BCL-2 family protein modulating compounds for cancer therapy

Abstract

The present invention relates to compounds, compositions and methods for treating cancer and cell proliferation diseases, and more particularly to the preparation and use of a compound that modulates a protein selected from Bcl-2, Mcl-1, Bcl-x L , Bcl-w, Bfl-1 and Bcl-B, in particular Bcl-2 protein, or BCl-x L and / or Mcl-1 protein; the compounds can be included in pharmaceutical compositions and used as therapeutic agents.

Description

Technical Field

[0001] The present invention relates to compounds, compositions and methods for treating cancer and cell proliferation diseases, and more particularly to the preparation and use of a molecule that regulates Bcl-2, Mcl-1, Bcl-x L At least one protein selected from Bcl-w, Bfl-1 and Bcl-B, in particular Bcl-2 protein or Bcl-x L and / or Mcl-1 protein; the compounds can be included in pharmaceutical compositions and used as therapeutic agents. Background Art

[0002] Cancer is the leading cause of death worldwide. Apoptosis, also known as programmed cell death, is a natural process used by multicellular organisms to eliminate old or damaged cells and is also involved in various physiological processes such as morphogenesis and tissue homeostasis. Apoptosis is a complex, highly regulated process involving many proteins. Some of these proteins promote cell death ("pro-apoptotic" proteins) and some prevent cell death ("anti-apoptotic" proteins). Cancer cells tend to overexpress anti-apoptotic genes. Overexpression of anti-apoptotic genes has been associated with tumor formation, metastatic growth and chemotherapy resistance, so there is a continuous need for therapeutic strategies that selectively kill cancer cells.

[0003] More specifically, defects in apoptosis control are frequently associated with chemoresistance in cancer cells in both hematologic malignancies and solid tumors, and dysregulated expression of Bcl-2 family members is one of the most common and important events. These proteins share the Bcl-2 homology domain (named BH domain). L ...) contain BH1 to BH4 domains, while pro-apoptotic proteins contain either BH1 to BH3 domains (multidomain members such as Bax and Bak) or only BH3 domains (BH3-only groups such as Bim, Puma, Bid, Bad, Noxa, and Hrk). Under cellular stress, BH3-only proteins initiate apoptosis by blocking the activity of anti-apoptotic members or directly activating multidomain pro-apoptotic members, which is mediated by the interaction of the BH3 domain of one protein with the hydrophobic pocket of the other protein.

[0004] Efforts have been made to inhibit the anti-apoptotic members such as Bcl-2 or Bcl-x L Among them, the development of highly efficient BH3 mimetic molecules is a promising avenue. These molecules bind to the BH3 binding groove in the anti-apoptotic proteins of the Bcl-2 family and promote cell death by releasing pro-apoptotic Bcl-2 family members.

[0005] In ovarian cancer, the inventors previously demonstrated that Bcl-x Land Mcl-1 cooperate to protect tumor cells from apoptosis, and their concomitant inhibition leads to massive apoptosis even in the absence of chemotherapy, whereas Bcl-x L Alternatively, Mcl-1 downregulation may still be ineffective in some cases. In this case, they also showed that Mcl-1 downregulation or inactivation is required to render ovarian cancer cells resistant to Bcl-x L -Sensitive to targeted BH3-mimetic molecules (such as ABT-737).

[0006] In hematological malignancies including lymphoma, the interest in targeting Bcl-2 family anti-apoptotic proteins has been widely documented. For example, the BCL-2 inhibitor Venetoclax (or Venclaxta, developed by Abbvie), approved by the US FDA, has successfully targeted Bcl-2 in vitro as well as in vivo, and BCL-2 inhibitors have been found to be clinically active against various hematological malignancies including multiple myeloma, acute myeloid leukemia, myelodysplastic syndrome, acute lymphoblastic leukemia, small lymphocytic lymphoma, etc.

[0007] Other studies have also shown that inhibition of Mcl-1 increases the expression of Bcl-2 or Bcl-x L Therapeutic efficacy of targeted BH3 mimetics in hematological malignancies In other lymphoid malignancies, such as aggressive mantle cell lymphoma (MCL), cell survival may or may only be dependent on BCL-2-associated Mcl-1.

[0008] Mcl-1 contains three BH domains (BH1-BH3) but lacks a well-defined BH4 domain at the NH2 terminus. Mcl-1 is localized to various intracellular membranes, especially the outer mitochondrial membrane, through its COOH-terminal transmembrane domain. L Mcl-1 can interact with Bak to inhibit mitochondrial-mediated apoptosis. L , Mcl-1 expression is rapidly induced by cytokines or growth factors. Increased Mcl-1 expression promotes cell viability in a variety of tumor cell types, including leukemia, hepatocellular carcinoma, melanoma, prostate cancer, and breast cancer cells. In addition, Mcl-1 plays a role in cell immortalization and tumorigenesis in a variety of cancers through somatic copy number amplification. Cancer cells carrying Mcl-1 amplification often rely on Mcl-1 for survival.

[0009] Mcl-1 is overexpressed in a variety of tumor cells, including ovarian cancer, hematological malignancies, breast cancer, colon cancer, and lung cancer, and its expression has also been associated with chemotherapy resistance. Importantly, the Mcl-1 locus is one of the most frequently amplified loci in human cancer, further indicating its central role in carcinogenesis and increasing its importance as a high-priority therapeutic target.

[0010] However, the development of several molecules inhibiting the Mcl-1 pathway has been abandoned recently due to cardiotoxicity reasons.

[0011] Among other promising therapeutic targets, the anti-apoptotic Bcl-x L The protein constitutes a target of choice. This is particularly true in the treatment of certain solid tumors, which overexpress this protein (including ovarian tumors) and whose expression is strongly associated with drug resistance. Although the pharmacological benefit of targeting this protein is currently not questioned, the development of molecules that inhibit this protein is currently hampered by the appearance of thrombocytopenia in patients.

[0012] Therefore, the object of the present invention is to provide a method for regulating the activity of Mcl-1 and / or Bcl-x L , especially degradation of Mcl-1 and / or Bcl-x L Proteins, alternative compounds that are non-toxic or low-toxic to the heart and platelets.

[0013] Another object of the present invention is to provide alternative compounds for treating cancer, in particular hematological malignancies such as lymphoma and / or solid tumors such as ovarian cancer, while having no or low toxicity to the heart and platelets. Summary of the invention

[0014] Therefore, the present invention relates in one aspect to compounds of the following formula (I):

[0015]

[0016] in:

[0017] Y1, Y2, Y3 and Y4 are each independently C or N, Y1 and Y2 are, for example, one is N and the other is C, Y3 and Y4 are, for example, one is N and the other is C;

[0018] Ar1 and Ar2 are each independently selected from (C6-C 10 )aryl, (5- to 7-membered)heteroaryl, -O-(C6-C 10 )aryl, -O-(5- to 7-membered)heteroaryl, -S-(C6-C 10 )aryl, -S-(5- to 7-membered)heteroaryl, -(C1-C6)alkyl-(C6-C 10)aryl, -(C1-C6)alkyl-(5- to 7-membered)heteroaryl and halogen, in particular Cl, said aryl and heteroaryl being optionally substituted by 1 to 3 R3 groups,

[0019] i and j are independently 0 or 1,

[0020] and

[0021] R1 and R2, when present at each time, are independently selected from (C1-C6) alkyl, (C6-C 10 )aryl, -(C1-C6)alkyl-(C6-C 10 )aryl, -(C2-C6)alkenyl-(C6-C 10 )aryl, -carbonyl-(C6-C 10 )aryl, -carbonyl-(C1-C6)alkyl-(C6-C 10 ) aryl, C(═O)H, COOH, OH, the alkyl is optionally substituted by OH, the aryl is optionally substituted by 1-3 R4 groups;

[0022] k and l are independently 0 or 1;

[0023] (R1) k and (R2) l At least one of them is, for example:

[0024] R1 is selected from (C6-C 10 )aryl, -(C1-C6)alkyl-(C6-C 10 )aryl, -(C2-C6)alkenyl-(C6-C 10 )aryl, -carbonyl-(C6-C 10 )aryl, -carbonyl-(C1-C6)alkyl-(C6-C 10 ) aryl, and k is 1; or

[0025] R2 is selected from (C6-C 10 )aryl, -(C1-C6)alkyl-(C6-C 10 )aryl, -(C2-C6)alkenyl-(C6-C 10 )aryl, -carbonyl-(C6-C 10 )aryl, -carbonyl-(C1-C6)alkyl-(C6-C 10 ) aryl, and l is 1;

[0026] R3, at each occurrence, is independently selected from (C1-C6)alkyl, (C1-C6)alkoxy, OH, C(=O)H, (CH2) m CONR a R b 、C(=O)R c 、(CH2)n CO2H, (CH2) n CO2R c 、(CH2) p CN, (CH2) q C(=N(OH))NH2、-SO2NR a R b 、I、Cl、Br、F、(C6-C 10 )aryl, (5- to 7-membered)heteroaryl, -(C1-C6)alkyl-(C6-C 10 )aryl, -(C2-C6)alkenyl-(C6-C10)aryl, the alkyl group is optionally substituted by OH, (C1-C6)alkoxy and / or -O-(C6-C 10 ) aryl substitution;

[0027] R4, at each occurrence, is independently selected from (C1-C6)alkyl, (CH2) n CO2H, (CH2) n CO2R c , I, Cl, Br, F, or two R4 form a naphthyl group with the aryl groups to which they are attached;

[0028] R a and R b Independently selected from H and (C1-C6)alkyl;

[0029] R c Selected from H and (C1-C6) alkyl;

[0030] m is 0, 1, 2, or 3;

[0031] n is 0, 1, 2, 3;

[0032] p is 0, 1, 2, 3;

[0033] q is 0, 1, 2, 3;

[0034] r is 0 or 1;

[0035] W and Y are each independently selected from a single bond, -CH=CH-, -C≡C-, -NH-, -NR5-, -O-, -S-, -C(=O)NH-, -NHC(=O)-, -C(=O)O-, -OC(=O)-;

[0036] R5, at each occurrence, is independently selected from (C1-C6)alkyl,

[0037] L1 and L2 are each independently selected from (C1-C 18 )alkyl, (C2-C 18 )alkenyl and (C2-C 18) alkynyl, which is optionally interrupted by one or more groups selected from -O-, -S-, -NR6-;

[0038] R6, at each occurrence, is independently selected from H and (C1-C6)alkyl,

[0039] X is selected from (5- to 7-membered) heteroaryl (especially triazole), -NH-, -NR7-, -O-, -S-, -C(=O)NH-, -NHC(=O)-, -C(=O)O-, -OC(=O)-;

[0040] R7 is selected from (C1-C6) alkyl,

[0041] Z is the E3 ubiquitin ligase ligand;

[0042] and pharmaceutically acceptable salts thereof.

[0043] According to one embodiment, Y1 and Y3 are N, and Y2 and Y4 are C.

[0044] According to one embodiment, Y1 and Y3 are C, and Y2 and Y4 are N.

[0045] According to one embodiment, Y1 and Y4 are N, and Y2 and Y34 are C.

[0046] According to one embodiment, when j is 0, then Y4 is N.

[0047] According to one embodiment, the compound of formula (I) defined above is of the following formula (II):

[0048]

[0049] According to one embodiment, the compound defined above has the following formula (IIIa) or (IIIb):

[0050]

[0051] The compound is specifically the following formula (IIIa1) or (IIIa2), (IIIb1) or (IIIb2):

[0052]

[0053]

[0054] In a particular aspect, it includes compounds as defined above, wherein at least one of Ar1 and Ar2 is (5-7 membered) heteroaryl, -O-(C6-C 10 )aryl, -O-(5- to 7-membered)heteroaryl, -S-(C6-C 10)aryl, -S-(5- to 7-membered)heteroaryl, -(C1-C6)alkyl-(C6-C 10 )aryl or -(C1-C6)alkyl-(5- to 7-membered)heteroaryl, especially a 5- to 7-membered heteroaryl containing a nitrogen atom, preferably pyridyl, especially 3-pyridyl.

[0055] In a particular aspect, it includes compounds as defined above, wherein at least one of Ar1 and Ar2 is (C6-C 10 ) aryl, especially phenyl.

[0056] According to one embodiment, Ar1 and / or Ar2 are selected from phenyl, pyridyl, pyrimidinyl, imidazolyl, pyrazolyl, phenylthio, triazolyl, in particular selected from phenyl, 3-pyridyl, 5-pyrimidinyl, 2-imidazolyl, 3-pyrazolyl, 2-phenylthio, 5-triazolyl.

[0057] According to one embodiment, Ar1 is selected from pyridyl, in particular from 3-pyridyl, and j is especially 0.

[0058] According to one embodiment, Ar1 is 3-pyridyl or phenyl.

[0059] According to one embodiment, Ar2 is 3-pyridyl or phenyl.

[0060] According to one embodiment, W is selected from -CH=CH- and -C≡C-, W is in particular -C≡C-, and j is in particular 0. According to one embodiment, R1 and R2 are independently selected from C1-C6 alkyl, in particular methyl; (C6-C 10 )aryl, especially phenyl or naphthyl; -(C1-C6)alkyl-(C6-C 10 )aryl, in particular -(C1-C6)alkyl-phenyl, more in particular phenethyl and -(C1-C6)alkyl-naphthyl, more in particular ethyl-naphthyl; and -(C2-C6)alkenyl-(C6-C 10 )aryl, in particular -(C2-C6)alkenyl-phenyl, more in particular styryl and -(C2-C6)alkenyl-naphthyl, more in particular -vinyl-naphthyl.

[0061] According to a particular embodiment, R1 is selected from C1-C6 alkyl and R2 is selected from (C6-C 10 )aryl, -(C1-C6)alkyl-(C6-C 10 )aryl and -(C2-C6)alkenyl-(C6-C 10 )aryl.

[0062] According to another particular embodiment, R2 is selected from C1-C6 alkyl and R1 is selected from (C6-C 10 )aryl, -(C1-C6)alkyl-(C6-C10 )aryl and -(C2-C6)alkenyl-(C6-C 10 )aryl.

[0063] According to one embodiment, R1 and R2 are independently selected from C1-C6 alkyl and (C6-C 10 )aryl(C2-C6)alkenyl, preferably selected from methyl, styryl and 2-(naphthalen-2-yl)vinyl.

[0064] According to a particular embodiment, R1 is selected from C1-C6 alkyl, in particular methyl, and R2 is selected from -(C1-C6)alkyl-(C6-C 10 )aryl, in particular styryl or 2-(naphthalen-2-yl)vinyl.

[0065] According to another particular embodiment, R2 is selected from C1-C6 alkyl and R1 is selected from (C6-C 10 ) aryl, (C6-C 10 )aryl, -(C1-C6)alkyl-(C6-C 10 )aryl and -(C2-C6)alkenyl-(C6-C 10 )aryl.

[0066] According to one embodiment, R1 is 5-methyl.

[0067] According to one embodiment, R2 is 5-phenylvinyl or 2-(naphthalen-2-yl)vinyl.

[0068] According to another embodiment, R2 is 5-methyl.

[0069] According to another embodiment, R1 is 5-phenylvinyl or 2-(naphthalen-2-yl)vinyl.

[0070] In other aspects, compounds of formula (I) are included, wherein the pharmaceutically acceptable salt is hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate, phosphate, acetate, citrate, propionate, fumarate, maleate, tartrate, glutamate, salicylate, oxalate, methanesulfonate, p-toluenesulfonate, succinate or benzoate.

[0071] In other aspects, compounds of formula (I) are included, wherein the pharmaceutically acceptable salt is a calcium salt, a magnesium salt, a lithium salt, a potassium salt or a sodium salt.

[0072] It will be appreciated that the compounds of the present invention may exist in various stereoisomeric forms. Thus, the compounds of the present invention include diastereomers, enantiomers and atropisomers. These compounds are typically prepared as racemates and can conveniently be used as racemates, but if desired, individual enantiomers may be separated or synthesized by conventional techniques. These racemates and individual enantiomers and mixtures thereof form part of the present invention.

[0073] How to prepare and separate such optically active forms is well known in the art. Specific stereoisomers can be prepared by stereospecific synthesis using pure enantiomers or enantiomer-enriched starting materials. Specific stereoisomers of starting materials or products can be separated and recovered by techniques known in the art, such as separation of racemic forms, normal phase, reverse phase and chiral chromatography, recrystallization, enzyme separation or fractional recrystallization of addition salts formed by reagents for this purpose. Eliel, EL; Wilen, SH Stereochemistry of Organic Compounds; Wiley: New York, 1994, and Jacques, J, et al. Enantiomers, Racemates, and Resolutions; Wiley: New York, 1981. Useful methods for separation and recovery of specific stereoisomers are all incorporated herein by reference in their entirety.

[0074] According to one embodiment, the compound defined above is of the following formula (IVa) or (IVb):

[0075]

[0076] In particular, it is the following formula (IVa1), (IVa2), (IVb1) or (IVb2):

[0077]

[0078]

[0079] Or the formula:

[0080]

[0081] in:

[0082] X1, X2, at each occurrence, are independently selected from C or N;

[0083] R1, R2, R3, at each occurrence, are independently as defined above.

[0084] According to one embodiment, the compound as defined above is one of the following formulae:

[0085]

[0086]

[0087]

[0088]

[0089] or a stereoisomeric form thereof, a mixture of stereoisomeric forms and a pharmaceutically acceptable salt form, in particular a sodium salt or a fumarate salt,

[0090] in:

[0091] R3 and R4, at each occurrence, are independently as defined above.

[0092] According to one embodiment, W is -NH-, L1 is (C1-C 18 )alkyl, r is 1, X is (5-7 membered) heteroaryl, in particular triazole, and L2 is (C1-C 18 ) alkyl, especially of the formula -CH2-(O-CH2-CH2) 1-6 -, and Y is a single bond, -NH- or -C(=O)NH-.

[0093] According to another embodiment, W is -NH-, L1 is interrupted by one or more -O- atoms (C1-C 18 ) alkyl, especially of the formula -CH2-CH2-(O-CH2-CH2) 1-6 -, r is 0, and Y is a single bond or -C(=O)NH-.

[0094] According to another embodiment, W is -C≡C-, and L1 is interrupted by one or more -O- atoms (C1-C 18 ) alkyl, especially of the formula -CH2-(O-CH2-CH2) 1-6 -, r is 0, and Y is a single bond or -C(=O)NH-.

[0095] According to one embodiment, Z is a VHL (von Hippel-Lindau), CRBN (cereblon), cIAP (inhibitor of apoptosis) or MDM2 (Mouse Double Minute 2) E3 ubiquitin ligase ligand, and Z is in particular a VHL E3 ubiquitin ligase ligand.

[0096] According to a specific embodiment, Z is selected from:

[0097]

[0098]

[0099] In other aspects, a compound of formula (I) is included, which is selected from the following:

[0100]

[0101]

[0102]

[0103]

[0104]

[0105]

[0106]

[0107]

[0108]

[0109]

[0110]

[0111]

[0112]

[0113] in:

[0114] R3 is H or as defined above, especially Cl;

[0115] R8 is (C1-C6)alkyl, especially propyl.

[0116] Preparation method of compound of formula (I)

[0117] The compounds of the present invention can be prepared by many methods known to those skilled in the art, including but not limited to those described below, or by modifying these methods using standard techniques known to those skilled in the art of organic synthesis. The reagents and starting materials are commercially available or easily synthesized by techniques known to those of ordinary skill in the art. Unless otherwise indicated, all substituents are as previously defined. All disclosed methods associated with the present invention can be envisioned to be implemented on any scale, including milligram, gram, multigram or commercial industrial scales.

[0118] As will be readily appreciated, the functional groups present on the compounds of formula I may contain protecting groups. Protecting groups themselves are known as chemical functional groups that can be selectively attached to or removed from functional groups such as hydroxyl and carboxyl. These groups are present in the compound so that the functional group is inert to the chemical reaction conditions to which the compound is exposed. The present invention may employ any of a variety of protecting groups. Other preferred protecting groups according to the present invention may be found in Greene, TWand Wuts, PGM, "Protective Groups in Organic Synthesis" 2d.Ed., Wiley & Sons, 1991, or PJ Kocienski, "Protecting Groups", 3rd.Ed., Thieme, Stuttgart, NY, 2004.

[0119] The compound thus prepared can be recovered from the reaction mixture by conventional methods. For example, the compound can be distilled from the solvent mixture after extraction, or if necessary, after distilling from the solvent mixture, the residue is poured into water, then extracted with a water-immiscible organic solvent and distilled from the solvent mixture. In addition, if necessary, the product can be further purified by various well-known techniques, such as recrystallization, precipitation or various chromatographic techniques, especially column chromatography or preparative thin layer chromatography, especially high performance liquid chromatography (HPLC).

[0120] In another object, the present invention relates to a process for preparing a compound of formula (I) as defined herein, wherein r is 1, the process comprising the steps of:

[0121]

[0122] Therein, Xa and Xb are chemical groups which after chemical reaction form the group X as defined above.

[0123] Xa and Xb are well known to those skilled in the art and are, for example, -N3 and -≡(alkynyl), respectively, which form triazoles via copper(I)-catalyzed Huisgen cycloaddition of azide-alkyne.

[0124] Compound (B) is commercially available, as described below, or can be obtained by:

[0125]

[0126] Therein, Ya and Yb are chemical groups which after chemical reaction form the group Y as defined above.

[0127] Ya and Yb are well known to those skilled in the art and are, for example, -COOH or an activated ester and -NH2, which form -CONH- by peptide coupling; or -NH2 and -F (-F is especially bound to an aromatic group), which form -NH- by nucleophilic aromatic substitution.

[0128] Compounds (B1) and (B2) are commercially available, as described below, or can be obtained by methods known to those skilled in the art.

[0129] In one embodiment, compound (A) is obtained as follows:

[0130]

[0131] Wherein Wa and Wb are chemical groups which, after chemical reaction, form the group W as defined above.

[0132] Wa and Wb are well known to those skilled in the art and are, for example, -F or -Cl and -NH2, wherein -F or -Cl and -NH2 form -NH- through nucleophilic aromatic substitution; or -COOH or an activated ester and -NH2, wherein -COOH or an activated ester and -NH2 form -CONH- through peptide coupling.

[0133] D is commercially available or obtained by methods known to those skilled in the art.

[0134] In one embodiment, compound (C) is obtained as follows, Ar1 is particularly selected from (C6-C 10 )aryl and (5-7 membered)heteroaryl:

[0135] When j is 1 and i is 0 or 1:

[0136]

[0137] Wherein, Hal2 is a halogen, especially Br,

[0138] or

[0139] When j is 1 and i is 1:

[0140]

[0141] Wherein, Hal1 is a halogen, especially Br,

[0142] or

[0143] When j=0 and i is 1:

[0144]

[0145] Wherein, Hal1 is a halogen, especially Br.

[0146] When j=0 and i=0, compound (C) can be obtained by methods known to those skilled in the art, in particular, by the method described in WO2015132727.

[0147] When Ar1 is Hal, compound (C) can be obtained similarly to compound (G) as described below.

[0148] When Ar1 is selected from -O-(C6-C 10 )aryl, -O-(5- to 7-membered)heteroaryl, -S-(C6-C 10 )aryl, -S-(5- to 7-membered)heteroaryl, -(C1-C6)alkyl-(C6-C 10 )aryl, -(C1-C6)alkyl-(5 to 7 membered)heteroaryl, compound (C) can be obtained as follows:

[0149] For Ar1=-O-(C6-C 10 )aryl, -O-(5-7 membered)heteroaryl, -S-(C6-C 10 )aryl or -S-(5-7 membered)heteroaryl, by contacting compound (G) (e.g., wherein Hal1 = Br) with an Ar-SH or Ar-OH compound via nucleophilic aromatic substitution;

[0150] For Ar1=-(C1-C6)alkyl-(C6-C 10 )aryl, -(C1-C6)alkyl-(5-7 membered)heteroaryl, from compound (G) (for example wherein Hal1=Br), in the presence of an Ar-alkyl-MgBr compound, in particular by Negishi coupling.

[0151] In one embodiment, when i is 1, compound (E) is obtained as follows:

[0152]

[0153] wherein Hal1 and Hal2 are halogens, such as I and Br, respectively.

[0154] J can be obtained by methods known to those skilled in the art, in particular the method described in WO2015132727.

[0155] When i is 0, (E) can be obtained by methods known to those skilled in the art, in particular the method described in WO2015132727.

[0156] Alternatively, compound (E) can be obtained as follows:

[0157]

[0158] wherein Hal5 is a halogen, in particular I, and Hal2 is, for example, Br.

[0159] In one embodiment, compound (G) is obtained as follows:

[0160]

[0161] wherein Hal3 is a halogen, in particular I, and Hal1 is, for example, Br.

[0162] Alternatively, compound (G) can be obtained as follows:

[0163]

[0164] wherein Hal4 is a halogen, in particular I, and Hal1 is, for example, Br.

[0165] (L) is commercially available or obtained by methods known to those skilled in the art.

[0166] (M) can be obtained by methods known to those skilled in the art, in particular, by the method described in WO2015132727.

[0167] (N) can be obtained by methods known to those skilled in the art, in particular, by the method described in WO2015132727.

[0168] In another object, the present invention relates to a process for preparing a compound of formula (I) as defined herein, wherein r is 0, comprising the steps of:

[0169]

[0170] The compound (B2) is as described above.

[0171] Compound (A') is as defined above for compound (A). Ya and Yb are well known to those skilled in the art, and are, for example, respectively -COOH or an activated ester and -NH2, -COOH or an activated ester and -NH2 forming -CONH- by peptide coupling, or -NH2 and -F (-F is particularly bound to an aromatic group), -NH2 and -F forming -NH- by nucleophilic aromatic substitution.

[0172] All substituents, such as the group R3, can be modified before, during or after the above steps. For example, the group R3=OH can be modified to R3=O-propyl by SN2 reaction.

[0173] When W is C≡C, the W-containing fragment can be introduced into the aromatic skeleton, for example, by Sonogashira coupling, which is well known to those skilled in the art.

[0174] In another object, the present invention relates to a pharmaceutical composition comprising a compound of formula (I) as defined above, in admixture with at least one pharmaceutically acceptable excipient or carrier.

[0175] All embodiments described above for the compounds of the formula (I) apply here either individually or in combination.

[0176] In one embodiment, the pharmaceutical composition defined above further comprises an additional anticancer agent, in particular selected from anti-EGFR, anti-Map kinase and anti-Akt compounds. Other exemplary additional anticancer agents include carboplatin, paclitaxel, doxorubicin, topotecan and functional derivatives thereof.

[0177] It will be apparent to one of ordinary skill in the art that the specific formulation of the pharmaceutical composition can be selected based on the type of cancer being treated. The compositions of the present invention can be formulated with materials that enhance their stability and / or provide controlled or sustained release in vivo for administration to a patient.

[0178] These pharmaceutical compositions may be prepared in a manner well known in the pharmaceutical art and administered by a variety of routes depending upon whether local or systemic treatment is desired and upon the area to be treated.

[0179] Administration can be topical (including dermal, ophthalmic and mucosal, including intranasal, vaginal and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal, intranasal, epidermal and transdermal), ocular, oral or parenteral.

[0180] In particular, the compounds and compositions of the present invention are administered orally or intravenously.

[0181] The compounds and pharmaceutical compositions of the present invention can be administered orally in any oral acceptable dosage form, including but not limited to capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, commonly used carriers include lactose and corn starch. Lubricants such as magnesium stearate are also commonly added. For oral administration in capsule form, useful diluents include lactose and dry corn starch. When an oral aqueous suspension is required, the active ingredient is combined with an emulsifier and a suspending agent. If desired, certain sweeteners, flavorings or coloring agents may also be added.

[0182] Methods of ocular delivery may include topical administration (eye drops), subconjunctival, periocular or intravitreal injection or introduction via a balloon catheter or an ocular insert surgically placed in the conjunctival sac. Parenteral administration includes intravenous, arterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial, such as intrathecal or intraventricular administration.

[0183] The pharmaceutical compositions can be formulated so as to provide controlled, quick, sustained or delayed release of the active ingredient after administration to the patient by employing procedures known in the art.

[0184] The pharmaceutical composition may also be formulated by techniques well known to those skilled in the art, for example by nanovectorization or by using micronized forms.

[0185] Pharmaceutical compositions typically include pharmaceutically acceptable inorganic or organic carriers, preservatives, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavoring agents, salts for changing osmotic pressure, buffers, masking agents or antioxidants. They may also contain other therapeutically valuable substances. These ingredients are selected based on the mode and route of administration. Suitable pharmaceutical ingredients and pharmaceutical necessities for use in pharmaceutical preparations are described in Remington's Pharmaceutical Sciences (EW Martin).

[0186] Pharmaceutically acceptable carriers that can be used in these pharmaceutical compositions include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as prolamin sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon dioxide, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, polyethylene glycol and wool fat. The therapeutic dose of the compounds of the present invention can vary, for example, depending on the specific use for which the treatment is intended, the mode of administration of the compounds, the health and condition of the patient, and the judgment of the attending physician. The proportion or concentration of the compounds of the present invention in the pharmaceutical composition can vary depending on many factors including dosage, chemical properties (e.g., hydrophobicity), and route of administration.

[0187] In another object, the invention therefore relates to compounds of formula (I) as defined above for use in the treatment of cancer, in particular hematological malignancies such as lymphoma, leukemia, multiple myeloma, and / or solid tumors such as ovarian cancer, mesothelioma, melanoma, pancreatic cancer, lung cancer, breast cancer, kidney cancer and liver cancer.

[0188] All embodiments described above for the compounds of the formula (I) apply here either individually or in combination.

[0189] In a particular embodiment, the present invention therefore relates to a compound of formula (I) as defined above for use in inducing the expression of a molecule selected from the group consisting of Bcl-2, Mcl-1, Bcl-x LAt least one protein selected from Bcl-w, Bfl-1 and Bcl-B, in particular Bcl-2 protein, or Bcl-x L and / or Mcl-1 protein, more particularly Bcl-2 protein, or Bcl-x L and Mcl-1 protein-mediated apoptosis to treat cancer.

[0190] Cancers suitable for the treatment method of the present invention may be selected from Bcl-2, Mcl-1, Bcl-x L At least one protein selected from Bcl-w, Bfl-1 and Bcl-B, in particular Bcl-2 protein, or Bcl-x L and / or Mcl-1 protein; thus, any cancer that is selected from Bcl-2, Mcl-1, Bcl-x L At least one protein among Bcl-w, Bfl-1 and Bcl-B, especially Bcl-2 protein, or Bcl-x L Cancers associated with malignant tumors and / or dysregulation (eg, overexpression or altered binding or activity) of the Mcl-1 protein are within the scope of the present invention.

[0191] Cancer or neoplastic diseases include, for example, but are not limited to, breast cancer, hematological cancers such as myeloma, leukemia and lymphoma (e.g., Burkitt lymphoma, non-Hodgkin lymphoma, Hodgkin lymphoma and acute T-cell leukemia), neural tumors such as brain tumors (e.g., gliomas, including astrocytomas or glioblastomas), melanoma, lung cancer, head and neck cancer, thyroid cancer, gastrointestinal tumors (such as stomach, colon or rectal cancer, etc.), liver cancer, pancreatic cancer, genitourinary tumors (such as ovarian cancer, vaginal cancer, vulvar cancer, endometrial cancer, bladder cancer, kidney cancer, testicular cancer, prostate cancer or penis cancer), bone tumors, vascular tumors, and skin cancer (such as basal cell carcinoma, squamous cell carcinoma and melanoma).

[0192] In another aspect, the compound of formula (I) is used as a L At least one protein selected from Bcl-w, Bfl-1 and Bcl-B, in particular Bcl-2 protein, or Bcl-x L and / or Mcl-1 protein, more particularly Bcl-2 protein, or Bcl-x L and a ligand for Mcl-1 protein.

[0193] In another aspect, the compound of formula (I) is used to induce a gene selected from Bcl-2, Mcl-1, Bcl-x L At least one protein selected from Bcl-w, Bfl-1 and Bcl-B, in particular Bcl-2 protein, or Bcl-x Land / or Mcl-1 protein, more particularly Bcl-2 protein, or Bcl-x L and Mcl-1 protein-mediated apoptosis.

[0194] A patient is effectively treated whenever a clinically beneficial outcome occurs. This may mean, for example, complete resolution of disease symptoms, a decrease in the severity of disease symptoms, or a slowing of disease progression.

[0195] An effective amount of any composition provided herein can be administered to an individual in need of treatment. The term "effective" as used herein refers to any amount that induces a desired response in a patient without inducing significant toxicity. Such an amount can be determined by evaluating the patient's response after administration of a known amount of a specific composition.

[0196] In another aspect, the present invention relates to a method for treating cancer, which method comprises administering to a patient in need thereof an effective amount of a pharmaceutical composition comprising a compound of formula (I) as defined above.

[0197] In another aspect, the present invention relates to a method for degrading a protein selected from the group consisting of Bcl-2, Mcl-1, Bcl-x L At least one protein selected from Bcl-w, Bfl-1 and Bcl-B, in particular Bcl-2 protein, or Bcl-x L and / or Mcl-1 protein, more particularly Bcl-2 protein, or Bcl-x L and Mcl-1 protein, which comprises contacting said protein with a compound of formula (I) as defined above.

[0198] In another aspect, the present invention relates to a method for degrading a protein selected from the group consisting of Bcl-2, Mcl-1, Bcl-x L At least one protein selected from Bcl-w, Bfl-1 and Bcl-B, in particular Bcl-2 protein or Bcl-x L and / or Mcl-1 protein, more particularly Bcl-2 protein, or Bcl-x L and Mcl-1 protein, the method comprising contacting the protein with the compound of formula (I) above, so as to degrade the protein through ubiquitination targeted by the compound of formula (I).

[0199] The present invention also relates to a method for killing cancer cells and regulating a protein selected from Bcl-2, Mcl-1, Bcl-x L At least one protein selected from Bcl-w, Bfl-1 and Bcl-B, in particular Bcl-2 protein, or Bcl-x LThe methods of treating the disease described herein can be combined with other cytotoxic treatments (eg, chemotherapy, hormone therapy, radiation therapy, antibody-based therapy, or targeted therapy for cell signaling pathways).

[0200] In a preferred embodiment, the compounds and compositions of the present invention can be used to prevent or reduce the incidence of Bcl-2, Mcl-1, Bcl-x L , Bcl-w, Bfl-1 and Bcl-B, especially Bcl-2, or Bcl-x L and / or Mcl-1 expressing cancer; more specifically, they can be used to increase the survival of such patients, increase the progression-free survival of such patients, increase the duration of response, thereby resulting in a statistically significant and clinically meaningful improvement in the treated patients, as measured by survival, progression-free survival, response rate or duration of response. In a preferred embodiment, the drug helps to increase the response rate of a group of patients.

[0201] As used herein, the term "alkyl" refers to a straight or branched alkyl group having 1 to 8 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, 1-ethylpropyl, 3-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, hexyl, octyl, etc. The alkyl portion of an alkyl-containing group (such as an alkoxy group and an alkoxycarbonyl group) has the same meaning as the alkyl group defined above. A preferred lower alkyl group is an alkyl group containing 1-4 carbons as defined above. Names such as "C1-C6 alkyl" refer to alkyl groups containing 1 to 6 carbon atoms. As used herein, the term "alkenyl" refers to a straight or branched hydrocarbon chain having 2 to 6 carbon atoms with at least one carbon-carbon double bond. Names such as "C2-C6 alkyl" refer to alkenyl groups containing 2 to 6 carbon atoms. Examples of the alkenyl group include ethenyl, propenyl, isopropenyl, 2,4-pentadienyl. "C2-C4 alkenyl" is particularly preferred.

[0202] As used herein, the term "alkoxy" refers to an alkyl-O- group, wherein alkyl is as described herein. Exemplary alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, and n-butoxy.

[0203] As used herein, the term "aryl" refers to a substituted or unsubstituted mono- or bicyclic hydrocarbon aromatic ring system having 6 to 10 ring carbon atoms. Examples include phenyl and naphthyl.

[0204] As used herein, the term "aralkyl" refers to an alkyl group substituted by an aryl group. Examples of aralkyl groups include, but are not limited to, benzyl, bromobenzyl, phenethyl, benzhydryl, diphenylmethyl, triphenylmethyl, diphenylethyl, naphthylmethyl.

[0205] As used herein, the term "arylalkenyl" refers to an alkenyl group substituted with an aryl group. Examples of arylalkenyl groups include, but are not limited to, styryl.

[0206] The term "arylcarbonyl" as used herein refers to an aryl-C(=O)- group wherein the aryl group is as described herein.

[0207] As used herein, the term "aralkylcarbonyl" refers to an aralkyl-C(=O)- group wherein the aralkyl group is as described herein.

[0208] As used herein, the term "heteroaryl" refers to an aromatic group containing 5 to 10, preferably 5 to 7, ring carbon atoms, wherein one or more ring carbon atoms are substituted with at least one heteroatom such as -O-, -N- or -S-. Examples of heteroaryl groups include pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxathiolate, oxadiazolyl, triazolyl, oxatriazolyl, furanyl, tetrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, indolyl, isoindolyl, indazolyl, benzofuranyl, isobenzofuranyl, purinyl, quinazolinyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzothiazolyl, benzophenylthio, thianaphthenyl, benzoxazolyl, benzisoxazolyl, cinnolinyl, phthalazinyl, naphthyridinyl, and quinoxalinyl. Included in the definition of "heteroaryl" are fused ring systems, including, for example, ring systems in which an aromatic ring is fused to a heterocycloalkyl ring. Examples of such fused ring systems include, for example, phthalamide, phthalic anhydride, indoline, isoindoline, tetrahydroisoquinoline, chroman, isochroman, chromene, and isochromene.

[0209] As used herein, the term "subject" refers to a warm-blooded animal, such as a mammal, preferably a human or a human child, that is suffering from or is likely to suffer from one or more of the diseases and disorders described herein.

[0210] As used herein, "therapeutically effective amount" refers to an amount of a compound of the present invention that is effective in preventing or treating a particular disease. Such diseases include, but are not limited to, those pathological and neurological diseases associated with abnormal activity of the receptors described herein, wherein treatment or prevention includes inhibiting, inducing or enhancing the activity of the receptor by contacting it with a compound of the present invention.

[0211] As used herein, the term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problematic complications commensurate with a reasonable benefit / risk ratio.

[0212] The meanings of all other terms used in the description of the present invention are well known in the art.

[0213] In another aspect, the present invention relates to pharmaceutically acceptable salts of the above compounds. As used herein, "pharmaceutically acceptable salts" include salts of the compounds of the present invention which are obtained by combining these compounds with non-toxic acid or base addition salts.

[0214] Acid addition salts include inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid and phosphoric acid, and organic acids such as acetic acid, citric acid, propionic acid, fumaric acid, maleic acid, tartaric acid, glutamic acid, salicylic acid, oxalic acid, methanesulfonic acid, p-toluenesulfonic acid, succinic acid and benzoic acid, and related inorganic and organic acids.

[0215] Base addition salts include those derived from inorganic bases such as ammonium and alkali and alkaline earth metal hydroxides, carbonates, bicarbonates and the like, and from basic organic amines such as aliphatic and aromatic amines, aliphatic diamines, hydroxyalkylamines and the like. Such bases useful in preparing the salts of this invention thus include ammonium hydroxide, potassium carbonate, sodium bicarbonate, calcium hydroxide, methylamine, diethylamine, ethylenediamine, cyclohexylamine, ethanolamine and the like.

[0216] In addition to pharmaceutically acceptable salts, the present invention also includes other salts that can be used as intermediates in the purification of compounds, the preparation of other salts, or the identification and characterization of compounds or intermediates. BRIEF DESCRIPTION OF THE DRAWINGS

[0217] Figure 1 According to Example 4, CRBN expression on the IGROV1-R10 ovarian cancer cell line is shown.

[0218] Figure 2 According to Example 4, the expression of Mcl-1 and Bcl-xL in IGROV1-R10 cells after exposure to 1 μM Pyridoclax or 1 μM CRBN-PBM1-001 for 24 h is shown, and the expression is quantified by density analysis of western blot signals. DETAILED DESCRIPTION

[0219] Example

[0220] Example 1: Synthesis of compound of formula (I)

[0221] Materials and methods are described below.

[0222] Material

[0223] Commercial reagents were used as received without additional purification. Melting points were determined on a Kofler heating table. IR spectra were recorded on a Perkin Elmer BX FT-IR spectrophotometer. Band positions are reported in reciprocal centimeters (cm -1 ). Recorded on a JEOL Lambda 400 spectrometer 1 H NMR (400 MHz) and 13 C NMR (100 MHz) spectrum. Chemical shifts are expressed in parts per million downfield with tetramethylsilane as internal standard, and coupling constants are expressed in Hertz. Chemical shifts are reported in parts per million (ppm) relative to the solvent resonance. Chromatography was performed on a column using fast silica gel 60 Merck (0.063-0.200 mm) as stationary phase. The elution solvent required for each purification was determined by thin layer chromatography (TLC) on 0.2 mm pre-coated silica gel 60 F-264 (Merck) plates, and the spots were observed with ultraviolet light. Elemental analysis was performed on new compounds, and the data for C, H, N were within ±0.4 of the theoretical values ​​for all final compounds.

[0224] method

[0225] The synthesis of the compounds of the present invention is as follows. The reaction is carried out according to the general procedure mentioned below in the order shown in brackets:

[0226]

[0227]

[0228]

[0229]

[0230]

[0231]

[0232]

[0233] General procedure for Suzuki-Miyaura cross-coupling reaction: To a reaction vessel containing a solution of the appropriate bromo or iodo pyridine derivative in 1,4-dioxane (0.2 M) was added the boronic acid reactant (1.1 eq.), tetrakis(triphenylphosphine)palladium(0) (5 mol%) and saturated aqueous K3PO4 (2.5 eq.) under an inert atmosphere. The mixture was refluxed to completion of the reaction (monitored by TLC). The mixture was cooled to room temperature, quenched with water, and extracted with ethyl acetate. The organic layer was collected, washed with brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to give a crude residue, which was purified by column chromatography.

[0234] Nucleophilic aromatic substitution SN Ar The general procedure:

[0235] The amine is dissolved in dry DMSO and DIPEA (N,N-diisopropylethylamine) (2-4 equivalents). Fluorine or chlorine (pyridyl) or (benzyl) compound (1.0 equivalent) is added, and the mixture is stirred at 90 ° C until the reaction is completed (monitored by TLC). After cooling to room temperature, the mixture is poured into half-saturated brine and extracted with EtOAc. The combined organic layer is further washed with saturated NH4Cl solution, 5% LiCl solution and brine. The organic layer is dried with Na2SO4, filtered and concentrated in vacuo.

[0236] General procedure for nucleophilic substitution SN2: To a reaction vessel containing phenol compound (1.0 eq.) in DMF, iodoalkane (2.0 eq.) and K2CO3 (4.0 eq.) were added at 0°C. The reaction mixture was stirred at room temperature until the reaction was complete (monitored by TLC). The mixture was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure to give a crude residue which was purified by column chromatography.

[0237] General procedure for the copper(I)-catalyzed azide-alkyne cycloaddition (CuACC):

[0238] Under an inert atmosphere, t To a mixture of azide (1.0 eq.) and alkyne (1.33 eq.) in BuOH-THF (1:1, v / v) was added CuSO4·5H2O (2.2 eq.) and sodium L-ascorbate (2.2 eq.) in water. The mixture was stirred at 55°C until the reaction was complete (monitored by TLC) and cooled to room temperature. It was then poured into water and extracted with DCM. The organic phase was washed with brine, dried over Na2SO4, filtered and evaporated to dryness. The crude product was purified by flash column chromatography on silica gel using DCM and MeOH as eluents.

[0239] General procedure for nitrile hydrolysis:

[0240] The benzonitrile compound (1.0 equivalent) was stirred in a mixture of NaOH (2N) and MeOH at 100°C until the reaction was complete (monitored by TLC). The reaction mixture was cooled at room temperature. The reaction mixture was acidified with AcOH until a precipitate appeared. The precipitate was filtered.

[0241] General procedure for peptide coupling reactions:

[0242] The carboxylic acid compound (1.0 equivalent) is dissolved in dry DMF, and DIPEA (3.0 equivalent) and HATU (1.1 equivalent) are added under an inert atmosphere. After stirring for 5 min, the amine derivative (1.1 equivalent) dissolved in dry DMF is added. The combined mixture is stirred at room temperature until the reaction is completed (monitored by TLC). Half-saturated brine is added and extracted with EtOAc. The combined organic layer is further washed with saturated NH4Cl solution, 5% LiCl solution and brine. The organic layer is dried with Na2SO4, filtered and concentrated in vacuo.

[0243] General procedure for coupling of BOC-deprotection with HATU:

[0244] The BOC protected joint conjugate (1.0 equivalent) is dissolved in dry CH2Cl2 and treated with trifluoroacetic acid. The mixture is stirred at room temperature until the reaction is completed. Remove the solvent and co-evaporate with dry CH2Cl2. The oily residue is further dried in a high vacuum. The deprotected acid derivative (1.0 equivalent) is dissolved in dry DMF, and DIPEA (3.0 equivalents) and HATU (1.1 equivalents) are added. After stirring for 5min, the amine derivative (1.1 equivalents) dissolved in dry DMF is added. The combined mixture is stirred at room temperature until the reaction is completed (monitored by TLC). Half-saturated brine is added and extracted with EtOAc. The combined organic layer is further washed with saturated NH4Cl solution, 5% LiCl solution and brine. The organic layer is dried with Na2SO4, filtered and concentrated in vacuo.

[0245] The above starting compound is obtained as follows:

[0246]

[0247] Similar to that described in WO2015132727.

[0248] When Ar1 is selected from -O-(C6-C 10 )aryl, -O-(5-7 membered)heteroaryl, -S-(C6-C 10 )aryl, -S-(5-7 membered)heteroaryl, -(C1-C6)alkyl-(C6-C 10)aryl, -(C1-C6)alkyl-(5-7 membered)heteroaryl, the Ar1 group can be introduced similarly as shown below:

[0249]

[0250] Nucleophilic aromatic substitution SN Ar General procedure: To a reaction vessel containing (E)-6-bromo-5-methyl-3'-phenylvinyl-3,2':5',3'-terpyridine (1.0 equiv) in DMSO, benzenethiol (1.0 equiv) and K2CO3 (2.0 equiv) were added. The reaction mixture was stirred at 110°C for 48 h, and the raw material conversion was only 50%. The reaction mixture was further charged with benzenethiol (1.0 equiv) and K2CO3 (2.0 equiv) and the reaction was complete after 24 h. The mixture was cooled to room temperature, quenched with water, and extracted with DCM. The combined organic layers were washed with brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to give a crude residue, which was purified by column chromatography (DCM / MeOH 95 / 5 to 9 / 1).

[0251] General procedure for Ullman-type coupling reaction: To a reaction vessel containing (E)-6-bromo-5-methyl-3'-phenylvinyl-3,2':5',3'-terpyridine (1.0 equiv) in DMF, phenol (1.0 equiv), copper (0) powder (10%) and Cs2CO3 (3.0 equiv) were added. The reaction mixture was stirred at 100°C for 24h. The mixture was cooled to room temperature, quenched with NaOH 1M solution, and extracted with DCM. The combined organic layers were washed with brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to give a crude residue, which was purified by column chromatography (DCM / MeOH 98 / 2).

[0252] General procedure for palladium-catalyzed Negishi cross-coupling reaction: To a reaction vessel containing (E)-6-bromo-5-methyl-3'-phenylvinyl-3,2':5',3'-terpyridine (1.0 equiv) in dry THF was added 0.5 M benzylzinc(II) bromide (1.0 equiv) and tetrakis(triphenylphosphine)palladium(0) (10%) under nitrogen atmosphere. The reaction mixture was stirred at room temperature for 48 h. The mixture was concentrated under reduced pressure to give a crude residue which was purified by column chromatography (DCM / MeOH 99 / 1 to 9 / 1).

[0253] Example 2: Biological activity of compounds of formula (I)

[0254] Morphological observation of cell nucleus

[0255] After treatment, detached and adherent cells were pooled after trypsinization, applied to poly-lysine coated slides by cell centrifugation, and fixed with ethanol / chloroform / acetic acid (6:3:1) solution. The preparations were then fixed with 1 solution. Then DAPI solution (Boehringer Mannheim-Roche, Mannheim, Germany) was incubated at room temperature for 15 min, washed in distilled water, mounted under a Mowiol (Calbiochem) cover slip, and analyzed under a fluorescence microscope (BX51, Olympus, Rungis, France). Condensed and fragmented nuclei indicate the induction of apoptosis.

[0256] Target expression and apoptosis induction analysis by Western blot

[0257] The cells were rinsed with ice-cold PBS, suspended in lysis buffer [RIPA: NaCl 150mM, Tris (pH 8) 50mM, Triton X100 1%, PMSF 4mM, EDTA 5mM, NaF 10mM, NaPPi 10mM, Na3OV4 1mM, aprotinin 0.5μL / ml and 4.6ml ultrapure water], and incubated on ice for 30min. Lysates were collected after centrifugation (13200g, 10min, 4°C) and protein concentration was determined using Bradford assay (Bio-Rad, Hercules, USA). In 4-15% gradient polyacrylamide TGX TM Equal amounts of protein (20 μg) were separated by SDS-PAGE on precast gels (Bio-Rad) and then transferred to PDVF membranes (Bio-Rad). The membranes were blocked with 5% (v / v) nonfat dry milk in TBS (T-TBS) containing 0.05% (v / v) Tween 20 for 1 h at room temperature. The membranes were then incubated with the appropriate antibodies listed above overnight at 4°C. The membranes were then washed with T-TBS and incubated with the appropriate secondary antibodies for 1 h. Enhanced chemiluminescence substrate (ECL) Prime Western Blot detection reagent (GE Healthcare Life Sciences) and Las4000Series (GE Healthcare Life Sciences) displays the signal and then uses The software performed quantification by pixel densitometry.

[0258] Antibodies used: Anti-Mcl-1 (#5453), Anti-Bcl-X L(#2764), anti-Caspase 3 (#9662), anti-PARP (#9542), (Cell Signaling Technology), anti-Actin (#MAB1501) (Merck Millipore) and anti-VHL (#GTX101087) (Genetex).

[0259] Quantification of apoptotic events using flow cytometry

[0260] Adherent and floating cells are collected, washed with phosphate buffered saline (PBS), and fixed with 70% ethanol. The cells are then centrifuged for 5 min at 2000 r.pm, and incubated for 30 min in PBS at 37 °C to allow the release of low molecular weight DNA. Using DNA Prep Coulter test kit (Beckman Coulter), cell mass is stained with propidium iodide. Samples are analyzed with Gallios flow cytometer (Beckman Coulter), and cell cycle distribution is measured with Gallios software (Beckman-Coulter).

[0261] Real-time cell activity analysis (impedance measurement, xCELLigence technology)

[0262] Real-time growth curve monitoring was performed using the xCELLigence system (ACEA, Ozyme, Saint Quentinen Yvelines, France) using a real-time cell analyzer multi-plate instrument. The system monitors cell events in real time by measuring the electrical impedance on the staggered microelectrodes integrated on the bottom surface of the 96-well E plate VIEW. These electrodes measure CI (cell index) based on impedance. CI is related to the area of ​​cells attached to the bottom of the plate. CI values ​​are shown in the figure. In short, cells were plated on a 96-well E plate View and placed on an RTCA MP in a tissue culture incubator. Cells were grown for 24 hours before treatment, and impedance was measured continuously until the end of treatment. The standard deviation of triplicate wells was analyzed using RTCA2.1.0 software.

[0263] Live cell imaging

[0264] The cells were seeded in 96-well plates in culture medium. The cells were maintained at 37°C and 5% CO2 and used S3 (Essen BioScience) was used for monitoring. Caspase-3 / 7 green cell apoptosis detection reagent (Essen Bioscience) was used, and baseline images were taken with a 10x objective. Fluorescence and phase contrast images were acquired in real time from 2 independent areas of each well every 1 hour. Caspase 3 / 7 reagent labeled dead cells that produced green fluorescence. Phase contrast images of live cells were used for Software was used to calculate confluence and provide morphological information. Each experiment was performed in triplicate, and the accumulation of Caspase 3 / 7 over time was normalized to the confluence of the cells.

[0265] Characterization of E3 ligase expression by immunohistochemistry

[0266] Immunohistochemistry with a Ventana Discovery XT autostainer was performed on 4 μm thick paraffin sections. Slides were dewaxed with EZPrep buffer and epitopes were uncovered by high temperature treatment in CC1 EDTA buffer for 15 min. Sections were incubated at 37°C with antibodies against various E3 ligases for 40 min, including anti-VHL (GTX101087) (Genetex). Secondary antibodies (Omnimap rabbit; Ventana Medical System Inc., Tucson, AZ, USA) were incubated at room temperature for 16 min. Immunodetection performed in the absence of primary antibodies was used as a control. After washing, staining was performed with DAB, and sections were counterstained with hematoxylin using Ventana reagent according to the manufacturer's protocol. The stained slides were then digitized using an Aperio ScanScope slide scanner (Aperio Technologies, Vista, CA, USA).

[0267] Protein quantification by ELISA

[0268] Colorimetric ELISA assays can quantitatively analyze target protein expression. The primary antibody targets the protein of interest. The secondary antibody recognizes the primary antibody and is coupled to HRP to allow colorimetric detection. Kits for ELISA assays can be used for Bcl-x L OKAG00550 for AMP and OKAG00858 for Mcl-1 and the application was according to the manufacturer's recommendations.

[0269] result

[0270] The compounds of the present invention induce Bcl-2, Mcl-1, Bcl-x L , Bcl-w, Bfl-1 or Bcl-B proteins, in particular Bcl-2 proteins, or proteolysis of Bcl-xL and / or Mcl-1 proteins.

[0271] Without being bound by any particular theory, it is believed that they recruit E3 ligases to the target protein, which enables ubiquitination and subsequent degradation of the target protein by the proteasome.

[0272] Example 3: Synthesis of other compounds of the present invention

[0273] As shown below, in certain exemplary embodiments, compounds are prepared according to the following general procedures. It should be understood that although the general methods describe the synthesis of certain compounds of the present invention, the following general methods and other methods known to those of ordinary skill in the art can be applied to all compounds and subclasses and species of each of these compounds, as described herein.

[0274] General Synthesis Methods

[0275] The following examples are intended to illustrate the present invention and should not be construed as limiting the present invention. Temperatures are expressed in degrees Celsius. If not otherwise mentioned, all evaporations are carried out under reduced pressure, preferably between about 15 mmHg and 100 mmHg (=20-133 mbar). The structures of the final products, intermediates and starting materials are confirmed by standard analytical methods such as microscopic analysis and spectral characteristics such as MS, IR, NMR. The abbreviations used are those conventional abbreviations in the art.

[0276] All starting materials, building blocks, reagents, acids, bases, dehydrating agents, solvents and catalysts used to synthesize the compounds of the present invention are either commercially available or can be produced by organic synthesis methods known to those of ordinary skill in the art. In addition, the compounds of the present invention can be produced by organic synthesis methods known to those of ordinary skill in the art, as shown in the following examples.

[0277] Unless otherwise stated, all reactions were performed under nitrogen or argon.

[0278] Analytical Instruments

[0279]

[0280] Unless otherwise stated, all solvents and chemicals were used as received.All NMR spectra were recorded on a Bruker Avance III 400 MHz or Avance III HD 500 MHz spectrometer. 1 H- and 13C NMR spectra are reported in chemical shifts (δ) in parts per million (ppm). Coupling constants (J) are reported in Hertz (Hz) to one decimal place. The following abbreviations are used to describe multiplicities: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), br (broad). Infrared spectra were recorded for pure compounds using a Perkin Elmer FT-IR spectrometer. Absorption is reported in wavenumbers (cm -1 ) and only the peaks of interest are reported. The melting points of the solids were measured with a Griffin instrument. LC-MS was performed using a Waters SQ detector as a mass spectrometer and a Waters Alliance 2695 as a separation module (chromatographic columns XBridge C18 2.5mM / 4.6 50mM) with the following gradient: A (H2O, containing 0.1% HCO2H) / B (CH3CN, containing 0.1% HCO2H) 95 / 5 to A / B 5 / 95 in 4.00min. This ratio was maintained for 1.50min, then returned to the initial conditions for 0.50min, and then maintained for 2.00min. The retention times (t R ) are expressed in min. Unless otherwise indicated, all reagents were obtained from commercial suppliers. Chromatography was performed using 60 Merck (0.063-0.200 mm) fast silica gel as the stationary phase. Flash chromatography was performed using Biotage Isolera One flash chromatography. The elution solvent required for each purification was determined by thin layer chromatography (TLC) using Merck 0.2 mm silica gel pre-coated plates and Kieselgel 60F264. The product was visualized using UV fluorescence (254 nm).

[0281] Building Block Synthesis Method

[0282]

[0283] 5-Bromo-3-iodopyridine-2-ol 1. To a round-bottom flask containing 5-bromopyridine-2-ol (5.0 g, 28.7 mmol) in acetonitrile (120 mL), N-iodosuccinimide (7.1 g, 31.6 mmol, 1.1 equivalents) was added. The solution was refluxed for 4 h and TLC was performed after the reaction. The suspension was cooled to room temperature, the solid was filtered and washed with MeOH to give 7.1 g of pink solid 1 (83% yield).

[0284] Chemical formula: C5H3BrINO; molecular weight: 299.89 g / mol; melting point: 63°C.

[0285] IR (KBr tablet): 3490, 1630, 1607, 1522, 1457, 1309, 863, 683, 628cm -1 .

[0286] 1 H NMR (CDCl3) δ 8.14 (d, J = 2.5 Hz, 1H), 7.59 (d, J = 2.5 Hz, 1H), 4.20 (br s, 1H, OH).

[0287] 13 C NMR (CDCl3) δ171.5, 146.8, 145.9, 100.7, 92.9.

[0288] MS (ESI+): m / z 300.25 [M+H + ],302.26[M + +2].

[0289] 5-Bromo-2-chloro-3-iodopyridine 2. 5-Bromo-3-iodopyridine-2-ol 1 (9.0 g, 30.0 mmol) was dissolved in 90% dichlorophenylphosphine (100 mL). The mixture was stirred and heated at 160 ° C for 4 h, and then subjected to TLC. The mixture was cooled to room temperature and quenched by slowly adding 1 liter of ice and 28% ammonia water (200 mL) into a flask. The mixture was extracted with ethyl acetate, the organic layer was collected, washed with brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to obtain a crude residue, which was purified by column chromatography (cyclohexane / ethyl acetate 9 / 1) to obtain 7.8 g of a white solid 2 (82% yield).

[0290] Chemical formula: C5H2BrClIN; molecular weight: 318.34 g / mol; melting point: 67°C.

[0291] IR (KBr tablet): 3081, 3038, 1525, 1391, 1352, 1136, 1012, 890, 711, 515cm -1 .

[0292] 1 H-NMR (CDCl3) δ 8.40 (d, J = 2.3 Hz, 1H), 8.29 (d, J = 2.3 Hz, 1H).

[0293] 13 C-NMR (CDCl3) δ153.2, 150.2, 149.6, 118.4, 95.3.

[0294] MS (ESI+): m / z 318.17 [M+H + ],320.14[M+ +2],322.15[M + +4].

[0295] (E)-5-bromo-2-chloro-3-phenylvinylpyridine 3. To a reaction vessel containing 5-bromo-2-chloro-3-iodopyridine 2 (13.0 g, 40.8 mmol) in n-butanol (300 mL) under nitrogen atmosphere, (E)-phenylvinylboronic acid (9.0 g, 61.2 mmol, 1.5 eq.), Pd(dppf)Cl2-DCM (166 mg, 0.21 mmol, 0.5 mol%) and K2CO3 (11.3 g, 81.6 mmol, 2.0 eq.) were added. The mixture was stirred at reflux overnight. The mixture was cooled to room temperature, quenched with water and extracted with ethyl acetate. The organic layer was collected, washed with brine, dried over anhydrous magnesium sulfate, filtered and then concentrated under reduced pressure to give a crude residue, which was purified by column chromatography (cyclohexane / ethyl acetate 99 / 1, then 98 / 2) to give 9.6 g of 3 as a white solid (80% yield).

[0296] Chemical formula: C 13 H9BrClN; molecular weight: 294.58 g / mol; melting point: 104°C.

[0297] IR (KBr tablet): 3036, 1627, 1530, 1494, 1447, 1389, 1152, 1116, 1063, 956, 897, 755, 719, 687, 491cm -1 .

[0298] 1 H-NMR(CDCl3)δ8.35(d,J=2.5Hz,1H,H Pyr ),8.11(d,J=2.5Hz,1H,H Pyr ),7.57(d,J=7.4Hz,2H,H Ar ),7.42(t,J=7.4Hz,2H,H Ar ),7.38–7.34(m,1H,H Ar ), 7.27 (d, J = 16.3 Hz, 1H, Ar-CH = CH-Pyr), 7.12 (d, J = 16.3 Hz, 1H, Ar-CH = CH-Pyr).

[0299] 13 C-NMR(CDCl3)δ148.6,148.5,136.7,135.9,134.6(Ar-CH=CH-Pyr),133.7,129.1,128.9(2C Ar H),127.1(2CAr H), 121.9 (Ar-CH=CH-Pyr), 119.3.

[0300] MS (ESI+): m / z 294.33 [M+H + ],296.30[M + +2],298.30[M + +4].

[0301]

[0302] (E)-6-chloro-5-phenylvinyl-3,3'-bipyridine 4. To a reaction vessel containing (E)-5-bromo-2-chloro-3-phenylvinylpyridine 3 (3.8 g, 12.7 mmol) in 1,4-dioxane under nitrogen atmosphere, pyridin-3-ylboronic acid (1.7 g, 14.0 mmol, 1.1 eq.), tetrakis(triphenylphosphine)palladium(0) (734 mg, 0.64 mmol, 5 mol%) and K3PO4 (6.7 g, 31.8 mmol, 2.5 eq.) were added. The mixture was refluxed for 2 h and then subjected to TLC. The crude residue was purified by column chromatography (cyclohexane / ethyl acetate 4 / 1 to 1 / 1) to give 3.0 g of 4 as a white solid (80% yield).

[0303] Chemical formula: C 18 H 13 ClN2; molecular weight: 292.77 g / mol; melting point: 159°C.

[0304] IR (KBr tablet): 3045, 3019, 2924, 2853, 1638, 1578, 1446, 1427, 1385, 1166, 1087, 1057, 1023, 951, 921, 804, 729, 714, 687cm -1 .

[0305] 1 H-NMR(CDCl3)δ8.90(d,J=2.4Hz,1H,H Pyr ),8.72(dd,J=4.9,1.6Hz,1H,H Pyr ),8.51(d,J=2.4Hz,1H,H Pyr ),8.16(d,J=2.4Hz,1H,H Pyr ),7.95(ddd,J=7.9,2.4,1.6Hz,1H,H Pyr ),7.61–7.59(m,2H,H Ar ),7.49–7.41(m,4H,H Pyr ,3HAr ,Ar-CH=CH-Pyr),7.38–7.34(m,1H,H Ar ), 7.21 (d, J=16.3Hz, 1H, Ar-CH=CH-Pyr).

[0306] 13 C-NMR3)δ150.0,149.5,147.9,146.1,136.2,136.1,134.7,134.0(Ar-CH=CH-Pyr),132.9,132.9,132.5,132.3,128.9(2C Ar H),127.1(2C Ar H), 123.9, 122.7 (Ar-CH=CH-Pyr).

[0307] MS (ESI+): m / z 293.35 [M+H + ],295.34[M + +2].

[0308] (E)-6-iodo-5-phenylvinyl-3,3'-bipyridine 5. To a solution of (E)-6-chloro-5-phenylvinyl-3,3'-bipyridine 4 (3.5 g, 12.1 mmol) in acetonitrile, NaI (4.5 g, 4.8 mmol, 2.5 eq.) and acetyl chloride (1.4 g, 18.1 mmol, 1.5 eq.) were added. The reaction mixture was refluxed for 16 h to give 4.3 g of a crude residue (80% of 5, obtained from 1 H-NMR confirmed), as a yellow solid, which was used in the next reaction without further purification.

[0309] Chemical formula: C 18 H 13 IN2; molecular weight: 384.22 g / mol; melting point: 160°C.

[0310] IR (KBr tablet): 3079, 3045, 3023, 2924, 1634, 1574, 1544, 1495, 1453, 1420, 1381, 1163, 1087, 1060, 1023, 959, 914, 805, 729, 712, 680cm -1 . 216

[0311] 1 H-NMR(CDCl3)δ8.89(dd,J=2.4,0.9Hz,1H,H Pyr ),8.71(dd,J=4.8,1.7Hz,1H,H Pyr),8.46(d,J=2.4Hz,1H,H Pyr ),7.95(d,J=2.4Hz,1H,H Pyr ),7.93(ddd,J=7.9,2.4,1.7Hz,1H,H Pyr ),7.61–7.59(m,2H,H Ar ),7.48–7.41(m,3H,2H Ar ,Pyr-H),7.38–7.34(m,2H,H Ar ), 7.29 (d, J = 16.1 Hz, 1H, Ar-CH = CH-Pyr), 7.11 (d, J = 16.1 Hz, 1H, Ar-CH = CH-Pyr).

[0312] 13 C-NMR3) δ149.1, 148.0, 144.8, 140.7, 135.4, 135.2, 134.1, 133.4, 133.2, 132.9, 128.7 (2C AR H),128.0,127.4,126.4(2C AR H), 124.0, 115.9.

[0313] MS (ESI+): m / z 385.41 [M+H + ],386.40[M + +1].

[0314] (E)-6-bromo-5-methyl-3'-phenylvinyl-3,2':5',3'-terpyridine 6. To a reaction vessel of (E)-6-iodo-5-phenylvinyl-3,3'-bipyridine 5 (1.5 g, 3.9 mmol) contained in 1,4-dioxane, (6-bromo-5-methylpyridin-3-yl)boric acid (930 mg, 4.3 mmol, 1.1 equiv), tetrakis(triphenylphosphine)palladium(0) (225 mg, 0.19 mmol, 5 mol%) and K3PO4 (2.0 g, 9.8 mmol, 2.5 equiv) were added under nitrogen atmosphere. The mixture was refluxed for 4 h and then subjected to TLC. The crude residue was purified by column chromatography (cyclohexane / ethyl acetate 95 / 5 to 4 / 1) to give 751 mg of yellow solid 6 (45% yield).

[0315] Chemical formula: C 24 H 18 BrN3; molecular weight: 428.33 g / mol; melting point: 149°C.

[0316] IR (KBr pellet): 3039, 1588, 1557, 1490, 1438, 1379, 1190, 1120, 1053, 1023, 970, 907, 804, 746, 708, 721, 686, 540 cm -1 。

[0317] 1 1H-NMR (CDCl3) δ 8.96 (dd, J = 2.4, 0.8 Hz, 1H, H Pyr ), 8.83 (d, J = 2.2 Hz, 1H, H Pyr ), 8.72 (dd, J = 4.8, 1.6 Hz, 1H, H Pyr ), 8.50 (d, J = 2.4 Hz, 1H, H Pyr ), 8.21 (d, J = 2.2 Hz, 1H, H Pyr ), 8.00 (dt, J = 7.9, 2.0 Hz, 1H, H Pyr ), 7.92 (d, J = 2.4 Hz, 1H, H Pyr ), 7.50–7.44 (m, 3H, 2H Ar , H Pyr ), 7.40–7.29 (m, 3H, H Ar ), 7.22 (d, J = 16.2 Hz, 1H, Ar-CH=CH-Pyr), 7.14 (d, J = 16.2 Hz, 1H, Ar-CH=CH-Pyr), 2.49 (s, 3H, CH3).

[0318] 13 13C-NMR (CDCl3) δ 152.8, 149.8, 148.4, 148.1, 147.0, 145.0, 139.9, 136.4, 135.3, 134.7, 134.6, 133.5, 133.2, 133.1, 132.8, 132.2, 129.0 (2C Ar H), 128.8, 127.0 (2C Ar H), 124.5, 124.0, 22.2 (CH3).

[0319] MS (ESI+): m / z 428.24 [M+H + , 430.29 [M + +2].

[0320]

[0321] (E)-5-bromo-2-iodo-3-phenylvinylpyridine 7. To a reaction vessel containing (E)-5-bromo-2-chloro-3-phenylvinylpyridine 3 in acetonitrile, NaI (6.1 g, 40.8 mmol, 2.5 eq.) and acetyl chloride (2.0 g, 24.5 mmol, 1.5 eq.) were added. The reaction mixture was refluxed for 12 h to give 5.22 g of a crude residue (80% of 7, obtained from 1 H-NMR confirmed), a yellow solid, which was used in the next reaction without further purification.

[0322] Chemical formula: C 13 H9BrIN; molecular weight: 386.03 g / mol; melting point: 78°C.

[0323] IR (KBr tablet): 3054, 3023, 1629, 1492, 1446, 1377, 1269, 1109, 1026, 952, 889, 751cm -1 .

[0324] 1 H-NMR(CDCl3)δ8.29(d,J=2.0Hz,1H,H Pyr ),7.83(d,J=2.0Hz,1H,H Pyr ),7.52(d,J=7.8Hz,2H,H Ar ),7.38(t,J=7.8Hz,2H,H Ar ),7.32(d,J=7.8Hz,1H,H Ar ), 7.09 (d, 1H, J = 16.1Hz, Ar-CH = CH-Pyr), 6.94 (d, J = 16.1Hz, 1H, Ar-CH = CH-Pyr).

[0325] 13 C-NMR(CDCl3)δ149.6,139.6,135.9,135.1,134.7,129.0,128.9(2C Ar H),128.1,127.1(2C Ar H), 121.1, 120.8.

[0326] MS (ESI+): m / z 386.31 [M+H + ],388.24[M + +2].

[0327] (E)-5,6'-dibromo-5'-methyl-3-phenylvinyl-2,3'-bipyridine 8. To a reaction vessel containing (E)-5-bromo-2-iodo-3-phenylvinylpyridine 7 (3.0 g, 7.7 mmol) in 1,4-dioxane was added (6-bromo-5-methylpyridin-3-yl)boric acid (1.8 g, 8.4 mmol, 1.1 eq.), tetrakis(triphenylphosphine)palladium(0) (445 mg, 0.38 mmol, 5 mol%) and K3PO4 (4.1 g, 19.3 mmol, 2.5 eq.) under nitrogen atmosphere. The mixture was refluxed for 4 h and then subjected to TLC. The crude residue was purified by column chromatography (cyclohexane / ethyl acetate 99 / 1 to 95 / 5) to give 1.8 g of yellow solid 8 (55% yield).

[0328] Chemical formula: C 19 H 14 Br2N2; molecular weight: 430.14 g / mol; melting point: 143°C.

[0329] IR (KBr tablet): 3031, 1393, 1111, 1050, 968, 898, 740, 685, 499cm -1 .

[0330] 1 H-NMR(CDCl3)δ8.65(d,J=1.9Hz,1H,H Pyr ),8,41(d,J=1.9Hz,1H,H Pyr ),8.17(d,J=1.9Hz,1H,H Pyr ),7.83(d,J=1.9Hz,1H,H Pyr ),7.42(d,J=7.8Hz,2H,H Ar ),7.36(t,J=7.8Hz,2H,H Ar ),7.31(d,J=7.8Hz,1H,H Ar ), 7.12 (d, J = 16.6Hz, 1H, Ar-CH = CH-Pyr), 6, 99 (d, J = 16.6Hz, 1H, Ar-CH = CH-Pyr), 2.46 (s, 3H, CH3).

[0331] 13 C-NMR(CDCl3)δ151.4,149.4,147.8,144.9,139.6,136.5,136.0,135.2,134.0,133.9,133.3,128.9(2C Ar H 128.8,126.9(2C ArH), 123.4, 120.4, 22.0 (CH3).

[0332] MS (ESI+): m / z 431.51 [M+H + ],433.53[M + +2].

[0333] (E)-5-bromo-6'-iodo-5'-methyl-3-phenylvinyl-2,3'-bipyridine 9. To a solution of (E)-5,6'-dibromo-5'-methyl-3-phenylvinyl-2,3'-bipyridine 8 (3.8 g, 8.9 mmol) in acetonitrile, NaI (3.3 g, 22.3 mmol, 2.5 eq.) and acetyl chloride (1.0 g, 13.4 mmol, 1.5 eq.) were added. The reaction mixture was refluxed for 8 h to give 4.0 g of a crude residue (90% of 9, obtained from 1 H-NMR confirmed), a yellow solid, which was used in the next reaction without further purification.

[0334] Chemical formula: C 19 H 14 BrIN2; molecular weight: 477.14 g / mol; melting point: 147°C.

[0335] IR (KBr tablet): 3029, 1576, 1449, 1428, 1389, 1112, 1089, 968, 900, 686, 498cm -1 .

[0336] 1 H-NMR(CDCl3)δ8.66(d,J=2.3Hz,1H,H Pyr ),8.38(d,J=2.3Hz,1H,H Pyr ),8.16(d,J=2.3Hz,1H,H Pyr ),7.73(d,J=2.3Hz,1H,H Pyr ),7.44(d,J=7.4Hz,2H,H Ar ),7.38(t,J=7.4Hz,2H,H Ar ),7.34–7.30(m,1H,H Ar ), 7.14 (d, J = 16.2Hz, 1H, Ar-CH = CH-Pyr), 6.98 (d, J = 16.2Hz, 1H, Ar-CH = CH-Pyr), 2.48 (s, 3H, CH3).

[0337] 13C-NMR(CDCl3)δ151.4,149.5,148.1,139.2,137.5,136.5,135.9,134.0,133.9,133.3,128.9(2C Ar H),128.8,126.9(2C Ar H),125.6,123.4,120.4,26.2(CH3).

[0338] MS (ESI+): m / z 477.48 [M+H + ],479.54[M + +2].

[0339] (E)-5-Bromo-5'-methyl-3-phenylvinyl-2,3':6',3"-terpyridine 10. To a reaction vessel containing (E)-5-bromo-6'-iodo-5'-methyl-3-phenylvinyl-2,3'-bipyridine 9 (2.0 g, 4.2 mmol) in 1,4-dioxane was added pyridin-3-ylboronic acid (570 mg, 4.6 mmol, 1.1 equiv), tetrakis(triphenylphosphine)palladium(0) (243 mg, 0.2 mmol, 5 mol%) and K3PO4 (2.3 g, 10.5 mmol, 2.5 equiv) under nitrogen atmosphere. The mixture was refluxed for 3 h and then subjected to TLC. The crude residue was purified by column chromatography (cyclohexane / ethyl acetate 8 / 2 to 1 / 1) to give 1.1 g of off-white solid 10 (60% yield).

[0340] Chemical formula: C 24 H 18 BrN3; molecular weight: 428.33 g / mol; melting point: 136-138°C.

[0341] IR (KBr tablet): 3032, 2917, 1635, 1414, 1384, 1118, 950, 740, 541cm -1 .

[0342] 1 H-NMR(CDCl3)δ8.88(dd,J=2.3,0.9Hz,1H,H Pyr ),8.78(dd,J=2.1,0.7Hz,1H,H Pyr ),8.69(d,J=2.2Hz,1H,H Pyr ),8.68(dd,J=4.9,1.7Hz,1H,H Pyr ),8.21(d,J=2.1Hz,1H,H Pyr ),7.97(ddd,J=7.8,2.3,1.7Hz,1H,HPyr ),7.95(dd,J=2.2,0.8Hz,1H,H Pyr ),7.45(m,3H,2H Ar ,H Pyr ),7.38–7.28(m,3H,H Ar ), 7.16 (d, J = 16.2Hz, 1H, Ar-CH = CH-Pyr), 7.11 (d, J = 16.2Hz, 1H, Ar-CH = CH-Pyr), 2.48 (s, 3H, CH3).

[0343] 13 C-NMR(CDCl3)δ155.5,152.4,149.9,149.6,149.3,148.1,139.9,136.9,136.6,136.3,133.9,133.8(Ar-CH=CH-Pyr),133.6,132.3,131.4,129.0(2C Ar H),128.9,127.1(2C Ar H), 123.8 (Ar-CH=CH-Pyr), 123.5, 120.4, 20.2 (CH3).

[0344] MS (ESI+): m / z 428.74 [M+H + ],430.62[M + +2].

[0345] HRMS (ESI): m / z calculated for [M+H] + :428.0762; Found value:428.0772.

[0346]

[0347] (E)-5-bromo-6'-(2-hydroxyphenyl)-5'-methyl-3-phenylvinyl-2,3'-bipyridine 11. To a reaction vessel containing (E)-5-bromo-6'-iodo-5'-methyl-3-phenylvinyl-2,3'-bipyridine 9 (356 mg, 0.75 mmol, 1.0 eq.) in 1,4-dioxane (20 mL) under nitrogen atmosphere, 2-hydroxyphenylboronic acid (120 mg, 0.83 mmol, 1.1 eq.), tetrakis(triphenylphosphine)palladium(0) (43 mg, 0.04 mmol, 5 mol%) and K3PO4 (398 mg, 1.88 mmol, 2.5 eq.) were added. The mixture was refluxed for 24 h and then subjected to TLC. The crude residue was purified by column chromatography (cyclohexane / ethyl acetate 100 / 0 to 90 / 10) to give 183 mg of 11 as an off-white solid (55% yield).

[0348] Chemical formula: C 25 H 19 BrN2O; molecular weight: 443.34 g / mol.

[0349] NMR 1 H (400MHz, CDCl3): δ (ppm) = 12.15 (s, 1H), 8.61 (dd, J = 9.6, 2.2Hz, 2H), 8.13 (d, J = 2.2Hz, 1H), 7.96 (d, J = 2.2Hz, 1H), 7.56 (dd, J = 7.9, 1.7Hz, 1H), 7.3 8(d,J=6.9Hz,1H),7.29(t,J=7.4Hz,1H),7.25(d,J=7.0Hz,1H),7.09(d,J =16.2Hz,1H),7.05–6.97(m,4H),6.88(td,J=7.6,1.3Hz,1H),2.59(s,3H).

[0350] NMR 13 C (75MHz, CDCl3): δ (ppm) = 157.98, 156.42, 151.84, 149.55, 144.98, 141.95, 136.62, 136.10, 133.78, 133.43 ,132.61,131.61,130.89,129.80,128.99,128.84,126.93,123.59,121.24,120.36,118.56,118.17,21.94.

[0351] (E)-5-bromo-6'-(2-propoxyphenyl)-5'-methyl-3-phenylvinyl-2,3'-bipyridine 12. To a reaction vessel containing (E)-5-bromo-6'-(2-hydroxyphenyl)-5'-methyl-3-phenylvinyl-2,3'-bipyridine 11 (163 mg, 0.37 mmol, 1.0 eq) in DMF (10 mL) was added 1-iodopropane (54 μL, 0.55 mmol, 1.5 eq) and K2CO3 (102 mg, 0.74 mmol, 2 eq). The reaction mixture was stirred at 50 °C for 17 h. The mixture was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure to give a crude residue, which was purified by column chromatography (cyclohexane / ethyl acetate 100 / 0 to 90 / 10) to give 147 mg of 12 as a yellow solid (82% yield).

[0352] Chemical formula: C 28 H 25 BrN2O; molecular weight: 485.43 g / mol.

[0353] NMR 1 H (400MHz, CDCl3): δ (ppm) = 8.65 (d, J = 2.1Hz, 1H), 8.62 (d, J = 2.2Hz, 1H), 8.14 (d,J=2.2Hz,1H),7.77(dd,J=2.2,0.9Hz,1H),7.39–7.35(m,2H),7.34–7.21(m ,5H),7.07(s,2H),7.00(td,J=7.5,1.0Hz,1H),6.91(dd,J=8.2,1.0Hz,1H),3. 86 (t, J = 6.5 Hz, 2H), 2.19 (s, 1H), 1.61 (h, J = 7.3 Hz, 2H), 0.81 (t, J = 7.4 Hz, 3H).

[0354] NMR 13 C (75MHz, CDCl3): δ (ppm) = 157.47, 156.16, 149.35, 147.06, 138.04, 136.33, 136.29, 133.37, 133.19, 132.86, 1 32.74,130.59,129.86,129.66,128.84,128.60,126.91,124.10,120.76,112.06,69.86,22.59,19.10,10.59.

[0355]

[0356] (E)-6'-(2-propoxyphenyl)-5'-methyl-5-(6-fluoropyridin-3-yl)-3-phenylvinyl-2,3':6',3"-terpyridine 13. Under nitrogen atmosphere, to a reaction vessel containing (E)-5-bromo-6'-(2-propoxyphenyl)-5'-methyl-3-phenylvinyl-2,3'-bipyridine 12 (147 mg, 0.303 mmol) in 1,4-dioxane (20 mL) was added commercially available 6-fluoro- Pyridin-3-ylboronic acid (47 mg, 0.333 mmol, 1.1 eq.), tetrakis(triphenylphosphine)palladium(0) (35 mg, 0.03 mmol, 10 mol%) and K3PO4 (225 mg, 1.06 mmol, 3.5 eq.). The mixture was refluxed for 26 h and then subjected to TLC. The crude residue was purified by column chromatography (dichloromethane / ethyl acetate 100 / 0 to 50 / 50) to give 17 mg of yellow oil 13 (13% yield).

[0357] Chemical formula: C 33 H 28 FN3O; molecular weight: 501.61 g / mol.

[0358] 1 H-NMR(CDCl3):8.76(d,J=2.2Hz,1H),8.73(dd,J=2.1,0.7Hz,1H),8.50(dt,J=2.7,0.9Hz,1H),8.13(d, J=2.3Hz,1H),8.04(ddd,J=8.5,7.5,2.7Hz,1H),7.85(dd,J=2.2,0.8Hz,1H),7.44–7.35(m,2H),7.22(s, 6H),7.14(d,J=16.2Hz,1H),7.06(ddd,J=8.4,3.0,0.7Hz,1H),7.01(td,J=7.4,1.0Hz,1H),6.92(d,J=8 .1Hz,1H),3.88(t,J=6.5Hz,2H),2.21(s,3H),1.63(dtd,J=13.9,7.4,6.5Hz,2H),0.83(t,J=7.4Hz,3H).

[0359] 13C-NMR(CDCl3)163.7(d,J=240.9Hz),157.5,156.2,154.4,147.2,146.6, 146.1(d,J=15.1Hz),139.9(d,J=8.1Hz),138.2,136.5,133.2,132.8,132 .4,131.9,131.6,131.4(d,J=4.7Hz),130.6,129.9,129.7,128.9,128.5, 126.9, 124.9, 120.8, 112.1, 110.1 (d, J = 37.5Hz), 69.9, 22.6, 19.1, 10.6.

[0360] 19 F NMR (376MHz, CDCl3) δ (ppm)-68.49.

[0361] The final compound was obtained from compound 12 as follows:

[0362]

[0363]

[0364] (E)-5'-Methyl-5-(6-fluoropyridin-3-yl)-3-phenylvinyl-2,3':6',3"-terpyridine 14. To a reaction vessel containing (E)-5-bromo-5'-methyl-3-phenylvinyl-2,3':6',3"-terpyridine 10 (214 mg, 0.5 mmol) in 1,4-dioxane under nitrogen atmosphere, commercially available 6-fluoropyridin-3-ylboronic acid (82 mg, 0.55 mmol, 1.1 equiv), tetrakis(triphenylphosphine)palladium(0) (58 mg, 0.05 mmol, 10 mol%) and K3PO4 (371 mg, 1.75 mmol, 3.5 equiv) were added. The mixture was refluxed for 12 h and then subjected to TLC analysis. The crude residue was purified by column chromatography (dichloromethane / ethyl acetate 100 / 0 to 20 / 80) to give 113 mg of 14 as an off-white solid (51% yield).

[0365] Chemical formula: C 29 H 21 FN4; molecular weight: 444.51 g / mol; melting point: 212°C.

[0366] NMR 1H(400MHz,CDCl3):δ(ppm)=8.85–8.83(m,1H),8.79(d,J=2.1Hz,1H),8.77(d,J=2.2Hz,1H),8.62(dd,J=4.9,1.7Hz,1H),8.53–8.48(m,1H),8.13(d,J=2.2Hz,1H),8.05(ddd,J=8.5,7.5,2.6Hz,1H),7.96(dd,J=2.2,0.8Hz,1H),7.92(dt,J=7.9,1.9Hz,1H),7.44–7.35(m,3H),7.33–7.27(m,2H),7.27–7.21(m,1H),7.18(d,J=2.2Hz,1H),7.06(ddd,J=8.5,3.1,0.7Hz,1H),2.44(s,3H)。

[0367] NMR 13 C(75MHz,CDCl3):δ(ppm)=163.75(d,J=241.2Hz),155.41,153.59,149.92,149.26,148.10,146.76,146.14(d,J=15.1Hz),139.99–139.75(m),136.53(d,J=35.2Hz),134.09,133.32,132.59,132.15,131.90,131.29,131.23,128.92,128.67,126.91,124.60,110.13(d,J=37.5Hz),20.07。

[0368] NMR 19 F(376MHz,CDCl3):δ(ppm)=-68.28。

[0369]

[0370] (E)-5'-Methyl-5-(6-(amino-propan-1-ol)pyridin-3-yl)-3-phenylvinyl-2,3':6',3"-terpyridine 15. 3-Aminopropan-1-ol (19 μl, 0.24 mmol, 2 equiv) was dissolved in dry DMF and DIPEA (N,N-diisopropylethylamine) (52 μ2, 0.3 mmol, 2.5 equiv). (E)-5'-Methyl-5-(6-fluoropyridin-3-yl)-3-phenylvinyl-2,3':6',3"-terpyridine 14 (53 mg, 0.12 mmol, 1.0 equiv) was added, and the mixture was stirred at 120 °C for 5 h under microwave irradiation. After cooling to room temperature, the mixture was diluted with EtOAc and washed with water and brine. The organic layer was dried over MgSO4, filtered, and concentrated in vacuo. The crude residue was purified by column chromatography (EtOAc / MeOH 100 / 0 to 90 / 10) to afford 36 mg of 15 as a light yellow solid (61% yield).

[0371] Appearance: Yellow powder

[0372] Chemical formula: C 32 H 29 N5O; molecular weight: 499.62 g / mol.

[0373] NMR 1 H (400MHz, CDCl3): δ (ppm) = 8.83 (d, J = 2.3Hz, 1H), 8.77 (d, J = 2.1Hz, 1H), 8.72 (d, J = 2.2Hz, 1H), 8.60 (dd, J = 4.9, 1.7Hz, 1H),8.36(d,J=2.5Hz,1H),8.07(d,J=2.3Hz,1H),7.94(d,J=2.1Hz,1H),7.91(dt,J=7.9,2.1Hz,1H),7.67(dd,J=8.6,2 .5Hz,1H),7.44–7.33(m,3H),7.32–7.26(m,2H),7.25–7.21(m,1H),7.18(d,J=15.9Hz,1H),7.15(d,J=16.5Hz,1H),6.4 9(d,J=8.6Hz,1H),4.87(t,J=6.4Hz,1H),3.64(t,J=5.5Hz,2H),3.61–3.50(m,2H),2.42(s,3H),1.75(p,J=5.8Hz,2H).

[0374] NMR 13C (75MHz, CDCl3): δ (ppm) = 158.94, 150.00, 149.24, 148.14, 146.20, 146.07, 139.85, 136.69, 136.60, 136.06, 1 33.42,132.71,131.22,131.17,128.86,128.43,126.86,125.05,123.28,108.72,58.84,38.09,33.34,20.05.

[0375]

[0376] 2-(2,6-dioxopiperidin-3-yl)-4-fluoro-2,3-dihydro-1H-isoindole-1,3-dione 16. Commercially available 3-fluorophthalic anhydride (171 mg, 1 mmol), commercially available 3-aminopiperidine-2,6-dione hydrochloride (169 mg, 1 mmol) and sodium acetate (121 mg, 1.2 mmol) were mixed in AcOH (30 mL) in a round-bottom flask. The resulting reaction mixture was heated and refluxed at 90 °C for 16 h. After cooling to room temperature, most of the AcOH was evaporated and the purple residue was dissolved in water (20 mL) and extracted with ethyl acetate (3×50 mL). The combined organic layers were dried (brine, then MgSO4) and concentrated under vacuum. The residue was purified by flash column chromatography using cyclohexane / ethyl acetate (50 / 50) to give the product as a light blue solid (yield: 68%).

[0377] Chemical formula: C 13 H9FN2O4; molecular weight: 276.22 g / mol.

[0378] NMR 1 H (400MHz, DMSO-d6): δ (ppm) = 11.15 (s, 1H, NH), 7.99–7.92 (m, 1H, H6), 7.80 (d, 3 J=7.2Hz,1H,H7),7.77-7.72(m,1H,H5),5.16(dd, 3 J ax-ax =12.8, 3 J ax-eq =5.4Hz,1H,H 3’ ),2.96–2.84(m,1H,H 4’ ),2.65–2.52(m,2H,H 4’ ,H 5’ ),2.11–2.02(m,1H,H 5’ ).

[0379] NMR13 C(75MHz,DMSO-d6):δ(ppm)=173.2(C 6’ ),170.2(C 2’ ),166.6(d, 3 J CF =2.9Hz,C3),164.4(C1),157.3(d, 1 J CF =262.4Hz,C4),138.5(d, 3 J CF =7.9Hz,C6),133.9(d, 3 J CF =1.2Hz,C 7a ),123.5(d, 2 J CF =19.6Hz,C5),120.5(d, 4 J CF =3.2Hz,C7),117.5(d, 2 J CF =12.7Hz,C 3a ),49.5(C 3’ ),31.4(C 4’ ),22.3(C 5’ )。

[0380] NMR 19 F(MHz,DMSO-d6):δ(ppm)=-114.7。

[0381] MS(ESI+):m / z:308.90[M+MeOH+H] + ,638.52[M+2MeCN+2H] 2+ ,552.59[2M+H] + 。

[0382] 2-(2,6-Dioxopiperidin-3-yl)-4-((2-(2-prop-2-yn-1-yloxy)ethoxy)ethyl)amino)isoindoline-1,3-dione 17. 4-Fluoro-thalidomide 16 (120 mg, 0.44 mmol), amine linker (75 μ5, 0.53 mmol, 1.2 eq.) and DIEA (150 Am, 0.87 mmol, 3 eq.) in dry DMF (5 mL) were stirred at 90 °C for 16 h in a round bottom flask. The reaction mixture was diluted with 20 mL of ethyl acetate and the organic phase was washed with water (5 x 10 mL), then dried (brine, then MgSO4) and evaporated under reduced pressure. The residue was purified by column chromatography using a gradient of ethyl acetate in cyclohexane (0%-70%) to give the product as a green solid (yield: 32%).

[0383] Chemical formula: C 20 H 21 N3O6; molecular weight: 399.40 g / mol.

[0384] NMR 1 H (400MHz, DMSO-d6): δ (ppm) = 11.16 (s, 1H, NH a ),7.64(dd, 3 J=8.5, 3 J=7.2Hz,1H,H6),7.20(d,J=8.6Hz,1H,H5),7.09(d,J=7.2Hz,1H,H7),6.66(t, 3 J=5.6Hz,1H,NH b ),5.11(dd, 3 J ax-ax =12.9, 3 J ax-eq =5.5Hz,1H,H 3’ ),4.19(d, 4 J=2.4Hz,2H,H 5” ),3.69–3.65(m,2H,H 2” ),3.65–3.60(m,4H,H 3” ,H 4” ),3.53(q, 3 J=5.6Hz,2H,H 1” ),3.46(t, 4 J=2.4Hz,1H,H 7” ),2.99–2.88(m,1H,H 4’ ),2.69–2.58(m,2H,H4',H 5’ ),2.13–2.04(m,1H,H5’ ).

[0385] NMR 13 C (75MHz, DMSO-d6): δ (ppm) = 172.9 (C 6’ ),170.2(C 2’ ),169.0(C3),167.3(C1),146.4(C4),136.3(C6),132.1(C7a),117.5(C5),110.7(C7),109.3(C 3a ),80.4(C 6” ),77.2(C 7” ),69.5(C 3” ),68.9(C 2” ),68.6(C 4’ ),57.6(C 5” ),48.6(C 3’ ),41.7(C 1” ),31.0(C 4’ ),22.2(C 5’ ).

[0386]

[0387] 3-Bromo-4-hydroxybenzoic acid methyl ester 18. To a solution of methyl 4-hydroxybenzoate (1 g, 6.57 mmol, 1.0 equiv) in dry CH2Cl2 (75 mL) was carefully added dibromide (0.33 mL, 65.7 mmol, 10.0 equiv) at room temperature. The mixture was stirred at room temperature for 18 h. Acetic acid (1 mL) was then added and the reaction was stirred for more than 3 h. The mixture was washed with saturated Na2S2O3 aqueous solution, brine, dried (MgSO4) and concentrated in vacuo. The resulting crude residue was purified by column chromatography (CH2Cl2 / MeOH 100 / 0 to 99 / 1) to give 592 mg of 18 as a white solid (39%).

[0388] Chemical formula: C8H7BrO3; molecular weight: 231.05g / mol; melting point: 109℃.

[0389] NMR 1 H (400MHz, CDCl3): δ (ppm) = 8.12 (d, J = 2.1Hz, 1H), 7.85 (dd, J = 8.5, 2.0Hz, 1H), 6.98 (d, J = 8.5Hz, 1H), 5.90 (s, 1H), 3.83 (s, 3H).

[0390] NMR 13C (75MHz, CDCl3): δ (ppm) = 165.6, 156.2, 134.0, 131.1, 124.1, 115.8, 110.1, 52.2.

[0391] Methyl 3-bromo-4-propoxybenzoate 19. To a solution of methyl 3-bromo-4-hydroxybenzoate 18 (462 mg, 2 mmol, 1.0 eq) in dry acetone (20 mL) was added 1-iodopropane (0.29 mL, 3 mmol, 1.5 eq) and K2CO3 (553 mg, 4 mmol, 2 eq) at room temperature. After cooling to room temperature, the mixture was diluted with EtOAc, washed with water, brine, dried (MgSO4) and concentrated in vacuo to give 492 mg of 19 as a light yellow oil (90%).

[0392] Chemical formula: C 11 H 13 BrO3; molecular weight: 273.13 g / mol.

[0393] NMR 1 H (400MHz, CDCl3): δ (ppm) = 8.14 (d, J = 2.1Hz, 1H), 7.86 (dd, J = 8.6, 2.1Hz, 1H), 6.79 (d, J = 8 .7Hz, 1H), 3.96 (t, J = 6.4Hz, 2H), 3.81 (s, 3H), 1.80 (h, J = 7.3Hz, 3H), 1.01 (t, J = 7.4Hz, 3H).

[0394] NMR 13 C (75MHz, CDCl3): δ (ppm) = 165.80, 159.09, 134.76, 130.51, 123.38, 111.83, 111.81, 70.76, 52.11, 22.38, 10.51.

[0395] 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-4-(propoxy)benzoic acid methyl ester 20. To a reaction vessel containing 3-bromo-4-propoxybenzoic acid methyl ester 19 (492 mg, 1.74 mmol, 1.0 equiv) in 1,4-dioxane (3.5 mL) under nitrogen atmosphere, commercially available bis(pinacol)diboron (664 mg, 2.62 mmol, 1.5 equiv), bis(triphenylphosphine)palladium(II) dichloride (250 mg, 0.35 mmol, 2 mol%) and KOAc (514 mg, 5.23 mmol, 3.0 equiv) were added. The mixture was refluxed for 24 h and then subjected to TLC. The crude residue was purified by column chromatography (cyclohexane / ethyl acetate 100 / 0-80 / 20) to give 285 mg of 20 as a white solid (51% yield).

[0396] Chemical formula: C 17 H 25 BO5; molecular weight: 320.19 g / mol; melting point: 87°C.

[0397] NMR 1 H (400MHz, CDCl3): δ (ppm) = 8.22 (d, J = 2.4Hz, 1H), 7.99 (dd, J = 8.7, 2.4Hz, 1H), 6.77 (d, J = 8.8Hz, 1H),3.91(t,J=6.2Hz,2H),3.80(s,3H),1.83–1.71(m,2H),1.19(s,12H),1.01(t,J=7.4Hz,3H).

[0398] NMR 13 C (75MHz, CDCl3): δ (ppm) = 167.2, 166.9, 138.1, 134.4, 121.8, 110.6, 83.5, 69.8, 51.8, 25.0, 22.6, 10.5.

[0399] 11 B NMR (128MHz, CDCl3) δ (ppm): 30.73.

[0400]

[0401] (E)-5'-methyl-5-(methyl 4-(propoxy)benzoate)-3-phenylvinyl-2,3':6',3"-terpyridine 21. To a reaction vessel containing (E)-5-bromo-5'-methyl-3-phenylvinyl-2,3':6',3"-terpyridine 10 (209 mg, 0.49 mmol, 1.0 equiv) in 1,4-dioxane (20 mL) under nitrogen atmosphere were added methyl 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4-(propoxy)benzoate 20 (235 mg, 0.73 mmol, 1.5 equiv), tetrakis(triphenylphosphine)palladium(0) (5.48 mg, 0.025 mmol, 5 mol%) and K3PO4 (363 mg, 1.71 mmol, 3.5 equiv). The mixture was refluxed for 19 h and then subjected to TLC. The crude residue was purified by column chromatography (cyclohexane / ethyl acetate 8 / 2 to 1 / 1) to give 45 mg of 21 as a white solid (yield 17%).

[0402] Chemical formula: C 35 H 31 N3O3; molecular weight: 541.24 g / mol.

[0403] NMR 1 H (400MHz, CDCl3): δ (ppm) = 8.85–8.83 (m, 1H), 8.81–8.77 (m, 2H), 8.64–8.59 (m, 1H), 8.20 (d, J = 2.1Hz, 1H), 8 .09(d,J=2.2Hz,1H),8.04(dd,J=8.7,2.2Hz,1H),7.97(dd,J=2.2,0.8Hz,1H),7.92(dt,J=7.8,1.9Hz,1H),7 .42–7.35(m,3H),7.32–7.26(m,2H),7.25–7.22(m,1H),7.20(d,J=15.8Hz,1H),7.12(d,J=16.2Hz,1H),7.00 (d,J=8.6Hz,1H),4.03(t,J=6.4Hz,2H),3.87(s,3H),2.43(s,3H),1.87–1.71(m,2H),0.99(t,J=7.4Hz,3H).

[0404]

[0405] Nucleophilic substitution :

[0406] 1,3-Diazidopropane 21a and 1,4-Diazidobutane 22b. Alkyldiazides were obtained by simple nucleophilic displacement reaction between appropriate alkyl dihalides and sodium azide according to the following steps. The appropriate dibromoalkyl compound (10.0 mmol, i.e. 2.02 g 1,3-dibromopropane) was dissolved in dry DMF (10 mL) under nitrogen and NaN3 (1.30 g, 20.0 mmol) was added. The mixture was kept at 60°C under magnetic stirring for 24 h. The reaction mixture was then poured into water (50 mL) and extracted five times with ether (50 mL each). The organic extract was washed with water (5 mL) and then dried (brine, then MgSO4). The reaction mixture was evaporated under reduced pressure to give a crude oily product.

[0407] 22a. Chemical formula: C3H6N6; Molecular weight: 126.12 g / mol; Appearance: colorless oil.

[0408] Yield: 55%.

[0409] NMR 1 H (400MHz, DMSO-d6): δ (ppm) = 3.42 (t, 3 J=6.7Hz,4H,H1,H3),1.77(p, 3 J = 6.7 Hz, 2H, H2).

[0410] NMR 13 C (75MHz, DMSO-d6): δ (ppm) = 48.5 (C1, C3), 28.1 (C2).

[0411] 22b. Chemical formula: C4H8N6; Molecular weight: 140.15 g / mol; Appearance: colorless oil.

[0412] Yield: 77%.

[0413] NMR 1 H (400MHz, DMSO-d6): δ (ppm) = 3.40–3.28 (m, 4H, H1, H4), 1.76–1.66 (m, 4H, H2, H3).

[0414] NMR 13 C (75MHz, DMSO-d6): δ (ppm) = 50.9 (C1, C4), 26.2 (C2, C3).

[0415] 3-Azidopropylamine 23a and 4-azidobutylamine 23b. Azidoalkylamines were obtained by regioselective Staudinger reduction of diazidoalkyl compound 22. To a mixture of diazidoalkyl compound 22 (10 mmol) cooled to 0°C, 1 M aqueous HCl solution (20 mL), ether (7 mL) and ethyl acetate (7 mL) were added. Triphenylphosphine was added in portions over 30 min. The mixture was warmed to room temperature and stirred for an additional 20 h, and then the organic layer was separated from the aqueous layer. The aqueous phase was washed with ether (2×50 mL) to remove triphenylphosphine oxide residues. The aqueous phase was basified to pH=13 with aqueous NaOH and then extracted with dichloromethane (3×100 mL). The combined dichloromethane layers were dried over MgSO4 and concentrated to give the derivative of 4-azidoalkylamine as an oil.

[0416] 23a. Chemical formula: C3H8N4; Molecular weight: 100.12 g / mol; Appearance: yellow oil.

[0417] Yield: 52%.

[0418] NMR 1 H (400MHz, DMSO-d6): δ (ppm) = 3.38 (t, 3 J=6.7Hz,2H,H1),2.59(t, 3 J=6.7Hz,2H,H3),1.59(p, 3 J = 6.7 Hz, 2H, H2).

[0419] NMR 13 C (75MHz, DMSO-d6): δ (ppm) = 48.9 (C3), 39.2 (C1), 32.6 (C2).

[0420] 23b, Chemical formula: C4H 10 N4; molecular weight: 114.15 g / mol; appearance: yellow oil.

[0421] Yield: 54%.

[0422] NMR 1 H (400MHz, DMSO-d6): δ (ppm) = 3.23 (t, 3 J=6.8Hz,2H,H1),2.67(t, 3 J=6.9Hz,2H,H3),1.65–1.53(m,2H,H2),1.51–1.44(m,2H,H4).

[0423] NMR 13C (75MHz, DMSO-d6): δ (ppm) = 51.4 (C4), 41.7 (C1), 30.8 (C2), 26.3 (C3).

[0424] Synthesis of one of the preferred compounds: CRBN-PBM1-001

[0425]

[0426] CRBN-PBM1-001 was obtained by Sonogashira cross-coupling reaction. Triethylamine (42 μL, 0.3 mmol, 3.0 equiv), copper (I) iodide (2 mg, 0.01 mmol, 10% mol), 2-(2,6-dioxapiperidin-3-yl)-4-((2-(2-(propan-2-yl-1-yloxy)ethoxy)ethyl)amino)isoindoline-1,3-dione 17 (44 mg, 0.11 mmol, 1.1 equiv) and tetrakis(triphenylphosphine)palladium(0) (8 mg, 0.005 mmol, 5% mol) were added to a solution of 10 (42.8 mg, 0.10 mmol, 1.0 equiv) in dry DMF (2 mL) under inert atmosphere. The reaction mixture was stirred at 50 °C for 6 h. The mixture was diluted with ethyl acetate. The aqueous layer was washed with 4 mL of saturated NaHCO3 solution and brine. The combined organic layers were dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by flash chromatography on a silica gel column (water / acetonitrile 100 / 0 to 0 / 100) to give 41 mg of CRBN-PBM1-001 as a yellow solid (55% yield).

[0427] Chemical formula: C 44 H 38 N6O6; molecular weight: 746.82 g / mol.

[0428] NMR 1H (400MHz, CDCl3): δ (ppm) = 8.82 (s, 1H), 8.75–8.66 (m, 2H), 8.64–8.58 (m, 3H), 8.14 (d, J = 2.2Hz, 1H) ,8.03(d,J=1.9Hz,1H),7.94–7.84(m,3H),7.69–7.60(m,1H),7.43–7.33(m,4H),7.31–7.20(m,3H), 7.09–6.99(m,3H),6.85(d,J=8.5Hz,1H),6.45(t,J=5.5Hz,1H),4.83(dd,J=12.1,5.4Hz,1H),4.44( s,1H),3.81–3.64(m,4H),3.43(q,J=5.5Hz,1H),2.82–2.57(m,2H),2.40(s,3H),2.06–1.97(m,1H).

[0429] NMR 13 C (75MHz, CDCl3): δ (ppm) = 171.2, 169.3, 168.5, 167.6, 155.4, 150.8, 149.9, 149.9, 149.5, 149.3, 14 9.3,148.0,147.9,146.8,139.9,139.8,136.9,136.69,136.65,136.5,136.4,136.2,136.1,135.8, 133.9,133.7,133.3,132.5,131.4,131.3,128.9,128.9,128.8,128.6,126.9,126.9,124.2,123.7,123.3,119.2,116.8,111.7,89.3,82.9,70.6,69.6,69.5,59.3,48.9,42.4,31.5,22.8,20.0,20.0.

[0430] MS (ESI+): m / z: 373.93 [M+2H] 2+ ,748.36[M+H] + .

[0431] Example 4: Further biological activities of the compounds of the present invention

[0432] Materials and methods

[0433] Cell culture and drug testing

[0434] Human platinum-resistant ovarian cancer cells IGROV1-R10 were used. The cell line was grown in RPMI 1640 medium supplemented with 2 mM Glutamax TM, 25 mM HEPES, 10% fetal bovine serum and 33 mM sodium bicarbonate (Fisher Scientific, Illkirch, France). The cells were maintained at 37°C in a humidified atmosphere with 5% CO2. The cells were seeded in 25 cm 2 flasks. After 24 h, exponentially growing cells were exposed to 1, 24 h of the compound. CRBN-PBM1-001 and Pyridoclax targeting Mcl-1 and CRBN were designed and synthesized by CERMN (Centre d'Etudes et de Recherche sur le Médicament de Normandie) and dissolved in dimethyl sulfoxide (DMSO, Sigma-Aldrich).

[0435] Target expression analysis by Western blot

[0436] The cells were rinsed with ice-cold PBS, suspended in lysis buffer [RIPA: NaCl 150 mM, Tris (pH = 8) 50 mM, Triton X100 1%, PMSF 4 mM, EDTA 5 mM, NaF 10 mM, NaPPi 10 mM, Na3VO4 1 mM, aprotinin 0.5 μL / mL and 4.6 mL ultrapure water] and incubated on ice for 30 min. Lysates were collected after centrifugation (13200 g, 10 min, 4 ° C) and protein concentration was determined using Bradford assay (Bio-Rad, Hercules, USA). TGX TM Equal amounts of protein (25 μg) were separated by SDS-PAGE on precast gels (Bio-Rad). The PVDF membrane was activated for 1 min in a 100% ethanol bath and then equilibrated with transfer buffer (3:1:1 ultrapure water, 100% ethanol, 5X transfer buffer [Bio-Rad]). The proteins were then transferred to the activated PDVF membrane (Bio-Rad). The membrane was blocked with 5% (v / v) nonfat dry milk in TBS (T-TBS) containing 0.05% (v / v) Tween20 for 1 h at room temperature. The membrane was then incubated with the appropriate antibodies listed below overnight at 4°C. The membrane was then washed with T-TBS and incubated with the appropriate secondary antibody for 1 h. The membrane was detected using enhanced chemiluminescence substrate (ECL Prime) Western Blot detection reagent (GE Healthcare Life Sciences) and Signals were visualized using Las4000 Series (GE Healthcare Life Sciences). The relative intensities of protein bands were quantified using ImageJ software, and the values ​​were normalized to the intensity of individual actin signals.

[0437] The following antibodies were used: anti-Actin (1 / 2000, #MAB1501, Merck Millipore), anti-CRBN (1 / 1000, ab230411, Abcam), anti-Mcl-1 (1 / 1000, #5453), anti-Bcl-x L (1 / 1000, #2764), anti-rabbit polyclonal secondary antibody (1 / 2000, #7074s) (Cell Signaling Technology), and anti-mouse polyclonal secondary antibody (1 / 2000, #NA931V-ML, Amersham).

[0438] result

[0439] The presence of CRBN was confirmed by western blot in IGROV1-R10 cells. Figure 1 shown.

[0440] The effect of the CRBN-PBM1-001 molecule on simultaneously targeting Mcl-1 and CRBN was characterized to demonstrate its ability to target and degrade Mcl-1. Western blot experiments showed that 24h exposure to Pyridoclax (1μM) as a single agent did not modulate the expression of Mcl-1 in IGROV1-R10 cells. In contrast, after 24h exposure to a 1μM CRBN-PBM1-001 concentration, CRBN-PBM1-001 was able to induce a 52% decrease in Mcl-1 expression, as shown by density analysis ( Figure 2 ). Furthermore, under these experimental conditions, the observed effects were specific to Mcl-1, since Bcl-x L Expression was not affected by exposure to CRBN-PBM1-001, whereas Bcl-2 was not expressed in these cells. Thus, these results suggest that CRBN-PBM1-001 is able to bind to Mcl-1 and initiate its degradation.

Claims

1. A compound of the following formula (I): in: Y1 is N and Y2 is C, Y3 is N and Y4 is C; Ar1 and Ar2 are each independently selected from (C6-C 10 )aryl, (5- to 7-membered)heteroaryl, -O-(C6-C 10 )aryl, -O-(5- to 7-membered)heteroaryl, -(C1-C6)alkyl-(C6-C 10 )aryl, -(C1-C6)alkyl-(5 to 7 membered)heteroaryl and halogen, or, Ar1, Ar2 are each independently selected from (C6-C 10 )aryl, (5- to 7-membered)heteroaryl, -O-(C6-C 10 )aryl, -O-(5- to 7-membered)heteroaryl, -(C1-C6)alkyl-(C6-C 10 )aryl, -(C1-C6)alkyl-(5- to 7-membered)heteroaryl and halogen, said aryl and said heteroaryl being substituted by 1 to 3 R3 groups, i and j are independently 0 or 1, as well as R1 is selected from (C1-C6) alkyl, (C6-C 10 )aryl, -(C1-C6)alkyl-(C6-C 10 )aryl, -(C2-C6)alkenyl-(C6-C 10 )aryl, -carbonyl-(C6-C 10 )aryl, -carbonyl-(C1-C6)alkyl-(C6-C 10 ) aryl; or, R1 is selected from (C1-C6) alkyl, (C6-C 10 )aryl, -(C1-C6)alkyl-(C6-C 10 )aryl, -(C2-C6)alkenyl-(C6-C 10 )aryl, -carbonyl-(C6-C 10 )aryl, -carbonyl-(C1-C6)alkyl-(C6-C 10 ) aryl, said aryl being substituted by 1 to 3 R4 groups, k is 0 or 1, l＝1； R2 is selected from -(C2-C6)alkenyl-(C6-C 10 )aryl, -carbonyl-(C6-C 10 )aryl, -carbonyl-(C1-C6)alkyl-(C6-C 10 ) aryl; or, R2 is selected from -(C2-C6)alkenyl-(C6-C 10 )aryl, -carbonyl-(C6-C 10 )aryl, -carbonyl-(C1-C6)alkyl-(C6-C 10 ) aryl, said aryl being substituted by 1 to 3 R4 groups, R3, at each occurrence, is independently selected from (C1-C6)alkyl, (C6-C 10 )aryl, (5- to 7-membered)heteroaryl, -(C1-C6)alkyl-(C6-C 10 )aryl, -(C2-C6)alkenyl-(C6-C10)aryl; R4, at each occurrence, is independently selected from (C1-C6)alkyl, (CH2) n CO2H, (CH2) n CO2R c , I, Cl, Br, F, or two R4 groups form a naphthyl group with the aryl groups to which they are attached; R c Selected from H and (C1-C6) alkyl; n is 0, 1, 2, 3; r is 0; W is -C≡C- and Y is selected from a single bond, -CH=CH-, -C≡C-, -NH-, -NR5-, -O-, -S-, -C(=O)NH-, -NHC(=O)-, -C(=O)O-, -OC(=O)-; R5, at each occurrence, is independently selected from (C1-C6)alkyl, L1 is selected from (C1-C 18 )alkyl; said group is interrupted by one or more -O- atoms, Z is an E3 ubiquitin ligase ligand selected from or a stereoisomeric form, a mixture of stereoisomeric forms and a pharmaceutically acceptable salt form of the compound of formula (I).

2. The compound according to claim 1, wherein The halogen is Cl.

3. The compound according to claim 1, wherein In R1 and R2, the alkyl group is substituted by OH.

4. The compound according to claim 1, wherein In R3, the alkyl group is replaced by OH, (C1-C6) alkoxy and / or -O-(C6-C 10 ) aryl substitution.

5. The compound according to claim 1, having the following formula (II):

6. The compound according to claim 1, having the following formula (IIIa1):

7. The compound according to claim 1, having the following formula (IVa): in: X1 and X2 are independently selected from C or N at each occurrence.

8. The compound according to claim 1, wherein the compound is selected from: in yes 9. A pharmaceutical composition comprising a compound of formula (I) as defined in any one of claims 1 to 8 in admixture with at least one pharmaceutically acceptable excipient or carrier.

10. A pharmaceutical composition comprising a compound of formula (I) as defined in any one of claims 1 to 8, further comprising an additional anticancer agent.

11. The pharmaceutical composition according to claim 10, wherein The additional anticancer agent is selected from anti-EGFR, anti-MAP kinase and anti-Akt compounds.

12. A compound of formula (I) according to any one of claims 1 to 8 for use in the treatment of cancer.

13. A compound of formula (I) according to any one of claims 1 to 8 for use in the treatment of hematological malignancies and / or solid tumors.

14. A compound of formula (I) according to any one of claims 1 to 8 for use in the treatment of lymphoma, leukemia, multiple myeloma.

15. A compound of formula (I) according to any one of claims 1 to 8 for use in the treatment of ovarian cancer, mesothelioma, melanoma, pancreatic cancer, lung cancer, breast cancer, kidney cancer and liver cancer.

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