without further purification.

[0135] Amide formation Dry CH 2 Cl 2 Carboxylic acid in (8 mL) 2(Haibin Gu, Polymer, 2018, 146, 275-290) (455 mg, 1.60 mmol, 1 equiv.) and crude amine 4 To a cooled solution (0 °C) of (1.85 mmol, 1.15 equiv.) of DMAP (215 mg, 1.76 mmol, 1.1 equiv.) and EDC HCl (337 mg, 1.76 mmol, 1.1 equiv.) were added. The resulting mixture was stirred at room temperature for 60 h and then cooled to 5°C. 2 Cl 2 (40 mL). The organic layer was washed with 1M HCl (40 mL). The aqueous layer was washed with CH 2 Cl 2 (3×40 mL). The combined organic layers were extracted with MgSO 4 The crude residue was purified by chromatography on a silica gel column (gradient elution CH 2 Cl 2 / acetone 95 / 5 → 80 / 20) to obtain the compound as a white solid. 5 (1.68 g, 86%)

[0136] R f : 0.65 (CH 2 Cl 2 / MeOH 95 / 5). 1 H NMR (400 MHz, CDCl 3 ) δ = 7.23 (s, 2H), 6.76 (bs, 1H), 5.00 (s, 1H), 4.81 (d, J = 2.3 Hz, 4H), 4.79 (d, J = 2.4 Hz, 2H), 3.83 - 3.54 (m, 12H), 3.49 (t, J = 5.1 Hz, 2H), 3.26 (m, 2H), 2.56 (t, J = 2.2 Hz, 2H), 2.46 (t, J = 2.4 Hz, 1H), 1.45 (s, 9H). 13 C NMR (100 MHz, CDCl 3) δ = 166.79, 156.13, 151.58, 140.06, 130.70, 108.20, 79.44, 78.92, 78.29, 77.36, 76.42, 75.68, 70.67, 70.60, 70.45, 70.36, 70.28, 70.04, 60.46, 57.43, 40.12, 28.55.

[0137] HRMS (ESI+): [M+Na] + C 29 H 38 N 2 NaO 9 Calculated value for 581.2470, found value 581.2487; [2M+Na] + C 58 H 76 N 4 NaO 18 Calculated value: 1139.5047, observed value: 1139.5085.

[0138] 2. Synthesis of Compound 7

[0139] [ka]

[0140] Cu(MeCN) 4 PF 6 (7.0 mg, 0.019 mmol, 0.12 equiv.) was dissolved in dry, degassed CH 2 Cl 2 Alkyne in (2 mL) 5 (89 mg, 0.159 mmol, 1 eq.) and azido-PEG10-amine 6 (285 mg, 0.541 mmol, 3.4 equiv.). The mixture was stirred at room temperature for 2 h. Upon completion, the resin QuadraPure® IDA (200 mg) was added to the mixture, stirred for an additional 2 h, and the resin was removed by filtration. The solvent was evaporated under reduced pressure and the crude product was purified by reversed phase chromatography on C18 grafted silica (gradient elution MeCN / H 2O 10 / 90→50 / 50 for 30 min) to give compound 1 as a colorless oil. 7 (203 mg, 60%)

[0141] R t : 5.72 minutes (method 1). 1 H NMR (400 MHz, MeOD) δ = 8.21 (s, 2H), 7.94 (s, 1H), 7.39 (s, 2H), 5.26 (s, 4H), 5.14 (s, 2H), 4.63 (t, J = 5.0 Hz, 4H), 4.54 (t, J = 5.0 Hz, 2H), 3.93 (t, J = 5.0 Hz, 4H), 3.85 (t, J = 5.0 Hz, 2H), 3.73 - 3.50 (m, 126H), 3.47 (t, J = 5.6 Hz, 3H), 3.19 (t, J = 5.6 Hz, 2H), 2.87 (t, J = 5.1 Hz, 6H), 1.42 (s, 9H).

[0142] 13 C NMR (100 MHz, MeOD) δ = 168.99, 158.38, 153.54, 145.22, 144.46, 141.53, 131.30, 126.57, 126.53, 108.48, 80.03, 72.09, 71.53, 71.48, 71.44, 71.23, 71.12, 70.64, 70.34, 66.97, 63.77, 51.50, 51.38, 41.75, 41.25, 41.19, 28.80.

[0143] HRMS (ESI+): [M+2H] 2+ C 95 H 178 N 14 O 39 Calculated for 1069.6182, Found for 1069.6170; [M+3H] 3+ C 95 H 179 N 14 O 39Calculated for 713.4145, Found 713.4138; [M+4H] 4+ C 95 H 180 N 14 O 39 Calculated value: 535.3127, measured value: 535.3125.

[0144] 3. Synthesis of Compound 9

[0145] [ka]

[0146] EDC HCl (134 mg, 0.7 mmol, 1 equiv.) was dissolved in dry CH 2 Cl 2 Carboxylic acid in (4.6 mL) 8 (100 mg, 0.7 mmol, 1 equiv.) and N-hydroxysuccinimide (80 mg, 0.7 mmol, 1 equiv.). The resulting mixture was stirred at room temperature for 60 h. Upon completion, the mixture was diluted with CH 2 Cl 2 (10 mL). The organic layer was washed with water (10 mL), then brine (10 mL) and MgSO 4 The crude residue was purified by chromatography on a silica gel column (gradient elution CH 2 Cl 2 / MeOH 100 / 0 → 99 / 1) to give the compound as a clear yellowish oil. 9 (135 mg, 80%)

[0147] R f : 033 (PE / AcOEt 70 / 30). 1 H NMR (400 MHz, CD 2 Cl 2 ) δ = 3.29 (t, J = 6.6 Hz, 2H), 2.78 (s, 4H), 2.61 (t, J = 7.2 Hz, 2H), 1.83 - 1.57 (m, 4H). 13C NMR (400 MHz, CD 2 Cl 2 ) δ = 169.24, 168.21, 50.73, 30.32, 27.78, 25.53, 21.75.

[0148] HRMS (ESI+): [M+H] + C 9 H 13 N 4 O 4 Calculated for 241.0931, found for 241.0936; [M+Na] + C 9 H 12 N 4 NaO 4 Calculated for: 263.0751, Found 263.0756; [2M+Na] + C 18 H 24 N 8 NaO 8 Calculated value: 503.1609, measured value 503.1618.

[0149] 4. Synthesis of Compound 10

[0150] [ka]

[0151] Et 3 N (13.7 μL, 0.098 mmol, 3.5 equiv.) in dry DMF (1 mL) 7 (60 mg, 0.028 mmol, 1 eq.) and NHS ester 9 (23.6 mg, 0.098 mmol, 3.5 equiv.) was added to a solution of 1,2-dichlorophenyl ether (1.0 mg, 0.098 mmol, 3.5 equiv.). The mixture was stirred at room temperature for 4 h. After completion, the solvent was removed under reduced pressure and the crude residue was purified by chromatography on a silica gel column (gradient elution CH 2 Cl 2 / MeOH 95 / 5→75 / 25) to obtain the compound as a colorless oil. 10 (54.8 mg, 78%).

[0152] R t : 11.00 pm (New Day 1)。 1 H NMR (400 MHz, CD 2 Cl 2 ) δ = 7.97 (s, 2H), 7.84 (s, 1H), 7.28 (s, 2H), 7.15 (bs, 1H), 6.32 (bs, 3H), 5.22 (s, 4H), 5.13 (s, 2H), 4.1 Hz = 5.4 (t (t, J= 5.2 Hz, 2H), 3.88 (t, J= 5.1 Hz, 4H), 3.84 (t, J= 5.3 Hz, 2H), 3.64 - 3.49 (m, 126H), 3.47 - 3.73 (m, 3. 2H), 3.47 - 3.73 (m. 3, 2H). J = 6.6 Hz, 6H), 3.21 (m, 2H), 2.60 (s, 1H), 2.17 (t, J= 7.1 Hz, 6H), 1.73 - 1.53 (m, 12H), 1.39 (s, 9H)。

[0153] 13 C NMR (400 MHz, CD 2 Cl 2 ) δ = 172.65, 166.88, 156.34, 152.62, 144.47, 143.71, 140.52, 130.84, 125.18, 125.01, 107.1.43, 79. 70.95, 70.89, 70.84, 70.79, 70.71, 70.67, 70.55, 70.32, 69.88, 69.81, 66.70, 63.41, 51.784, 50.74, 40.50, 39.65, 36.12, 28.87, 28.63, 11.26pm。

[0154] HRMS (ESI+): [M+Na] + C 110 H 197 N 23 O 42Calculated for Na: 2535.3879, found 2535.3893; [M+2Na] 2+ C 110 H 197 N 23 O 42 Na 2 Calculated for: 1279.1885, Found 1279.1877; [M+3Na] 3+ C 110 H 197 N 23 O 42 Na 3 Calculated for: 860.4554, Found 860.4543; [M+4Na] 4+ C 110 H 197 N 23 O 42 Na 4 Calculated value: 651.0889, measured value 651.0888.

[0155] 5. Synthesis of Compound 11

[0156] [ka]

[0157] TFA (160 μL) was dissolved in dry CH 2 Cl 2 Carbamate in (700 μL) 10 (70 mg, 0.02788 mmol, 1 equiv.) was added to a cooled (0° C.) solution of 1,2-dichlorophenyl ether (1,2-dichlorophenyl ether). The mixture was stirred at 0° C. for 30 min and then at room temperature for an additional 30 min. After completion, the solvent was removed under reduced pressure and the crude mixture was purified by reversed phase chromatography on C18 grafted silica (gradient elution MeCN / H 2 The compound was purified by distillation in 20 / 80 to 100 / 0 (0.05% TFA) for 30 min to give a colorless oil. 11 (65.5 mg, 91%)

[0158] R t : 8.66 minutes (method 1). 1 H NMR (400 MHz, CD2 Cl 2 ) δ = 8.07 (bs, 3H), 7.99 (bs, 3H), 7.87 (s, 1H), 7.35 (s, 2H), 6.50 (bs, 3H), 5.21 (s, 4H), 5.14 (s, 2H), 4.55 (t, J = 4.5 Hz, 4H), 4.52 - 4.42 (m, 2H), 3.88 (t, J = 4.6 Hz, 4H), 3.86 - 3.80 (m, J = 4.9 Hz, 2H), 3.71 (s, 4H), 3.69 - 3.45 (m, 131H), 3.44 - 3.33 (m, J = 4.5 Hz, 6H), 3.27 (t, J = 6.2 Hz, 6H), 3.14 (s, 3H), 2.18 (t, J = 7.0 Hz, 6H), 1.78 - 1.43 (m, 12H).

[0159] 13 C NMR (400 MHz, CD 2 Cl 2 ) δ = 172.89, 167.28, 152.54, 144.38, 143.77, 140.32, 130.68, 125.39, 125.19, 107.65, 71.00, 70.92, 70.83, 70.81, 70.69, 70.47, 70.39, 69.85, 69.79, 67.40, 66.41, 63.19, 51.76, 50.85, 50.62, 40.47, 40.25, 39.67, 36.12, 28.89, 23.30.

[0160] HRMS (ESI+): [M+H] + C 105 H 189 N 23 O 40 Hについての calculated value: 2413.3535, measured value 2413.3482; [M+2Na] 2+ C 105 H 189 N 23 O 40 Na 2Calculated for: 1229.1623, Found 1229.1575; [M+2H] 2+ C 105 H 189 N 23 O 40 H 2 Calculated for: 1207.1804, Found 1207.1776; [M+H+Na] 2+ C 105 H 189 N 23 O 40 Calculated for HNa: 1218.1714, found 1218.1685; [M+3H] 3+ C 105 H 189 N 23 O 40 H 3 Calculated for: 805.1227 Found: 805.1186; [M+3Na] 3+ C 105 H 189 N 23 O 40 Na 3 Calculated for: 827.1046, Found 827.1007; [M+H+2Na] 3+ C 105 H 189 N 23 O 40 HNa 2 Calculated for: 812.4500 Found: 812.4480; [M+2H+Na] 3+ C 105 H 189 N 23 O 40 H 2 Calculated for Na: 819.7773 Found 819.7746.

[0161] 6. Synthesis of Compound 13

[0162] [ka]

[0163] H 2 LiOH H in O (6 mL) 2A cooled (0 °C) solution of O (31.6 mg, 0.75 mmol, 8.75 equiv.) was added to the protected glucuronide derivative (PAPOT S. et al., Chem. Sci., 2017, 8, 3427-3433) in MeOH (6 mL). 12 A cooled solution (-5 °C) of (109 mg, 0.086 mmol, 1 equiv.) was added dropwise. The mixture was stirred at -5 °C until the disappearance of the starting material. Hydrolysis was then carried out using IRC-50 acidic resin. After filtration, the solvent was removed under reduced pressure and the crude material was purified by reversed phase chromatography on C18 grafted silica (gradient elution MeCN / H 2 20 / 80→80 / 20 for 30 min) to give compound 1H as a white solid. 13 (70 mg, 72%)

[0164] R t : 11.49 - 11.80 minutes (method 1). 1 H NMR (400 MHz, DMSO-d6) δ = 8.59 - 8.27 (m, 0.5H), 8.10 - 8.04 (m, 1H), 7.94 - 7.82 (m, 1H), 7.80 - 7.58 (m, 1.5H), 7.45 - 7.41 (m, 1H), 7.36 - 7.21 (m, 4H), 7.19 - 7.15 (m, 1H), 5.79 - 5.58 (m, J = 29.3 Hz, 1H), 5.31 - 5.14 (m, 1H), 4.82 - 4.54 (m, 1H), 4.54 - 4.15 (m, 2.5H), 4.09 - 3.87 (m, 3H), 3.78 (d, J = 9.4 Hz, 0.5H), 3.67 - 3.35 (m, 2H), 3.35 - 2.54 (m, 20H), 2.46 - 2.16 (m, 2H), 2.16 - 1.92 (m, 3H), 1.88 - 1.64 (m, 3H), 1.61 - 1.40 (m, 2H), 1.31 (s, 1H), 1.09 - 0.95 (m, 6.5H), 0.94 - 0.72 (m, 16H), 0.72 - 0.48 (m, 3.5H).

[0165] 19 F NMR (376 MHz, DMSO-d6) δ = -75.63. 13 C NMR (400 MHz, DMSO-d6) δ = 172.38, 172.32, 169.92, 169.86, 169.78, 169.76, 169.65, 168.74, 168.71, 158.52, 158.14, 154.56, 154.54, 154.38, 148.76, 148.68, 143.67, 139.84, 134.20, 132.63, 127.81, 127.74, 126.74, 126.66, 126.48, 126.41, 122.90, 116.19, 100.11, 85.45, 81.64, 77.68, 75.82, 75.43, 74.79, 73.36, 72.74, 71.16, 63.20, 62.88, 60.93, 60.29, 58.67, 58.17, 57.16, 54.15, 49.76, 49.17, 47.22, 46.25, 43.76, 43.21, 31.58, 30.09, 29.70, 29.49, 25.91, 25.37, 24.34, 23.12, 18.89, 18.85, 18.77, 18.60, 18.57, 18.37, 15.46, 15.28, 15.00, 10.42, 10.36, 10.32.

[0166] HRMS (ESI+): [M+Na] + C 56 H 82 N 4 NaO 18 Calculated value: 1149.5578, observed value: 1149.5635.

[0167] 7. Synthesis of Compound 14

[0168] [ka]

[0169] CuSO 4 (5.7 mg, 0.0356 mmol, 3.5 equiv.) and sodium ascorbate (7.0 mg, 0.0356 mmol, 3.5 equiv.) were dissolved in t-BuOH (6 mL) / H 2 Alkyne in a degassed mixture of O (10 mL) 13 (34 mg, 0.030 mmol, 3 equiv.) and azide 11 (30 mg, 0.01187 mmol, 1.2 equiv.). The resulting mixture was stirred at room temperature under Ar for 1.5 h. Upon completion, the resin QuadraPure® IDA (350 mg) was added to the mixture, stirred for an additional 3 h, and filtered off. The solvent was evaporated under reduced pressure and the crude residue was purified by reversed phase chromatography on C18 grafted silica (gradient elution MeCN / H 2 HO (0.05% TFA) (20 / 80→60 / 40) for 30 min to give compound 1 as a white solid. 14 (24 mg, 41%)

[0170] R t : 11.97-12.09 minutes (method 1). 1H NMR (400 MHz, DMSO-d6) δ = 8.73 - 7.11 (m, 39H), 5.99 - 5.76 (m, 3H), 5.29 - 5.20 (m, 3H), 5.18 (s, 4H), 5.03 (s, 2H), 4.83 - 4.59 (m, 1H), 4.56 (t, J = 5.1 Hz, 4H), 4.51 - 3.89 (m, 30H), 3.83 (t, J = 5.2 Hz, 4H), 3.81 - 3.72 (m, 3H), 3.69 - 3.35 (m, 148H), 3.34 - 3.10 (m, 44H), 3.10 - 2.69 (m, 17H), 2.62 - 2.53 (m, 1H), 2.47 - 2.35 (m, 5H), 2.31 - 2.19 (m, 2H), 2.19 - 1.89 (m, 14H), 1.89 - 1.62 (m, 15H), 1.61 - 1.11 (m, 16H), 1.09 - 0.93 (m, 21H), 0.92 - 0.71 (m, 45H), 0.71 - 0.51 (m, 12H), 0.44 (d, J = 6.4 Hz, 3H)。

[0171] 19 F NMR (376 MHz, DMSO-d6) δ = -74.53。 13C NMR (400 MHz, DMSO-d6) δ = 172.36, 172.29, 171.74, 169.89, 169.84, 168.71, 165.46, 158.27, 157.92, 151.60, 148.60, 143.65, 143.21, 142.50, 139.24, 129.64, 127.79, 127.73, 126.72, 126.64, 126.44, 126.39, 126.34, 124.78, 124.70, 123.01, 122.62, 106.83, 99.64, 85.42, 81.61, 75.81, 75.42, 74.77, 72.72, 71.13, 69.75, 69.61, 69.55, 69.15, 69.04, 68.68, 66.68, 65.53, 62.41, 60.91, 60.27, 58.65, 58.13, 57.14, 54.99, 54.18, 49.74, 49.47, 49.31, 48.91, 47.20, 46.23, 43.74, 43.18, 38.65, 38.46, 34.40, 31.57, 29.41, 29.28, 25.34, 24.33, 23.10, 22.13, 22.00, 18.71, 18.59, 17.97, 15.61, 15.43, 15.27, 14.99, 10.39, 10.34.

[0172] HRMS (ESI+): [M+7H] 7+ C 273 H 442 N 41 O 94 Calculated for: 828.4417, Found 828.4419; [M+6H] 6+ C 273 H 441 N 41 O 94 Calculated for: 966.3475, Found 966.3468; [M+5H] 5+ C 273 H 440 N 41 O 94 Calculated value: 1159.4155, measured value 1159.4154.

[0173] 8. Synthesis of RC-Alb

[0174] [ka]

[0175] Et 3 N (0.94 μL, 0.00677 mmol, 4 equiv.) was dissolved in 1 mL of dry DMSO to give 14 (10 mg, 0.00169 mmol, 1 eq.) and NHS ester 15 (0.57 mg, 0.00186 mmol, 1.1 equiv.). The mixture was stirred at room temperature for 7 h. Upon completion, the solvent was evaporated under reduced pressure and the crude residue was purified by reversed phase chromatography on C18 grafted silica (gradient elution MeCN / H 2 Purification by O 2 (0.05% TFA) 20 / 80→80 / 20 for 30 min gave RC-Alb (9.9 mg, 98%, purity >98%) as a white solid.

[0176] R t : 12.80-12.92 minutes (method 1). HRMS (ESI+): [M+6H] 6+ C 283 H 452 N 42 O 97 Calculated for 998.5263, found 998.5261; [M+5H] 5+ C 283 H 451 N 42 O 97 Calculated for: 1198.0301, Found 1198.0306; [M+4H] 4+ C 283 H 450 N 42 O 97 Calculated value: 1497.2858, measured value 1497.2857.

[0177] biology Cells: The MIA PaCa-2 human pancreatic cell line was obtained from ATCC (American Type Culture Collection) and stably transfected to express the luciferase gene by the INSERM Trichet team, Nantes, France.

[0178] In vivo experimental procedures: Female nude mice aged 6-8 weeks were purchased from Charles River Laboratories. They were acclimated for 7 days in the laboratory before the experiment and were kept in sterile filter-stopped cages inside racks with controlled ventilation and had free access to food and water. All experimental procedures involving animals were verified by the local ethical committee (CECCO No. 3) and were carried out in accordance with the French Ministry of Agriculture and Forestry guidelines (Ministerial Decree 2013-118) and the European Community (ECC) Directive guidelines (2010 / 63 / UE). Throughout the study, mice were examined for at least three weeks for clinical signs, distress, physical activity and weight loss as indicators of health status.

[0179] In vivo efficacy with respect to orthotopic models: Taking into account obvious ethical issues, the evaluation of new therapies and the elucidation of biological mechanisms must be performed from animal models. These represent the proof-of-concept (POC) stage for molecules that can be tested in phase I and II clinical trials. A crucial step in this preclinical process is the use of appropriate models that take into account clinical reality. The lack of clinical reality leads to a large lack of predictability in the results obtained in vivo in humans, which leads to risky extrapolations. To evaluate the efficacy of antitumor therapies, it is necessary to perform these experiments in orthotopic models. Orthotopic models have the advantage of being better predictive of human tumor development. Models of tumors growing in the original primary tumor tissue closely resemble the clinical pathological state. Given the complexity of the processes involved, including interactions with the tumor microenvironment, no reliable transposition methods are available.

[0180] Human pancreatic cancer xenografts from the pancreatic cancer cell line MIA PaCa2-luc were established in Swiss nude mice by orthotopic implantation. Mice were anesthetized by inhalation of 1.5% isoflurane with air. The abdomen of the mouse was prepared with a solution of povidone-iodine (betadine). A small incision was made in the left lateral flank through the skin and peritoneum. The tip of the pancreatic tail was gently grasped and exteriorized laterally to fully expose the pancreas / spleen. A needle was inserted into the tip of the pancreatic tail and placed in the head region of the pancreas. Inoculum (2 × 10 in 10 μL of PBS) was added to the pancreas. 6 MIA PaCa2-luc cells) were slowly injected using a Hamilton syringe with a 27-gauge needle. The spleen was then returned to the appropriate position in the abdomen, the peritoneum was sutured using 7-0 sutures, and the skin was sutured using 4-0 sutures.

[0181] Pancreatic Tumor Treatment Mice (8 animals per group) were inoculated with 5% DMSO and 95% PBS mix (vehicle group), 2 mg / kg BR-Alb, 4.32 mg / kg RC-Alb or 6.48 mg / kg RC-Alb on days 37, 44, 50 and 64. Tumor volumes were determined by ultrasound imaging.

[0182] Ultrasound ImagingMice were anesthetized with inhalation of 1.5% isoflurane with air and placed on a thermostatically controlled heating pad with limbs taped onto ECG electrodes attached to a table. ECG-derived respiratory gating served to avoid artifacts due to the animal's respiratory movements. Animal body temperature was recorded using an internal temperature probe. Warmed water-based ultrasound gel (purchased from Supragel) was applied to the overlying skin to optimize visualization of internal tissues. Tumor tissue was imaged using a Vevo LAZR system (FUJIFILM Visualsonics Inc.). A transducer with a central frequency of 40 MHz, providing an axial resolution of 40 μm with a field of view of 14.1 × 15 mm, was used for imaging of small tumors. A transducer with a central frequency of 21 MHz, providing an axial resolution of 75 μm with a field of view of 23.1 × 36 mm, was used for imaging of larger tumors. 3D scans of ultrasound images were digitally recorded and reviewed. Tumor area in the coronal plane was measured by manually delineating the borders using Vevo LAB1.7.2. software (FUJIFILM Visualsonics Inc.). The software was then used to measure tumor area by 0.5 mm depending on the location of the tumor. 3 ~ 1.5 mm 3 Corresponding tumor volumes were calculated from each coronal slice segment with a detection threshold ranging from 0.01 to 0.01.

[0183] As explained above, RC-Alb can vectorize three molecules of MMAE to enter inside the microenvironment of solid tumors. RC-Alb consists of 1) a maleimide that allows the formation of a covalent bond with plasma albumin in the bloodstream, 2) a gallic acid structure that builds a ring (link) between the maleimide function and three glucuronylation units, and 3) three linkers that selectively release the activator into the tumor microenvironment through the action of β-glucuronidase.

[0184] The therapeutic activity of RC-Alb was evaluated in mice as described above (Figure 1). The results show that RC-Alb exerts significant anticancer activity without any side effects. Moreover, the use of RC-Alb at 6.48 mg / kg resulted in complete regression of tumor masses in 38% of treated animals.

[0185] The therapeutic activity of RC-Alb was also compared to that of the corresponding monomeric BR-Alb (carrying a single MMAE molecule) (Figure 2). Both compounds were administered at the same dose (1.1 × 10 -6 When administered at 100 mg / kg, RC-Alb caused an 18-fold reduction in tumor volume compared to BR-Alb.

[0186] Other compounds according to the invention 1. Synthesis of YG-Alb and AB-Alb With the aim of evaluating the effect of the maleimide moiety on the toxicity of the trimeric glucuronide prodrugs, two compounds, YG-Alb and AB-Alb, were prepared as follows:

[0187] [ka]

[0188] 1.1 Preparation of YG-Alb 5-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-fluorobenzoic acid 15 The synthesis of was accomplished using published protocols (WO 2016054315). a) 2,5-dioxopyrrolidin-1-yl = 5-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-fluorobenzoic acid ester 16

[0189] [ka]

[0190] 5-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-fluorobenzoic acid in anhydrous DMF (3 mL) 15 To a stirred solution of (602 mg, 2.56 mmol, 1.0 equiv) was added NHS (352 mg, 3.06 mmol, 1.2 equiv) and EDC hydrochloride (590 mg, 3.08 mmol, 1.2 equiv). The reaction mixture was left stirring at room temperature overnight. The solvent was removed under vacuum. The crude material was taken up in DCM (15 mL) and washed successively with 0.1 N aqueous HCl (10 mL) and water (2×10 mL). The organic layer was dried over magnesium sulfate, filtered and evaporated. The crude material was purified by flash silica gel column chromatography using a DCM / AcOEt gradient 50 / 50→0 / 100 to give the activated ester as a beige solid. 16 (468 mg, 1.41 mmol) was obtained. Yield: 55%.

[0191] b) YG-Alb Compound in anhydrous DMSO (2 mL) 14 To a stirred solution of (10.3 mg, 1.78 μmol, 1.0 equiv.) of 2,5-dioxopyrrolidin-1-yl 5-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-fluorobenzoate 16 (1.8 mg, 5.33 μmol, 3.0 equiv.) and triethylamine (1.5 μL, 10.8 μmol, 6.0 equiv.) were added. The reaction mixture was allowed to stir at room temperature overnight. The solvent was removed under vacuum. The crude material was purified by distillation with H 2 After purification by chromatography using a gradient of O+0.05% TFA / ACN 90 / 10→40 / 60 and lyophilization, compound YG-Alb (5.1 mg, 0.854 μmol) was obtained. Yield: 48%. HRMS m / z [M+4H] 4+ Calculated: 1504.0271, Found: 1504.0273;[M+5H] 5+ Calculated: 1203.2225, Found: 1203.2224; [M+6H] 6+ Calculated value: 1003.0205, Measured value: 1003.0204.

[0192] 1.2 Preparation of AB-Alb 3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-4,4,4-trifluorobutanoic acid 17 The synthesis of was accomplished using published protocols Bioconjugate. Chem., 2015, 26, 14-152.

[0193] a) 2,5-dioxopyrrolidin-1-yl = 3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-4,4,4-trifluorobutanoate 18 Synthesis of

[0194] [ka]

[0195] 3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-4,4,4-trifluorobutanoic acid in anhydrous DMF (3 mL) 17 To a stirred solution of (500 mg, 2.51 mmol, 1.0 equiv) was added NHS (291 mg, 2.53 mmol, 1.2 equiv) and EDC hydrochloride (485 mg, 3.08 mmol, 1.2 equiv). The reaction mixture was left stirring at room temperature overnight. The solvent was removed under vacuum. The crude material was purified by flash silica gel chromatography using a DCM / MeOH gradient 100 / 0 to 75 / 25 to give the activated ester as a white solid. 18 (487 mg, 1.46 mmol) was obtained. Yield: 58%.

[0196] b) AB-Alb Compound in anhydrous DMSO (2 mL) 14 To a stirred solution of (10.5 mg, 1.78 μmol, 1.0 equiv.) of 2,5-dioxopyrrolidin-1-yl 3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-4,4,4-trifluorobutanoate 18(1.8 mg, 5.33 μmol, 3.0 equiv.) and triethylamine (1.5 μL, 10.8 μmol, 6.0 equiv.) were added. The reaction mixture was left stirring at room temperature overnight. The solvent was removed under reduced pressure. The crude material was purified by distillation with H 2 Purification by flash reversed-phase (C18) silica gel column chromatography using a gradient of 90 / 10 to 40 / 60 O+0.05% TFA / ACN gave, after lyophilization, compound AB-Alb (6.4 mg, 1.06 μmol). Yield: 60%. HRMS m / z [M+4H] 4+ Calculated value: 1504.5263, Measured value: 1504.5269, [M+5H] 5+ Calculated value: 1203.6219, Found value: 1203.6224, [M+6H] 6+ Calculated value: 1003.3533, Measured value: 1003.3529.

[0197] 2. MTD research Maximum Tolerated Dose Study (MTD): MTD study was performed on BALB / c nude mice (n=3). A single dose of the prodrugs RC-Alb, YG-Alb and AB-Alb (9.72 mg / kg or 12.96 mg / kg) was administered intravenously. Toxicity of the prodrugs was evaluated by the maximum weight loss or gain expressed as a percentage of the animals' initial weight. A dose was considered toxic if it caused a relative weight loss >20% of the initial weight.

[0198] The results are shown in Figures 3 and 4.

[0199] Comparative MTD studies performed with RC-Alb, YG-Alb and AB-Alb showed that the structure of the maleimide moiety influenced the toxicity of the glucuronide prodrug in mice. 3 The prodrugs YG-Alb and AB-Alb, bearing electron-withdrawing groups such as , were shown to be less toxic than RC-Alb, which lacks this type of chemical functionality.

Claims

1. A compound having the following formula (I): 【Chemistry 1】 [In the ceremony: R a is optionally interrupted by H or one or several oxygen atoms, (C 1 ~C 12 ) represents an alkyl group; L represents a group having the formula (II): 【Chemistry 2】 Where: A is an anticancer drug, monomethylauristatin E (MMAE); Y is an electron withdrawing group selected from the group consisting of halogen, NO2, and CF3; X is -O-; G is a glucuronyl group or a derivative thereof selected from glucuronide ester radicals; L 1 represents a linker represented by the following formula (III): -A 1 -A 2 -A 3 -A 4 -A 5 -A 6 -A 7 - (III) where A 1 (C 1 ~C 6 ) an alkylene group; A 2 is a triazole group; A 3 (C 1 ~C 6 ) an alkylene group; A 4 is -C(=O)-NR b -, -C(=S)-NR b -, -NR b -C(=O)-, -NR b -C(=S)- and NR b R is selected from the group consisting of b is H or (C 1 ~C 12 ) an alkyl group; A 5 is interrupted by at least one oxygen atom (C 1 ~C 32 ) an alkylene group; A 6 is a triazole group; A 7 (C 1 ~C 6 ) an alkylene group; L' represents a group having the formula (IV): -A 8 -A 9 -L'' (IV) where A 8 is interrupted by at least one oxygen atom (C 1 ~C 6 ) an alkylene group; A 9 HA-NR c -, -O-, and S-; R c is H or (C 1 ~C 12 ) alkyl group; and L″ is a maleimidocaproyl group or a group having one of the following formulas: 【Chemistry 3】 or a pharma- ceutically acceptable salt thereof.

2. L is of the following formula (VII): 【Chemistry 4】 (Wherein, A, Y and L 1 is as defined in claim 1) 2. The compound of claim 1 having the formula:

3. L is of the following formula (VIII): 【Chemistry 5】 (In the formula, A, A 1 , A 3 , A 4 , A 5 and A 7 is as defined in claim 1) 2. The compound of claim 1 having the formula:

4. A 5 is of the formula -CH 2 -(CH 2 -O-CH 2 ) n -CH 2 - group, and n is an integer from 1 to 12, inclusive.

5. L is of the following formula (IX): 【Chemistry 6】 wherein i is an integer from 1 to 6, inclusive; j is an integer from 1 to 10, inclusive; n is an integer from 1 to 12, inclusive; k is an integer from 1 to 6, inclusive. The compound according to any one of claims 1 to 4, having the formula:

6. A 8 is of the formula -CH 2 - (CH 2 -O-CH 2 ) m -CH 2 - group and m is an integer from 1 to 12 inclusive.

7. L' is a group represented by the following formula (X): 【Chemistry 7】 wherein m is an integer from 1 to 6, inclusive; and p is an integer from 1 to 6, inclusive. The compound according to any one of claims 1 to 12, having the formula:

8. The following formula (XI), (XI') or (XI''): 【Chemistry 8】 【Chemistry 9】 【Chemistry 10】 The compound according to any one of claims 1 to 7, having the formula:

9. A compound according to any one of claims 1 to 8 for use as a drug.

10. A pharmaceutical composition comprising a compound according to any one of claims 1 to 8, or a pharma- ceutically acceptable salt thereof, and also at least one pharma- ceutically acceptable excipient.

11. A compound according to any one of claims 1 to 8 for use in the treatment and / or prevention of cancer.

Images