EP4340887A1 - Bifunctional protac-type compounds targeting pxr, method for preparing same, and therapeutic use thereof - Google Patents

Abstract

The present application relates to novel bifunctional PROTAC-type compounds simultaneously binding the target protein PXR and E3-ubiquitin ligase, to a method for preparing same, and to uses thereof for treating cancers overexpressing PXR.

Description

DESCRIPTION

TITLE: BIFUNCTIONAL COMPOUNDS OF THE PROTAC TYPE TARGETING PXR, THEIR PREPARATION METHOD AND THEIR USE

IN THERAPEUTIC

The present invention relates to the treatment of cancer and more particularly cancers overexpressing the PXR nuclear receptor, such as colorectal cancer.

Colorectal cancer (CRC) is the third most common cancer and is the third leading cause of cancer death. Current treatments include surgery, radiation therapy, and chemotherapy, sometimes in combination with targeted therapies that demonstrate minor improvement. However, the efficacy of these treatments is seriously compromised by the frequent appearance of resistance which leads to relapse in patients after stopping treatment (50% of patients). In recent years, cancer cell subpopulations, cancer stem cells (CSCs), have been shown to be involved in tumor initiation, metastatic development and drug resistance, thus leading to tumor recurrence. .

The inventors have now demonstrated that the nuclear receptor PXR (NR112) is preferentially activated in cancer stem cells and that the extinction of its expression by interference with RNA (shRNA) sensitizes this cell population, normally resistant to chemotherapy, and significantly delays tumor recurrence in mice. Inhibition of the PXR nuclear receptor (NR112) therefore makes it possible to sensitize cancer stem cells to current treatments.

The PXR antagonists identified to date (L-sulforaphane, ketoconazole and SAP-70) are, however, either non-specific and/or toxic at the concentrations necessary for PXR inactivation, or not yet validated for clinical use.

PROTACs (“Proteolysis Targeting Chimera”) are bifunctional molecules that simultaneously bind a target protein and an E3-ubiquitin ligase. This causes poly-ubiquitination of the target protein which is thus degraded into small peptides and amino acids by the proteasome complex. The PROTAC approach is therefore a chemical protein knock-down strategy

It is therefore desirable to provide bi-functional chimeric ligands capable of inducing the targeted proteolysis of PXR according to the PROTAC strategy. According to a first object, the present invention relates to the bifunctional compounds corresponding to the general formula (I):

L(PXR)-Linker-L(E3 ligase)

(l) in which:

L(PXR) is a ligand capable of binding to the PXR nuclear receptor,

L(E3 ligase) represents a ligand of the E3 ubiquitin ligase, and

Linker represents a group that makes it possible to covalently bind L(PXR) to

L(E3 ligase).

The ubiquitin-proteasome (UPP) pathway is an essential cellular pathway that regulates key regulatory proteins and degrades misfolded or abnormal proteins. UPP is central to several cellular processes. If it is faulty or unbalanced, it leads to the pathogenesis of various diseases. The covalent attachment of ubiquitin to specific protein substrates is achieved by the action of ubiquitin ligases E3. These ligases include more than 500 different proteins and are classified into several classes defined by the structural element of their functional activity E3.

The E3 ligase ligand, which is a functional modality of the present compounds, binds to an E3 ubiquitin ligase. The ligase catalyzes the covalent attachment of ubiquitin to the target protein, which in turn induces the degradation of the target protein by native proteasomes. Thus, the compounds of the present invention are designed in a manner that exploits native cellular degradative processes but where the degradative action is directed to undesirable target proteins that are implicated in disease etiology.

Unlike conventional chemical inhibitors, the PROTACs according to the invention act as degradation enzymes with a super-stoichiometric action capacity.

The advantages of the compounds according to the invention are therefore multiple:

1) they are active at lower concentrations than the inhibitor alone which requires high levels of systemic exposure in order to achieve target saturation,

2) they can carry out multiple cycles of degradation leading to the degradation of the targeted protein,

3) compared to the rapid dissociation kinetics of inhibitors on their target, the restoration of protein function after PROTAC-induced degradation requires de novo synthesis of the protein by the cell, which takes much longer and thus increases the duration effects of PROTACs. L(PXR) which is a functional modality of the present compounds which binds to PXR. In certain embodiments, the targeting ligand is an analog of the ligands of

According to one embodiment, L(PXR) can be chosen from the groups of formula

(II): where I represents the group's attachment to Linker; or a pharmaceutically acceptable salt.

Thus, the compounds according to the invention can correspond to the following formula (1-1): where Linker, L(E3 ligase) are as defined above or below, or a pharmaceutically acceptable salt.

According to one embodiment, the E3 ligase ligand binds to cereblon. L(E3 ligase) can in particular be chosen from: - the groups of formula (NIA): and

- formula groups (INB): or a pharmaceutically acceptable salt, in which formulas (NIA) and (INB):

X is NH;

X' is -C(O)- or -CH ₂ -;

Y represents H or a C1-C6 alkyl group; represents the group's attachment to Linker.

Thus, the compounds according to the invention may in particular correspond to the formula (I-) or (1-3): where Linker, L(PXR), L(E3 ligase), X, X′, Y are as defined above or below; or a pharmaceutically acceptable salt.

More particularly, the compounds according to the invention can correspond to one of the following formulas

(1-5) in which L(PXR), Linker are defined above or below; or a pharmaceutically acceptable salt.

Linker provides covalent attachment of the targeting ligand with the E3 ligase ligand. According to one embodiment, Linker represents a C1 -C20 alkylene group, optionally interrupted or optionally ending at one and/or the other of the two ends, by one of the groups -0-, -S-, -N (R') -, -C(O)-, -C(0)0-, - OC(O) -, -0C(0)0 -, - C(NOR')-, -C(0 )N(R')-, -C(0)N(R')C(0)-, -C(0)N(R')C(0)N(R')-, -N(R' )C(0)-, -

N(R')C(0)N(R')-, -N(R')C(0)0-, -0C(0)N(R')-, -C(NR')-, - N(R')C(NR')-, -C(NR')N(R') -, - N(R')C(NR')N(R')-, -S(0) ₂ - , -OS(O)-, -S(0)0-, -S(O)-, -0S(0) ₂ -, -N(R')S(0) ₂ -, -S(0) ₂ N(R')-, -N(R')S-, -S(0)N(R')-, -N(R')S(0) ₂ N(R') -, -N(R ')S(0)N(R')-, C3 to C12 cycloalkylene, heterocyclene of 3 to 12 members and comprising 1, 2 or 3 heteroatoms chosen from N, O, S, heteroarylene of 5 to 12 members and comprising 1 , 2 or 3 heteroatoms chosen from N, O, S, or any combination thereof, and in which identical or different R′ represent H or a C1-C6 alkyl group. Thus, according to a particular embodiment, Linker can be chosen from C4-C20 alkylene groups, optionally interrupted by and/or ending with one or more groups chosen from -NH-, -O-, -C(O) -; piperidinyl, piperazinylene.

More particularly, Linker can be represented among the groups of formula

(IV): where Li and L ₂ , identical or different, independently represent an alkylene group of 1 to 12 carbon atoms optionally interrupted or ending in a heterocyclene of 3 to 12 members and comprising 1, 2 or 3 heteroatoms chosen from N,

BONE ;

Li is bound is linked to L(E3 ligase);

Z represents H or a C1-C6 alkyl group.

According to a more particular embodiment, Li is a C7-alkylene group

(-C7H14-).

According to a more particular embodiment, L ₂ is a (C2 to C8)-alkylene group optionally interrupted by a piperidinyl group.

According to one embodiment, the compounds according to the invention may correspond to the following formula (V): in which L ₂ represents a linear C2-C8 alkylene group optionally interrupted by a piperidinyl group, and L(E3 ligase) is as defined above or below.

The formulas (I), (II), (NIA), (INB), (IV), (V) represented here also cover the pharmaceutically acceptable salts thereof, their isotopic derivatives and their stereoisomers.

As used above and below and unless specifically stated: "Alkyl" means an aliphatic hydrocarbon group which may be linear or branched having about 1 to about 20 carbon atoms in the chain. Preferred alkyl groups have 1 to about 12 carbon atoms in the chain, especially 1 to 6 carbon atoms. Branched means that one or more lower alkyl groups, such as methyl, ethyl or propyl, are attached to a straight alkyl chain. "Lower alkyl" means from about 1 to about 4 carbon atoms in the chain which may be straight or branched. The alkyl may be substituted with one or more "alkyl group substituents", which may be the same or different and include halo, cycloalkyl, hydroxy, alkoxy, amino, acylamino, aroylamino, carboxy, alkoxycarbonyl, aralkoxycarbonyl, heteroaralkoxycarbonyl or Y ¹ Y ² NCO-, wherein Y ¹ and Y ² are, independently, hydrogen, optionally substituted alkyl, optionally substituted aryl, optionally substituted aralkyl or optionally substituted heteroaralkyl, or Y ¹ and Y ² , taken together in conjunction with N through which Y ¹ and Y ² are linked, form a 4 to 7 membered heterocyclyl. Typical examples of alkyl groups include methyl, trifluoromethyl, cyclopropylmethyl, cyclopentylmethyl, ethyl, n-propyl, 17-propyl, n-butyl, t-butyl, n-pentyl, 3- pentyl, methoxyethyl, carboxymethyl, methoxycarbonylethyl, benzyloxycarbonylmethyl, pyridylmethyloxycarbonylmethyl.

“Alkylene” designates a bivalent alkyl group as defined above. Preferred alkylene groups are lower alkylene groups having 1 to about 6 carbon atoms. Typical examples of alkylene groups include methylene and ethylene.

"Cycloalkyl" means a mono- or multi-cyclic non-aromatic ring system of about 3 to about 10 carbon atoms, preferably about 5 to about 10 carbon atoms. Preferred ring sizes of the rings of the ring system include about 5 to about 6 ring atoms, optionally substituted with one or more substituents. Exemplary monocyclic cycloalkyls include cyclopentyl, cyclohexyl, cycloheptyl, and the like. Exemplary multicyclic cycloalkyls include 1-decalin, norbornyl, adamant-(1 or 2-)yl, and the like.

“Cycloalkylene” means a saturated, divalent cycloalkyl group as defined above, such as in particular cyclohexylene.

"Heterocyclyl" means a non-aromatic saturated monocyclic or multicyclic ring system of about 3 to about 10 carbon atoms, preferably about 5 to about 10 carbon atoms, wherein one or more of the carbon atoms in the system ring is/are hetero element(s) different from carbon, for example nitrogen, oxygen or sulphur. Preferred ring sizes of the rings of the ring system include about 5 to about 6 ring atoms. The designation of aza, oxa or thia as a prefix before heterocyclyl defines that at least one nitrogen, oxygen or sulfur atom is present, respectively, as a ring atom. The heterocyclyl may be optionally substituted with one or more substituents, which may be the same or different, and are as defined herein. The nitrogen atom of a heterocyclyl may be a basic nitrogen atom. The nitrogen or sulfur atom of the heterocyclyl can also be optionally oxidized to the corresponding N-oxide, S-oxide or S,S-dioxide. Exemplary monocyclic heterocyclyl rings include piperidyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiazolidinyl, 1,3-dioxolanyl, 1,4-dioxanyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like.

The term “heterocyclene” denotes a bivalent heterocyclyl radical as defined above.

"Heteroaryl" means an aromatic monocyclic or multicyclic ring system of about 5 to about 14 carbon atoms, preferably about 5 to about 10 carbon atoms, wherein one or more of the carbon atoms in the ring system is/are heteroelement(s) other than carbon, for example nitrogen, oxygen or sulphur. Preferred ring sizes of the rings of the ring system include about 5 to about 6 ring atoms. "Heteroaryl" can also be substituted with one or more substituents. The designation of aza, oxa or thia as a prefix before heteroaryl defines that at least one nitrogen, oxygen or sulfur atom is present respectively as a ring atom. A nitrogen atom of a heteroaryl can be a basic nitrogen atom and can also be optionally oxidized to the corresponding N-oxide. Exemplary heteroaryl and substituted heteroaryl groups include pyrazinyl, thienyl, isothiazolyl, oxazolyl, pyrazolyl, furazanyl, pyrrolyl, 1,2,4-thiadiazolyl, pyridazinyl, quinoxalinyl, phthalazinyl, l imidazo[1,2-a]pyridine, imidazo[2,1-b]thiazolyl, benzofurazanyl, azaindolyl, benzimidazolyl, benzothienyl, thienopyridyl, thienopyrimidinyl, pyrrolopyridyl, imidazopyridyl, benzoazaindole, 1,2,4-triazinyl, benzthiazolyl, furanyl, imidazolyl, indolyl, indolizinyl, isoxazolyl, isoquinolinyl, isothiazolyl, oxadiazolyl, pyrazinyl, pyridazinyl , pyrazolyl, pyridyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolinyl, 1,3,4-thiadiazolyl, thiazolyl, thienyl and triazolyl. Preferred heteroaryl groups include pyrazinyl, theinyl, pyridyl, pyrimidinyl, isoxazolyl and isothiazolyl.

"Heteroarylene" denotes a bivalent heteroaryl radical as defined above. “Substituents” denotes one or more identical or different groups chosen from halogen, cyano, cycloalkyl, hydroxy, alkoxy, amino, alkylamino, dialkylamino, aroylamino, carboxy, alkoxycarbonyl, aralkoxycarbonyl, heteroaralkoxycarbonyl.

The compounds of the present invention may be in the form of a free acid or a free base, or a pharmaceutically acceptable salt.

The term "pharmaceutically acceptable salts" refers to the relatively non-toxic, inorganic and organic acid addition salts, and base addition salts, of the compounds of the present invention. These salts can be prepared in situ during the final isolation and purification of the compounds. In particular, acid addition salts can be prepared by separately reacting the purified compound in its purified form with an organic or inorganic acid and isolating the salt thus formed. Examples of acid addition salts include hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate salts. , maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptanate, lactobionate, sulfamates, malonates, salicylates, propionates, methylenebis-b-hydroxynaphthoates, gentisic acid, isethionates, di-p-toluoyltartrates, methanesulfonates, ethanesulfonates, benzenesulfonates, p- toluenesulfonates, cyclohexyl sulfamates and quinateslaurylsulfonate, and the like. (See, for example, SM Berge et al. "Pharmaceutical Salts" J. Pharm. Sci, 66:p.1-19 (1977) which is incorporated herein by reference). Acid addition salts can also be prepared by separately reacting the purified compound in its acid form with an organic or inorganic base and isolating the salt thus formed. Acid addition salts include amine and metal salts. Suitable metal salts include sodium, potassium, calcium, barium, zinc, magnesium and aluminum salts. Sodium and potassium salts are preferred. Suitable basic inorganic addition salts are prepared from metallic bases which include sodium hydride, sodium hydroxide, potassium hydroxide, calcium hydroxide, aluminum hydroxide, lithium hydroxide, magnesium hydroxide, zinc hydroxide . Suitable basic amine addition salts are prepared from amines which have sufficient alkalinity to form a stable salt, and preferably include amines which are often used in medicinal chemistry due to their low toxicity and acceptability. for medical use: ammonia, ethylenediamine, N-methyl-glucamine, lysine, arginine, ornithine, choline, N,N'-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine, diethylamine, piperazine, tris(hydroxymethyl)- aminomethane, tetramethylammonium hydroxide, triethylamine, dibenzylamine, ephenamine, dehydroabietylamine, N-ethylpiperidine, benzylamine, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, ethylamine, basic amino acids, for example lysine and arginine, and dicyclohexylamine, and the like.

Compounds of the present invention may have at least one chiral center and therefore may be in the form of a stereoisomer, which as used herein encompasses all isomers of individual compounds which differ only in orientation. of their atoms in space. The term stereoisomer includes mirror image isomers (enantiomers that include the (R-) or (S-) configurations of compounds), mixtures of mirror image isomers (physical mixtures of enantiomers and racemates or racemic mixtures ) of geometric compounds (cis/trans or E/Z, R/S isomers) of compounds and isomers of compounds with more than one chiral center that are not mirror images of each other (diastereoi somers). The chiral centers of compounds can undergo epimerization in vivo; thus, for these compounds, administration of the compound in its (R~) form is considered equivalent to administration of the compound in its (S-) form. Accordingly, the compounds of the present invention can be made and used as individual isomers and substantially free of other isomers, or as a mixture of various isomers, eg, racemic mixtures of stereoisomers.

In certain embodiments, the following compounds are suitable for binding Cereblon and PXR:

[Table 1]

Very particularly, the compounds according to the invention can be chosen from the compounds corresponding to one of the following formulas: According to another object, the present invention also relates to the process for the preparation of a compound according to the invention.

The compounds of general formula (I) can be prepared by applying or adapting any method known per se to and/or within the reach of those skilled in the art, in particular those described by Larock in Comprehensive Organic Transformations, VCH Pub., 1989 , or by application or adaptation of the methods described in the examples which follow.

According to the invention, said method comprises the coupling of a compound of formula (B) and a compound of formula (C):

L(PXR)-T T’-L(E3 ligase)

(B) (C) such that L(PXR) and L(E3 ligase) are as defined above, and T and T' are two precursor groups of Linker, that is to say the coupling of which makes it possible to drive to the Linker group, such that they each respectively possess a complementary reactive terminal function.

Here, the term "complementary reactive functions" designates two functions capable of reacting together to form a function providing a covalent bond between T and T'. Thus, typically, T and T' are such that T has a terminal function of the amine type and T' has a terminal function of the carboxylic acid type.

Thus, typically, T represents a group of formula (T-B):

-LI-NH2 (TB) and T' represents a group

(TC) in which Li and L ₂ are as defined above. Said coupling can advantageously be carried out in the presence of a peptide coupling agent such as BOP (benzotriazol-l -yloxytris (dimethylamino) phosphonium hexafluorophosphate, typically in the presence of an organic base such as Hünig base (N, /V- diisopropylethylamine (DIPEA or DIEA).

According to one embodiment, compound (B) corresponds to formula (A):

According to one embodiment, the compound (C) corresponds to the formula (C-1): ligase)

(C-1) in which L ₂ and L(E3 ligase) are as defined above.

Optionally, said method may also comprise the step consisting in isolating the product of formula (I) obtained.

In the reactions described below, it may be necessary to protect reactive functional groups, for example hydroxy, amino, imino, thio, carboxy groups, when they are desired in the final product, to avoid their undesirable participation in the reactions. Traditional protecting groups can be used according to standard practice, for examples see TW Green and PGM Wuts in Protective Groups in Organic Chemistry, John Wiley and Sons, 1991; JFW McOmie in Protective Groups in Organic Chemistry, Plenum Press, 1973. The compound thus prepared can be recovered from the reaction mixture by conventional means. For example, compounds can be recovered by distilling the solvent from the reaction mixture or if necessary after distilling the solvent from the solution mixture, pouring the remainder into water followed by extraction with an organic solvent immiscible in the solution. water, and distilling the solvent from the extract. Further, the product can, if desired, be further purified by various techniques, such as recrystallization, reprecipitation or the various chromatography techniques, including column chromatography or preparative thin layer chromatography. It will be appreciated that compounds useful according to the present invention may contain asymmetric centers. These asymmetric centers may be independently in the R or S configuration. It will be apparent to those skilled in the art that certain compounds which are useful according to the invention may also exhibit geometric isomerism. It is to be understood that the present invention includes individual geometric isomers and stereoisomers and mixtures thereof, including racemic mixtures, of compounds of formula (I) above. This type of isomers can be separated from their mixtures, by the application or adaptation of known methods, for example chromatography techniques or recrystallization techniques, or they are prepared separately from the appropriate isomers of their intermediates. The basic products or reagents used are commercially available and/or can be prepared by the application or adaptation of known methods, for example methods as described in the Reference Examples or their obvious chemical equivalents. The process according to the invention can implement the intermediate of formula (A) which is new.

According to another subject, the present invention therefore also relates to the compound of formula (A): The compound of formula (A) can be prepared by coupling the following compounds:

This coupling can typically be achieved by applying or adapting the procedure described in example 1.

According to the present invention, the compounds of formula (I) are capable of inducing the targeted proteolysis of PXR. The compounds of formula (I) are therefore useful in the treatment and/or prevention of cancers, in particular cancers overexpressing PXR. A subject of the present invention is therefore also pharmaceutical compositions comprising a compound according to the invention with a pharmaceutically acceptable excipient.

Preferably, said composition contains an effective amount of the compound according to the invention.

According to another object, the present invention also relates to a compound of general formula (I) for the treatment and/or prevention of cancers, in particular cancers overexpressing PXR.

Cancers overexpressing PXR include colorectal cancer, and cancers of the pancreas, liver and breast.

Typically, the compounds according to the invention can be used in combination with an anti-cancer agent. Such anticancer agents can be chosen in particular from 5-Fluorouracil (5-FU), Irinotecan (CPT11), Oxaliplatin, Cisplatin, Tamoxifen, Paclitaxel, Doxorubicin, Vonblastine, Cyclophosphamide (CPA), Isophosphamide (IFO).

Preferably, said composition is administered to a patient in need thereof. Said patient is in particular a patient resistant to the aforementioned anti-cancer agents. The type of formulation of the pharmaceutical compositions of the invention depends on the mode of administration, which may include enteral (e.g. oral), parenteral (e.g. subcutaneous (sc), intravenous (iv), intramuscular (im) injection. and intrasternal, or infusion techniques, intravenous or intravenous), arterial, intramedullary, intrathecal, intraventricular, transdermal, interdermal, rectal, intravaginal, intraperitoneal, topical mucosa, nasal, buccal, sublingual, intratracheal instillation, bronchial instillation and/or inhalation . In general, the most appropriate route of administration depends on a variety of factors, including the nature of the agent (e.g., its stability in the digestive tract environment) and/or the condition of the subject (e.g., whether subject is able to tolerate oral administration). In some embodiments, the compositions are formulated for oral or intravenous administration (eg, systemic intravenous injection).

The term "pharmaceutically acceptable carrier", as known in the art, means a pharmaceutically acceptable material, composition or vehicle suitable for administering compounds of the present invention to mammals. Suitable carriers can include, for example, liquids (both aqueous and non-aqueous and combinations thereof), solids, encapsulating materials, gases and combinations thereof (e.g. semi-solids) , which function to carry or transport the compound from one organ or body part to another organ or body part. A carrier is "acceptable" in the sense that it is physiologically inert and compatible with the other components of the formulation and is non-toxic to the subject or patient. Depending on the type of formulation,

Therefore, the compounds of formula I can be formulated into solid compositions (eg powders, tablets, dispersible granules, capsules, cachets and suppositories), liquid compositions (eg solutions in which the compound is dissolved, suspensions in which the particles of the compound are dispersed, emulsions and solutions containing liposomes, micelles or nanoparticles, syrups and elixirs); semi-solid compositions (for example, gels, suspensions and creams); and gases (eg propellants for aerosol compositions). Compounds can also be formulated for rapid, intermediate or sustained release.

The excipients which are suitable for solid administration are derivatives of cellulose or microcrystalline cellulose, alkaline-earth carbonates, magnesium phosphate, starches, modified starches, lactose for the forms solid. For parenteral use, water, aqueous solutions, saline, isotonic solutions are the most conveniently used vehicles.

The dosage may vary within significant limits depending on the therapeutic indication and the route of administration, as well as the age and weight of the subject.

tricks

[Fig 1] Figure 1 depicts the PXR affinity of pre-PROTAC JMV6944 measured by RT-FRET.

[Fig 2] Figure 2 illustrates the activation of PXR by the pre-PROTAC JMV6944 and the resulting PROTACs measured using a luciferase reporter gene placed under the control of the CYP3A4 promoter, the target gene of PXR.

[Fig 3] Figures 3A and 3B represent the induction of a PXR target gene (i.e. CYP3A4) by pre-PROTAC JMV6944 and subsequent PROTACs measured by RT-qPCR.

[Fig 4] Figures 4A and 4B illustrate and represent the effect of PROTACs JMV7048 and JMV7965 on the induction of CYP34 by Western blot.

[Fig 5] Figures 5A and 5B illustrate the effects of PROTACs on cell viability in different cell lines (LS174T, FIT29) and primary culture (CRC1) from colon cancer.

[Fig 6] Figures 6A-E represent the effects of PROTACs on the degradation of the PXR protein in LS174T cells, measured by Western blot.

[FIG 7] Figures 7A and 7B respectively represent the effects of PROTACs on the degradation of the PXR protein in FIEPG2 (7A) and ASPC1 (7B) cells, measured by Western blot.

[Fig 8] Figures 8A-B illustrate the importance of the proteasome pathway in the effects of PROTACs on PXR protein degradation measured by Western blot.

[Fig 9] Figures 9A-C represent the effect of JMV7048 on the degradation of the PXR protein in vivo, on xenografts of LS174T cells in SCID mice, respectively.

[Fig 10] Figures 10A-D represent respectively the effects of PROTACs on the cancer stem cell population: inhibition of ALDFI activity (10A), inhibition of their self-renewal capacity (10B) and sensitization to chemotherapy (10C and 10D)

[Fig 11] Figure 11 illustrates the mode of interaction of JMV6944 with the LBD of hPXR. (11 A) Whole structure of the complex. The F112 activating helix is indicated. The arrow symbolizes the extension of the PROTACs synthesized thereafter. (11 B) Zoom on the JMV6944 exit pathway and superposition with the structure of the hPXR-LBD/SR12813 complex. The end of the H2' helix, residues 206 to 209, rearranges in the presence of the ligand. (11C) Interactions of JMV6944 with hPXR binding pocket residues and representation of ligand electron density (omit type difference map).

The following examples illustrate the invention, without however limiting it. The starting products used are known products or products prepared according to known procedures.

The compounds of the present invention will be better understood in conjunction with the synthetic schemes described in various working examples and which illustrate non-limiting methods by which the compounds of the invention may be prepared. The percentages are expressed by weight, unless otherwise stated.

Example 1: Synthesis of JMV6944

Step 1: N1-benzyl-4-nitrobenzene-1.2-diamine

To a solution containing 2-fluoro-5-nitroaniline (5 g, 32.03 mmol) and benzylamine (7.01 ml, 64.05 mmol) in DMF (50 ml) and add K2CO3 (13.28 g, 96.08 mmol). The reaction medium is stirred for 24 hours at 100°C. The reaction medium is diluted in an ethyl acetate/H20 mixture. The organic phase is washed successively with water, 1N KHSO4, saturated NaCl and dried over magnesium sulphate. After evaporation, the product is triturated in diethyl ether and drained. Compound 1 N1 - benzyl-4-nitrobenzene-1,2-diamine is obtained in the form of a yellow solid with a mass of 7.5 g (96% yield). ESI: M+H 244.1 .

Step 2: (9H-fluoren-9-yl)methyl N-f2-[4-(1-benzyl-5-nitro-1H-1,3-benzodiazol-2-vDbutoxylethvDcarbamate

To a solution containing N1-benzyl-4-nitrobenzene-1,2-diamine (0.747 g, 3.07 mmol) and (9H-fluoren-9-yl)methyl N-[8-(1H-1,2, 3-benzotriazol-1-yl)-8-oxooctyl]carbamate (1.63 g, 3.37 mmol) in a toluene/DMF (9/1) mixture (45ml/5 ml) and added TFA (0.91 ml, 12.28 mmol). The reaction medium is stirred for 6 hours at 60°C. The reaction medium is cooled to room temperature and then to 0°C. The solid is drained and then washed twice with diethyl ether. The powder is dissolved in acetic acid and heated at 100° for 18 hours. After evaporation the compound 2 (9FI-fluoren-9-yl)methyl N-{2-[4-(1-benzyl-5-nitro-1 FI-1,3-benzodiazol-2-yl)butoxy]ethyl}carbamate is obtained in the form of a yellow oil 0.55 g (30% yield). ESI: M+FI 589.2.

Step 3: (9FI-fluoren-9-yl)methyl N-[7-(5-amino-1-benzyl-1H-1,3-benzodiazol-2-yl)heptyllcarbamate

To a solution containing (9FI-fluoren-9-yl)methyl N-{2-[4-(1-benzyl-5-nitro-1H-1,3-benzodiazol-2-yl)butoxy]ethyl}carbamate (0.75 g, 1.27 mmol) in ethanol (30 ml) and added SnCl2 (1.2 g, 6.34 mmol). The reaction medium is stirred for 2 hours at 80°C. The reaction medium is diluted in an ethyl acetate/NaFIC03 sat mixture and filtered through celite. The organic phase is collected and dried over MgS04. After evaporation, the compound 3 (9FI-fluoren-9-yl)methyl N-[7-(5-amino-1-benzyl-1H-1,3-benzodiazol-2-yl)heptyl]carbamate is obtained in the form of yellow powder 0.55 g (yield 77%). ESI: M+H 559.3. Step 4: N-[2-(7-aminoheptyl)-1-benzyl-1H-1.3-benzodiazol-5-yl1-2.4.6-trimethylbenzene-1-sulfonamide To a solution containing (9H-fluoren-9-yl)methyl N-[7-(5-amino-1-benzyl-1H-1,3-benzodiazol-2-yl)heptyl]carbamate (0.386 g, 0.69 mmol) in a pyridine / DCM (1/1) mixture (5 ml / 5 ml) at 0°C and added 2-mesitylenesulfonyl chloride (0.166 g, 0.75 mmol) per portion. The reaction medium is brought to room temperature and stirred for 18 h. Diethylamine (2ml) is added to the reaction medium and stirred for 2 hours. The solution is concentrated under reduced pressure. The oil obtained is purified by preparative HPLC. After lyophilization, a yellow powder is obtained 0.201 g (yield 56%). ESI: M+H 519.4. ¹ H NMR (600 MHz, DMSO-d6): d 10.53 (s, 1 H), 7.77 (m, 3H), 7.64 (d, J = 8.92 Hz, 1 H), 7.33 (m, 4H), 7.20 ( d, J= 6.81 Hz, 2H), 7.10 (dd, J = 1.79, 8.88 Hz, 1 H), 7.01 (s, 2H), 5.63 (s, 2H), 3.08 (m, 2H), 2.75 (m, 2H), 2.58 (s, 6H), 2.21 (s, 3H), 1.66 (m, 2H), 1.48 (m, 2H), 1.26 (m, 6H). 13C NMR (125 MHz, DMSO-d6) d 155.3, 142.7, 139.2, 135.8, 135.4, 133.9, 132.3, 129.4,

129.3, 129.3, 128.5, 127.3, 117.7, 113.7, 47.7, 39.4, 39.2, 28.6, 28.4, 27.3, 26.5.

Example 2: Synthesis of JMV7048

Step 1: 2-(2.6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione

The reaction medium containing 4-Fluorophthalic anhydride (2.43 g, 14.63 mmol) and 3-aminopiperidine-2,6-dione (2.38 g, 14.63 mmol) and sodium acetate (2.4 g, 29.26 mmol) in the acetic acid (50 ml) is heated at 100° C. for 24 hours. After cooling to room temperature add water (150ml) to the reaction mixture, drain the mixture and wash with ether several times. Put in a desiccator overnight at 50°C, the compound 1 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione is obtained in the form of a pink solid with a mass of 4 g ( Yield 99%) ESI: M+H 277.2. ¹ H NMR (600 MHz, DMSO-cfe): d 11.15 (s, 1 H), 8.03-8.00 (dd, J = 4.59, 8.02 Hz, 1 H), 7.87-7.85 (dd, J =

2.29, 8.02 Hz, 1 H), 7.75-7.71 (t, J= 2.29, 4.59, 8.02 Hz, 1 H), 5.19-5.16 (dd, J= 5.51, 13.03, 1 H), 2.94-2.87 (m, 1H), 2.64-2.59 (m, 1H), 2.58-2.51 (m, 1H), 2.10-2.05 (m,1H); ¹³ C NMR (125 MHz, DMSO-CFE) D 173.2, 173.2, 170.2, 170.1, 167.4, 166.6, 166.6, 166.3, 165.4, 134.7, 134.6, 127.9, 126.7, 126.7, 122.3, 122.1, 112.0, 111.8 31.3, 22.4. Step 2: tert-butyl 4-(2-(2.6-dioxopiperidin-3-yl)-1.3-dioxoisoindolin-5-yl)piperazine-1-carboxylate

The 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione compound (500 mg, 1.81 mmol) is dissolved in NMP (7 ml) at ambient temperature. DIEA (0.89 ml, 5.43 mmol) and t-butyl 1 -piperazinecarboxylate (370.9 mg, 1.99 mmol) are added and the mixture is stirred at 140° C. for 24 hours. The solution is diluted in water (100 ml). Extract with ethyl acetate twice and the organic phase is washed with saturated NaCl and dried over magnesium sulphate. After evaporation, the oil obtained is purified on silica gel with a petroleum ether/ethyl acetate (3/1) eluent. tert-butyl 4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazine-1-carboxylate is obtained as a yellow solid with a mass of 655 mg (yield 82%). ESI: M+H 443.1 . ¹ H NMR (600 MHz, DMSO-d6): d 11.09 (s, 1H), 7.70 (d, J= 8.56 Hz, 1H), 7.35 (d, J= 2.08 Hz, 1 H), 7.26-7.24 (dd , J= 2.08, 8.56 Hz, 1H), 5.08 (m, 1H), 3.47 (s, 8H), 2.93-2.86 (m, 1H), 2.61-2.48 (m, 2H), 2.03 (m, 1H), 1.43 (s, 9H). 13C NMR (125 MHz, DMSO-d6) d 173.2,

170.5, 167.9, 167.4, 155.4, 154.3, 134.3, 125.3, 119.0, 118.3, 108.5, 79.6, 49.2, 47.0, 31.4,

28.5, 22.6.

Step 3: 2-(2,6-dioxopiperidin-3-yl)-5-(piperazin-1-yl)isoindoline-1,3-dione

The compound tert-butyl 4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazine-1-carboxylate (464 mg, 1.04 mmol) is dissolved in a solution of 4N HCl in dioxane (4ml) and the reaction medium is stirred at room temperature for 2 hours then concentrated and triturated with ether. The solid obtained in the form of a yellow powder with a mass of 323 mg (90% yield). ESI: M+H 343.1 . ¹ H NMR (600 MHz, DMSO-d6): d 11.09 (s, 1H), 9.71 (m, 2H), 7.73 (d, J= 8.61 Hz, 1H), 7.44 (d, J= 2.08 Hz, 1 H ), 7.32 (dd, J= 2.08, 8.61 Hz, 1H), 5.09 (m, 1H), 3.73 (m, 4H), 3.19 (m, 4H), 2.89 (m, 1H), 2.61-2.48 (m, 2H), 2.03 (m, 1H). 13C NMR (125 MHz, DMSO-d6) d 173.2, 170.4, 167.8, 167.3, 154.8, 134.2, 125.4, 120.0, 119.0, 109.2, 49.2, 44.5, 42.4, 31.4, 22.6. Step 4: 6-(4-(2-(2.6-dioxopiperidin-3-yl)-1.3-dioxoisoindolin-5-yl)piperazin-1-vDhexanoiaue acid The 2-(2,6-dioxopiperidin-3-yl)-5-(piperazin-1-yl)isoindoline-1,3-dione compound (100 g, 0.29 mmol) is dissolved in acetonitrile (5 ml). 6-Bromohexanoic acid (152 mg, 0.73 mmol) and DIEA (0.193 ml, 1.16 mmol) are added and the mixture is stirred at 60° C. for 24 hours. The solution is concentrated under reduced pressure. The oil obtained is purified by preparative HPLC. After lyophilization, a yellow powder is obtained with a mass of 90 mg (65% yield). ESI: M+H 457.3. ¹ H NMR (600 MHz, DMSO-d6): d 12.08 (m, 1 H), 11.04 (s, 1 H), 9.73 (m, 1 H), 7.77 (d, J = 8.50 Hz), 7.50 (d, J= 1.90 Hz), 7.37 (dd, J= 1.90, 8.50 Hz), 5.10 (m, 1H), 4.23 (m, 2H), 3.59 (m, 2H), 3.25 (m , 2H), 3.14 (m, 4H), 2.90 (m, 1H), 2.59 (m, 2H), 2.25 (m, 2H), 2.04 (m, 1H), 1.69 (m, 2H), 1.55 (m, 2H), 1.33 (m, 2H). 13C NMR (125 MHz, DMSO-d6) d 174.7, 173.2, 170.4, 167.8, 167.3, 154.6, 134.2, 125.4, 120.4, 119.2, 109.4, 55.7, 50.7, 49.3, 44.8, 25.7, 31.7 , 23.4.

To a solution containing N-[2-(7-aminoheptyl)-1-benzyl-1H-1,3-benzodiazol-5-yl]-2,4,6-trimethylbenzene-1-sulfonamide (41 mg, 0.079 mmol) (example 1, JMV6944), 6-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)hexanoic acid (34 mg , 0.079 mmol) and DIEA (0.039 ml, 0.237 mmol) in DMF (5 ml) is added BOP (52 mg, 0.12 mmol). The reaction medium is stirred for two hours at room temperature. The solution is concentrated under reduced pressure. The oil obtained is purified by preparative HPLC. After lyophilization, a yellow powder is obtained with a mass of 52 mg (66% yield). ESI: M+H 958.0. ¹ H NMR (600 MHz, DMSO-d6): d 11.02 (s, 1 H),

10.42 (m, 1 H), 9.78 (m, 1 H), 7.69 (d, J= 8.49 Hz, 1 H), 7.65 (m, 1 H), 7.54 (d, J = 8.89 Hz, 1H), 7.41 (d, J=2.01Hz, 1H), 7.29-7.20 (m, 5H), 7.11 (m, 2H), 7.00 (dd, J=2.01, 8.89Hz, 1H), 6.93 ( s, 2H), 5.02 (dd, J= 5.53, 13.14Hz, 1H)

Example 3: Synthesis of JMV7505

Step 1: 7-{4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperazin-1-yljheptanoic acid

The compound 2-(2,6-dioxopiperidin-3-yl)-5-(piperazin-1-yl)isoindoline-1,3-dione (100 mg, 0.29 mmol) (example 2, step 3) is dissolved in the acetonitrile (5ml). 7-bromoheptanoic acid (155 mg, 0.73 mmol) and DIEA (0.193 ml, 1.16 mmol) are added and the mixture is stirred at 60° C. for 24 hours. The solution is concentrated under reduced pressure. The oil obtained is purified by preparative HPLC. After freeze-drying, a yellow powder is obtained with a mass of 93 mg (65% yield). ESI: M+H 471 .3. 2nd step :

To a solution containing N-[2-(7-aminoheptyl)-1-benzyl-1H-1,3-benzodiazol-5-yl]-2,4,6-trimethylbenzene-1-sulfonamide (41 mg, 0.079 mmol) (example 1, JMV6944), 6-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)heptanoic acid (34 mg , 0.079 mmol) and DIEA (0.039 ml, 0.237 mmol) in DMF (5 ml) is added BOP (52 mg, 0.12 mmol). The reaction medium is stirred for two hours at ambient temperature. The solution is concentrated under reduced pressure. The oil obtained is purified by preparative HPLC. After lyophilization, a white powder is obtained with a mass of 52 mg (68% yield). ESI: M+H 971.5. ¹ H NMR (600 MHz, DMSO-d6): d 11.02 (s, 1 H), 10.42 (m, 1 H), 9.78 (m, 1 H), 7.69 (d, J= 8.49 Hz, 1 H), 7.65 (m, 1H), 7.54 (d, J=8.89Hz, 1H), 7.41 (d, J=2.01Hz, 1H), 7.29-7.20 (m, 5H), 7.11 (m, 2H), 7.00 (dd, J= 2.01 , 8.89 Hz, 1H), 6.93 (s, 2H), 5.53 (s, 2H), 5.02 (dd, J= 5.53, 13.14 Hz, 1H) Example 4: Synthesis of JMV7506

Step 1: 8-{4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperazin-1-yljoctanoic acid

The compound 2-(2,6-dioxopiperidin-3-yl)-5-(piperazin-1-yl)isoindoline-1,3-dione (100 mg, 0.29 mmol) (example 2, step 3) is dissolved in the acetonitrile (5ml). 8-bromooctanoic acid (160 mg, 0.73 mmol) and DIEA (0.193 ml, 1.16 mmol) are added and the mixture is stirred at 60° C. for 24 h. The solution is concentrated under reduced pressure. The oil obtained is purified by preparative HPLC. After freeze-drying, a yellow powder is obtained with a mass of 101 mg (67% yield). ESI: M+H 485.6.

2nd step :

To a solution containing N-[2-(7-aminoheptyl)-1-benzyl-1H-1,3-benzodiazol-5-yl]-2,4,6-trimethylbenzene-1-sulfonamide (41 mg, 0.079 mmol) (example 1, JMV6944), 6-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)octanoic acid (36 mg , 0.079 mmol) and DIEA (0.039 ml, 0.023 mmol) in DMF (5 ml) is added BOP (52 mg, 0.12 mmol). The reaction medium is stirred for two hours at room temperature. The solution is concentrated under reduced pressure. The oil obtained is purified by preparative HPLC. After lyophilization, a white powder is obtained with a mass of 45 mg (61% yield). ESI: M+H 985.5.

1 H NMR (600 MHz, DMSO-d6): d 11.02 (s, 1 H), 10.42 (m, 1 H), 9.78 (m, 1 H), 7.69 (d, J= 8.49 Hz, 1 H), 7.65 (m, 1H), 7.54 (d, J=8.89Hz, 1H), 7.41 (d, J=2.01Hz, 1H), 7.29- 7.20 (m, 5H), 7.11 (m, 2H), 7.00 (dd, J= 2.01 , 8.89 Hz, 1H), 6.93 (s, 2H), 5.54 (s, 4H), 5.02 (dd, J= 5.53, 13.14 Hz, 1H) Example 5: Synthesis of JMV7965

Step 1: tert-butyl 4-({4-[2-(2,6-dioxopiperidin-3-yl),3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperazin-1- yl}methyl)piperidine-1-carboxylate

To a solution containing 2-(2,6-dioxopiperidin-3-yl)-5-(piperazin-1-yl)isoindoline-1,3-dione (100 mg, 0.29 mmol) (example 2, step 3) and tert -butyl 4-formylpiperidine-1-carboxylate (112 mg, 0.53 mmol) in DCE is added 3 ml of MeOH. The reaction medium is stirred at room temperature for 30 minutes. Add the sodium triacetoxyborohydride in portions and stir the reaction medium at room temperature for 18 h. Concentrate the reaction medium and carry out a preparative HPLC. After freeze-drying, a yellow powder is obtained with a mass of 85 mg (53% yield). ESI: M+H 540.2.

Step 2: 2-(2,6-dioxopiperidin-3-yl)-5-{4-[(piperidin-4-yl)methyl]piperazin-1-yl}-2,3-dihydro-1H-isoindole- 1,3-dione The tert-butyl compound 4-({4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperazin-1 -yl}methyl)piperidine-1-carboxylate (100 mg, 0.19 mmol) is dissolved in DCM (50ml). TFA (5ml) is added drop by drop to the reaction medium and stirred at room temperature for 5 hours. The solution is concentrated under reduced pressure. The oil obtained (75mg, yield 92%) is used as is in step 3. ESI: M+FI

440.3.

Step 3: tert-butyl 2-[4-({4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl ]piperazin-1 -yl}methyl)piperidin-1 -yljacetate

The compound 2-(2,6-dioxopiperidin-3-yl)-5-{4-[(piperidin-4-yl)methyl]piperazin-1 -yl}-2,3-dihydro-1H-isoindole-1 ,3-dione (128 mg, 0.29 mmol) is dissolved in DCM in the presence of DIEA (0.14 ml, 0.88 mmol). Add tert-butyl bromoacetate (0.043 ml, 0.29 mmol) drop by drop and stir at room temperature for 18 h. Concentrate the reaction medium and carry out a preparative HPLC. After freeze-drying, a yellow powder is obtained with a mass of 85 mg (53% yield). ESI: M+H 554.4.

Step 4: 2-[4-({4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperazin acid -1 -yl}methyl)piperidin-1 -yljacetic

The tert-butyl compound 2-[4-({4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl] piperazin-1-yl}methyl)piperidin-1-yljacetate (83 mg, 0.19 mmol) is dissolved in DCM (25ml). TFA (5ml) is added drop by drop to the reaction medium and stir at room temperature for 18 h. The solution is concentrated under reduced pressure. The oil obtained (70 mg, yield 93%) is used as it is in step 3. ESI: M+H 498.3. Step 5:

To a solution containing N-[2-(7-aminoheptyl)-1-benzyl-1H-1,3-benzodiazol-5-yl]-2,4,6-trimethylbenzene-1-sulfonamide (21 mg, 0.0402 mmol) (Example 1, JMV6944), 2-[4-({4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol- 5-yl]piperazin-1 - yl}methyl)piperidin-1-yl]acetic acid (20 mg, 0.0402 mmol) and DIEA (0.020 ml, 0.12 mmol) in DMF (5ml) is added BOP (27 mg , 0.0603 mmol). The reaction medium is stirred for two hours at room temperature. The solution is concentrated under reduced pressure. The oil obtained is purified by preparative HPLC. After lyophilization, a yellow powder is obtained with a mass of 25 mg (62% yield). ESI: M+H 998.3.

Example 6: Synthesis of JMV7605

JMV7605

Step 1: tert-butyl 4-{4-[(2,6-dioxopiperidin-3-yl)carbamoyl]phenyl}piperazine-1-carboxylate To a solution containing 4-[4-(tert-butoxycarbonyl)piperazino]benzoic acid

(0.775 g, 2.53 mmol), 3-Aminopiperidine-2,6-dione HCl (0.50 g, 3.03 mmol) and DIEA (1.25 ml, 7.59 mmol) in DMF (50ml) is added the BOP (1.11 g, 2.53 mmol). The reaction medium is stirred for two hours at room temperature. Add water to the reaction medium and extract with ethyl acetate. Wash the organic phase successively with 1N HCl, NaHCO3 sat and NaCl sat. Dry the organic phase with MgS04, filter and concentrate under reduced pressure. A white powder is obtained with a mass of 0.4 g (38% yield). ESI: M+H 417.3.

Step 2: N-(2.6-dioxopiperidin-3-vl)-4-(piperazin-1-vDbenzamide

The compound tert-butyl 4-{4-[(2,6-dioxopiperidin-3-yl)carbamoyl]phenyl}piperazine-1 - carboxylate (0.4 g, 0.96 mmol) is dissolved in a solution of 4N HCl in dioxane (6ml) and the reaction medium is stirred at room temperature for 2 hours then concentrated and triturated with ether. The solid obtained in the form of a white powder with a mass of 0.285 mg (94% yield).

The reaction medium is stirred for two hours at ambient temperature. The solution is concentrated under reduced pressure. The oil obtained is purified by preparative HPLC. After lyophilization, a white powder is obtained with a mass of 45 mg (52% yield). ESI: M+H 317.3.

Step 3: 7-(4-{4-[(2,6-dioxopiperidin-3-yl)carbamoyl]phenyl}piperazin-1 - yl)heptanoic acid

The N-(2,6-dioxopiperidin-3-yl)-4-(piperazin-1-yl)benzamide compound (50 mg, 0.15 mmol) is dissolved in DMF (5 ml). 7-bromoheptanoic acid (66 mg, 0.31 mmol) and DIEA (0.078 ml, 0.47 mmol) are added and the mixture is stirred at 100° C. for 24 hours. The solution is concentrated under reduced pressure. The oil obtained is purified by preparative HPLC. After freeze-drying, a yellow powder is obtained with a mass of 38 mg (55% yield). ESI: M+H 445.1.

Step 4:

JMV7605

To a solution containing N-[2-(7-aminoheptyl)-1-benzyl-1H-1,3-benzodiazol-5-yl]-2,4,6-trimethylbenzene-1-sulfonamide (35 mg, 0.067 mmol) (example 1, JMV6944), 7- (4-{4-[(2,6-dioxopiperidin-3-yl)carbamoyl]phenyl}piperazin-1-yl)heptanoic acid (30 mg, 0.067 mmol) and DIEA (0.033 ml, 0.20 mmol) in DMF (5ml) is added BOP (44 mg, 0.101 mmol). The reaction medium is stirred for two hours at room temperature. The solution is concentrated under reduced pressure. The oil obtained is purified by preparative HPLC. After lyophilization, a white powder is obtained with a mass of 41 mg (65% yield). ESI: M+H 945.8.

Example 7: Synthesis of JMV7159 (comparative example) Step 1: 5-fluoro-2-(1-methvl-2.6-dioxopiperidin-3-vl)-2.3-dihvdro-1H-isoindole-1,3-dione

The 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione compound (250mg, 0.90 mmol) is dissolved in anhydrous DMF (5ml) and the reaction medium is stirred and brought to 0. °C. Add the NaH in portions and leave to stir for 20 minutes. Add the iodomethane and stir for 2 hours. Stop the reaction with a solution of NH4Cl. Extract with ethyl acetate and wash twice with NaCl sat the organic phase. Dry over MgS04, filter and concentrate under reduced pressure. The compound 5-fluoro-2-(1-methyl-2,6-dioxopiperidin-3-yl)-2,3-dihydro-1H-isoindole-1,3-dione is obtained in the form of a white powder with a mass of 253 mg (96% yield). ESI: M+H 291 .1 .

Step 2: tert-butyl 4-[2-(1-methyl-2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1 H-isoindol-5-yl]piperazine- 1-carboxylate

The compound 5-fluoro-2-(1-methyl-2,6-dioxopiperidin-3-yl)-2,3-dihydro-1H-isoindole-1,3-dione (250 mg, 0.86 mmol) is dissolved in NMP (4 ml) at room temperature. DIEA (0.42 ml, 2.58 mmol) and t-butyl 1-piperazinecarboxylate (176 mg, 0.94 mmol) are added and the mixture is stirred at 140° C. for 24 hours. The solution is diluted in water (100 ml). Extract with ethyl acetate twice and the organic phase is washed with saturated NaCl and dried over magnesium sulphate. After evaporation, the oil obtained is purified on silica gel with a petroleum ether/ethyl acetate (3/1) eluent. tert-butyl 4-[2-(1-methyl-2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperazine-1-carboxylate is obtained as a yellow solid with a mass of 338 mg (86% yield). ESI: M+H 457.3.

Step 3: 2 (1-methyl-2,6-dioxopiperidin-3-yl)-5-(piperazin-1-yl)-2,3-dihydro-1H-isoindole-

1,3-dione The tert-butyl compound 4-[2-(1-methyl-2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperazine-1 -carboxylate (250 mg, 0.54 mmol) is dissolved in a solution of 4N HCl in dioxane (4 ml) and the reaction medium is stirred at room temperature for 2 hours then concentrated and triturated with ether. The solid obtained as a yellow powder with a mass of 175 mg (90% yield). ESI: M+H 357.3.

Step 4: 6-{4-[2-(1-methyl-2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperazin acid -1-yl}hexanoic

The compound 2-(1-methyl-2,6-dioxopiperidin-3-yl)-5-(piperazin-1-yl)-2,3-dihydro-1H-isoindole-1,3-dione (100 mg, 0.28 mmol) is dissolved in acetonitrile (5 ml). 6-bromohexanoic acid (136 mg, 0.70 mmol) and DIEA (0.139 ml, 0.84 mmol) are added and the mixture is stirred at 60° C. for 24 hours. The solution is concentrated under reduced pressure. The oil obtained is purified by preparative HPLC. After lyophilization, a yellow powder is obtained with a mass of 85 mg (65% yield). ESI: M+H 471 .3.

To a solution containing N-[2-(7-aminoheptyl)-1-benzyl-1H-1,3-benzodiazol-5-yl]-2,4,6-trimethylbenzene-1-sulfonamide (28 mg, 0.055 mmol) (Example 1, JMV6944), 6-{4-[2-(1-methyl-2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol- 5-yl]piperazin-1 - ylhexanoic acid (26 mg, 0.055 mmol) and DIEA (0.165 ml, 0.165 mmol) in DMF (5ml) is added BOP (36 mg, 0.082 mmol). The reaction medium is stirred for two hours at room temperature. The solution is concentrated under reduced pressure. The oil obtained is purified by preparative HPLC. After freeze-drying, a white powder is obtained with a mass of 30 mg (56% yield). ESI: M+H 971 .6.

Example 8: Biochemistry and crystalloaraphy

The human PXR receptor ligand-binding domain (hPXR-LBD, residues 130-434), was produced as a recombinant protein in E. coli BL21-DE3 bacteria. The protein was purified on an affinity column then by size exclusion chromatography. After concentration, hPXR-LBD was crystallized in the presence of the JMV6944 ligand. The structure of the hPXR-LBD/JMV6944 complex was determined by X-ray crystallography by the molecular replacement method, then reconstructed and refined on the basis of the electron density (diffraction data collected at the ESRF synchrotron, Grenoble). The structure is shown in Figure 11 . In A, the entire structure of the complex shows the binding mode of JMV6944. The originality of JMV6944 lies in the position of the extension added to the parent molecule JMV6845 and its way out of the protein domain. Unlike the known PROTACs for other nuclear receptors which are all based on the modification of an antagonist ligand, the extension grafted onto the agonist JMV6845 does not extend towards the H12 helix but points in the opposite direction between the helices H2', H6, H7 and the S1 strand to finally reach the outer surface of the LBD. The presence of the alkyl/NH arm (circled in B) induces a conformational change at the end of H2' which allows this ligand to extract itself from the binding pocket and to interact specifically with a surface residue, C207 (VS). Within the ligand binding pocket, JMV6944 also establishes hydrogen bonds with IΉ407 and S247, as well as hydrophobic interactions with L411 and F428 on the one hand, and the residues of the “p-trap” region. on the other hand (F288, W299, Y306).

Example 9: Biological results 9.1 Measurement of prePROTAC/PXR affinity

The binding affinity between the JMV6944 molecule (prePROTAC) and the ligand binding domain (LBD) of PXR was quantified by FRET using the LanthaScreen TR-FRET PXR Competitive binding assay Kit (Invitrogen). The molecules were incubated for 1 hour 30 minutes at ambient temperature with the LBD of PXR in the presence of a fluorescent reference ligand. The displacement of the fluorescent ligand caused by the prePROTAC or the PXR ligand SR12813 was measured by reading the emissions at 520nm and 495nM after excitation at 337 nM on a PHERA-Star device (BMG LABTECH). The results are illustrated in Figure 1 which shows that the JMV6944 molecule is a PXR ligand with an affinity of 18.38 nM.

9.2 Measurement of the effects of PROTACs on the transcriptional activity of PXR (reporter gene)

Treatment of LS174T cells stably transfected with an expression vector encoding the PXR protein, a Luciferase reporter gene placed under the control of the CYP3A4 promoter (target gene of PXR) and an expression cassette encoding the GFP protein placed under CMV promoter control for signal normalization. The cells were treated for 48 h with 5 mM of the molecules JMV6944 (prePROTACs), the PROTACs JMV7048 and JMV7605, as well as rifampicin (5 mM, PXR ligand). At the end of the treatments, the transcriptional activity of PXR is measured by the ratio of the luciferase/GFP signals measured on a PHERA-Star device (BMG LABTECH). Figure 2 shows that only prePROTAC JMV6944 and rifampicin are able to activate PXR transcriptional activity.

9.3 Measurement of the effects of PROTACs on the transcriptional activity of PXR (expression of CYP3A4 mRNA)

The LS174T cells were treated for 48 hours with 5 mM of the molecules JMV6944 (prePROTACs), JMV7048, JMV7505, or JMV5159 (inactive equivalent of JMV7048 following the addition of a methyl group to the CNBR ubiquitin ligase ligand) in the presence or in the absence of rifampicin (ligand of PXR) at 5 mM final. After lysis of the cells and purification of the total RNAs (Qiagen RNAeasy), the complementary DNAs were prepared (SuperScript II in the presence of random primers of 6 nucleotides, Invitrogen). The expression of CYP3A4 mRNAs and housekeeping genes RPLO and actin was measured by RT-qPCR on an LC480 device (Roche) in the presence of SyberGreen (Millipore). Relative expression levels were calculated using the RQ=Relative quantification=2-AACt method, with untreated cells serving as a calibrator set at 1. Figures 3A and 3B show that while prePROTAC and inactivated PROTAC (JMV7159) have an additive effect on CYP3A4 mRNA expression, PROTACs JMV7048 and JMV7965 significantly decrease rifampicin-mediated CYP3A4 induction.

9.4 Measurement of the effects of PROTACs on the transcriptional activity of PXR (expression of CYP3A4) The LS174T cells were treated for 48 hours with 5 mM JMV7048 in the presence or absence of rifampicin (ligand of PXR) at a final 5 mM. After cell lysis (RIPA + antiproteases), the proteins were purified and assayed before being loaded (QOmV) on a 10% SDS-PAGE gel. After gel migration, they were transferred to a nitrocellulose membrane (GE Healthcare) before being revealed with antibodies directed against CYP3A4 (sc-53850, Santa Cruz) and beta-actin (A5441, Sigma or Ab-253283 , AbCAm) then secondary antibodies coupled to peroxidase (anti-mouse HRP, Santa Cruz). The signal intensities were measured by a camera (BioRad MP Touch). Figures 4A and 4B show that the PROTACs JMV7048 and JMV7965 decrease the induction of the CYP3A4 enzyme mediated by rifampicin.

9.5 Measurement of the effects of PROTACs on cell viability

The effect of PROTACs on cell viability was tested on different CRC1, HT29 and LS174T lines. The cells were incubated in the presence of increasing concentrations of molecules for 72 hours before being fixed and labeled with Sulforhodamine B (Sigma). After cell washes and lysis, the incorporated dye released by the cells is directly proportional to the cell biomass. It is measured at 565 nM by a spectrophotometer for 96-well plates (Técan). The signal obtained for the untreated cells is set at 100%. Figure 5A illustrates the absence of toxicity of PROTACs JMV7048, JMV7505 and JMV7605 on the LS174T line. In Figure 5B, it is seen that PROTAC JMV7048 does not affect the viability of HT29 cells or CRC1 protoculture (from a patient with colon cancer.

9.6 Measurement of the effects of PROTACs on PXR degradation in LS174T cells by western-blot in vitro

The effect of PROTACs on the level of expression of the PXR protein was studied by Western-blot. The LS174T cells were transfected in the absence or presence of 50 nM of an siRNA targeting PXR (siPXR: NR112 Silencer, Thermofischer) or treated with PROTACs. After lysis of the cells (RIPA+antiproteases), the proteins were purified and assayed before being deposited (9C^g) on a 10% SDS-PAGE gel. After gel migration, they were transferred to a nitrocellulose membrane (GE Healthcare) before being revealed with antibodies directed against PXR (sc-48340, Santa Cruz), GAPDH (sc-32233, Santa Cruz) and beta -actin (A5441, Sigma or Ab-253283, AbCAm) then secondary antibodies coupled to peroxidase (anti-mouse HRP, Santa Cruz). The signal intensities were measured by a camera (BioRad MP Touch). Figures 6A-C show that after 24h of treatment at 5mM, the PROTACs JMV7048, JMV7505, JMV7506, JMV7605 and JMV7965 significantly reduce the level of expression of the PXR protein, unlike an inactive mutant of JMV7048 (ie JMV7159). Figures 6D and 6E illustrate the effect of JMV7048 on the level of expression of PXR according to the treatment time (maximum effect reached after 3 hours of treatment) and the concentration used (decrease dependent on the dose, with a maximum effect observed from 500 nM).

9.7 Measurement of the effects of PROTACs on PXR degradation in HEPG2 and ASPC1 cells by western-blot in vitro

The effect of PROTACs (5mM, 24h of treatment) on the level of expression of the PXR protein in HepG2 cells (Hepatocellular Carcinoma, ATCC #HB-8065 ^™ ) or ASPC1 (Human Pancreatic Cancer Cell Line, ATCC #CRL- 1682) were studied by Western-blot. After lysis of the cells (RIPA + antiproteases), the proteins were purified and assayed before being deposited (9C^g) on a 10% SDS-PAGE gel. After gel migration, they were transferred to a nitrocellulose membrane (GE Healthcare) before being revealed with antibodies directed against PXR (sc-48340, Santa Cruz), GAPDH (sc-32233, Santa Cruz) and beta -actin (A5441, Sigma or Ab-253283, AbCAm) then secondary antibodies coupled to peroxidase (anti-mouse HRP, Santa Cruz). Figures 7A and 7B illustrate the effect of PROTACs JMV7048 and JMV7965 on the level of expression of PXR in hepatic (7A) or pancreatic (7B) cancer cells

9.8 Importance of the proteasome pathway on the effects of PROTACs on PXR degradation.

The involvement of the proteasome pathway on the effects of PROTACs on the level of expression of the PXR protein was studied by Western-blot. LS174T cells were treated for 24 hours with JMV7048 in the presence or absence of CNBR ubiquitin ligase (MLN4924) or proteasome inhibitor (Bortezomib). Figures 8A and 8B confirm the importance of the proteasome pathway on the decrease in the level of expression of the PXR protein induced by PROTAC JMV7048: the decrease in the level of expression of PXR induced by JMV7048 is reversed by inhibitors ubiquitin ligase CRBN (MLN4924, Figure 8A) or by a 26S proteasome inhibitor (Bortezomib, Bz; Figure 8B), whereas the JMV7048 mutant (i.e. JMV7159, not allowing CNBR recruitment) does not cause any decrease in PXR expression level.

9.9 Measurement of the effects of PROTACs on western-blot PXR degradation in vivo

The effect of PROTACs on the level of expression of the PXR protein was studied in vivo by Western-blot after xenografts of LS174T cells in SCID mice. When the tumors reached 100mm3, 10 mice were treated by IV with 5% EtOH solvent, 20% solutol in D5W or PROTACS (25mg/kg) every 24 hours for 4 days. The mice were weighed daily. Four hours after the last treatment, the tumors were resected before being lysed in RIPA buffer using ceramic beads (Lysing matrix D, MP-Bio) with a Fast-Prep 24 device (MP-Bio). The proteins were purified and assayed before being loaded (£^g) on a 10% SDS-PAGE gel. After gel migration, they were transferred to a nitrocellulose membrane (GE Healthcare) before being revealed with antibodies directed against PXR (sc-48340, Santa Cruz), GAPDH (sc-32233, Santa Cruz) and beta -actin (A5441, Sigma or Ab-253283, AbCAm) then secondary antibodies coupled to peroxidase (anti-mouse HRP, Santa Cruz). The signal intensities were measured by a camera (Biorad MP Touch). It can be seen in FIG. 9A that a treatment at 25 mk/kb for 4 days does not significantly modify the weight of the mice. Figures 9B and 9C confirm that this treatment is capable of inducing a significant drop in the level of expression of the PXR protein within the tumours.

9.10 Measurement of the effects of PROTACs on self-renewal and chemoresistance of colon cancer stem cells

The effects of PROTACs on the survival and self-renewal of colon cancer stem cells were studied in vitro on the HT29 line or cancer cells isolated from patients (CRC1). The cells were treated with or without 5mM PROTACs for 48 hours before being analyzed: Aldefluor labeling, enzymatic activity preferentially present in cancer stem cells (Figure 10A); formation of tumorspheres under aseric and non-adherent conditions, (FIG. 10B), and finally resistance to chemotherapies (FIG. 10C and 10D).

FIG. 10A shows that the PROTACs JMV7048, JMV 7505, JMV7506 and JMV7965 significantly reduce the percentage of ALDH-positive cells after dissociation of CRC1 cells and labeling with Aldefluor™ (STEMCELL Technologies) compared to untreated cells. FIG. 10B shows that the JMV7048 and JMV7965 molecules significantly reduce the number of HT29 cells capable of surviving anoikis () and of inducing the formation of T umorphers (Sphere Forming Cells). Tumorphers of more than 50 mM in diameter were counted 10 days after treatment and culturing 200 cells per well (previously treated with poly2Hema in order to prevent any cell adhesion) in 100 mί medium devoid of calf serum. These culture conditions only allow cancer stem cells to survive. Figures 10C and 10D show the effects of PROTACs JMV7048 and JMV7965 on survival (10C) and tumorphere-forming capacity (10D) of HT29 cells maintained in the presence of different concentrations of 5-FU and SN38 (Folfiri 1X= dqmV 5-FU + 500nM SN38) and cultured in 100mI_ of medium devoid of calf serum in dishes previously treated with poly2Flema to prevent cell adhesion. Tumorphers over 50 mM in diameter were counted 10 days after seeding 200 cells/well. Thus, FIGS. 10A-D show that a treatment at 5 mM for 2 days with the PROTACs JMV7048 and JMV7965 significantly decreases the survival and the chemoresistance of stem cells in colon cancer cell lines.

Claims

1 . Bifunctional compound corresponding to the general formula (I): L(PXR)-Linker-L(E3 ligase) (I) wherein: L(PXR) is a ligand capable of binding to the PXR nuclear receptor, L(E3 ligase) represents a ligand for the E3 ubiquitin ligase, and Linker represents a group that allows L(PXR) to be covalently linked to L(E3 ligase). 2. Bifunctional compound according to claim 1 such that L(PXR) is a group of formula (II): where represents the attachment of the group to Linker; or a pharmaceutically acceptable salt. 3. Compound according to claim 1 or 2, such that L(E3 ligase) is chosen from: - the groups of formula (NIA): and - formula groups (INB): or a pharmaceutically acceptable salt, wherein: X is NH; X' is -C(O)- or -CH2-; Y represents H or a C1-C6 alkyl group; represents the group's attachment to Linker. 4. Compound according to any one of the preceding claims, such as Linker represents a C1 -C20 alkylene group, optionally interrupted or optionally terminating at one and/or the other of the two ends, by one of the groups - 0-, -S-, -N (R') -, -C(O)-, -C(0)0-, -OC(O) -, -0C(0)0 -, -C(NOR')-, -C(0)N(R') -, - C(0)N(R')C(0)-, -C(0)N(R')C(0)N(R')-, -N(R')C(0)- , -N(R')C(0)N(R')-, -N(R')C(0)0-, - 0C(0)N(R')-, -C(NR')- , -N(R')C(NR')-, -C(NR')N(R') -, -N(R')C(NR ^, )N(R')-, -S(0) ₂ -, -OS(O)- , -S(0)0-, -S(O)-, -0S(0) ₂ -, -N(R')S(0) ₂ -, -S(0) ₂ N(R')-, -N(R')S-, -S(0)N(R , - N(R')S(0) ₂ N(R') -, -N(R')S(0)N(R' ) -, C3 to C12 carbocyclene, heterocyclene of 3 to 12 members and comprising 1, 2 or 3 heteroatoms chosen from N, O, S, heteroarylene of 5 to 12 members and comprising 1, 2 or 3 heteroatoms chosen from N, O , S, or any combination thereof, and in which the same or different R' represents H or a C1-C6 alkyl group. 5. Compound according to any one of the preceding claims, such that Linker represents a group of where Li and L- ₂ , identical or different, independently represent an alkylene group of 1 to 12 carbon atoms optionally interrupted or ending in a heterocyclene of 3 to 12 members and comprising 1, 2 or 3 heteroatoms chosen from N, O, S; Li is bound is linked to L(E3 ligase); Z represents H or a C1-C6 alkyl group. 6. Compound according to any one of the preceding claims, such that it corresponds to one of the following formulas (I-4) and (I-5): (I-4) L(PXR)-Linker (I-5) in which L(PXR), Linker are defined according to any one of claims 1 to 5; or a pharmaceutically acceptable salt. 7. Compound according to any one of the preceding claims, such that it corresponds to the following formula (V): wherein L_ ₂ represents a linear C2-C8 alkylene group optionally interrupted by a piperidinyl group, and L(E3 ligase) is as defined in any one of claims 1 to 6. 8. Compound according to any one of the preceding claims, such that it corresponds to one of the following formulas: 9. Process for the preparation of a compound according to any one of the preceding claims comprising the coupling of a compound of formula (B) and a compound of formula (C): L(PXR)-T T’-L(E3 ligase) (B) (C) such that L(PXR) and L(E3 ligase) are as defined according to any one of claims 1 to 7, and T and T' are two Linker precursor groups, as they possess each respectively a complementary reactive terminal function. 10. Process according to claim 9, such that compound (B) corresponds to formula (A): and the compound (C) corresponds to the formula (C-1): ligase) wherein L2 and L(E3 ligase) are as defined in any one of claims 1 to 7. 11. Compound of formula (A): 12. Pharmaceutical composition comprising a compound according to any one of claims 1 to 7, and at least one pharmaceutically acceptable excipient. 13. Compound according to any one of claims 1 to 7 for its use for the treatment and/or prevention of cancer overexpressing the PXR nuclear receptor. 14. A compound for use according to claim 13 in combination with an anti-cancer agent. 15. A compound for use according to any of claims 13 or 14 for administration in patients resistant to the anticancer agent.