New therapeutic vectors and prodrugs for treating cancer

Abstract

The present invention relates to a compound having the following formula (I) and a corresponding prodrug, and a pharmaceutical composition comprising the compound, in particular for use in treating cancer. #imgabs0#

Description

[0001] The present invention relates to novel therapeutic carriers allowing the immobilization of anticancer agents, and their corresponding prodrugs. It also relates to the use of said carriers and prodrugs in the treatment of cancer.

[0002] Cancer is one of the most common pathologies currently. In particular, in France and in most industrialized countries, cancer is one of the leading causes of contemporary death. Death is often caused by the development of metastases and the lack of treatments, at least for the most common tumor localizations: breast, colorectal, prostate, pancreatic and lung cancer. In this late stage of the disease, cancer chemotherapy remains the most powerful treatment and often the only available treatment. However, chemotherapy is not completely effective against many common solid tumor types. Most anticancer drugs lack any intrinsic antitumor selectivity. In turn, chemotherapy is often associated with severe side effects due to the destruction of normal tissues. Therefore, the amount of drug that can be administered is often insufficient to deliver lethal concentrations of anticancer agents at the tumor site. In addition, the lack of selectivity of cytotoxic drugs significantly increases the risk of tumor cells developing cellular resistance. Therefore, given the nonspecific toxicity of most anticancer drugs, the development of more selective chemotherapeutic methods represents a major interest in the fight against cancer.

[0003] Within this framework, much research effort has focused on developing self-responsive chemical systems that are programmed to selectively deliver potent cytotoxicity at tumor sites. Such systems are typically complex molecular assemblies that have built into their structure (1) targeting units that are able to recognize tumor-associated specificities and (2) enzymatic or chemical triggers that can be activated specifically in cancer tissues to induce the release of the drug in a tightly controlled manner.

[0004] The vast majority of drug delivery systems developed so far are designed to target specificities on the surface of cancer cells (e.g., membrane receptors or antigens). In this approach, the molecular assembly includes monoclonal antibodies or low molecular weight ligands that show high affinity for the corresponding tumor-associated cell surface markers. When the cancer cell surface is detected by the targeting unit, the entire system is internalized via receptor-mediated endocytosis. Once inside the cell, activation of the trigger leads to the selective release of the active drug in the intracellular medium. Currently, several drug delivery systems of this type are under clinical evaluation for various applications in oncology.

[0005] However, the "Achilles' heel" of drug delivery systems designed to target cell surface specificity depends on the heterogeneity of cancer tissues. In fact, all cells of a tumor mass are not the same, and they show different concentrations of given cell surface markers. Therefore, only cancer cells that express the selected tumor-related markers at sufficient levels will be directly affected by such targeted systems. In this case, the use of enzyme-reactive prodrugs that can be selectively activated by the corresponding enzymes naturally overexpressed in the tumor microenvironment provides a valuable alternative to this targeted approach. In this case, the anticancer agent is released in the extracellular medium and can further passively penetrate into various types of surrounding malignant cells, regardless of their membrane characteristics.

[0006] Therefore, it is an object of the present invention to provide a system for selectively releasing anticancer agents in the tumor microenvironment.

[0007] Therefore, the present invention relates to a compound having the following formula (I):

[0008]

[0009] in:

[0010] R a Indicates H or (C1-C 12 ) alkyl, optionally interrupted by one or more oxygen atoms, R a Preferably H;

[0011] L represents a group having the following formula (II):

[0012]

[0013] in:

[0014] A is an anticancer agent;

[0015] Y is an electron withdrawing group;

[0016] X is -O-;

[0017] G is glucuronic acid or its derivative;

[0018] L1 represents a linker represented by the following formula (III):

[0019] -A1-A2-A3-A4-A5-A6-A7- (III)

[0020] in:

[0021] A1 is (C1-C6)alkylene;

[0022] A2 is a group accessible by click chemistry;

[0023] A3 is (C1-C6)alkylene;

[0024] A4 is selected from the following: -C(=O)-NR b -、-C(=S)-NR b -、-NR b -C(=O)-, -NR b -C(=S)- and NR b , R b Indicates H or (C1-C 12 )alkyl;

[0025] A4 is preferably -C(=O)-NR b -group, more preferably -C(=O)-NH-,

[0026] A5 is interrupted by at least one oxygen atom (C1-C 32 )alkylene, preferably (C1-C6)alkylene interrupted by at least one oxygen atom, more preferably a polyoxyalkylene group,

[0027] A6 is a group accessible by click chemistry; and

[0028] A7 is (C1-C6)alkylene;

[0029] -L' represents a group having the following formula (IV):

[0030] -A8-A9-L"(IV)

[0031] in:

[0032] A8 is a (C1-C6)alkylene group interrupted by at least one oxygen atom, preferably a polyoxyalkylene group.

[0033] A9 is selected from the following: -NR c -, -O- and -S-, R c Indicates H or (C1-C 12 ) alkyl; and

[0034] L" is a group capable of reacting with an amino, hydroxyl or thiol functional group;

[0035] or a pharmaceutically acceptable salt thereof, or a racemate, diastereomer or enantiomer thereof.

[0036] The compound of the present invention is a compound having a gallic acid structure that allows three anticancer agent molecules (A) to be fixed via a linker L.

[0037] The compounds described herein may have asymmetric centers. Compounds of the invention containing asymmetrically substituted atoms may be isolated in optically active or racemic forms. It is well known in the art how to prepare optically active forms, such as by resolving racemic forms or by synthesizing from optically active starting materials. Unless stereochemical forms or isomeric forms are specifically indicated, all chiral, diastereoisomer, racemic forms and all geometric isomeric forms of the compound are intended.

[0038] The term "pharmaceutically acceptable salt" refers to a salt that retains the biological effectiveness and properties of the compounds of the present invention and is not biologically or otherwise undesirable. In many cases, the compounds of the present invention are capable of forming acid salts and / or base salts due to the presence of amino and / or carboxyl groups or groups similar thereto. Pharmaceutically acceptable acid addition salts can be prepared from inorganic acids and organic acids, while pharmaceutically acceptable base addition salts can be prepared from inorganic bases and organic bases. For a review of pharmaceutically acceptable salts, see Berge et al. ((1977) J. Pharm. Sd, Vol. 66, 1). The expression "non-toxic pharmaceutically acceptable salt" refers to a non-toxic salt formed with a non-toxic, pharmaceutically acceptable inorganic or organic acid or inorganic or organic base. For example, these salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and the like, and salts 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, and toluenesulfonic acid, and the like.

[0039] As mentioned above, A is an anticancer agent.

[0040] According to one embodiment, the anticancer agent is selected from: cytostatics, antimetabolites, DNA intercalating substances, topoisomerase I and II inhibitors, tubulin inhibitors, alkylating agents, neocarcinomatins, calicheamicin, dynemicin or esperamycin A, ribosomal inhibitors, tyrosine phosphatase inhibitors, compounds that induce cell differentiation, histone deacetylase inhibitors, small molecule immunomodulators or small molecules targeting cancer stem cells.

[0041] Even more specifically, the anticancer agent according to the invention is selected from: cytostatics and antimetabolites, such as 5-fluorouracil, 5-fluorocytidine, 5-fluorouracil nucleoside, cytarabine or methotrexate, from DNA intercalating substances, such as doxorubicin, daunomycin, idarubicin, epirubicin or mitoxantrone, from topoisomerase I and II inhibitors, such as camptothecin, etoposide or m-AMSA, from tubulin inhibitors, such as vincristine, vinblastine, vindesine, paclitaxel, nocodazole or colchicine, from alkylating agents, such as cyclophosphamide, mitomycin C, rachelmycin, cisplatin, mustard gas phosphoramide, milfoil, squalane ... Blue, bleomycin, N-bis(2-chloroethyl)-4-hydroxyaniline, or selected from neocarcinomatitis, calicheamicin, danemycin or esperamicin A, or selected from ribosome inhibitors, such as verrucosporin A, selected from tyrosine phosphokinase inhibitors, such as quercetin, genistein, genistein, tyrosine phosphorylation inhibitors or rohitukin and its derivatives, selected from compounds that induce cell differentiation, such as retinoic acid, butyric acid, phorbol esters or aclarubicin, selected from histone deacetylase inhibitors, such as CI-994 or MS275, selected from immunomodulators, such as imiquimod, and selected from small molecules targeting cancer stem cells, such as hedgehog inhibitors, such as cyclopamine derivatives.

[0042] According to a preferred embodiment, A is an anticancer agent comprising at least one primary or secondary amine functional group.

[0043] Preferably, according to the present invention, the anticancer agent A is modified to be able to bind to the -C(=O)-O- of the L group of the above formula (II).

[0044] According to one embodiment, A is a group of the Aplysia family or its derivatives. In the context of the present invention, a "derivative" of the Aplysia family refers to a compound that is structurally very related and retains equivalent biological properties (particularly the ability to inhibit tubulin polymerization) to ultimately inhibit cell mitosis. This may be a problem especially for substitution or deletion derivatives.

[0045] The Aplysia family represents a class of compounds having a structure of at least 4 amino acids, at least 3 of which are specific, ie different from the 20 most common amino acids in nature.

[0046] Reference may especially be made to document WO 2004 / 010957, which describes those compounds suitable for the present invention.

[0047] In a particularly preferred embodiment of the present invention, A represents a group derived from Aplysia 10, derived from rioocetin PE, derived from rioocetin E, derived from monomethylrioocetin E and derivatives thereof, preferably a group derived from monomethylrioocetin E or a derivative thereof.

[0048] The structural difference between Aplysia 10 and the synthetic compounds of the rioxetine subfamily lies in, inter alia, the replacement of the aminothiazole phenethyl group at the C-terminal position of Aplysia 10 by a norephedrine unit in the case of rioxetine PE, rioxetine E or monomethylrioxetine.

[0049] For the purposes of the present invention, the derivatives of Aplysia 10, Reoxetine PE, Reoxetine E or Monomethyl Reoxetine E have a chemical structure that is very relevant to at least one of its active agents and have the anti-mitotic properties attributed to Aplysia family compounds. The structural differences may be, in particular, substitutions on at least one side chain of at least one of the four amino acids of its composition. Such substitutions may be made to include or represent linear, cyclic and / or branched alkyl, aryl, heterocyclic or carbocyclic rings. Such structural differences may also include modifications to Aplysia 10, Reoxetine PE or Reoxetine E molecules, for example at the level of the tertiary amine at its N-terminal position, so that the function is compatible with the covalent bond established by the considered connecting arm.

[0050] According to a preferred embodiment, the anticancer agent A is monomethyloxetine E (MMAE) or a derivative thereof.

[0051] Preferably, A is represented by the following formula (A-1):

[0052]

[0053] In the context of the present invention, "electron withdrawing group" refers to the electron withdrawing property of an atom or group of atoms.

[0054] Preferably, in formula (II), Y is an electron withdrawing group selected from: halogen, NO2 and CF3. More preferably, Y is NO2.

[0055] As described above, in formula (II), G is a glucuronic acid group or a derivative thereof.

[0056] In the context of the present invention, G is dedicated to the enzymatic removal in order to thereby provide for the intramolecular rearrangement thereof connecting the linker and the Aplysia family molecule and thus leading to the release of the active molecule (A group).

[0057] Furthermore, the glucuronyl groups according to the invention are enzymatically hydrolyzable, which may impart tissue and / or cell specificity to the conjugates and prodrugs according to the invention.

[0058] As is well known, β-glucuronidase is a high concentration enzyme naturally present near many tumors. Therefore, during prodrug monotherapy (or PMT), the conjugates and prodrugs of the present invention comprising a glucuronic acid group can be advantageously activated at the extracellular level. In the context of the present invention, the term "activation" refers to the release of, for example, a group of the Aplysia family at the tumor site, thereby being able to exert its anti-mitotic biological activity.

[0059] In the context of the present invention, "derivatives" of glucuronyl groups refer to compounds that are structurally closely related and retain equivalent biological properties (in particular the ability to be an enzymatic substrate for β-glucuronidase). This may be a problem in particular for derivatives with substitution or deletion of one or more hydroxyl (-OH) or carboxyl (-COOH) groups.

[0060] As the derivative of glucuronic acid group, glucuronic acid ester group may be mentioned.

[0061] As described in the above formula (II), the linker L also includes another divalent linker L1, which is directly bonded to the oxygen atom of the gallic acid structure of the compound of the present invention.

[0062] In other words, each A7 of the linker L1 corresponds to the terminal group of each L group that is attached to each oxygen atom of the gallic acid structure. Thus, A7 is attached to the oxygen atom that carries the L group.

[0063] In this application, the term "alkyl" refers to a saturated or unsaturated aliphatic hydrocarbon group, which may be straight or branched, and unless otherwise specified, has 1 to 12 carbon atoms in the chain. Preferably, the alkyl group has 1 to 6 carbon atoms in the chain. "Branched" refers to one or more lower alkyl groups (such as methyl, ethyl or propyl) attached to a linear alkyl chain. "Lower alkyl" refers to a chain (which may be straight or branched) having 1 to 4 carbon atoms.

[0064] Unless otherwise specified, the term "alkylene" as used herein refers to a divalent group containing 1 to 6 carbon atoms. The group can be represented by the formula (CH2) n , where n is an integer from 1 to 6.

[0065] According to the present invention, in formula (III), A2 and A6 are groups obtainable by click chemistry. Therefore, these groups are obtained by a click chemistry reaction.

[0066] These click chemistry reactions include, in particular, cycloadditions of unsaturated compounds, among which there may be mentioned the Diels-Alder reaction between dienophiles and dienes, in particular the azide-alkyne 1,3-dipolar cycloaddition, preferably the copper-catalyzed azide-alkyne cycloaddition (CuAAC).

[0067] Other click chemistry reactions include reactions involving thiol functionalities (e.g., thioether formation from alkenes and mixed disulfides), as well as reactions involving non-aldol electrophilic carbonyl groups, such as the formation of oxime ethers from hydroxylamines, hydrazones from hydrazines, or thiosemicarbazones from thiosemicarbazine.

[0068] As click chemistry reactions, there can also be mentioned reactions involving thiocarboxylic acids or thioesters to result in the formation of thioesters and amides to result in the formation of, and reactions between azides and phosphines (such as Staudinger ligation).

[0069] Preferably, groups A2 and A6 are obtained by a reaction between two reactive functional groups selected from:

[0070] -Reaction between azides and alkynes.

[0071] -Reaction of aldehydes or ketones with hydrazides.

[0072] -Reaction between aldehydes or ketones and hydroxylamine.

[0073] -Reaction between azide and phosphine.

[0074] -Reaction between alkenes and tetrazines.

[0075] - the reaction between isonitrile and tetrazine, and

[0076] - Reaction between thiols and olefins (thiol-ene reaction).

[0077] According to a preferred embodiment, A2 and A6 are triazolyl.

[0078] Preferably, A2 of formula (III) is a triazole group, preferably a group having the following formula (V).

[0079]

[0080] Preferably, A6 of formula (III) is a triazole group, preferably a group having the following formula (VI).

[0081]

[0082] According to one embodiment, L has the following formula (VII):

[0083]

[0084] A, Y and L1 are as defined above.

[0085] Preferably, in formula (VII), Y is a nitro group.

[0086] Preferably, in formula (VII), A is monomethylrioxetine E (MMAE) or a derivative thereof, more preferably formula (A-1) having the above definition.

[0087] According to one embodiment, L has the following formula (VIII):

[0088]

[0089] A, A1, A3, A4, A5 and A7 are as defined above.

[0090] Preferably, in formula (VII), A is monomethylrioxetine E (MMAE) or a derivative thereof, more preferably formula (A-1) having the above definition.

[0091] According to one embodiment, in formula (VIII), A4 is -C(=O)-NH-.

[0092] According to one embodiment, in formula (VIII), A5 represents a compound of the formula -CH2-(CH2-O-CH2) n -CH2- group, wherein n is an integer from 1 to 12.

[0093] The present invention relates to compounds of formula (I) as defined above, wherein L has the following formula (IX):

[0094]

[0095] i is an integer from 1 to 6.

[0096] j is an integer from 1 to 10.

[0097] n is an integer from 1 to 12, and

[0098] k is an integer from 1 to 6.

[0099] Preferably, in formula (IX), i=1.

[0100] Preferably, in formula (IX), j=4.

[0101] Preferably, in formula (IX), n=10.

[0102] Preferably, in formula (IX), k=1.

[0103] As mentioned above, in formula (I), L" (bound to the gallic acid structure via a linker L') is a group capable of reacting with an amino, hydroxyl or thiol functional group.

[0104] In the context of the present invention, a "group capable of reacting with an amino, hydroxyl or thiol functional group" refers to a group, generally a hydrocarbon group, having a chemical functional group or unit capable of interacting with a free secondary amino, hydroxyl or thiol functional group, thus being able to establish a covalent bond between the conjugated molecule and a unique chemical entity carrying such a functional group (compatible with the generation of such a covalent functional group). In the context of the present invention, such a unique chemical entity is more particularly a macromolecule naturally present in an organism, advantageously an endogenous albumin molecule, such as human serum albumin.

[0105] According to a preferred embodiment, L" comprises a maleimide group.

[0106] According to a preferred embodiment, L" has the following formula:

[0107]

[0108] Where L'" is (C1-C 12 )alkylene, which is optionally substituted with an electron withdrawing group, in particular a halo(C1-C6)alkyl group, such as CF3, or a phenylene group, which is optionally substituted with an electron withdrawing group, in particular a halogen.

[0109] According to a preferred embodiment, L" is a maleimidocaproyl group.

[0110] According to a preferred embodiment, L" is a group having one of the following formulae:

[0111]

[0112] According to one embodiment, in formula (IV), A8 represents the formula -CH2-(CH2-O-CH2) m -CH2- radical, m is an integer from 1 to 12, in particular equal to 3.

[0113] The present invention relates to compounds of formula (I) as defined above, wherein L' has the following formula (X):

[0114]

[0115] m is an integer from 1 to 6, preferably 3, and

[0116] p is an integer of 1 to 6, and preferably 5.

[0117] According to one embodiment, the compound of the present invention, which may also be named conjugate, has formula (I) as defined above, wherein the linker L has formula (VII), preferably (VIII), more preferably (IX) as defined above, and the linker L' has formula (X) as defined above.

[0118] Preferred compounds of the present invention have the following formula (XI):

[0119]

[0120] Another preferred compound of the present invention has the following formula (XI'):

[0121]

[0122] Another preferred compound of the present invention has the following formula (XI"):

[0123]

[0124] The present invention also relates to a prodrug, which comprises a compound of formula (I) as defined above linked to albumin or a derivative or fragment thereof via a covalent bond.

[0125] According to the present invention, the term "prodrug" refers to a molecule that can transport an inactivated form of an anticancer agent (A) in a living body and release the compound in a specific targeted organ, tissue or cell under the action of β-glucuronidase.

[0126] More specifically, such prodrugs conveniently correspond to the following general formula (XII):

[0127]

[0128] Among them, L and R a As defined above.

[0129] As for the L'1 units themselves, they are derived in one aspect from a reaction between L' groups comprising units capable of reacting with: free amino, hydroxyl or thiol functions, in particular free thiol functions carried by a macromolecule, advantageously an albumin molecule, even more advantageously serum albumin.

[0130] In the present application, the prodrug can be formed in vivo or in vitro with a macromolecule, preferably with an albumin molecule.

[0131] Thus, fragments of endogenous or exogenous albumin, in particular human serum albumin, recombinant albumin or other albumins may be designed.

[0132] According to one embodiment, the covalent bond between the conjugate molecule of the present invention and the endogenous albumin molecule, in particular the human serum albumin molecule or a derivative thereof, is carried out in vivo.

[0133] In one embodiment, the prodrug according to the invention comprises at least one conjugate molecule according to the invention linked to the sulfur of cysteine ​​at position 34 of an endogenous albumin molecule via a thioether bond.

[0134] Indeed, it has been shown that covalent bonds are established spontaneously in vivo, for example, in one aspect between a compound carrying a group capable of reacting with a thiol function and the thiol function of cysteine ​​at position 34 of human serum albumin (Kratz et al., 2002, J. Med. Chem.).

[0135] 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):

[0136]

[0137] m and p are defined as above.

[0138] According to another embodiment, the prodrug according to the invention can also be formed in vitro by at least one binding molecule of formula (I) linked via a covalent bond to an albumin molecule, a recombinant albumin molecule or an albumin molecule fragment or a derivative thereof.

[0139] For the purposes of the present invention, it is important that "fragment of an albumin molecule" means a fragment of an albumin molecule which is large enough to ensure that the prodrug produced thereby has satisfactory bioavailability, penetration into tumor tissue and impermeability to the endothelial barrier of healthy tissue.

[0140] In this particular embodiment, the in vitro coupling between the conjugate of general formula (I) via its L' group and the albumin molecule, recombinant albumin molecule or albumin molecule fragment can be carried out using free and complementary reactive functional groups present on the albumin molecule, recombinant albumin molecule or albumin molecule fragment.

[0141] In a particular embodiment, said fragment of the albumin molecule may comprise a cysteine ​​corresponding to the cysteine ​​at position 34 in the endogenous albumin sequence.

[0142] Surprisingly, the coupling of the conjugate of general formula (I) and the albumin molecule does not in any way affect the ability of the prodrug thus formed:

[0143] - are transported and specifically targeted to the microenvironment of the tissue to be treated.

[0144] - cleavage by β-glucuronidase in the microenvironment of the tissue to be treated, and

[0145] -After cleavage of the glucuronyl group, the linker arm rearranges to release the anticancer agent.

[0146] Furthermore, the conjugate of general formula (I) does not in any way affect the ability of the anticancer agent released thereby to exert its biological activity by coupling its L' group with the amino, hydroxyl or thiol functional group of the albumin molecule, especially the endogenous albumin molecule.

[0147] Finally, the conjugates of general formula (I) limit the elimination of the prodrug by the kidneys through the coupling between their L'group and the amino, hydroxyl or thiol functional groups of the albumin molecule, in particular the endogenous albumin molecule. Thus, the half-life of the prodrug according to the invention in the blood is increased compared to the prodrugs represented by anticancer agents functionalized with glucuronidyl groups.

[0148] In another embodiment of the invention, the albumin molecule or the albumin fragment of the prodrug may also be modified, in particular by glycosylation or by pegylation.

[0149] The present invention also relates to a compound or a conjugate according to the invention as defined above, having formula (I) as defined above, or a prodrug as defined above, for use as a medicament.

[0150] The present invention also relates to a pharmaceutical composition comprising a compound of formula (I) as defined above, or a prodrug as defined above, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

[0151] Although the compounds of the present invention having formula (I) can be administered alone, it is preferred that they are presented in the form of a pharmaceutical composition. According to the present invention, the pharmaceutical composition for veterinary and human use comprises at least one compound having formula (I) as defined above, and one or more pharmaceutically acceptable carriers and optionally other therapeutic ingredients.

[0152] In certain preferred embodiments, the active ingredients required for combination therapy may be combined in a single pharmaceutical composition for simultaneous administration.

[0153] As used herein, the term "pharmaceutically acceptable" and its grammatical variations, when they refer to compositions, carriers, diluents and agents, are used interchangeably and indicate that these materials are capable of being administered to or on mammals without producing adverse physiological effects, such as nausea, dizziness, stomach upset, etc.

[0154] 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 injections of liquid solutions or suspensions; however, solid forms suitable for solution or suspension in liquid prior to use may also be prepared. The preparation may also be emulsified. In particular, the pharmaceutical composition may be formulated into solid dosage forms such as capsules, tablets, pills, powders, dragees or granules.

[0155] The selection of vehicle and the content of active substance in the vehicle are generally determined according to the solubility and chemical characteristics of the active compound, the specific mode of administration and the regulations to be followed in pharmaceutical practice. For example, excipients such as lactose, sodium citrate, calcium carbonate, dicalcium phosphate and disintegrants such as starch, alginic acid and some composite silicates are used in combination with lubricants such as magnesium stearate, sodium lauryl sulfate and talcum powder to prepare tablets. In order to prepare capsules, it is advantageous to use lactose and high molecular weight polyethylene glycol. When using aqueous suspensions, they can contain emulsifiers or agents that promote suspension. Diluents such as sucrose, ethanol, polyethylene glycol, propylene glycol, glycerol and chloroform or mixtures thereof can also be used.

[0156] The compounds of formula (I) or conjugates, prodrugs or pharmaceutical compositions according to the present invention may be administered orally, parenterally (subcutaneously, intravenously or intramuscularly) or topically (by topical application) to the skin and mucosa.

[0157] The conjugates, prodrugs or pharmaceutical compositions according to the invention can be administered in particular alone or in combination with chemotherapy or radiotherapy, and also in combination with other therapeutic agents, in particular anticancer agents and antimitotic agents, but also in combination with anti-inflammatory agents.

[0158] The dosage suitable for the present invention can be determined according to conventional methods commonly used in the art. The adjustment of the dosage is obviously part of the general ability of those skilled in the art.

[0159] In fact, it depends inter alia on the weight, age and sex of the individual to be treated, as well as on the advancement of the disease to be treated.

[0160] The present invention also relates to compounds of formula (I) or prodrugs as defined above for use in the treatment and / or prevention of cancer.

[0161] In the context of the present invention, the terms "treating" or "treatment" as used herein refer to reversing, alleviating, inhibiting or preventing the disorder or condition to which such term applies, or one or more symptoms of such disorder or condition.

[0162] The present invention also relates to a method for treating cancer comprising administering a conjugate of formula (I), a prodrug or a pharmaceutical composition as defined above according to the present invention, in combination with another treatment selected from the group consisting of chemotherapy, radiotherapy, treatment with at least one anti-inflammatory agent and combinations thereof.

[0163] According to one embodiment, the cancer is selected from solid tumors.

[0164] As solid tumors, the following may be mentioned: neuroblastoma, glioblastoma, osteosarcoma, retinoblastoma, soft tissue sarcomas, cancers of the central nervous system, Wilms' tumor, lung cancer, breast cancer, prostate cancer, colorectal cancer, thyroid cancer, cervical cancer, endometrial cancer, ovarian cancer, kidney cancer, liver cancer, brain cancer, testicular cancer, pancreatic cancer, bone cancer, skin cancer, small intestine cancer, stomach cancer, pleural cancer, esophageal cancer, laryngeal cancer and bladder cancer.

[0165] In a specific embodiment, the solid tumor is selected from the group consisting of pancreatic cancer, lung cancer and breast cancer, preferably pancreatic cancer.

[0166] Attached photos

[0167] Figure 1 MIA PaCa2 was used in the presence of vehicle and RC-AIb (compound of formula (XI) of the present invention) (4.32 and 6.48 mg.kg -1 ) were used to treat tumor growth. Injections were performed on days 37, 44, 50, and 64.

[0168] The solid curve with black circles corresponds to the vehicle, and the dashed curve with white circles corresponds to 4.32 mg.kg -1 The dashed curve with black circles corresponds to 6.48 mg.kg -1 RC-AIb.

[0169] Figure 2 .MIA PaCa2 was used in the presence of Br-AIb (2 mg.kg -1 , 1.1x10 -6 mol.kg -1 ), RC-AIb(4.32mg.kg -1 , 0.7x10 -6 mol.kg -1 ) and RC-AIb (6.48 mg.kg -1 , 1.1x10 -6 mol.kg -1 ) were used to treat tumor growth. Injections were performed on days 37, 44, 50, and 64.

[0170] The solid curve with black squares corresponds to BR-AIb, and the dashed curve with white circles corresponds to 4.32 mg.kg -1 The dashed curve with black circles corresponds to 6.48 mg.kg -1 RC-AIb.

[0171] Figure 3 .Use 9.72mg.kg -1 or 12.96mg.kg -1 Body weight of mice treated with a single intravenous injection of RC-AIb, YG-AIb and AB-AIb.

[0172] Figure 4 The survival curves show the percentage of survival as a function of time (in days). Example

[0173] Chemical

[0174] General experimental methods

[0175] All reactions were performed under argon. Unless otherwise stated, solvents used were of HPLC quality. Chemicals were of analytical grade from commercial sources and used without further purification. The progress of the reaction was monitored on SIL G / UV254. (0.2 mm silica gel 60). Spots were visualized under 254 nm UV light and / or by immersing the TLC plate in a solution of phosphomolybdic acid (3 g) in ethanol (100 mL) and then heating with a heat gun.

[0176] Automatic chromatography is used The analysis was performed using a RF 200I TELEDYNE ISCO instrument equipped with UV and ESLD detectors using a fast column Silica gel 15 or 50 μm for normal phase chromatography, HPC18 GOLD 4g or 15.5g is used for reverse phase chromatography.

[0177] The 400MHz, 376MHz and 100MHz frequencies were recorded on a Bruker 400Avance III instrument at 400MHz, 376MHz and 100MHz, respectively. 1 H. 19 F and 13 C NMR spectra, the instrument was equipped with an ultra-shielded magnet and a BBFO 5 mm broadband probe. Chemical shifts (δ) are reported in parts per million (ppm) from low field to high field and are referenced to residual solvent. Coupling constants (J) are reported in Hertz (Hz).

[0178] The exact masses of all derivatives were determined by injection on high-resolution ESI mass spectrometers at the CBM / ICOA FR2708 of the University of Orleans and the IC2MP Center for Organic Analysis of the University of Poitiers.

[0179] Analytical RP-HPLC was performed on a Dionex Ultimate 3000 system at 30 °C and 1 ml.min -1 The system was equipped with a UV / visible variable wavelength detector and a reverse phase column chromatography MACHEREY-NAGEL (150 / 4.6, RP18, 5μm).

[0180] Method 1 used a linear gradient consisting of A (0.2% TFA in water) and B (CH3CN), starting from A / B = 80 / 20 v / v and reaching A / B = 0 / 100 v / v in 30 minutes. All chromatograms were recorded at 254 nm.

[0181] Synthetic strategy for obtaining RC-AIb

[0182]

[0183] 1. Synthesis of compound 5

[0184]

[0185] Azide reduction

[0186] PPh3 (730 mg, 2.78 mmol, 1.5 eq) was added to a solution of azide 3 (590 mg, 1.85 mmol, 1 eq) in THF (8 mL). The mixture was stirred at room temperature for 20 hours and at 50 ° C for 5 hours. After completion, the mixture was cooled to room temperature, and then water (2.5 mL) was added and the mixture was stirred for 18 hours. The solvent was removed under reduced pressure and the crude amine 4 was used in the next step without further purification.

[0187] Amide formation

[0188] To a cooled solution (0 ° C) of carboxylic acid 2 (Haibin Gu, Polymer, 2018, 146, 275-290) (455 mg, 1.60 mmol, 1 eq.) and crude amine 4 (1.85 mmol, 1.15 eq.) in dry CH2Cl2 (8 mL) was added DMAP (215 mg, 1.76 mmol, 1.1 eq.) and EDC.HCl (337 mg, 1.76 mmol, 1.1 eq.). The mixture was stirred at room temperature for 60 hours and diluted with CH2Cl2 (40 mL). The organic layer was washed with 1M HCl (40 mL). The aqueous layer was extracted with CH2Cl2 (3x40 mL). The combined organic layers were dried over magnesium sulfate and concentrated. The crude residue was purified by silica gel column chromatography (gradient elution CH2Cl2 / acetone 95 / 5 to 80 / 20) to give compound 5 (1.68 g, 86%) as a white solid.

[0189] R f :0.65(CH2Cl2 / MeOH 95 / 5)

[0190] 1 H NMR (400MHz, CDCl3) δ = 7.23 (s, 2H), 6.76 (bs, 1H), 5.00 (s, 1H), 4.81 (d, J = 2.3Hz, 4H), 4.79 (d, J = 2.4Hz, 2H), 3. 83-3.54(m,12H),3.49(t,J=5.1Hz,2H),3.26(m,2H),2.56(t,J=2.2Hz,2H),2.46(t,J=2.4Hz,1H),1.45(s,9H).

[0191] 13 C NMR(100MHz, CDCl3)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.

[0192] HRMS(ESI+):[M+Na] + C 29 H 38 The calculated value of N2NaO9 is: 581.2470, the measured value is 581.2487; [2M+Na] + C 58 H 76 N4NaO 18The calculated value is: 1139.5047, the measured value is 1139.5085.

[0193] 2. Synthesis of compound 7

[0194]

[0195] Cu(MeCN)4PF6 (7.0 mg, 0.019 mmol, 0.12 eq) was added to a dry and degassed CH2Cl2 (2 mL) solution of alkyne 5 (89 mg, 0.159 mmol, 1 eq) and azido-PEG10-amine 6 (285 mg, 0.541 mmol, 3.4 eq). The mixture was stirred at room temperature for 2 h. After completion, the resin IDA (200 mg) was added to the mixture, stirred for another 2 hours and removed by filtration. The solvent was evaporated under reduced pressure and the crude product was purified by reverse phase chromatography on C18 grafted silica (gradient elution MeCN / H2O 10 / 90 to 50 / 50 in 30 minutes) to give compound 7 (203 mg, 60%) as a colorless oil.

[0196] R t :5.72min(Method 1)

[0197] 1 H NMR (400MHz, 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 = 5.0Hz, 4H), 4.54 (t, J = 5.0Hz, 2H), 3.93 (t ,J=5.0Hz,4H),3.85(t,J=5.0Hz,2H),3.73-3.50(m,126H),3.47(t,J=5.6Hz,3H),3.19(t,J=5.6Hz,2H),2.87(t,J=5.1Hz,6H),1.42(s,9H).

[0198] 13 C NMR(100MHz,MeOD)δ=168.99,158.38,153.54,145.22,144.46,141.53,131.30,126.57,126.53,108.48,80.03,7 2.09,71.53,71.48,71.44,71.23,71.12,70.64,70.34,66.97,63.77,51.50,51.38,41.75,41.25,41.19,28.80.

[0199] HRMS(ESI+):[M+2H] 2+ C 95 H 178 N 14 O 39 Calculated value: 1069.6182, measured value: 1069.6170; [M+3H] 3+ C 95 H 179 N 14 O 39 Calculated value: 713.4145, measured value: 713.4138; [M+4H] 4+ C 95 H 180 N14O 39 The calculated value is: 535.3127, the measured value is 535.3125.

[0200] 3. Synthesis of compound 9

[0201]

[0202] EDC.HCl (134 mg, 0.7 mmol, 1 eq.) was added to a dry CH2Cl2 (4.6 mL) solution of carboxylic acid 8 (100 mg, 0.7 mmol, 1 eq.) and N-hydroxysuccinimide (80 mg, 0.7 mmol, 1 eq.). The mixture was stirred at room temperature for 60 hours. After completion, the mixture was diluted with CH2Cl2 (10 mL). The organic layer was washed with water (10 mL), then with brine (10 mL), dried over magnesium sulfate and concentrated under vacuum. The crude residue was purified by silica gel column chromatography (gradient elution CH2Cl2 / MeOH 100 / 0 to 99 / 1) to give compound 9 (135 mg, 80%) as a translucent yellow oil.

[0203] R f :033(PE / AcOEt 70 / 30)

[0204] 1 H NMR (400MHz, CD2Cl2) δ = 3.29 (t, J = 6.6Hz, 2H), 2.78 (s, 4H), 2.61 (t, J = 7.2Hz, 2H), 1.83-1.57 (m, 4H).

[0205] 13 C NMR (400MHz, CD2Cl2) δ = 169.24, 168.21, 50.73, 30.32, 27.78, 25.53, 21.75.

[0206] HRMS(ESI+):[M+H]+ C9H 13 The calculated value of N4O4 is 241.0931, and the measured value is 241.0936; [M+Na] + C9H 12 The calculated value of N4NaO4 is 263.0751, and the measured value is 263.0756; [2M+Na] + C 18 H 24 The calculated value of N8NaO8 is: 503.1609, and the measured value is 503.1618.

[0207] 4. Synthesis of compound 10

[0208]

[0209] To a solution of triamine 7 (60 mg, 0.028 mmol, 1 eq.) and NHS ester 9 (23.6 mg, 0.098 mmol, 3.5 eq.) in dry DMF (1 mL) was added Et3N (13.7 μL, 0.098 mmol, 3.5 eq.). The mixture was stirred at room temperature for 4 hours. After completion, the solvent was removed under reduced pressure and the crude residue was purified by silica gel column chromatography (gradient elution CH2Cl2 / MeOH 95 / 5 to 75 / 25) to give compound 10 (54.8 mg, 78%) as a colorless oil.

[0210] R t :11.00min(Method 1)

[0211] 1 H NMR(400MHz, CD2Cl2)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),4.55(t,J=5.1Hz,4H),4.49(t,J=5.2Hz,2H),3.88(t,J=5.1Hz,4H ),3.84(t,J=5.3Hz,2H),3.64-3.49(m,126H),3.47-3.43(m,2H),3.37(m,6H),3.27(t,J=6. 6Hz, 6H), 3.21 (m, 2H), 2.60 (s, 1H), 2.17 (t, J = 7.1Hz, 6H), 1.73-1.53 ​​(m, 12H), 1.39 (s, 9H).

[0212] 13C NMR (400MHz, CD2Cl2)δ=172.65,166.88,156.34,152.62,144.47,143.71 ,140.52,130.84,125.18,125.01,107.46,79.23,71.35,71.00,70.95,70 .89,70.84,70.79,70.71,70.67,70.55,70.32,69.88,69.81,66.70,63.4 1,51.74,50.77,50.54,40.84,40.50,39.65,36.12,28.87,28.63,23.26.

[0213] HRMS(ESI+):[M+Na] + C 110 H 197 N 23 O 42 The calculated value of Na is: 2535.3879, the measured value is 2535.3893; [M+2Na] 2+ C 110 H 197 N 23 O 42 The calculated value of Na2 is 1279.1885, and the measured value is 1279.1877; [M+3Na] 3+ C 110 H 197 N 23 O 42 The calculated value of Na3 is 860.4554, and the measured value is 860.4543; [M+4Na] 4+ C 110 H 197 N 23 O 42 The calculated value of Na4 is: 651.0889, and the measured value is 651.0888.

[0214] 5. Synthesis of compound 11

[0215]

[0216] TFA (160 μL) was added to a cooled (0°C) solution of carbamate 10 (70 mg, 0.02788 mmol, 1 eq) in dry CH2Cl2 (700 μL). The mixture was stirred at 0°C for 30 minutes and at room temperature for another 30 minutes. After completion, the solvent was evaporated under reduced pressure and the crude mixture was purified by reverse phase chromatography on C18 grafted silica (gradient elution MeCN / H2O (0.05% TFA) 20 / 80 to 100 / 0 over 30 minutes) to give compound 11 (65.5 mg, 91%) as a colorless oil.

[0217] R t :8.66min(Method 1)

[0218] 1 H NMR (400MHz, CD2Cl2)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=4.5Hz,4H),4.52-4.42(m,2H),3.88(t,J=4.6H z,4H),3.86-3.80(m,J=4.9Hz,2H),3.71(s,4H),3.69-3.45(m,131H),3.44-3.33(m,J=4 .5Hz,6H),3.27(t,J=6.2Hz,6H),3.14(s,3H),2.18(t,J=7.0Hz,6H),1.78-1.43(m,12H).

[0219] 13 C NMR (400MHz, CD2Cl2)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.

[0220] HRMS(ESI+):[M+H] + C 105 H 189 N 23 O 40 H calculated value: 2413.3535, measured value: 2413.3482; [M+2Na]2+ C 105 H 189 N 23 O 40 Calculated value of Na2: 1229.1623, measured value: 1229.1575; [M+2H] 2+ C 105 H 189 N 23 O 40 H2 calculated value: 1207.1804, measured value: 1207.1776; [M+H+Na] 2+ C 105 H 189 N 23 O 40 HNa calculated value: 1218.1714, measured value: 1218.1685; [M+3H] 3+ C 105 H 189 N 23 O 40 H3 calculated value: 805.1227, measured value: 805.1186; [M+3Na] 3+ C 105 H 189 N 23 O 40 The calculated value of Na3 is 827.1046, and the measured value is 827.1007; [M+H+2Na] 3+ C 105 H 189 N 23 O 40 The calculated value of HNa2 is 812.4500, and the measured value is 812.4480; [M+2H+Na] 3+ C 105 H 189 N 23 O 40 The calculated value of H2Na is 819.7773, and the measured value is 819.7746.

[0221] 6. Synthesis of compound 13

[0222]

[0223] A cooled solution (0°C) of LiOH.H2O (31.6 mg, 0.75 mmol, 8.75 equiv) in H2O (6 mL) was added dropwise to a cooled solution (-5°C) of MeOH (6 mL) of protected glucuronide derivative (PAPOT S. et al., Chem. Sci., 2017, 8, 3427-3433) 12 (109 mg, 0.086 mmol, 1 equiv). The mixture was stirred at -5°C until the starting material disappeared (30 min). It was then hydrolyzed with IRC-50 acidic resin. After filtration, the solvent was removed under reduced pressure and the crude product was purified by reverse phase chromatography on C18 grafted silica (gradient elution MeCN / H2O (0.05% TFA) 20 / 80 to 80 / 20 in 30 minutes) to give compound 13 (70 mg, 72%) as a white solid.

[0224] R t :11.49-11.80min(Method 1)

[0225] 1 H NMR (400MHz, 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,1 H),7.36-7.21(m,4H),7.19-7.15(m,1H),5.79-5.58(m,J=29.3Hz,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=9.4Hz,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).

[0226] 19 F NMR(376MHz,DMSO-d6)δ=-75.63

[0227] 13C NMR (400MHz, DMSO-d6)δ=172.38,172.32,169.92,169.86,169.78,169.76,169.65,168.74,168.71,158.52,158.14,154.56,154.54,154.38,1 48.76,148.68,143.67,139.84,134.20,132.63,127.81,127.74,126.7 4,126.66,126.48,126.41,122.90,116.19,100.11,85.45,81.64,77.68 ,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.

[0228] HRMS(ESI+):[M+Na] + C 56 H 82 N4NaO 18 The calculated value is: 1149.5578, the measured value is 1149.5635.

[0229] 7. Synthesis of compound 14

[0230]

[0231] CuSO4 (5.7 mg, 0.0356 mmol, 3.5 eq) and sodium ascorbate (7.0 mg, 0.0356 mmol, 3.5 eq) were added to a solution of alkyne 13 (34 mg, 0.030 mmol, 3 eq) and azide 11 (30 mg, 0.01187 mmol, 1.2 eq) in a degassed t-BuOH (6 mL) / H2O (10 mL) mixture. The mixture was stirred at room temperature under Ar atmosphere for 1.5 h. After completion, the resin was IDA (350 mg) was added to the mixture, stirred for another 3 hours and removed by filtration. The solvent was evaporated under reduced pressure and the crude residue was purified by reverse phase chromatography on C18 grafted silica (gradient elution MeCN / H2O (0.05% TFA) 20 / 80 to 60 / 40 in 30 minutes) to give compound 14 (24 mg, 41%) as a white solid.

[0232] R t :11.97-12.09min(Method 1)

[0233] 1 H NMR (400MHz, 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),4.83-4.5 9(m,1H),4.56(t,J=5.1Hz,4H),4.51-3.89(m,30H),3.83(t,J=5.2Hz,4H),3.81-3.72(m,3H),3.69-3.35(m,148H),3.3 4-3.10(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=6.4Hz,3H).

[0234] 19 F NMR (376MHz, DMSO-d6) δ = -74.53.

[0235] 13C NMR (400MHz, 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.

[0236] HRMS(ESI+):[M+7H] 7+ C 273 H 442 N 41 O 94 Calculated value: 828.4417, measured value: 828.4419; [M+6H] 6+ C 273 H 441 N 41 O 94 Calculated value: 966.3475, measured value: 966.3468; [M+5H] 5+ C 273 H 440 N 41 O 94 The calculated value is: 1159.4155, the measured value is 1159.4154.

[0237] 8. Synthesis of RC-AIb

[0238]

[0239] Et3N (0.94 μL, 0.00677 mmol, 4 eq) was added to a solution of amine 14 (10 mg, 0.00169 mmol, 1 eq) and NHS ester 15 (0.57 mg, 0.00186 mmol, 1.1 eq) in dry DMSO (1 mL). The mixture was stirred at room temperature for 7 hours. After completion, the solvent was evaporated under reduced pressure and the crude residue was purified by reverse phase chromatography on C18 grafted silica (gradient elution MeCN / H2O (0.05% TFA) 20 / 80 to 80 / 20 in 30 minutes) to give RC-Alb (9.9 mg, 98%, purity>98%) as a white solid.

[0240] R t :12.80-12.92min(Method 1)

[0241] HRMS(ESI+):[M+6H] 6+ C 283 H 452 N 42 O 97 Calculated value: 998.5263, measured value: 998.5261; [M+5H] 5+ C 283 H 451 N 42 O 97 Calculated value: 1198.0301, measured value: 1198.0306; [M+4H] 4+ C 283 H 450 N 42 O 97 The calculated value is: 1497.2858, and the measured value is 1497.2857.

[0242] Biological experiments

[0243] Cells: The MIA PaCa-2 human pancreatic cell line was obtained from the American Type Culture Collection and stably transfected with the luciferase gene by Trichet et al., INSERMNantes, France.

[0244] In vivo experimental procedures: Female nude mice aged 6 to 8 weeks were purchased from Charles River Laboratories. Before the experiments, mice were acclimated in the laboratory for 7 days and housed in sterile filter-intercept cages in controlled ventilation racks with ad libitum access to food and water. All experimental procedures involving animals were validated by the Regional Ethics Committee (CECCO n°3) and performed in accordance with the guidelines of the French Ministry of Agriculture and Forestry (Decree 2013-118) and the European Community Council Directive (2010 / 63 / UE). Throughout the study, mice were examined at least 3 times a week for clinical symptoms, distress, decreased physical activity, and body weight as indicators of health status.

[0245] In vivo efficacy on orthotopic models:

[0246] Considering the obvious ethical issues, the evaluation of new therapies and the understanding of biological mechanisms must be obtained from animal models. These animal studies correspond to the proof-of-concept phase for molecules that can be tested in clinical phases I and II. A critical 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 significant lack of predictability and a risky extrapolation of the results obtained in humans. In order to evaluate the efficiency of antitumor treatments, it is necessary to perform these studies on orthotopic models. Orthotopic models have the advantage of being more predictive of human tumor development. Tumors grow in original primary tumor tissue and these models are closer to the clinical pathological situation. Given the complexity of the processes involved, including the interaction with the tumor microenvironment, there are no reliable alternatives.

[0247] Human pancreatic cancer xenografts derived 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 and air. The abdomen of the mouse was prepared with povidone-iodine (Betadine) solution. A small transverse incision was made in the left flank through the skin and peritoneum. Gently grasping the tip of the pancreatic tail, the pancreas / spleen was exteriorized laterally for complete exposure. The needle was inserted into the pancreatic tail and positioned in the pancreatic head region. The inoculum (2x10 in 10 μL PBS) was slowly injected using a 27-gauge needle of a Hamilton syringe. 6 MIA PaCa2-luc cells). The spleen was then returned to its proper position in the abdomen, and the peritoneum was sutured with 7-0 sutures and the skin was sutured with 4-0 sutures.

[0248] Treatment of pancreatic tumors: Mice (8 per group) received intravenous injections of 5% DMSO and 95% PBS mixture (vehicle group), 2 mg / kg BR-AIb, 4.32 mg / kg RC-AIb, or 6.48 mg / kg RC-AIb on days 37, 44, 50, and 64. Tumor volume was determined by ultrasound imaging.

[0249] Ultrasound imaging: Mice were anesthetized by inhalation of 1.5% isoflurane in air and placed on a thermostatically controlled electric heating pad with their paws tied to electrocardiogram (ECG) electrodes fixed to a table. ECG-derived respiratory gating avoided artifacts due to respiratory movements of the animals. The temperature of the animals was recorded with an internal temperature probe. Aqueous warming ultrasound gel (purchased from Supragel) was applied to the skin to cover the skin to optimize visualization of internal organs. Tumors were imaged with a Vevo LAZR system (FUJIFILM Visualsonics Inc.). A transducer with a center frequency of 40 MHz, providing an axial resolution of 40 μm and a field of view of 14.1x15 mm was used to image smaller tumors. A transducer with a center frequency of 21 MHz, providing an axial resolution of 75 μm and a field of view of 23.1x36 mm was used to image larger tumors. 3D scans of the ultrasound images were digitally recorded and reviewed. The tumor area in the coronal plane was measured by manually outlining the margins using Vevo LAB 1.7.2 software (FUJIFILM Visualsonics Inc.). The software then calculated the corresponding volume for each coronal slice with a detection threshold of 0.5 mm. 3 Up to 1.5mm 3 , depending on the location of the tumor.

[0250] As mentioned above, RC-AIb is able to vectorize three MMAE molecules and introduce them into the microenvironment of solid tumors. RC-AIb includes:

[0251] 1) Maleimide, which allows covalent bonding with plasma albumin and entry into the bloodstream;

[0252] 2) a gallic acid structure, which provides a connection between the maleimide functional group and the three glucuronidated units; and

[0253] 3) The three linkers selectively release the active agent into the tumor microenvironment through the action of β-glucuronidase.

[0254] The therapeutic activity of RC-AIb in mice was evaluated as described above ( Figure 1 The results showed that RC-AIb had significant anticancer activity without any side effects. In addition, the use of 6.48 mg / kg of RC-AIb caused complete regression of tumor masses in 38% of the treated animals.

[0255] The therapeutic activity of RC-AIb was also compared with that of the corresponding monomeric BR-AIb (carrying a single MMAE molecule) ( Figure 2 When the two compounds were administered at the same dose (1.1x10 -6When administered at 100 mol / kg, RC-AIb resulted in an 18-fold reduction in tumor volume compared with BR-AIb.

[0256] Other compounds according to the invention

[0257] 1. Synthesis of YG-AIb and AB-AIb

[0258] To evaluate the effect of the maleimide moiety on the toxicity of the trimeric glucuronide prodrugs, two compounds, YG-AIb and AB-AIb, were prepared as follows:

[0259]

[0260] 1.1. Preparation of YG-AIb

[0261] The synthesis of 5-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-fluorobenzoic acid 15 was achieved using a published protocol (WO 2016054315).

[0262] a) 2,5-dioxopyrrolidin-1-yl 5-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-fluorobenzoate 16

[0263]

[0264] NHS (352 mg, 3.06 mmol, 1.2 equiv) and EDC hydrochloride (590 mg, 3.08 mmol, 1.2 equiv) were added to a solution of 5-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)-2-fluorobenzoic acid 15 (602 mg, 2.56 mmol, 1.0 equiv) in anhydrous DMF (3 μL) stirring. The reaction mixture was stirred at room temperature overnight. The solvent was removed under vacuum. The crude product was dissolved in DCM (15 μL) and washed with 0.1N HCl aqueous solution (10 μL) and water (2x10 μL) successively. The organic layer was dried over magnesium sulfate, filtered and evaporated. The crude product was purified by flash silica gel column chromatography using a gradient of DCM / AcOEt 50 / 50 to 0 / 100 to obtain an active ester 16 (468 mg, 1.41 mmol) as a beige solid. Yield: 55%.

[0265] b)YG-AIb

[0266] To a stirred solution of compound 14 (10.3 mg, 1.78 μmol, 1.0 equiv) in anhydrous DMSO (2 μL) was added 5-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-fluorobenzoic acid 2,5-dioxopyrrolidin-1-yl ester 16 (1.8 mg, 5.33 μmol, 3.0 equiv) and triethylamine (1.5 μL, 10.8 μmol, 6.0 equiv). The reaction mixture was stirred at room temperature overnight. The solvent was removed under vacuum. The crude product was purified by flash reverse phase (C18) silica gel column chromatography using a gradient of H2O+0.05% TFA / ACN 90 / 10 to 40 / 60 to give compound YG-AIb (5.1 mg, 0.854 μmol) after freeze drying. Yield: 48%. HRMS m / z[M+4H] 4+ Calculated value: 1504.0271, measured value: 1504.0273, [M+5H] 5+ Calculated value: 1203.2225, measured value: 1203.2224, [M+6H] 6+ Calculated value: 1003.0205, measured value: 1003.0204

[0267] Preparation of AB-AIb

[0268] The synthesis of 3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-4,4,4-trifluorobutyric acid 17 was achieved using a published protocol Bioconjugate. Chem., 2015, 26, 145-152.

[0269] a) 3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-4,4,4-trifluorobutyric acid 2,5-dioxopyrrolidin-1-yl ester 18

[0270]

[0271] To a solution of 3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-4,4,4-trifluorobutyric acid 17 (500 mg, 2.51 mmol, 1.0 equiv) in anhydrous DMF (3 mL) 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 stirred at room temperature overnight. The solvent was removed under vacuum. The crude product was purified by flash silica gel column chromatography using a gradient of DCM / MeOH 100 / 0 to 75 / 25 to give the active ester 18 (487 mg, 1.46 mmol) as a white solid. Yield: 58%.

[0272] b)AB-AIb

[0273] To a stirred solution of compound 14 (10.5 mg, 1.78 μmol, 1.0 equiv) in anhydrous DMSO (2 mL) was added 3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-4,4,4-trifluorobutyric acid 2,5-dioxopyrrolidin-1-yl ester 18 (1.8 μg, 5.33 μmol, 3.0 equiv) and triethylamine (1.5 μL, 10.8 μmol, 6.0 equiv). The reaction mixture was stirred at room temperature overnight. The solvent was removed under vacuum. The crude product was purified by flash reverse phase (C18) silica gel column chromatography using a gradient of H2O+0.05% TFA / ACN 90 / 10 to 40 / 60 to give compound AB-AIb (6.4 mg, 1.06 μmol) after freeze drying. Yield: 60%. HRMS m / z[M+4H] 4+ Calculated value: 1504.5263, measured value: 1504.5269, [M+5H] 5+ Calculated value: 1203.6219, measured value: 1203.6224, [M+6H] 6+ Calculated value: 1003.3533, measured value: 1003.3529

[0274] 2. MTD studies

[0275] Maximum tolerated dose study (MTD): MTD studies were conducted on BALB / c nude mice (n=3). Single administration of prodrugs RC-AIb, YG-AIb and AB-AIb (9.72 mg / kg or 12.96 mg / kg) was performed by intravenous injection. The toxicity of the prodrugs was evaluated by maximum body weight loss or gain, expressed as a percentage of the initial body weight of the animals.

[0276] If the relative weight loss was greater than 20% of the initial body weight, the dose was considered toxic.

[0277] The results are shown in Figure 3 and Figure 4 middle.

[0278] Comparative MTD studies of RC-AIb, YG-AIb, and AB-AIb showed that the structure of the maleimide moiety had an effect on the toxicity of the glucuronide prodrugs in mice. Thus, prodrugs YG-AIb and AB-AIb with electron-withdrawing groups such as F and CF3 were less toxic than RC-AIb, which lacked this chemical functional group.

Claims

1. A compound having the following formula (I): in: R a Indicates H or (C1-C 12 )alkyl, optionally interrupted by one or more oxygen atoms; L represents a group having the following formula (II): in: A is an anticancer agent, and the anticancer agent is monomethyl oxetine E (MMAE); Y is an electron withdrawing group selected from the group consisting of halogen, NO2 and CF3; X is -O-; G is a glucuronic acid group or a derivative thereof selected from a glucuronide group; L1 represents a linker represented by the following formula (III); -A1-A2-A3-A4-A5-A6-A7-(III) in: A1 is (C1-C6)alkylene; A2 is triazolyl; A3 is (C1-C6)alkylene; A4 is selected from: -C(=O)-NR b -、-C(=S)-NR b -、-NR b -C(=O)-, -NR b -C(=S)- and NR b , R b Indicates H or (C1-C 12 )alkyl; A5 is interrupted by at least one oxygen atom (C1-C 32 ) alkylene; A6 is triazolyl; A7 is (C1-C6)alkylene; L' represents a group having the following formula (IV): -A8-A9-L"(IV) in: A8 is a (C1-C6)alkylene group interrupted by at least one oxygen atom; A9 is selected from the following: -NR c -, -O- and -S-, R c Indicates H or (C1-C 12 ) alkyl; and L" is a maleimidocaproyl group or a group having one of the following formulae: or a pharmaceutically acceptable salt thereof.

2. The compound according to claim 1, wherein L has the following formula (VII): A, Y and L1 are as defined in claim 1.

3. The compound according to claim 1, wherein L has the following formula (VIII): A, A1, A3, A4, A5 and A7 are as defined in claim 1.

4. The compound according to claim 3, wherein A5 is represented by -CH2-(CH2-O-CH2) n -CH2- group, wherein n is an integer from 1 to 12.

5. The compound according to claim 1, wherein L has the following formula (IX): i is an integer from 1 to 6; j is an integer from 1 to 10; n is an integer from 1 to 12, and k is an integer from 1 to 6.

6. The compound according to claim 1, wherein A8 is represented by -CH2-(CH2-O-CH2) m -CH2- group, and m is an integer of 1 to 12.

7. The compound according to claim 1, wherein L' has the following formula (X): m is an integer from 1 to 6, and p is an integer of 1-6.

8. The compound according to claim 1, having the following formula (XI), (XI') or (XI"):

9. The compound of claim 1 for use as a medicament.

10. A pharmaceutical composition for treating pancreatic cancer, comprising the compound of claim 1, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

11. Use of the compound according to claim 1 in the preparation of a medicament for treating and / or preventing pancreatic cancer.

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