Improved 2,4-dianilinopyrimidine-based aurora-a kinase degradation-inducing compound

Abstract

The present invention relates to an improved 2,4-dianilinopyrimidine-based Aurora-A kinase (AURKA) degradation-inducing compound, a preparation method therefor, and a use thereof. The AURKA degradation-inducing compound of the present invention has optimized AURKA protein degradation and related efficacy by linking an AURKA binding moiety and a CRBN E3 ubiquitin ligase moiety through a rigid ring-mediated linker, and thus can be effectively used for preventing or treating AURKA-associated diseases such as cancer.

Description

Improved 2,4-dianilinopyrimidine-based Aurora A kinase degradation-inducing compounds

[0001] The present invention relates to an improved 2,4-dianilinopyrimidine-based Aurora A kinase degradation-inducing compound, a method for preparing the same, and a use thereof.

[0002] Aurora A kinase (AURKA) is a protein belonging to the serine / threonine kinase family that is essential for cell proliferation. Selective inhibitors of AURKA are considered potential antitumor agents; therefore, compounds based on the 2,4-dianilinopyrimidine structure with selective AURKA inhibitory activity have been studied as potential anticancer candidates (Aliagas-Martin, Ignacio, et al. "A class of 2,4-dianilinopyrimidine Aurora A inhibitors with unusually high selectivity against Aurora B." Journal of medicinal chemistry 52.10 (2009): 3300-3307.).

[0003] However, small molecule compound inhibitors have limitations, such as the difficulty in sufficiently controlling the non-catalytic function of AURKA due to the nature of drugs that temporarily inhibit the catalytic function of the disease target protein. Therefore, AURKA protein degradation-inducing compounds applying the PROTEOlysis-Targeting Chimera (PROTAC) modality have been proposed as an alternative. For example, International Patent Publication WO2022 / 270987 discloses an early form of AURKA degradation-inducing PROTAC compound utilizing CRBN ligands of the IMiD series, such as thalidomide. Subsequently, International Patent Publication WO2024 / 155112 of the same applicant discloses improved PROTAC compounds through structural modifications of the PROTAC compound according to its modality.

[0004] Unlike small-molecule compounds, the structure and behavior of three-dimensional complexes of PROTACs, multifunctional compounds, are generally known to be very difficult to predict. Developing novel PROTAC compounds to improve their pharmacological efficacy, such as cell killing, while maintaining target resolution is a challenging task.

[0005] The purpose of the present invention is to provide an aurora A kinase degradation-inducing compound having improved aurora A kinase degradation and cell death effects, a method for producing the same, and a use thereof.

[0006] To solve the above technical problems, the present invention provides a 2,4-dianilinopyrimidine-based Aurora A kinase cleavage-inducing PROTAC compound having an improved ring-mediated rigid linker, a method for preparing the same, and a use thereof.

[0007] AURKA degradation-inducing compounds

[0008] In one aspect, the present invention provides a novel compound represented by the following chemical formula I:

[0009] [Chemical Formula I]

[0010]

[0011] In the above chemical formula I,

[0012] PTM is an Aurora kinase A (AURKA) protein binding moiety represented by the following chemical formula II,

[0013] [Chemical Formula II]

[0014]

[0015] {In the above chemical formula II,

[0016] R1 is hydrogen or halo;

[0017] R2 is hydrogen, halo or CF3;

[0018] R3 is CH, N or CF;

[0019] R4 is -CONH- or -NHCO-;

[0020] is phenyl, pyridyl or pyrimidinyl, which may be substituted with 1 to 2 halo or OCH3.

[0021] ULM is an E3 ubiquitin ligase binding moiety represented by the following chemical formula III,

[0022] [Chemical Formula III]

[0023]

[0024] {In the above chemical formula III,

[0025] U is a covalent bond or -NH-;

[0026] Silver has 1 to 3 halos, C 1-3 Alkyl or OC 1-3 phenyl, pyridyl or pyrimidinyl which may be substituted with alkyl}

[0027] Linker is a group that connects ULM and PTM, represented by the following chemical formula IV.

[0028] [Chemical Formula IV]

[0029]

[0030] {In the above chemical formula IV,

[0031] X1 to X4 are each independently CH or N;

[0032] X5 is hydrogen or CH3;

[0033] L M is -CH2CH2-, -CH2NH-, -CH2N(CH3)-, -CH2CHOH- or -CH2CH2CH2-;

[0034] L P is -NHCO-, -CO- or -NHCH2-;

[0035] X1 and L P are each {Covalently linked to ULM and PTM via

[0036]

[0037] The compound represented by the above chemical formula I is a bispecific compound implementing a PROTAC (Proteolysis targeting chimera) modality. The compound is characterized by having a structure in which a moiety (PTM) that binds to the target protein AURKA and an E3 ubiquitin ligase binding moiety (ULM) are connected by a linker molecule. The compound can effectively induce inhibition and degradation of the AURKA protein by the ubiquitin-proteasome system (UPS) in the cell by positioning the intracellular AURKA protein and E3 ubiquitin ligase in close proximity and inducing artificial ubiquitination of the AURKA protein.

[0038] In the present invention, Aurora A kinase (AURKA) is a serine / threonine protein kinase that plays a key role in the G2 / M phase transition and mitosis of the cell cycle, and can be encoded by the AURKA gene (e.g., NCBI Reference Sequence: NP_001310234.1). AURKA is involved in the replication and maturation of centrosomes, the formation and stabilization of spindles, and the alignment and segregation of chromosomes, and is essential for regulating the normal progression of cell division. In normal cells, its expression and activity are strictly regulated according to the cell cycle, but in various cancer cells, it is known to be overexpressed or its function abnormally activated, causing abnormal cell division, genomic instability, and continuous proliferation of cancer cells.

[0039] In the present invention, the AURKA protein binding moiety represented by the above chemical formula II may be a moiety represented by the following chemical formulas II-1 to II-4.

[0040] [Chemical Formula II-1]

[0041]

[0042] {In the above chemical formula II-1,

[0043] R2 and R5 are each independently hydrogen, F or Cl;

[0044] R3 is CH, N or CF}

[0045] In one embodiment, the moiety represented by formula II-1 is a moiety having the following structure:

[0046]

[0047]

[0048]

[0049] [Chemical Formula II-2]

[0050]

[0051] {In the above chemical formula II-2,

[0052] R1 is hydrogen or F;

[0053] R2 is hydrogen, F or CF3;

[0054] R5 is hydrogen, F or OCH3}

[0055] In one embodiment, the moiety represented by formula II-2 is a moiety having the following structure:

[0056]

[0057]

[0058]

[0059] [Chemical Formula II-3]

[0060]

[0061] {In the above chemical formula II-3,

[0062] R1 is hydrogen or F;

[0063] R5 is hydrogen or F;

[0064] R 6A and R 6B are each independently CH or N}

[0065] In one embodiment, the moiety represented by formula II-3 is a moiety having the following structure:

[0066]

[0067]

[0068]

[0069] [Chemical Formula II-4]

[0070]

[0071] {In the above chemical formula II-4,

[0072] R1 is hydrogen or F;

[0073] R 5A and R 5B are each independently hydrogen, F or OCH3}

[0074] In one embodiment, the moiety represented by formula II-4 is a moiety having the following structure:

[0075]

[0076]

[0077]

[0078] In the present invention, E3 ubiquitin ligase is a protein that promotes ubiquitin transfer to a target substrate protein, and the ULM moiety described above in the compound of the present invention can bind to CRBN, i.e., Cereblon E3 ubiquitin ligase. CRBN forms an E3 ubiquitin ligase complex together with DDB1, Cul4A, and ROC1, wherein CRBN is a substrate recognition subunit of the complex.

[0079] In the present invention, the E3 ubiquitin ligase binding moiety (ULM) represented by chemical formula II is a glutarimide series compound, which provides chemical and functional advantages over existing IMiD series (e.g., thalidomide, lenalidomide, and pomalidomide, etc.) in PROTAC design.

[0080] In the present invention, the E3 ubiquitin ligase binding moiety represented by the above chemical formula III may be a moiety represented by the following chemical formulas III-1 to III-4.

[0081] [Chemical Formula III-1]

[0082]

[0083] {In the above chemical formula III-1,

[0084] U2 and U3 are each independently hydrogen, F or Cl}

[0085] In one embodiment, the moiety represented by formula III-1 is a moiety having the following structure:

[0086]

[0087]

[0088] [Chemical Formula III-2]

[0089]

[0090] {In the above chemical formula III-2,

[0091] U2 is hydrogen, F or Cl respectively;

[0092] U 4A and U 4B {One of them is CH and the other is N}

[0093] In one embodiment, the moiety represented by formula III-2 is a moiety having the following structure:

[0094]

[0095]

[0096] [Chemical Formula III-3]

[0097]

[0098] {In the above chemical formula III-3,

[0099] U2 and U3 are each independently hydrogen, F or OCH3}

[0100] In one embodiment, the moiety represented by formula III-3 is a moiety having the following structure:

[0101]

[0102]

[0103] [Chemical Formula III-4]

[0104]

[0105] {In the above chemical formula III-4,

[0106] U2 is hydrogen, F or Cl;

[0107] U 4A and U 4B {One of them is CH and the other is N}

[0108] In one embodiment, the moiety represented by formula III-1 is a moiety having the following structure:

[0109]

[0110] In the compound of the present invention, the linker is a group that connects an AURKA binding moiety (PTM) and an E3 ubiquitin ligase binding moiety (ULM), thereby allowing the E3 ubiquitin ligase protein targeted by the ULM moiety and the AURKA protein targeted by the PTM moiety to interact with each other within an appropriate physical distance, thereby inducing ubiquitination of the target AURKA protein. The linker may be, for example, the following moiety, and the definition of the substituent is as described above.

[0111] In the present invention, the linker represented by Chemical Formula IV has a unique structure in which two six-membered rings in a para-linked form are mediated, imparting rigidity to the entire linker molecule and fixing the relative directionality and distance between the PTM and ULM moieties. This can improve the efficacy and physicochemical profile of PROTAC drugs by forming a ternary complex mediated by the PROTAC compound, contributing to increased proteolytic efficiency, improved resistance to metabolic enzymes, and increased intracellular half-life, thereby contributing to increased drug efficacy.

[0112] In the present invention, the linker represented by the chemical formula IV may be a group represented by the following chemical formulas IV-1 to IV-4 {wherein, the group and {Covalently linked to ULM and PTM, respectively}.

[0113] [Chemical Formula IV-1]

[0114]

[0115] {In the above chemical formula IV-1,

[0116] X5 is hydrogen or CH3;

[0117] X6 is hydrogen or OH}

[0118] In one embodiment, the group represented by Chemical Formula IV-1 is a group having the following structure.

[0119]

[0120]

[0121] [Chemical Formula IV-2]

[0122]

[0123] {In the above chemical formula IV-2,

[0124] X7 is -CH2- or -CO-;

[0125] In one embodiment, the group represented by Chemical Formula IV-2 is a group having the following structure.

[0126]

[0127] [Chemical Formula IV-3]

[0128]

[0129] {In the above chemical formula IV-3,

[0130] X4 is CH or N,

[0131] X6 is hydrogen or CH3}

[0132] In one embodiment, the group represented by Chemical Formula IV-3 is a group having the following structure.

[0133]

[0134] [Chemical Formula IV-4]

[0135]

[0136] {In the above chemical formula IV-4,

[0137] X5 is hydrogen or CH3}

[0138] In one embodiment, the group represented by Chemical Formula IV-4 is a group having the following structure.

[0139]

[0140] In one embodiment of the present invention, the compound represented by formula I is selected from the group consisting of the following compounds.

[0141] [Table 1]

[0142]

[0143]

[0144]

[0145]

[0146]

[0147]

[0148]

[0149]

[0150]

[0151]

[0152]

[0153]

[0154]

[0155] Compounds 1 to 61 described above are characterized by an overall improved killing effect on blood cancer cells and lung cancer cells compared to the comparative compounds described in Table 2 below among the compounds represented by the chemical formula I (Experimental Examples 1 and 2).

[0156] In the present invention, some or all of the comparative compounds described in Table 2 below (see International Application No. PCT / KR2024 / 000878; International Patent Publication No. WO2024 / 155112) may be excluded from the scope of the patent claims of the present invention.

[0157] [Table 2]

[0158]

[0159]

[0160]

[0161]

[0162]

[0163]

[0164]

[0165]

[0166]

[0167]

[0168] Additionally, the following compounds in the present invention may be excluded from the scope of the claims of the present invention.

[0169]

[0170]

[0171] The following compounds (Examples 1, 2, 6) are included in the present invention, but may be excluded from the scope of the present invention as needed.

[0172]

[0173]

[0174]

[0175] The compounds of the present invention may include, in addition to compounds explicitly represented by formula I, tautomers, optical isomers (including racemic mixtures), specific enantiomers or enantiomerically enriched mixtures thereof.

[0176] The compound of the present invention may be in the form of a salt thereof, preferably a pharmaceutically acceptable salt, in addition to the compound explicitly represented by Chemical Formula I. The pharmaceutically acceptable salt refers to any organic or inorganic acid addition salt having a concentration that is relatively non-toxic and harmless to the patient and has an effective effect, and the side effects caused by the salt do not diminish the beneficial effects of the compound represented by Chemical Formula I.

[0177] The compounds of the present invention may exist in the form of chimeric molecules conjugated to a functional macromolecule via an additional chemical linker to the compound explicitly represented by Formula I. For example, the macromolecule may be a biomolecule comprising a nucleic acid, an aptamer, a carbohydrate, a peptide or a fragment thereof, or an antibody or a fragment thereof.

[0178]

[0179] Method for producing AURKA decomposition-inducing compounds

[0180] The compounds of the present invention can be prepared by synthetic methods known in the field of organic chemistry or by modification techniques apparent to those skilled in the art, for example, by the following reaction schemes 1 to 3.

[0181] [Reaction Formula 1]

[0182]

[0183] [Reaction Formula 2]

[0184]

[0185] [Reaction Formula 3]

[0186]

[0187] In the above reaction schemes 1 to 3, PTM, Linker and ULM are the groups defined above or their reactive derivatives, and RG 1 , RG 2 , RG 2a , RG 2b , RG 3 , RG 3a , RG 3b and RG 4 is a moiety comprising a suitable reactive group capable of linking together PROTAC compound intermediates represented by formula I through covalent bond formation in the field of organic synthesis. The covalent bond formation may be formed through synthetic reactions such as, but not limited to, amide formation, ester formation, carbamate formation, urea formation, ether formation, amine formation, and various carbon-carbon single bond, double bond formation, click chemistry, etc., depending on the specific reactive group.

[0188] In the above reaction scheme, each step may comprise one or multiple synthetic steps. Isolation and purification of the product can be accomplished by standard procedures known to those skilled in the art of organic chemistry.

[0189]

[0190] Uses of AURKA degradation-inducing compounds

[0191] The compound represented by Chemical Formula I of the present invention can induce AURKA protein degradation. After treating a cell line with an example compound of the present invention, a luciferase assay experiment comparing the AURKA protein level with that of a control cell line that was not treated with the compound confirmed the effect of inducing degradation of the target protein (Experimental Example 1; Tables 3 and 4).

[0192] The compound represented by Chemical Formula I of the present invention exhibits a potent cancer cell killing effect through AURKA protein degradation. As a result of an experiment measuring the apoptotic effect after treating AURKA-expressing cancer cells with the example compound of the present invention, it was confirmed that the compound effectively exhibits a cell killing effect in various cancer cells (Experimental Example 2; Tables 5 and 6).

[0193] First, the compounds of the present invention were confirmed to have excellent cancer cell killing effects in a cell killing effect experiment conducted on human bone marrow-derived cancer cell lines HEL92.1.7, SET-2, and K-562 (Table 5).

[0194] Furthermore, it was confirmed that the exemplary compounds of the present invention exhibited an improved and excellent cancer cell killing effect in a cell killing effect experiment measured against a small cell lung cancer cell line (NCI-H446) (Table 6).

[0195] Because the compound of the present invention exhibits potent cancer cell killing effects, it can be utilized as a payload for antibody-drug conjugates (ADCs). ADCs enable the delivery of cancer-specific cytotoxic payloads, thereby achieving maximum efficacy in cancer cells while minimizing undesirable effects in non-cancerous cells. Therefore, a degrader-antibody conjugate (DAC) utilizing the PROTAC of the present invention can be a useful strategy for significantly improving the cell permeability and bioavailability shortcomings of PROTAC compounds while also enhancing tissue and cell-type selectivity.

[0196] In one embodiment, the invention provides a pharmaceutical composition comprising a PROTAC compound of the invention, wherein the PROTAC compound is conjugated to an antibody or an antigen-binding fragment thereof via an additional linker. The additional linker may be covalently linked to a functional group, such as an amine group, of an ULM of the PROTAC compound or an internal linker of the PROTAC compound. In certain embodiments, the additional linker is a cleavable or non-cleavable linker.

[0197] In one embodiment, the present disclosure provides an antibody-drug conjugate comprising an antibody and an antigen-binding fragment thereof and a PROTAC of the present invention, wherein the PROTAC compound is conjugated to the antibody or antigen-binding fragment thereof via a linker.

[0198] Because the PROTAC of the present invention eliminates rather than inhibits the target protein, it exhibits superior therapeutic efficacy compared to the small molecule AURKA inhibitor from which it is derived. Accordingly, the PROTAC of the present invention, particularly when conjugated to a DAC payload, can be utilized for the treatment of AURKA-associated disorders or conditions in which abnormal expression of the AURKA protein is involved in the onset and / or progression of the disease.

[0199] In the present invention, the term "AURKA-related protein-related disease" refers to any disease or condition that can be treated, alleviated, delayed, inhibited, or prevented by inducing degradation or inhibiting the activity of the AURKA protein, such as cancer. The term "cancer" includes all types of cancer that can exhibit preventive or therapeutic efficacy due to inhibition of the activity of the AURKA protein. For example, the cancer may be a solid cancer or a blood cancer, and may be at least one selected from the group consisting of squamous cell carcinoma, small cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, squamous cell carcinoma of the lung, peritoneal cancer, skin cancer, cutaneous or intraocular melanoma, rectal cancer, anal cancer, esophageal cancer, small intestine cancer, endocrine cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, chronic or acute leukemia, lymphocytic lymphoma, hepatocellular carcinoma, gastric cancer, stomach cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, large intestine cancer, endometrial or uterine cancer, salivary gland cancer, kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, head and neck cancer, brain cancer, and osteosarcoma, but is not limited thereto. The cancer includes not only primary cancer but also metastatic cancer.

[0200] The pharmaceutical composition of the present invention can be formulated through a conventional method in the field of pharmaceutical science, and can be administered orally or parenterally depending on the intended method, and the dosage can be determined by taking into consideration the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of the disease.

[0201] The compounds of the present invention can be used to degrade AURKA proteins in a sample by treating the sample in vivo or in vitro. The sample may be a cell, a cell culture, a bodily fluid, or tissue of a mammal, including a human. The method can be used for diagnostic or therapeutic purposes.

[0202] The present invention provides a method for preventing or treating an AURKA protein-related disease, comprising administering to a patient a therapeutically effective amount of a compound represented by Chemical Formula I, preferably conjugated to a DAC payload. The AURKA protein-related disease includes cancer.

[0203] The compound of the present invention effectively degrades the degradation of Aurora A kinase (AURKA) and has a high cell killing effect on various cancers, and thus can be usefully utilized for AURKA-related diseases.

[0204] Hereinafter, the composition and effects of the present invention will be described in more detail through examples and experimental examples. These examples and experimental examples are intended solely to illustrate the present invention and are not intended to limit the scope of the present invention. All references cited throughout this application are expressly incorporated herein by reference in their entirety.

[0205] Specific example compounds of the present invention were synthesized according to the following manufacturing method and their structures were analyzed.

[0206] <Example>

[0207] The compound of the present invention was purified and its structure was analyzed according to the following method.

[0208] machinery and tools

[0209] LCMS: Shimadzu LCMS-2020

[0210] HPLC: Agilent 1260 II LC, Agilent 1200 / G6410B

[0211] NMR: BRUKER AVANCE / 400 MHZ

[0212] LCMS analysis

[0213] LCMS data were recorded on a Shimadzu LCMS-2020 equipped with an ESI (electron spray ionization) device. 0.0375% TFA in water (solvent A) and 0.01875% TFA in ACN (solvent B) were used as mobile phases. The columns used were Kinetex EVO C18 (2.1 × 30 mm, 5 μm) or HALO C18 (3.0 × 30 mm, 2.7 μm).

[0214] HPLC analysis

[0215] For HPLC analysis, an Agilent 1260 II LC or Agilent 1200 / G6410B was used. 0.0375% TFA in water (solvent A) and 0.01875% TFA in ACN (solvent B) were used as mobile phases. The columns used were Zobrax Eclipse Plus C18 (4.6 × 150 mm, 3.5 μm) or YMC ODS A (4.6 × 150 mm, 3 μm).

[0216] NMR Analysis

[0217] 1 H NMR spectra were recorded with a Bruker AVANCE III 400 MHz / 5 mm Probe (BBO).

[0218] Example 1. Synthesis of N-(2-chlorophenyl)-4-((2-((4-((4-(2-((2R)-4-(4-(2,6-dioxopiperidin-3-yl)phenyl)-2-methylpiperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)benzamide (Compound 1)

[0219]

[0220] Step 1. Synthesis of tert-butyl (R)-4-(4-(2,6-bis(benzyloxy)pyridin-3-yl)phenyl)-2-methylpiperazine-1-carboxylate (2)

[0221] To a solution of 2,6-bis(benzyloxy)-3-(4-bromophenyl)pyridine (1 g, 2.17 mmol) and tert-butyl (R)-2-methylpiperazine-1-carboxylate (435.25 mg, 2.17 mmol) in toluene (10 mL) were added Cs2CO3 (2.12 g, 6.52 mmol) and RuPhos Pd G3 (181.76 mg, 217.33 μmol) under N2. The mixture was degassed and purged with N2 three times. The mixture was stirred at 100 °C under N2 for 16 h. The peak of the target mass was identified by LCMS. Water (50 mL) was added to the reaction mixture and extracted with EtOAc (50 mL x 2). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (Biotage; 12 g SepaFlash® silica flash column, eluent 8% EtOAc / petroleum ether, gradient @ 30 mL / min) to give the title compound (1.01 g, 1.77 mmol, 81.33% yield, 99% purity) as a white oil. MS (M+H) + =566.4

[0222] Step 2. Synthesis of tert-butyl (2R)-4-(4-(2,6-dioxopiperidin-3-yl)phenyl)-2-methylpiperazine-1-carboxylate (3)

[0223] To a solution of Pd / C (188.10 mg, 176.75 μmol, 10% purity, wet) and Pd(OH)2 / C (100 mg, 20% purity, wet) in THF (20 mL) were added AcOH (106.14 mg, 1.77 mmol) and tert-butyl (R)-4-(4-(2,6-bis(benzyloxy)pyridin-3-yl)phenyl)-2-methylpiperazine-1-carboxylate (1.01 g, 1.77 mmol) under Ar. The mixture was degassed and purged three times with H2. The mixture was stirred at 26°C under H2 (50 Psi) for 48 h. The target mass peak (97%) was identified by LCMS. The reaction mixture was filtered. The filter cake was washed with THF (3 × 30 mL). The mixed organic layer was concentrated under reduced pressure to obtain the title compound as a white solid (680 mg, 1.58 mmol, 89.36% yield, 90% purity). MS (M-56+H) + =332.1.

[0224] Step 3. Synthesis of 3-(4-((R)-3-methylpiperazin-1-yl)phenyl)piperidine-2,6-dione (4)

[0225] A solution of tert-butyl (2R)-4-(4-(2,6-dioxo-3-piperidyl)phenyl)-2-methyl-piperazine-1-carboxylate (680 mg, 1.58 mmol) in HCl / dioxane (10 mL, 2 M) was stirred at 20 °C for 3 h. The peak of the desired mass was identified by LCMS. The reaction mixture was concentrated under reduced pressure to obtain the title compound (790 mg, crude, HCl) as a yellow solid. MS (M+H) + =288.1.

[0226] Step 4. Synthesis of tert-butyl (4-(2-((2R)-4-(4-(2,6-dioxopiperidin-3-yl)phenyl)-2-methylpiperazin-1-yl)ethyl)piperidin-1-yl)carbamate (5)

[0227] To a solution of 3-(4-((R)-3-methylpiperazin-1-yl) phenyl) piperidine-2,6-dione (370 mg, 1.14 mmol, HCl) in DCM (10 mL) was added TEA (563.75 mg, 5.57 mmol), followed by a solution of tert-butyl (4-(2-oxoethyl)piperidin-1-yl)carbamate (300 mg, 1.11 mmol) in DCM (2 mL) and stirred at 20 °C for 30 min. At 20 °C, NaBH(OAc)3 (708.47 mg, 3.34 mmol) was added and the mixture was stirred at 20 °C for 16 h. The peak of the target mass (60%) was confirmed by LCMS. The reaction mixture was added saturated aqueous NaHCO3 (30 mL) and extracted with DCM (30 mL x 2). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (Biotage; 12 g SepaFlash® silica flash column, eluent 50% EtOAc / MeOH, gradient @ 60 mL / min) to give the title compound (196 mg, 366.30 μmol, 32.87% yield, 96% purity) as a yellow oil. MS (M+H) + =514.4.

[0228] Step 5. Synthesis of 3-(4-((R)-4-(2-(1-aminopiperidin-4-yl)ethyl)-3-methylpiperazin-1-yl)phenyl)piperidine-2,6-dione (6)

[0229] To a solution of tert-butyl (4-(2-((2R)-4-(4-(2,6-dioxopiperidin-3-yl)phenyl)-2-methylpiperazin-1-yl)ethyl)piperidin-1-yl)carbamate (196 mg, 366.30 μmol) in DCM (6 mL) was added TFA (2 mL) at 20 °C. The mixture was stirred at 20 °C for 1 h. The peak of the target mass (83%) was confirmed by LCMS. The reaction mixture was concentrated under reduced pressure to obtain the title compound (320 mg, crude, TFA) as a yellow oil. MS (M+H)+ =414.4.

[0230] Step 6. Synthesis of N-(2-chlorophenyl)-4-((2-((4-((4-(2-((2R)-4-(4-(2,6-dioxopiperidin-3-yl)phenyl)-2-methylpiperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)benzamide (Compound 1)

[0231] To a solution of 3-(4-((R)-4-(2-(1-aminopiperidin-4-yl)ethyl)-3-methylpiperazin-1-yl)phenyl)piperidine-2,6-dione (320 mg, 606.55 μmol, TFA) in DMF (10 mL) were added DIPEA (175.79 mg, 1.36 mmol), HOBt (73.52 mg, 544.08 μmol), EDCI (104.30 mg, 544.08 μmol), and 4-((4-((4-((2-chlorophenyl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-2-yl)amino)benzoic acid (130 mg, 272.04 μmol) at 20 °C. The mixture was stirred at 20 °C for 16 h. The reaction was confirmed to be complete by LCMS. The reaction mixture was diluted with EtOAc (80 mL), then added with water (30 mL x 3) and saturated NaHCO3 aqueous solution. (50 mL). The mixed organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The mixture was purified by flash silica gel chromatography (Biotage; 12 g SepaFlash® silica flash column, eluent 50% EtOAc / MeOH, gradient @ 30 mL / min). The residue was repurified by prep-HPLC (column: Phenomenex Luna C18 150*25mm*10um; mobile phase: [water(TFA)-ACN]; gradient: 13%-43% B over 12 min) and lyophilized to obtain the title compound (47.0 mg, 38.40 μmol, 14.12% yield, 90% purity, 2TFA) as a white solid. MS (M+H) + =873.5.

[0232] 1 H NMR (400 MHz, DMSO-d6) δ = 10.79 (s, 1H), 9.94 (s, 1H), 9.78 (s, 1H), 9.68 - 9.60 (m, 2H), 9.51 (br s, 1H), 8.26 - 8.23 ​​(m, 1H), 8.04 - 8.00 (m, 4H), 7.81 - 7.65 (m, 4H), 7.57 (d,J= 6.8 Hz, 1H), 7.40 (t,J= 7.6 Hz, 1H), 7.32 - 7.28 (m, 1H), 7.13 - 7.11 (m, 2H), 7.00 - 6.95 (m, 2H), 3.90 - 3.57 (m, 4H), 3.42 - 3.30 (m, 2H), 3.21 - 2.96 (m, 4H), 2.90 - 2.61 (m, 4H), 2.50 - 2.41 (m, 2H), 2.21 - 2.12 (m, 1H), 2.02 - 1.98 (m, 1H), 1.77 - 1.55 (m, 4H), 1.40 - 1.33 (m, 6H).

[0233]

[0234] Example 2. Synthesis of N-(4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)phenyl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 2)

[0235]

[0236] To a solution of 3-(4-(piperazin-1-yl)phenyl)piperidine-2,6-dione (87.76 mg, 283.30 μmol, HCl) in DMSO (3 mL) was added NaOAc (38.73 mg, 472.17 μmol), followed by 2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)-N-(4-(2-oxoethyl)piperidin-1-yl)benzamide (100 mg, 157.39 μmol), and the mixture was stirred at 20 °C for 30 min. At 20 °C, NaBH(OAc)3 (66.72 mg, 314.78 μmol) was added, and the mixture was stirred at 20 °C for 16 h. The peak of the target mass was confirmed by LCMS. Water (20 mL) was added to the reaction mixture, and the formation of a brown solid was confirmed. The mixture was filtered and washed with water (10 mL x 3). The filter cake was collected and dried to obtain the crude product. The crude product was recrystallized with NMP / ACN (1 mL / 7 mL) at 60 °C to obtain the product (80 mg, 83% purity). The product was further purified by prep-HPLC (column: Phenomenex luna C18 150*25mm* 10um; mobile phase: [water(TFA)-ACN]; gradient: 8%-38% B over 10 min) and lyophilized to obtain the title compound (29.67 mg, 27.25 μmol, 35% yield, 2TFA) as a white solid. MS (M+H) + = 861.4.

[0237] 1H NMR (400 MHz, DMSO-d6) δ = 10.79 (s, 1H), 10.42 (s, 1H), 9.70 - 9.65 (m, 1H), 9.59 - 9.53 (m, 2H), 9.16 (br s, 1H), 8.18 (d,J= 3.6 Hz, 1H), 7.81 (br d,J= 14.0 Hz, 1H), 7.76 - 7.66 (m, 4H), 7.62 - 7.56 (m, 1H), 7.44 (br d,J= 4.0 Hz, 2H), 7.38 - 7.32 (m, 2H), 7.12 (br d,J=8.8 Hz, 2H), 6.98 (br d,J= 8.8 Hz, 2H), 3.83 (br d,J= 12.4 Hz, 2H), 3.76 (dd,J= 11.2, 4.8 Hz, 1H), 3.60 (br d,J= 11.6 Hz, 2H), 3.20 - 2.89 (m, 9H), 2.68 - 2.61 (m, 3H), 2.47 - 2.45 (m, 1H), 2.21 - 2.10 (m, 1H), 2.01 - 1.98 (m, 1H), 1.71 - 1.64 (m, 4H), 1.35 - 1.30 (m, 2H).

[0238]

[0239] Example 3. Synthesis of N-(4-((2-((4-(4-(3-(4-(4-(2,6-dioxopiperidin-3-yl)phenyl)piperazin-1-yl)propyl)piperidine-1-carbonyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 3)

[0240]

[0241] Step 1. Synthesis of tert-butyl 4-(3-oxopropyl)piperidine-1-carboxylate (2)

[0242] To a solution of DMSO (3.21 mL, 41.09 mmol) in DCM (30 mL) was added (COCl)2 (1.44 mL, 16.44 mmol) at -65 °C under N2, and the mixture was stirred at -65 °C for 1 h. A solution of tert-butyl 4-(3-hydroxypropyl)piperidine-1-carboxylate (2 g, 8.22 mmol) in DCM (5 mL) was slowly added and stirred at -65 °C for 1 h. TEA (5.72 mL, 41.09 mmol) was added to the mixture and stirred at -65 °C for 1 h. TLC (PE:EtOAc=0:1) confirmed that the starting material was consumed and a new main spot appeared. Water (100 mL) was added to the reaction mixture, extracted with DCM (50 mL x 3), and the combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the title compound (3.2 g, crude) as a yellow oil. The product was used directly in the next reaction.

[0243] Step 2. Synthesis of tert-butyl 4-(3-(4-(4-(2,6-dioxopiperidin-3-yl)phenyl)piperazin-1-yl)propyl)piperidine-1-carboxylate (4)

[0244] The title compound (762 mg, 1.44 mmol, 55% yield, 94% purity) as a brown oil was obtained by a similar method to step 4 of Example 1. MS (M+H) + = 499.4.

[0245] Step 3. Synthesis of 3-(4-(4-(3-(piperidin-4-yl)propyl)piperazin-1-yl)phenyl)piperidine-2,6-dione (5)

[0246] The title compound (764 mg, crude, 2TFA) as a yellow oil was obtained by synthesis in a similar manner to step 5 of Example 1. MS (M+H) + = 399.3

[0247] Step 4. Synthesis of N-(4-((2-((4-(4-(3-(4-(4-(2,6-dioxopiperidin-3-yl)phenyl)piperazin-1-yl)propyl)piperidine-1-carbonyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 3)

[0248] The title compound (50.7 mg, 56.00 μmol, 16% yield, 93% purity) was obtained as a yellow solid by a similar method to step 6 of Example 1. MS (M+H) + = 842.3.

[0249] 1 H NMR (400 MHz, DMSO-d6) δ = 10.77 (s, 1H), 10.39 (s, 1H), 9.43 (s, 2H), 8.12 (d,J= 3.6 Hz, 1H), 7.78 - 7.66 (m, 7H), 7.61 - 7.56 (m, 1H), 7.38 - 7.32 (m, 2H), 7.25 (d,J= 8.4 Hz, 2H), 7.04 (d,J= 8.8 Hz, 2H), 6.87 (d,J= 8.8 Hz, 2H), 3.72 (dd,J= 10.8, 4.8 Hz, 1H), 3.14 - 3.02 (m, 4H), 2.95 - 2.75 (m, 2H), 2.67 - 2.58 (m, 1H), 2.46 - 2.43 (m, 7H), 2.25 (t,J= 7.2 Hz, 2H), 2.17 - 2.07 (m, 1H), 2.03 - 1.96 (m, 1H), 1.75 - 1.65 (m, 2H), 1.52 - 1.36 (m, 3H), 1.23 - 1.19 (m, 2H), 1.10 - 1.02 (m, 2H).

[0250]

[0251] Example 4. Synthesis of N-(4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)-4-((5-fluoro-4-((4-((2-fluorophenyl)carbamoyl)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 4)

[0252]

[0253] Step 1. Synthesis of 4-((5-fluoro-2-((4-((4-(2-hydroxyethyl)piperidin-1-yl)carbamoyl)phenyl)amino)pyrimidin-4-yl)amino)-N-(2-fluorophenyl)benzamide (2)

[0254] To a solution of 4-((5-fluoro-4-((4-((2-fluorophenyl)carbamoyl)phenyl)amino)pyrimidin-2-yl)amino)benzoic acid (1 g, 1.95 mmol), 2-(1-amino-4-piperidyl)ethanol (550.58 mg, 2.54 mmol, 2HCl) and DIPEA (1.26 g, 9.75 mmol, 1.70 mL) in DMF (10 mL) were added EDCI (486.09 mg, 2.54 mmol) and HOBt (342.63 mg, 2.54 mmol) at 20 °C, and the reaction mixture was stirred at 20 °C for 12 h. LCMS confirmed the presence of a small amount of acid material and the peak of the target mass (40%). Water (40 mL) was added to the reaction mixture, filtered, and washed with water (20 mL). The filter cake was collected and dried to obtain the crude product. The crude product was dissolved in a DMF / ACN (1 mL / 8 mL) mixture and stirred at 50 °C for 14 h. The product was filtered and washed with ACN (3 mL). The filter cake was collected and dried to obtain the title compound (500 mg, 765.80 μmol, 39.26% yield, 90% purity) as a white solid.

[0255] Step 2. Synthesis of 4-((5-fluoro-2-((4-((4-(2-oxoethyl)piperidin-1-yl)carbamoyl)phenyl)amino)pyrimidin-4-yl)amino)-N-(2-fluorophenyl)benzamide (3)

[0256] To a solution of 4-((5-fluoro-2-((4-((4-(2-hydroxyethyl)piperidin-1-yl)carbamoyl)phenyl)amino)pyridin-4-yl)amino)-N-(2-fluorophenyl)benzamide (500 mg, 765.80 μmol) in DCM (5 mL) and DMSO (5 mL) was added TEA (309.96 mg, 3.06 mmol). After adding Py·SO3 (243.77 mg, 1.53 mmol) at 20 °C, the reaction mixture was stirred at 20 °C for 14 h. Complete consumption of the starting material and a peak of the target mass were confirmed by LCMS. Water (40 mL) was slowly added to the reaction mixture, which was filtered and washed with ACN (5 mL). The filter cake was collected and dried to obtain the title compound (500 mg, crude) as a white solid, which was used directly in the next reaction. MS(M+H) + = 586.3.

[0257] Step 3. Synthesis of N-(4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)-4-((5-fluoro-4-((4-((2-fluorophenyl)carbamoyl)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 4)

[0258] The title compound (85.56 mg, 71.88 μmol, 14.03% yield, 93% purity, 2TFA) was obtained as a white solid by a similar method as in Example 2. MS (M+H) + = 879.5.

[0259] 1H NMR (400 MHz, DMSO-d6) δ = 10.89 (s, 1H), 10.01 (s, 1H), 9.76 (s, 1H), 9.67 (s, 1H), 9.65 - 9.45 (m, 2H), 8.25 (d,J= 3.6 Hz, 1H), 8.05 - 7.97 (m, 3H), 7.82 - 7.75 (m, 2H), 7.75 - 7.69 (m, 2H), 7.67 - 7.60 (m, 1H), 7.33 - 7.18 (m, 4H), 7.02 (br dd,J= 7.8, 11.8 Hz, 1H), 4.00 (br dd,J= 4.6, 12.7 Hz, 1H), 3.65 - 3.52 (m, 4H), 3.28 - 3.00 (m, 8H), 2.91 - 2.64 (m, 3H), 2.57 - 2.54 (m, 1H), 2.25 - 2.15 (m, 1H), 2.01 - 1.93 (m, 1H), 1.79 - 1.70 (m, 2H), 1.69 - 1.58 (m, 2H), 1.47 - 1.30 (m, 3H).

[0260]

[0261] Example 5. Synthesis of N-(2-chlorophenyl)-4-((2-((4-((4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)benzamide (Compound 5)

[0262]

[0263] Step 1. Synthesis of 2-(1-aminopiperidin-4-yl)ethan-1-ol (2)

[0264] To a solution of tert-butyl (4-(2-hydroxyethyl)piperidin-1-yl)carbamate (2 g, 8.19 mmol) in dioxane (5 mL) was added HCl / dioxane (2 M, 15 mL) at 20 °C, and the mixture was stirred at 20 °C for 2 h. LCMS confirmed complete consumption of the starting material and the target mass. The reaction mixture was concentrated under reduced pressure to obtain the title compound (2 g, 7.83 mmol, 95.65% yield, 85% purity, 2HCl) as a white solid, which was used directly in the next reaction. MS (M+H) + = 145.2.

[0265] Step 2. Synthesis of N-(2-chlorophenyl)-4-((5-fluoro-2-((4-((4-(2-hydroxyethyl)piperidin-1-yl)carbamoyl)phenyl)amino)pyrimidin-4-yl)amino)benzamide (4)

[0266] The title compound (400 mg, 595.95 μmol, 43.13% yield, 90% purity) was obtained as a yellow solid by a similar method to step 1 of Example 4. MS (M+H) + = 604.2.

[0267] Step 3. Synthesis of N-(2-chlorophenyl)-4-((5-fluoro-2-((4-((4-(2-oxoethyl)piperidin-1-yl)carbamoyl)phenyl)amino)pyrimidin-4-yl)amino)benzamide (5)

[0268] The title compound (300 mg, 458.43 μmol, 76.92% yield, 92% purity) was obtained as a white solid by a similar method to step 2 of Example 4. MS (M+H) + = 602.3.

[0269] Step 4. Synthesis of N-(2-chlorophenyl)-4-((2-((4-((4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)benzamide (Compound 5)

[0270] The title compound (38.61 mg, 34.37 μmol, 29.98% yield, 100% purity, 2TFA) was obtained as a white solid by a similar method to Example 2. MS (M+H) + = 895.3.

[0271] 1 H NMR (400 MHz, DMSO-d6) δ = 10.88 (s, 1H), 9.93 (s, 1H), 9.76 (s, 1H), 9.65 (s, 1H), 9.55 - 9.44 (m, 2H), 8.25 (d,J= 3.6 Hz, 1H), 8.05 - 7.99 (m, 3H), 7.81 - 7.76 (m, 2H), 7.73 - 7.69 (m, 2H), 7.65 (dd,J= 1.4, 8.0 Hz, 1H), 7.57 (dd,J= 1.4, 8.1 Hz, 1H), 7.40 (dt,J= 1.4, 7.7 Hz, 1H), 7.34 - 7.27 (m, 1H), 7.22 (dd,J= 6.8, 13.0 Hz, 1H), 7.02 (dd,J= 7.4, 11.7 Hz, 1H), 4.00 (br dd,J= 4.8, 12.7 Hz, 1H), 3.63 - 3.55 (m, 4H), 3.26 - 3.16 (m, 4H), 3.15 - 3.02 (m, 4H), 2.85 - 2.68 (m, 3H), 2.56 - 2.54 (m, 1H), 2.24 - 2.15 (m, 1H), 2.00 - 1.93 (m, 1H), 1.79 - 1.69 (m, 2H), 1.68 - 1.59 (m, 2H), 1.42 - 1.30 (m, 3H).

[0272]

[0273] Example 6. Synthesis of N-(4-((2-((4-((4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 6)

[0274]

[0275] Step 1. Synthesis of 2-(1-aminopiperidin-4-yl)ethan-1-ol (2)

[0276] A solution of tert-butyl (4-(2-hydroxyethyl)piperidin-1-yl)carbamate (8 g, 32.74 mmol) in HCl / dioxane (2 M, 50 mL) was stirred at 20 °C for 2 h. The reaction mixture was concentrated under reduced pressure to obtain the title compound (7.6 g, crude, 2HCl) as a yellow oil. The product was used directly in the next reaction.

[0277] Step 2. Synthesis of 2-fluoro-N-(4-((5-fluoro-2-((4-((4-(2-hydroxyethyl)piperidin-1-yl)carbamoyl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)benzamide (4)

[0278] To a solution of 4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzoic acid (15 g, 31.53 mmol) in DMF (200 mL) were added DIPEA (20.56 g, 159.10 mmol), HOBt (8.60 g, 63.64 mmol), EDCI (12.20 g, 63.64 mmol), and 2-(1-aminopiperidin-4-yl)ethan-1-ol (7.6 g, 35.00 mmol, 2HCl) at 20 °C. The mixture was stirred at 20 °C for 16 h. The major peak of the target mass was identified by LCMS. Water (300 mL) was added to the reaction mixture, and it was confirmed that a large amount of pink solid was formed. The product was filtered and washed with water (50 mL). The filter cake was collected and dried to obtain the title compound as a white solid (17 g, 24.59 mmol, 77% yield, 85% purity). MS(M+H) + = 588.3.

[0279] Step 3. Synthesis of 2-fluoro-N-(4-((5-fluoro-2-((4-((4-(2-oxoethyl)piperidin-1-yl)carbamoyl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)benzamide (5)

[0280] To a solution of 2-fluoro-N-(4-((5-fluoro-2-((4-((4-(2-hydroxyethyl)piperidin-1-yl)carbamoyl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)benzamide (7 g, 10.13 mmol) in DCM (70 mL) and DMSO (70 mL) were added TEA (4.10 g, 40.50 mmol) and SO3·Py (3.22 g, 20.25 mmol) at 0 °C. The mixture was stirred at 20 °C for 2 h. SO3·Py (1.61 g, 10.13 mmol) was further added and stirred at 20 °C for 16 h. The major peak of the target mass (83%) was confirmed by LCMS. Water (300 mL) was poured into the reaction mixture and filtered. The filter cake was washed with water (30 mL x 3), dried and filtered to obtain the title compound (6.6 g, 9.58 mmol, 94% yield, 85% purity) as a light yellow solid. MS(M+H) + =586.2.

[0281] Step 4. Synthesis of N-(4-((2-((4-((4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 6)

[0282] To a solution of 3-(2,5-difluoro-4-(piperazin-1-yl)phenyl)piperidine-2,6-dione (2.61 g, 6.83 mmol, 2HCl) in DMSO (40 mL) was added NaOAc (986.19 mg, 12.02 mmol), followed by 2-fluoro-N-(4-((5-fluoro-2-((4-((4-(2-oxoethyl) piperidin-1-yl)carbamoyl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)benzamide (4 g, 5.46 mmol), and the mixture was stirred at 25 °C for 30 min. At 25 °C, NaBH(OAc)3 (1.74 g, 8.20 mmol) was added, and the mixture was stirred at 25 °C for 16 h. The main peak of the target mass (61%) was confirmed by LCMS. Water (500 mL) was slowly added to the reaction mixture, and it was confirmed that a large amount of brown solid was generated. The reaction mixture was adjusted to pH = 2-3 with 1 M HCl aqueous solution and filtered. The filter cake was washed with water (30 mL x 3). The filtrate was extracted with EtOAc (150 mL x 2) to remove impurities. The aqueous phase was adjusted to pH = 8 with solid NaHCO3, filtered, and washed with water (30 mL x 3). The filter cake was collected and dried to obtain the crude product (~5.5 g, 72% purity). NMP (15 mL) was added to the crude product, and the temperature was increased to 60 °C and stirred for 1 h. The mixture was filtered, ACN (120 mL) was slowly added at 60 °C, and the mixture was stirred at 60 °C for 1 h. The mixture was slowly cooled to 20 °C and stirred for 16 h. The mixture was filtered, and the filter cake was washed with ACN (10 mL x 2) and dried to obtain a residue (~5 g, 80% purity). The same procedure was repeated four times to obtain the crude product (~4.2 g, 88% purity).The crude product was mixed with another batch (900 mg, 89% purity) and purified by prep-HPLC (column: Phenomenex luna C18 (250*70mm, 10 um); mobile phase: [water(TFA)-ACN]; gradient: 5%-35% B over 20 min). The product was lyophilized to obtain the title compound (3 g, 2.60 mmol, 44% yield, 96% purity, 2TFA) as a light yellow solid. MS(M+H). + = 879.3.

[0283] 1 H NMR (400 MHz, DMSO-d6) δ = 10.89 (s, 1H), 10.44 (s, 1H), 9.75 - 9.73 (m, 1H), 9.61 (br s, 2H), 9.54 (br s, 1H), 8.16 (d,J= 4.0 Hz, 1H), 7.79 - 7.57 (m, 9H), 7.39 - 7.33 (m, 2H), 7.22 (dd,J= 13.2, 7.2 Hz, 1H), 7.02 (br dd,J= 11.2, 7.2 Hz, 1H), 4.00 (dd,J= 12.8, 5.2 Hz, 1H), 3.62 - 3.50 (m, 4H), 3.30 - 3.16 (m, 6H), 3.09 - 3.03 (m, 2H), 2.85 - 2.83 (m, 2H), 2.79 - 2.66 (m, 1H), 2.55 - 2.52 (m, 1H), 2.26 - 2.16 (m, 1H), 1.98 - 1.95 (m, 1H), 1.80 - 1.70 (m, 2H), 1.70 - 1.65 (m, 2H), 1.42 - 1.30 (m, 3H).

[0284]

[0285] Example 7. Synthesis of N-(4-((2-((4-((4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)picolinamide (Compound 7)

[0286]

[0287] Step 1. Synthesis of tert-butyl (4-(picolinamido)phenyl)carbamate (2)

[0288] To a solution of tert-butyl N-(4-aminophenyl)carbamate (10 g, 48.02 mmol), pyridine-2-carboxylic acid (6.50 g, 52.82 mmol), and DIPEA (18.62 g, 144.05 mmol) in DMF (150 mL) were added EDCI (11.05 g, 57.62 mmol) and HOBt (7.79 g, 57.62 mmol), and the mixture was stirred at 25 °C for 14 h. The main peak of the desired mass was identified by LCMS. Water (400 mL) was poured into the mixture while stirring. The reaction mixture was filtered and washed with water (100 mL). The filter cake was collected and dried to give the title compound (15 g, 47.87 mmol, 99.69% yield) as a brown solid.

[0289] Step 2. Synthesis of N-(4-aminophenyl)picolinamide (3)

[0290] To a solution of tert-butyl (4-(picolinamido)phenyl)carbamate (15 g, 47.87 mmol) in dioxane (50 mL) was added HCl / dioxane (2 M, 150 mL) at 25 °C, and the reaction mixture was stirred at 25 °C for 14 h. The main peak of the target mass was confirmed by LCMS. The reaction mixture was concentrated under reduced pressure to obtain the title compound (15 g, crude, 2HCl) as a gray solid. MS (M+H) + =214.1.

[0291] Step 3. Synthesis of N-(4-((2-chloro-5-fluoropyrimidin-4-yl)amino)phenyl)picolinamide (4)

[0292] To a solution of N-(4-aminophenyl)picolinamide (10 g, 40.05 mmol, HCl) and 2,4-dichloro-5-fluoro-pyrimidine (6.69 g, 40.05 mmol) in i-PrOH (150 mL) was added DIPEA (20.70 g, 160.19 mmol) at 25 °C, and the mixture was stirred at 70 °C for 2 h. The main peak of the desired mass was identified by LCMS. After cooling to 25 °C, the mixture was filtered and washed with MTBE (50 mL). The filter cake was collected and dried to give the title compound (9 g, 26.18 mmol, 65.38% yield) as a yellow solid.

[0293] Step 4. Synthesis of methyl 4-((5-fluoro-4-((4-(picolinamido)phenyl)amino)pyrimidin-2-yl)amino)benzoate (5)

[0294] A solution of N-(4-((2-chloro-5-fluoropyrimidin-4-yl)amino)phenyl)picolinamide (9 g, 26.18 mmol), methyl 4-aminobenzoate (3.96 g, 26.18 mmol), and TsO·H2O (9.96 g, 52.36 mmol) in dioxane (150 mL) was stirred at 100 °C for 14 h. LCMS confirmed complete consumption of the starting material. After cooling to 25 °C, the mixture was filtered and washed with MTBE (100 mL). The filter cake was collected and dried to give the title compound (13 g, crude) as a yellow solid, which was used directly in the next reaction. MS(M+H) + = 459.2.

[0295] Step 5. Synthesis of 4-((5-fluoro-4-((4-(picolinamido)phenyl)amino)pyrimidin-2-yl)amino)benzoic acid (6)

[0296] To a solution of methyl 4-((5-fluoro-4-((4-(picolinamido)phenyl)amino)pyrimidin-2-yl)amino)benzoate (7 g, 15.27 mmol) in THF (40 mL) and MeOH (30 mL) was added a solution of NaOH (1.83 g, 45.81 mmol) in water (20 mL) at 25 °C, and the mixture was stirred at 45 °C for 14 h (the reaction mixture turned into a black solution). LCMS confirmed complete consumption of the starting material. After cooling to 25 °C, 1 N HCl solution was slowly added to oxidize the mixture to pH = 6. (The black solution turned into a yellow mixture, and a large amount of yellow solid was produced). The mixture was filtered. The filter cake was collected and dried to obtain the title compound as a yellow solid (4.5 g, 9.11 mmol, 59.68% yield, 90% purity).

[0297] Step 6. Synthesis of N-(4-((5-fluoro-2-((4-((4-(2-hydroxyethyl)piperidin-1-yl)carbamoyl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)picolinamide (8)

[0298] The title compound (600 mg, 893.77 μmol, 79.44% yield, 85% purity) as a pink solid was obtained by a similar method to step 1 of Example 4. MS (M+H) + = 571.3.

[0299] Step 7. Synthesis of N-(4-((5-fluoro-2-((4-((4-(2-oxoethyl)piperidin-1-yl)carbamoyl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)picolinamide (9)

[0300] The title compound (200 mg, crude) as a pink solid was obtained by a similar method to step 3 of Example 6, and was used directly in the following reaction. MS(M+H) + = 569.2.

[0301] Step 8. Synthesis of N-(4-((2-((4-((4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)picolinamide (Compound 7)

[0302] The title compound (9.3 mg, 10.79 μmol, 3.07% yield, 100% purity) was obtained as a white solid by a similar method to Example 2. MS (M+H) + = 862.6.

[0303] 1 H NMR (400 MHz, DMSO-d6) δ = 10.86 (s, 1H), 10.67 (s, 1H), 9.50 (s, 1H), 9.44 (s, 1H), 9.11 (s, 1H), 8.76 (d, J = 4.8 Hz, 1H), 8.19 (d,J= 7.8 Hz, 1H), 8.14 (d,J= 3.5 Hz, 1H), 8.08 (dt,J= 1.5, 7.7 Hz, 1H), 7.92 (d,J= 8.9 Hz, 2H), 7.83 - 7.72 (m, 4H), 7.71 - 7.63 (m, 3H), 7.18 - 7.09 (m, 1H), 6.90 - 6.81 (m, 1H), 4.01 - 3.93 (m, 1H), 3.11 - 2.92 (m, 6H), 2.75 - 2.65 (m, 3H), 2.54 - 2.52 (m, 4H), 2.38 - 2.31 (m, 2H), 2.24 - 2.14 (m, 1H), 2.00 - 1.91 (m, 1H), 1.73 - 1.63 (m, 2H), 1.47 - 1.38 (m, 2H), 1.32 - 1.20 (m, 4H).

[0304]

[0305] Example 8. Synthesis of N-(4-((2-((4-((4-(2-((2R)-4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)-2-methylpiperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 8)

[0306]

[0307] The title compound (117 mg, 100.20 μmol, 24% yield, 96% purity, 2TFA) was obtained as a white solid by a similar method to step 4 of Example 6. MS (M+H) + = 893.4.

[0308] 1 H NMR (400 MHz, DMSO-d6) δ = 10.89 (s, 1H), 10.44 (s, 1H), 9.87 (br s, 1H), 9.72 - 9.71 (m, 1H), 9.65 (s, 1H), 9.56 (br s, 1H), 8.17 (d,J= 4.0 Hz, 1H), 7.79 - 7.56 (m, 9H), 7.39 - 7.33 (m, 2H), 7.24 - 7.19 (m, 1H), 7.04 - 6.96 (m, 1H), 4.00 (dd,J= 12.4, 4.4 Hz, 1H), 3.80 - 3.37 (m, 6H), 3.22 - 3.08 (m, 5H), 2.94 - 2.88 (m, 2H), 2.79 - 2.70 (m, 1H), 2.57 - 2.54 (m, 1H), 2.26 - 2.16 (m, 1H), 1.97 - 1.95 (m, 1H), 1.82 - 1.75 (m, 2H), 1.71 - 1.61 (m, 2H), 1.40 - 1.35 (m, 6H).

[0309]

[0310] Example 9. Synthesis of N-(4-(2-((2R)-4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)-2-methylpiperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 9)

[0311]

[0312] Step 1. Synthesis of tert-butyl (R)-4-(4-bromo-2,5-difluorophenyl)-2-methylpiperazine-1-carboxylate (2)

[0313] A solution of 1-bromo-2,5-difluoro-4-iodo-benzene (4.2 g, 13.17 mmol), tert-butyl (2R)-2-methylpiperazine-1-carboxylate (2.64 g, 13.17 mmol), Pd2(dba)3 (603.04 mg, 658.55 μmol), Xantphos (762.09 mg, 1.32 mmol), and Cs2CO3 (8.58 g, 26.34 mmol) in dioxane (60 mL) was degassed and purged three times with N2, and the mixture was stirred at 100 °C under N2 for 16 h. The target mass peak (24%) was identified by LCMS. The reaction mixture was filtered and washed with EtOAc (50 mL). The filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (20 g SepaFlash® silica flash column, eluent 0–6% EtOAc / petroleum ether, gradient @ 80 mL / min) to give the title compound (2.3 g, 5.00 mmol, 37% yield, 85% purity) as a yellow oil. MS (M+H) + = 391.0.

[0314] Step 2. Synthesis of tert-butyl (R)-4-(4-(2,6-bis(benzyloxy)pyridin-3-yl)-2,5-difluorophenyl)-2-methylpiperazine-1-carboxylate (4)

[0315] A solution of tert-butyl (R)-4-(4-bromo-2,5-difluorophenyl)-2-methylpiperazine-1-carboxylate (2.3 g, 5.88 mmol), 2,6-dibenzyloxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (2.45 g, 5.88 mmol), Pd(dppf)Cl2 (430.14 mg, 587.86 μmol), and Cs2CO3 (3.83 g, 11.76 mmol) in dioxane (40 mL) and water (10 mL) was purged three times with N2, and the mixture was stirred at 100 °C under N2 for 16 h. The target mass peak (44%) was confirmed by LCMS. The reaction mixture was filtered and concentrated under reduced pressure to remove the organic phase. The residue was diluted with water (50 mL) and washed with EtOAc (50 mL × 3). The combined organic layer was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (20 g SepaFlash® silica flash column, eluent 0–6% EtOAc / petroleum ether, gradient @ 80 mL / min) to give the title compound (3.5 g, 5.00 mmol, 85% yield, 86% purity) as a yellow solid. MS (M+H) + = 602.2.

[0316] Step 3. Synthesis of tert-butyl (2R)-4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)-2-methylpiperazine-1-carboxylate (5)

[0317] To a solution of tert-butyl (R)-4-(4-(2,6-bis(benzyloxy)pyridin-3-yl)-2,5-difluorophenyl)-2-methylpiperazine-1-carboxylate (3.5 g, 5.82 mmol) and AcOH (349.32 mg, 5.82 mmol, 333.01 μL) in THF (30 mL) and MeOH (30 mL) were added Pd / C (619.05 mg, 581.70 μmol, 10% purity) and Pd(OH)2 / C (408.46 mg, 581.70 μmol, 20% purity) under N2. The mixture was degassed and purged with H2 three times. The mixture was stirred at 30 °C under H2 (50 psi) for 48 h. The target mass peak (81%) was confirmed by LCMS. The reaction mixture was filtered through Celite and concentrated under reduced pressure. The residue was triturated with MTBE (30 mL) at 15 °C for 10 min to obtain the title compound (1.9 g, 4.40 mmol, 75% yield, 98% purity) as a gray solid. MS (M+H) + = 424.2.

[0318] Step 4. Synthesis of 3-(2,5-difluoro-4-((R)-3-methylpiperazin-1-yl)phenyl)piperidine-2,6-dione (6)

[0319] To a solution of tert-butyl (2R)-4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)-2-methylpiperazine-1-carboxylate (1.9 g, 4.49 mmol) in dioxane (10 mL) was added HCl / dioxane (2 M, 20 mL), and the mixture was stirred at 20 °C for 12 h. LCMS confirmed the presence of a small amount of starting material and the main peak of the target mass. The reaction mixture was concentrated under reduced pressure to obtain the title compound (1.8 g, crude, 2HCl) as a white solid. MS (M+H) + = 324.1.

[0320] Step 5. Synthesis of N-(4-(2-((2R)-4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)-2-methylpiperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 9)

[0321] The title compound (122.39 mg, 102.08 μmol, 20.54% yield, 95% purity, 2TFA) was obtained as a white solid by a similar method to step 4 of Example 6. MS (M+H) + = 911.3.

[0322] 1 H NMR (400 MHz, DMSO-d6) δ = 10.88 (s, 1H), 10.42 (s, 1H), 9.68 (s, 1H), 9.60 - 9.37 (m, 2H), 9.16 - 9.06 (m, 1H), 8.18 (d,J= 3.5 Hz, 1H), 7.76 - 7.72 (m, 4H), 7.68 (br t,J= 7.3 Hz, 1H), 7.62 - 7.55 (m, 1H), 7.47 - 7.39 (m, 2H), 7.39 - 7.31 (m, 2H), 7.25 - 7.18 (m, 1H), 7.05 - 6.95 (m, 1H), 4.01 - 3.98 (m, 1H), 3.65 - 3.56 (m, 2H), 3.52 - 3.34 (m, 3H), 3.31 - 2.99 (m, 6H), 2.94 - 2.84 (m, 1H), 2.72 - 2.61 (m, 3H), 2.26 - 2.16 (m, 1H), 2.01 - 1.92 (m, 1H), 1.79 - 1.68 (m, 2H), 1.67 - 1.53 (m, 2H), 1.43 - 1.26 (m, 6H).

[0323]

[0324] Example 10. Synthesis of N-(4-(2-((3S)-4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)-3-methylpiperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 10)

[0325]

[0326] Step 1. Synthesis of tert-butyl (S)-4-(4-bromo-2,5-difluorophenyl)-3-methylpiperazine-1-carboxylate (2)

[0327] The title compound (830 mg, 1.48 mmol, 9.47% yield, 70% purity) was obtained as a light yellow solid by a similar method to step 1 of Example 9. MS (M+H) + = 391.1.

[0328] Step 2. Synthesis of tert-butyl (S)-4-(4-(2,6-bis(benzyloxy)pyridin-3-yl)-2,5-difluorophenyl)-3-methylpiperazine-1-carboxylate (4)

[0329] To a solution of 2,6-bis(benzyloxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (545.03 mg, 1.31 mmol), tert-butyl (S)-4-(4-bromo-2,5-difluorophenyl)-3-methylpiperazine-1-carboxylate (730 mg, 1.31 mmol), and Cs2CO3 (1.28 g, 3.2 mmol) in dioxane (10 mL) and water (1 mL) was added Pd(dppf)Cl2·CH2Cl2 (106.66 mg, 130.61 μmol) at 25 °C under N2. After purging and degassing with N2 for 3 min, the mixture was stirred at 100 °C for 16 h. The target mass was confirmed as a major peak (47%) by LCMS. The reaction mixture was filtered and washed with EtOAc (20 mL x 2). The filtrate was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (Biotage; 20 g SepaFlash® silica flash column, eluent 8% EtOAc / petroleum ether, gradient @ 60 mL / min) to give the title compound (550 mg, 776.99 μmol, 59.49% yield, 85% purity) as a light yellow oil. MS (M+H) + = 602.2.

[0330] Step 3. Synthesis of tert-butyl (3S)-4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)-3-methylpiperazine-1-carboxylate (5)

[0331] The title compound (300 mg, 637.62 μmol, 82.06% yield, 90% purity) was obtained as a yellow solid by a similar method to step 3 of Example 9. MS (M+H) + = 424.2.

[0332] Step 4. Synthesis of 3-(2,5-difluoro-4-((S)-2-methylpiperazin-1-yl)phenyl)piperidine-2,6-dione (6)

[0333] The title compound (372 mg, crude, HCl) as a yellow solid was obtained by a similar method to step 3 of Example 1, and was used directly in the next reaction.

[0334] Step 5. Synthesis of N-(4-(2-((3S)-4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)-3-methylpiperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 10)

[0335] The title compound (138.31 mg, 104.86 μmol, 32.86% yield, 95% purity, 3TFA) was obtained as a white solid by a similar method to step 4 of Example 6. MS (M+H) + = 911.3.

[0336] 1H NMR (400 MHz, DMSO-d6) δ = 10.90 (d,J= 13.6 Hz, 1H), 10.43 (s, 1H), 9.71 (s, 1H), 9.54 (s, 1H), 9.39 - 9.21 (m, 1H), 8.18 (d,J= 3.6 Hz, 1H), 7.81 (br d,J= 14.0 Hz, 1H), 7.76 - 7.72 (m, 4H), 7.70 - 7.66 (m, 1H), 7.62 - 7.56 (m, 1H), 7.44 (br d,J= 4.4 Hz, 2H), 7.38 - 7.32 (m, 2H), 7.30 - 6.99 (m, 2H), 4.09 - 3.98 (m, 1H), 3.64 - 3.52 (m, 2H), 3.45 - 3.32 (m, 2H), 3.26 - 3.01 (m, 6H), 2.89 - 2.67 (m, 4H), 2.56 - 2.53 (m, 1H), 2.26 - 2.18 (m, 1H), 1.99 - 1.97 (m, 1H), 1.74 - 1.70 (m, 2H), 1.67 - 1.61 (m, 2H), 1.36 - 1.29 (m, 3H), 1.12 - 0.93 (m, 3H).

[0337]

[0338] Example 11. Synthesis of N-(4-((2-((4-((4-(2-((3S)-4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)-3-methylpiperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 11)

[0339]

[0340] The title compound (83.74 mg, 64.41 μmol, 20.19% yield, 95% purity, 3TFA) was obtained as a white solid by a similar method to step 4 of Example 6. MS (M+H) + = 893.4.

[0341] 1H NMR (400 MHz, DMSO-d6) δ = 10.90 (d,J= 13.2 Hz, 1H), 10.43 (s, 1H), 9.78 - 9.76 (m, 1H), 9.62 (br s, 1H), 9.54 (br s, 1H), 8.17 (d,J= 4.0 Hz, 1H), 7.79 - 7.68 (m, 9H), 7.62 - 7.57 (m, 1H), 7.39 - 7.33 (m, 2H), 7.30 - 6.99 (m, 2H), 4.07 - 3.98 (m, 1H), 3.64 - 3.52 (m, 2H), 3.45 - 3.31 (m, 2H), 3.26 - 3.04 (m, 6H), 2.92 - 2.66 (m, 4H), 2.58 - 2.55 (m, 1H), 2.27 - 2.16 (m, 1H), 2.01 - 1.98 (m, 1H), 1.77 - 1.73 (m, 2H), 1.68 - 1.63 (m, 2H), 1.42 - 1.39 (m, 3H), 1.12 - 0.93 (m, 3H).

[0342]

[0343] Example 12. Synthesis of N-(4-((2-((4-((4-((1-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperidin-4-yl)methyl)(methyl)amino)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 12)

[0344]

[0345]

[0346] Step 1. Synthesis of tert-butyl 4-(((benzyloxy)carbonyl)(methyl)amino)piperidine-1-carboxylate (2)

[0347] To a solution of tert-butyl 4-(methylamino)piperidine-1-carboxylate (5 g, 23.33 mmol) in NaOH (2 M, 70 mL) was slowly added CbzCl (5.57 g, 32.66 mmol, 4.66 mL) at 0 °C, and the resulting mixture was stirred at 20 °C for 16 h. LCMS confirmed complete consumption of the starting material and the peak of the target mass (43%). The reaction mixture was filtered, washed with petroleum ether (50 mL x 3), and concentrated under reduced pressure to give the title compound (7.5 g, 21.52 mmol, 92.26% yield) as a white solid. MS(M-Boc+H) + = 249.0.

[0348] Step 2. Synthesis of benzyl methyl(piperidin-4-yl)carbamate (3)

[0349] To a solution of tert-butyl 4-(((benzyloxy)carbonyl)(methyl)amino)piperidine-1-carboxylate (7.5 g, 21.52 mmol) in DCM (10 mL) was added HCl / dioxane (2 M, 80 mL) at 20 °C, and the resulting mixture was stirred at 20 °C for 2 h. LCMS confirmed complete consumption of the starting material and the peak of the target mass (74%). The reaction mixture was concentrated under reduced pressure to obtain the title compound (6.2 g, crude, HCl salt) as a white solid. MS (M+H) + =249.0.

[0350] Step 3. Synthesis of benzyl methyl(1-nitrosopiperidin-4-yl)carbamate (4)

[0351] To a solution of benzyl N-methyl-N-(4-piperidyl)carbamate (6.2 g, 21.77 mmol, HCl salt) in water (70 mL) was added NaNO2 (4.51 g, 65.31 mmol) at 0 °C. AcOH (5.23 g, 87.08 mmol, 4.99 mL) was slowly added at 0 °C, and the resulting mixture was stirred at 20 °C for 12 h. LCMS confirmed complete consumption of the starting material and a peak of the target mass (59%). The reaction mixture was diluted with water (30 mL) and washed with EtOAc (100 mL x 2). The combined organic layer was washed with saturated aqueous NaHCO3 solution (100 mL x 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the title compound (6.4 g, crude) as a yellow oil. MS(M+H) + = 278.0.

[0352] Step 4. Synthesis of benzyl (1-aminopiperidin-4-yl)(methyl)carbamate (5)

[0353] To a solution of benzyl methyl(1-nitrosopiperidin-4-yl)carbamate (3.5 g, 12.62 mmol) in THF (40 mL) and water (10 mL) was added NH4Cl (2.03 g, 37.86 mmol) at 0 °C. Zn (4.2 g, 64.23 mmol) was added at 0 °C, and the resulting mixture was stirred at 20 °C for 2 h. LCMS confirmed complete consumption of the starting material and a peak of the target mass (77%). The reaction mixture was filtered and washed with THF (50 mL x 3). The filtrate was concentrated under reduced pressure to give the title compound (3.4 g, crude) as a yellow oil. MS(M+H) + = 264.0.

[0354] Step 5. Synthesis of benzyl (1-((tert-butoxycarbonyl)amino)piperidin-4-yl)(methyl)carbamate (6)

[0355] To a solution of benzyl (1-aminopiperidin-4-yl)(methyl)carbamate (3.4 g, 12.91 mmol) in THF (60 mL) and water (20 mL) were added NaOH (1.55 g, 38.73 mmol) and (Boc)2O (5.64 g, 25.82 mmol, 5.93 mL) at 20 °C. The resulting mixture was stirred at 20 °C for 12 h. LCMS confirmed that 25% of the starting material remained and a peak of the target mass (33%) was formed. NaOH (2.07 g, 51.65 mmol) and (Boc)2O (2.82 g, 12.91 mmol, 2.97 mL) were further added and the resulting mixture was stirred at 20 °C for 12 h. LCMS confirmed that the starting material was completely consumed and a peak of the target mass (33%) was formed. The reaction mixture was diluted with water (200 mL) and extracted with EtOAc (100 mL x 3). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (40 g SepaFlash® silica flash column, eluent 5–50% EtOAc / petroleum ether, gradient @ 200 mL / min) to give the title compound (1.4 g, 3.85 mmol, 29.83% yield) as a white solid. MS (M+H) + = 364.1.

[0356] Step 6. Synthesis of tert-butyl (4-(methylamino)piperidin-1-yl)carbamate (7)

[0357] Benzyl (1-((tert-butoxycarbonyl)amino)piperidin-4-yl)(methyl)carbamate (0.4 g, 1.10 mmol) was added to a solution of Pd / C (0.1 g, 10% purity) in CF3CH2OH (10 mL) under N2. The suspension was degassed and purged three times with H2. The mixture was stirred under H2 (50 Psi) at 20 °C for 12 h. LCMS confirmed complete consumption of the starting material and the peak of the target mass. The reaction mixture was filtered and washed with MeOH (200 mL). The filtrate was concentrated under reduced pressure to give the title compound (0.6 g, crude) as a yellow oil. MS (M+H) + = 230.1.

[0358] Step 7. Synthesis of tert-butyl (4-(((1-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperidin-4-yl)methyl)(methyl)amino)piperidin-1-yl)carbamate (9)

[0359] To a solution of tert-butyl (4-(methylamino)piperidin-1-yl)carbamate (0.2 g, 872.15 μmol) and 1-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperidine-4-carbaldehyde (410.67 mg, 1.22 mmol) in DCM (8 mL) and DMF (2 mL) were added TEA (176.50 mg, 1.74 mmol, 242.78 μL) and MgSO4 (157.47 mg, 1.31 mmol) at 20 °C. After stirring for 0.5 h, NaBH(OAc)3 (554.53 mg, 2.62 mmol) was added at 20 °C, and the resulting mixture was stirred at 20 °C for 12 h. LCMS confirmed complete consumption of the starting material and the peak of the target mass (40%). The reaction mixture was diluted with water (20 mL) and extracted with DCM (20 mL x 3). The combined organic layer was washed with saturated aqueous NaHCO3 solution (20 mL x 2), dried over Na2SO4, filtered, and concentrated. The residue was purified by flash silica gel chromatography (10 g SepaFlash® silica flash column, eluent 100–100% EtOAc / petroleum ether to 10–10% MeOH / DCM, gradient @ 180 mL / min) to give the title compound (141 mg, 256.53 μmol, 29.41% yield) as a white solid. MS (M+H) + = 550.3.

[0360] Step 8. Synthesis of 3-(4-(4-(((1-aminopiperidin-4-yl)(methyl)amino)methyl)piperidin-1-yl)-2,5-difluorophenyl)piperidine-2,6-dione (10)

[0361] To a solution of tert-butyl (4-(((1-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperidin-4-yl)methyl)(methyl)amino)piperidin-1-yl)carbamate (141 mg, 256.53 μmol) in DCM (1 mL) was added TFA (351.00 mg, 3.08 mmol, 228.66 μL) at 20 °C, and the resulting mixture was stirred at 20 °C for 16 h. LCMS confirmed complete consumption of the starting material and the peak of the target mass (78%). The reaction mixture was concentrated under reduced pressure to obtain the title compound (145 mg, crude, TFA salt) as a yellow oil. MS (M+H) + = 450.1.

[0362] Step 9. Synthesis of N-(4-((2-((4-((4-((1-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperidin-4-yl)methyl)(methyl)amino)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 12)

[0363] To a solution of 3-(4-(4-(((1-aminopiperidin-4-yl)(methyl)amino)methyl)piperidin-1-yl)-2,5-difluorophenyl)piperidine-2,6-dione (145 mg, 257.29 μmol, TFA salt) and 4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzoic acid (130.59 mg, 283.02 μmol) in DMF (4 mL) were added EDCI (73.99 mg, 385.94 μmol), HOBt (52.15 mg, 385.94 μmol) and DIPEA (266.03 mg, 2.06 mmol, 358.53 μL) at 20 °C, and the resulting mixture was stirred at 20 °C. The mixture was stirred for 12 hours. LCMS confirmed that the starting material was completely consumed and the peak of the target mass (25%) was observed. The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (20 mL x 3). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Phenomenex Luna C18 150*25 mm*10 μm; mobile phase: [H2O (0.1%TFA) -ACN]; gradient: 3%-33% B over 15 min) and lyophilized to obtain the title compound (74.8 mg, 63.39 μmol, 24.64% yield, 95% purity, 2TFA salt) as a white solid. MS (M+H) + = 893.2.

[0364] 1H NMR (400 MHz, CD3OD) δ = 8.07 (d,J= 4.9 Hz, 1H), 7.82 - 7.71 (m, 5H), 7.69 - 7.56 (m, 5H), 7.39 - 7.25 (m, 2H), 7.06 - 6.98 (m, 1H), 6.87 - 6.78 (m, 1H), 3.96 (dd,J= 5.0, 12.5 Hz, 1H), 3.54 - 3.42 (m, 2H), 3.26 - 3.16 (m, 4H), 3.09 - 3.00 (m, 1H), 2.94 - 2.85 (m, 3H), 2.82 - 2.63 (m, 6H), 2.28 - 2.23 (m, 1H), 2.16 - 1.93 (m, 7H), 1.88 - 1.85 (m, 1H), 1.63 - 1.46 (m, 2H).

[0365]

[0366] Example 13. Synthesis of N-(4-((2-((4-((1r,4r)-4-(((1-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperidin-4-yl)methyl)(methyl)amino)cyclohexyl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 13)

[0367]

[0368] Step 1. Synthesis of methyl 1-(4-bromo-2,5-difluorophenyl) piperidine-4-carboxylate (2)

[0369] The title compound (5 g, 14.96 mmol, 71.42% yield) was obtained as a yellow solid by a similar method to step 1 of Example 10. MS (M+H) + =334.1

[0370] Step 2. Synthesis of methyl 1-(4-(2,6-bis(benzyloxy)pyridin-3-yl)-2,5-difluorophenyl)piperidine-4-carboxylate (3)

[0371] The title compound (7.4 g, crude) was obtained as a yellow solid by a similar method to step 2 of Example 10. MS (M+H) + =545.3

[0372] Step 3. Synthesis of (1-(4-(2,6-bis(benzyloxy)pyridin-3-yl)-2,5-difluorophenyl)piperidin-4-yl)methanol (4)

[0373] To a solution of LiAlH4 (2.5 M, 3.3 mL) in THF (10 mL) was added methyl 1-(4-(2,6-bis(benzyloxy)pyridin-3-yl)-2,5-difluorophenyl)piperidine-4-carboxylate (3 g, 5.51 mmol) in THF (20 mL) at 0 °C, and the mixture was stirred at 0 °C for 0.5 h. The peak of the target mass (96%) was confirmed by LCMS. The reaction mixture was quenched at 0 °C with water (0.3 mL), aqueous NaOH solution (15%, 0.3 mL), and water (0.9 mL), the mixture was filtered, and the filtrate was extracted with EtOAc (15 mL x 3). The combined organic layer was washed with water (10 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain the title compound (2.8 g, crude) as a yellow oil. MS (M+H) + = 517.3.

[0374] Step 4. Synthesis of 3-(2,5-difluoro-4-(4-(hydroxymethyl)piperidin-1-yl)phenyl)piperidine- 2,6-dione (5)

[0375] To a solution of Pd(OH)2 / C (0.5 g, 20% purity) in THF (50 mL), (1-(4-(2,6-bis(benzyloxy)pyridin-3-yl)-2,5-difluorophenyl)piperidin-4-yl)methanol (2.8 g, 5.42 mmol) in THF (50 mL), MeOH (50 mL) was added under N2, and the mixture was stirred at H2 (45 Psi) and 30 °C for 30 h. The peak of the target mass (90%) was confirmed by LCMS. The mixture was filtered and washed with THF (500 mL). The filtrate was concentrated under reduced pressure to give the title compound (1.7 g, crude) as a white solid. MS(M+H) + = 339.2.

[0376] Step 5. Synthesis of 1-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperidine-4-carbaldehyde (6)

[0377] To a solution of 3-(2,5-difluoro-4-(4-(hydroxymethyl)piperidin-1-yl)phenyl)piperidine-2,6-dione (0.5 g, 1.48 mmol) in DCM (10 mL) was added DMP (815 mg, 1.92 mmol, 595.33 μL), and the mixture was stirred at 20 °C for 1.5 h. The peak of the target mass (85%) was identified by LCMS. The mixture was diluted with saturated aqueous NaHCO3 solution (20 mL) and extracted with DCM (20 mL x 3). The combined organic layer was dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (12 g SepaFlash® silica flash column, eluent 50% EtOAc / petroleum ether, gradient @ 100 mL / min) to give the title compound (660 mg, crude) as a yellow oil. MS(M+H) + = 337.2.

[0378] Step 6. Synthesis of tert-butyl ((1r,4r)-4-(((1-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperidin-4-yl)methyl)(methyl)amino)cyclohexyl)carbamate (7)

[0379] The title compound (140 mg, 247.51 μmol, 35.32% yield, 97% purity) was obtained as a yellow solid by a similar method to step 7 of Example 12. MS (M+H) + = 549.4.

[0380] Step 7. Synthesis of 3-(4-(4-(((1r,4r)-4-aminocyclohexyl)(methyl)amino)methyl)piperidin-1-yl)-2,5-difluorophenyl)piperidine-2,6-dione (8)

[0381] The title compound (150 mg, crude, TFA salt) was obtained as a yellow solid by a similar method to step 8 of Example 12. MS (M+H) + = 449.3.

[0382] Step 8. Synthesis of N-(4-((2-((4-((1r,4r)-4-(((1-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperidin-4-yl)methyl)(methyl)amino)cyclohexyl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2fluorobenzamide (Compound 13)

[0383] The title compound (55.7 mg, 51.49 μmol, 25.11% yield, 93% purity, TFA salt) was obtained as a white solid by a similar method to step 9 of Example 12. MS (M+H) + = 892.6.

[0384] 1H NMR (400 MHz, DMSO-d6) δ = 10.85 (s, 1H), 10.42 (s, 1H), 9.49 (br d,J= 11.9 Hz, 2H), 8.79 - 8.66 (m, 1H), 8.14 (d,J= 3.8 Hz, 1H), 8.02 (br d,J= 7.9 Hz, 1H), 7.82 - 7.65 (m, 9H), 7.62 - 7.54 (m, 1H), 7.39 - 7.29 (m, 2H), 7.12 (dd,J= 6.9, 13.1 Hz, 1H), 6.87 (dd,J= 7.4, 12.0 Hz, 1H), 4.00 - 3.88 (m, 1H), 3.85 - 3.70 (m, 1H), 3.48 - 3.36 (m, 2H), 3.25 - 3.11 (m, 2H), 2.98 - 2.86 (m, 1H), 2.83 - 2.62 (m, 6H), 2.54 (br d,J= 3.4 Hz, 1H), 2.25 - 2.13 (m, 1H), 2.08 - 1.74 (m, 8H), 1.71 - 1.56 (m, 2H), 1.49 - 1.30 (m, 4H).

[0385] Example 14. Synthesis of N-(4-((2-((4-((1-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperazin-1-yl)ethyl)piperidin-4-yl)amino)methyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 14)

[0386]

[0387] Step 1. Synthesis of tert-butyl (1-(2-hydroxyethyl)piperidin-4-yl)carbamate (2)

[0388] To a solution of tert-butyl N-(4-piperidyl)carbamate (5 g, 24.97 mmol) and 2-bromoethanol (3.43 g, 27.46 mmol, 1.95 mL) in ACN (80 mL) was added K2CO3 (10.35 g, 74.90 mmol) at 20 °C, and the mixture was stirred at 80 °C for 16 h. LCMS confirmed complete consumption of the starting material and a peak of the target mass (64%). The mixture was diluted with water (300 mL) and extracted with EtOAc (100 mL x 3). The combined organic layers were washed with brine (100 mL x 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the title compound (5.1 g, 20.87 mmol, 83.61% yield) as a yellow oil. MS(M-Boc+H) + = 145.1.

[0389] Step 2. Synthesis of tert-butyl (1-(2-chloroethyl)piperidin-4-yl)carbamate (3)

[0390] To a solution of tert-butyl (1-(2-hydroxyethyl)piperidin-4-yl)carbamate (5.1 g, 20.87 mmol) in DCM (300 mL) was added Py (6.60 g, 83.49 mmol, 6.74 mL) at 0 °C. SOCl2 (7.45 g, 62.62 mmol, 4.55 mL) was slowly added at 0 °C, and the resulting mixture was stirred at 20 °C for 2 h. TLC (SiO2, DCM:MeOH=10:1) confirmed complete consumption of the starting material and the appearance of a new spot. The reaction mixture was diluted with water (300 mL) and adjusted to pH~8 with saturated aqueous NaHCO3 solution at 0 °C. The aqueous phase was extracted with DCM (200 mL x 3). The mixed organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a residue. The residue was diluted with hexane (100 mL) and concentrated under reduced pressure at 40–50 °C to remove pyridine to obtain the title compound (5.5 g, crude) as a yellow solid.

[0391] Step 3. Synthesis of tert-butyl (1-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperazin-1-yl)ethyl)piperidin-4-yl)carbamate (5)

[0392] To a solution of 3-(2, 5-difluoro-4-piperazin-1-yl-phenyl)piperidine-2, 6-dione (300 mg, 969.90 μmol, 2HCl salt) and tert-butyl (1-(2-chloroethyl)piperidin-4-yl)carbamate (410 mg, 1.56 mmol) in DMF (6 mL) were added KI (161.01 mg, 969.90 μmol) and DIPEA (752.11 mg, 5.82 mmol, 1.01 mL) at 20 °C, and the resulting mixture was stirred at 80 °C for 12 h. LCMS confirmed complete consumption of the starting material and the peak of the target mass (67%). The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (20 mL x 3). The organic phase was washed with brine (20 mL x 3), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (10 g SepaFlash® silica flash column, eluent 100–100% EtOAc / petroleum ether to 10–10% MeOH / DCM, gradient @ 180 mL / min) to give the title compound (178 mg, 332.32 μmol, 34.26% yield) as a yellow solid. MS (M+H) + = 536.2.

[0393] Step 4. Synthesis of 3-(4-(4-(2-(4-aminopiperidin-1-yl)ethyl)piperazin-1-yl)-2,5-difluorophenyl)piperidine-2,6-dione (6)

[0394] The title compound (183 mg, crude, TFA salt) was obtained as a yellow oil by a similar method to step 8 of Example 12. MS (M+H) + = 436.2.

[0395] Step 5. Synthesis of N-(4-((2-((4-((1-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl) piperazin-1-yl)ethyl)piperidin-4-yl)amino)methyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 14)

[0396] To a solution of 3-(4-(4-(2-(4-aminopiperidin-1-yl)ethyl)piperazin-1-yl)-2,5-difluorophenyl)piperidine-2,6-dione (173 mg, 314.81 μmol, TFA salt) and 2-fluoro-N-(4-((5-fluoro-2-((4-formylphenyl)amino)pyrimidin-4-yl)amino)phenyl)benzamide (182.29 mg, 409.26 μmol) in DMSO (4 mL) was added NaOAc (206.60 mg, 2.52 mmol) at 20 °C. After stirring for 0.5 h, NaBH(OAc)3 (200.17 mg, 944.44 μmol) was further added at 20 °C, and the resulting mixture was stirred at 20 °C for 29 h. LCMS confirmed that the starting material was completely consumed and the peak of the target mass (26%) was obtained. The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (20 mL x 3). The organic phase was washed with brine (10 mL x 3), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain the crude product. The crude product was dissolved in a mixed solution (DMF:TFA=1:1, 5 mL) and purified by prep-HPLC (column: Phenomenex Luna C18 150*25 mm*10um; mobile phase: [H2O (0.1%TFA)-ACN]; gradient: 0%-22% B over 15 min), and lyophilized to obtain the title compound (46.3 mg, 41.51 μmol, 13.19% yield, 98% purity, 2TFA salt) as a white solid. MS(M+H) + = 865.6.

[0397] 1H NMR (400 MHz, DMSO-d6) δ = 10.89 (s, 1H), 10.41 (s, 1H), 9.44 (br d,J= 18.6 Hz, 2H), 9.03 - 8.87 (m, 1H), 8.13 (d,J= 3.8 Hz, 1H), 7.82 - 7.75 (m, 4H), 7.74 - 7.65 (m, 3H), 7.63 - 7.46 (m, 1H), 7.40 - 7.31 (m, 4H), 7.22 - 7.10 (m, 1H), 6.95 (br dd,J= 7.4, 11.8 Hz, 1H), 4.12 (br d,J= 5.4 Hz, 2H), 3.99 (br dd,J= 4.9, 13.2 Hz, 1H), 3.51 - 3.39 (m, 3H), 3.28 - 3.00 (m, 12H), 2.88 - 2.68 (m, 4H), 2.30 - 2.14 (m, 3H), 2.03 - 1.90 (m, 1H), 1.85 - 1.68 (m, 2H).

[0398]

[0399] Example 15. Synthesis of N-(4-((2-((4-((4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperazin-1-yl)-1-hydroxyethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 15)

[0400]

[0401] Step 1. Synthesis of tert-butyl 4-(oxiran-2-yl)piperidine-1-carboxylate (2)

[0402] To a solution of TMSOI (6.19 g, 28.13 mmol) in DMSO (50 mL) was added NaH (1.22 g, 30.48 mmol, 60% purity) under N2 at 0 °C. After stirring at 20 °C for 1 h, tert-Butyl 4-formylpiperidine-1-carboxylate (5 g, 23.44 mmol) in DMSO (20 mL) was slowly added under N2 at 20 °C, and the resulting mixture was stirred at 20 °C for 15 h. TLC (SiO2, petroleum ether: EtOAc = 3:1) confirmed complete consumption of the starting material and a new main spot. The reaction was quenched with saturated aqueous NH4Cl solution (150 mL) and extracted with EtOAc (100 mL x 3). The organic layer was washed with brine (100 mL x 3), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (40 g SepaFlash® silica flash column, eluent 5–12% EtOAc / petroleum ether, gradient @ 200 mL / min) to afford the title compound (1.8 g, 7.92 mmol, 33.78% yield) as a yellow oil.

[0403] Step 2. Synthesis of tert-butyl 4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperazin-1-yl)-1-hydroxyethyl)piperidine-1-carboxylate (4)

[0404] To a solution of 3-(2,5-difluoro-4-(piperazin-1-yl)phenyl)piperidine-2,6-dione (2 g, 5.23 mmol, 2HCl salt) and tert-butyl 4-(oxiran-2-yl)piperidine-1-carboxylate (1.43 g, 6.28 mmol) in i-PrOH (30 mL) was added NaHCO3 (2.20 g, 26.16 mmol, 1.02 mL) at 20 °C, and the resulting mixture was stirred at 80 °C for 64 h. LCMS confirmed the complete consumption of 3-(2,5-difluoro-4-piperazin-1-yl-phenyl)piperidine-2,6-dione and the peak of the target mass (59%). The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (20 g SepaFlash® silica flash column, eluent 33–100% EtOAc / petroleum ether, gradient @200 mL / min). The product was repurified by prep-HPLC (column: Phenomenex luna C18 150*25 mm* 10 um; mobile phase: [H2O (0.1%TFA) -ACN]; gradient: 18%–48% B over 15 min) and lyophilized to give the title compound (1.4 g, 2.15 mmol, 41.12% yield, TFA salt) as a white solid. MS (M+H) + = 537.2.

[0405] Step 3. Synthesis of 3-(2,5-difluoro-4-(4-(2-hydroxy-2-(piperidin-4-yl)ethyl)piperazin-1-yl)phenyl)piperidine-2,6-dione (5)

[0406] To a solution of tert-butyl 4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl)piperazin-1-yl)-1-hydroxyethyl)piperidine-1-carboxylate (1.4 g, 2.15 mmol, TFA salt) in DCM (10 mL) was added HCl / dioxane (2 M, 30 mL) at 20 °C, and the resulting mixture was stirred at 20 °C for 12 h. LCMS confirmed complete consumption of the starting material and the main peak of the target mass. The reaction mixture was concentrated under reduced pressure to obtain the title compound (1 g, crude, HCl salt) as a white solid. MS (M+H) + = 437.2.

[0407] Step 4. Synthesis of 3-(2,5-difluoro-4-(4-(2-hydroxy-2-(1-nitrosopiperidin-4-yl)ethyl)piperazin-1-yl)phenyl)piperidine-2,6-dione (6)

[0408] To a solution of 3-(2,5-difluoro-4-(4-(2-hydroxy-2-(piperidin-4-yl)ethyl)piperazin-1-yl)phenyl)piperidine-2,6-dione (0.6 g, 1.27 mmol, HCl salt) in THF (10 mL) were added TEA (385.11 mg, 3.81 mmol, 529.73 μL) and t-BuONO (392.46 mg, 3.81 mmol, 452.66 μL) at 20 °C, and the resulting mixture was stirred at 60 °C for 60 h. LCMS confirmed that 65% of 3-(2,5-difluoro-4-(4-(2-hydroxy-2-(piperidin-4-yl)ethyl)piperazin-1-yl)phenyl)piperidine-2,6-dione remained and the peak of the target mass (23%) was obtained. t-BuONO (392.46 mg, 3.81 mmol, 452.66 μL) was further added to the mixture at 20 °C, and the resulting mixture was stirred at 60 °C for 12 h. LCMS confirmed that 62% of 3-(2,5-difluoro-4-(4-(2-hydroxy-2-(piperidin-4-yl)ethyl)piperazin-1-yl)phenyl)piperidine-2,6-dione remained and the peak of the target mass (28%) was obtained. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Phenomenex luna C18 150*25 mm* 10um; mobile phase: [H2O (0.1% TFA) -ACN]; gradient: 8%-38% B over 15 min) and lyophilized to obtain the title compound (0.3 g, 631.59 μmol, 49.79% yield, 98% purity) as a yellow solid. MS (M+H) + = 466.2.

[0409] Step 5. Synthesis of 3-(4-(4-(2-(1-aminopiperidin-4-yl)-2-hydroxyethyl)piperazin-1-yl)-2,5-difluorophenyl)piperidine-2,6-dione (7)

[0410] To a solution of 3-(4-(4-(2-(1-aminopiperidin-4-yl)-2-hydroxy)piperazin-1-yl)-2,5-difluorophenyl)piperidine-2,6-dione (250 mg, 537.07 μmol) in THF (3 mL) and water (1 mL) was added NH4Cl (114.91 mg, 2.15 mmol) at 20 °C. Zn (280.95 mg, 4.30 mmol) was added at 20 °C, and the resulting mixture was stirred at 20 °C for 1 h. LCMS confirmed complete consumption of the starting material and the peak of the target mass (88%). The reaction mixture was filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Phenomenex Luna C18 150*25 mm*10um; mobile phase: [H2O (0.1% TFA) -ACN]; gradient: 0%-10% B over 15 min) and lyophilized to obtain the title compound (258 mg, 456.21 μmol, 84.94% yield, TFA salt) as a yellow solid. MS (M+H) + = 452.2.

[0411] Step 6. Synthesis of N-(4-((2-((4-((4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-2,5-difluorophenyl) piperazin-1-yl)-1-hydroxyethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 15)

[0412] The title compound (25.2 mg, 21.54 μmol, 12.18% yield, 96% purity, 2TFA salt) was obtained as a white solid by a similar method to step 9 of Example 12. MS (M+H) + = 895.3.

[0413] 1H NMR (400 MHz, DMSO-d6) δ = 10.88 (s, 1H), 10.44 - 10.39 (m, 1H), 9.70 - 9.37 (m, 2H), 8.18 - 8.12 (m, 1H), 7.80 - 7.66 (m, 9H), 7.63 - 7.56 (m, 1H), 7.42 - 7.16 (m, 4H), 7.05 - 6.95 (m, 1H), 3.99 (br dd,J= 4.7, 12.3 Hz, 1H), 3.80 - 3.72 (m, 1H), 3.62 - 3.47 (m, 4H), 3.36 - 3.21 (m, 4H), 3.19 - 3.05 (m, 4H), 2.81 - 2.69 (m, 3H), 2.60 - 2.55 (m, 2H), 2.27 - 2.16 (m, 1H), 2.00 - 1.91 (m, 1H), 1.85 - 1.73 (m, 1H), 1.63 - 1.59 (m, 1H), 1.54 - 1.17 (m, 3H).

[0414]

[0415] Example 16. Synthesis of N-(4-((2-((4-((4-(2-(4-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluoro-5-methoxyphenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 16)

[0416]

[0417] Step 1. Synthesis of 3-((4-(4-(2-(1-aminopiperidin-4-yl)ethyl)piperazin-1-yl)-5-fluoro-2-methoxyphenyl)amino)piperidine-2,6-dione (2)

[0418] The title compound (880 mg, crude, 2TFA) as a yellow oil was obtained by a similar method to step 8 of Example 12 and was used directly in the following reaction. MS(M+H) + =463.3.

[0419] Step 2. Synthesis of N-(4-((2-((4-((4-(2-(4-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluoro-5-methoxyphenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 16)

[0420] The title compound (45.8 mg, 48.53 μmol, 18% yield, 96% purity) was obtained as a white solid by a similar method to step 9 of Example 12. MS (M+H) + = 906.4.

[0421] 1 H NMR (400 MHz, DMSO-d6) δ = 10.84 (br s, 1H), 10.42 (s, 1H), 9.47 (s, 1H), 9.44 (s, 1H), 9.08 (s, 1H), 8.14 (d,J= 3.6 Hz, 1H), 7.79 - 7.64 (m, 10H), 7.61 - 7.56 (m, 1H), 7.38 - 7.32 (m, 2H), 6.61 (d,J= 8.4 Hz, 1H), 6.54 (d,J= 14.4 Hz, 1H), 5.08 (d,J= 6.8 Hz, 1H), 4.28 - 4.22 (m, 1H), 3.79 (s, 3H), 2.98 - 2.91 (m, 6H), 2.85 - 2.75 (m, 1H), 2.71 - 2.67 (m, 2H), 2.56 - 2.52 (m, 3H), 2.36 - 2.33 (m, 3H), 2.15 - 2.12 (m, 1H), 1.96 - 1.86 (m, 1H), 1.75 - 1.65 (m, 2H), 1.46 - 1.38 (m, 2H), 1.36 - 1.20 (m, 3H).

[0422]

[0423] Example 17. Synthesis of N-(4-(2-(4-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluoro-5-methoxyphenyl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 17)

[0424]

[0425] The title compound (67.6 mg, 70.97 μmol, 28% yield, 97% purity) was obtained as a white solid by a similar method to step 9 of Example 12. MS (M+H) + =924.4.

[0426] 1 H NMR (400 MHz, DMSO-d6) δ = 10.84 (br s, 1H), 10.42 (s, 1H), 9.65 (s, 1H), 9.54 - 9.46 (m, 1H), 8.87 (s, 1H), 8.17 (d,J= 3.6 Hz, 1H), 7.78 (br d,J= 14.4 Hz, 1H), 7.76 - 7.72 (m, 4H), 7.68 (br t,J= 7.2 Hz, 1H), 7.61 - 7.56 (m, 1H), 7.42 - 7.32 (m, 4H), 6.60 (d,J= 8.4 Hz, 1H), 6.54 (br d,J= 14.4 Hz, 1H), 5.08 (br d,J= 6.8 Hz, 1H), 4.28 - 4.22 (m, 1H), 3.78 (s, 3H), 3.31 (s, 1H), 2.98 (br d,J= 9.2 Hz, 2H), 2.90 - 2.76 (m, 5H), 2.62 - 2.55 (m, 4H), 2.43 - 2.23 (m, 4H), 2.14 - 2.11 (m, 1H), 1.96 - 1.86 (m, 1H), 1.74 - 1.53 (m, 2H), 1.42 - 1.35 (m, 2H), 1.25 - 1.38 (m, 3H).

[0427]

[0428] Example 18. Synthesis of N-(4-((2-((4-((4-(2-(4-(4-((2,6-dioxopiperidin-3-yl)amino)-2,5-difluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 18)

[0429]

[0430] Step 1. Synthesis of tert-butyl 4-(4-((2,6-dioxopiperidin-3-yl)amino)-2,5-difluorophenyl)piperazine-1-carboxylate (2)

[0431] To a solution of tert-butyl 4-(4-amino-2,5-difluorophenyl)piperazine-1-carboxylate (3 g, 9.57 mmol) and 3-bromopiperidine-2,6-dione (3.68 g, 19.15 mmol) in ACN (20 mL) was added NaHCO3 (4.02 g, 47.87 mmol) at 20 °C, and the resulting mixture was stirred at 80 °C for 36 h. LCMS confirmed that half of the starting material remained. TBAI (1.06 g, 2.87 mmol) was added, and the resulting mixture was stirred at 80 °C for 16 h. LCMS confirmed that the reaction had not progressed. The reaction mixture was filtered and washed with ACN (10 mL) and water (15 mL). LCMS confirmed that a small amount of product was formed in the filtrate. The filter cake was collected and dried to give the title compound as a black solid (3 g, 6.36 mmol, 66.44% yield, 90% purity). MS(M+H) + = 425.1.

[0432] Step 2. Synthesis of 3-((2,5-difluoro-4-(piperazin-1-yl)phenyl)amino)piperidine-2,6-dione (3)

[0433] To a solution of tert-butyl 4-(4-((2,6-dioxopiperidin-3-yl)amino)-2,5-difluorophenyl)piperazine-1-carboxylate (1 g, 2.36 mmol) in DCM (5 mL) was added HCl / dioxane (2 M, 10 mL) at 20 °C, and the resulting mixture was stirred at 20 °C for 2 h. The peak of the desired mass was confirmed by LCMS. The reaction mixture was concentrated to obtain the title compound (1 g, crude, 2HCl) as a black solid. MS (M+H) + = 325.2.

[0434] Step 3. Synthesis of tert-butyl (4-(2-(4-(4-((2,6-dioxopiperidin-3-yl)amino)-2,5-difluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamate (5)

[0435] To a solution of 3-((2,5-difluoro-4-(piperazin-1-yl)phenyl)amino)piperidine-2,6-dione (400 mg, 1.01 mmol, 2HCl) and tert-butyl N-(4-(2-oxoethyl)-1-piperidyl)carbamate (450.45 mg, 1.21 mmol) in DCM (10 mL) was added TEA (305.67 mg, 3.02 mmol), and the mixture was stirred at 20 °C for 1 h. NaBH(OAc)3 (426.82 mg, 2.01 mmol) was added, and the resulting mixture was stirred at 20 °C for 1 h. The peak of the target mass was confirmed by LCMS. The reaction was quenched with NaHCO3 solution (10 mL) and extracted with DCM (10 mL x 2). The mixed organic layer was dried over Na2SO4 and concentrated. The crude product was purified by flash silica gel chromatography (4 g silica gel column, eluent 80% EtOAc / petroleum ether) to obtain the title compound (150 mg, 258.79 μmol, 25.70% yield, 95% purity) as a blue solid. MS (M+H) + = 551.3.

[0436] Step 4. Synthesis of 3-((4-(4-(2-(1-aminopiperidin-4-yl)ethyl)piperazin-1-yl)-2,5-difluorophenyl)amino)piperidine-2,6-dione (6)

[0437] The title compound (150 mg, crude, 2HCl) was obtained as a yellow solid by a similar method to step 4 of Example 9, and was used directly in the following reaction. MS(M+H) + = 451.3.

[0438] Step 5. Synthesis of N-(4-((2-((4-((4-(2-(4-(4-((2,6-dioxopiperidin-3-yl)amino)-2,5-difluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 18)

[0439] The title compound (2.6 mg, 2.53 μmol, 1.02% yield, 98% purity, TFA) was obtained as a white solid by a similar method to step 8 of Example 12. MS (M+H) + = 894.6.

[0440] 1H NMR (400 MHz, DMSO-d6) δ = 10.79 (s, 1H), 10.40 (s, 1H), 9.47 (br d,J= 15.0 Hz, 2H), 9.40 - 9.17 (m, 2H), 8.14 (d,J= 3.6 Hz, 1H), 7.80 - 7.65 (m, 8H), 7.62 - 7.54 (m, 1H), 7.40 - 7.30 (m, 2H), 7.03 - 6.92 (m, 1H), 6.78 (dd,J= 8.3, 14.4 Hz, 1H), 5.72 - 5.52 (m, 1H), 4.41 - 4.34 (m, 1H), 3.38 - 3.29 (m, 3H), 3.27 - 3.10 (m, 5H), 3.10 - 3.02 (m, 2H), 3.01 - 2.92 (m, 2H), 2.78 - 2.69 (m, 3H), 2.62 - 2.58 (m, 1H), 2.11 - 1.98 (m, 2H), 1.78 - 1.67 (m, 2H), 1.67 - 1.56 (m, 2H), 1.40 - 1.23 (m, 3H).

[0441]

[0442] Example 19. Synthesis of N-(4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-3-fluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 19)

[0443]

[0444] Step 1. Synthesis of tert-butyl 4-(4-bromo-3-fluorophenyl)piperazine-1-carboxylate (2)

[0445] The title compound (4 g, 10.02 mmol, 60.31% yield, 90% purity) was obtained as a yellow solid by a similar method to step 1 of Example 9. MS (M+H) + = 359.0.

[0446] Step 2. Synthesis of tert-butyl 4-(4-(2,6-bis(benzyloxy)pyridin-3-yl)-3-fluorophenyl)piperazine-1-carboxylate (3)

[0447] The title compound (5 g, 8.34 mmol, 95.09% yield, 95% purity) was obtained as a yellow solid by a similar method to step 2 of Example 9. MS (M+H) + = 570.2.

[0448] Step 3. Synthesis of tert-butyl 4-(4-(2,6-dioxopiperidin-3-yl)-3-fluorophenyl)piperazine-1-carboxylate (4)

[0449] The title compound (3.5 g, 8.49 mmol, 96.78% yield, 95% purity) was obtained as a white solid by a similar method to step 3 of Example 9. MS (M+H) + = 392.1.

[0450] Step 4. Synthesis of 3-(2-fluoro-4-(piperazin-1-yl)phenyl)piperidine-2,6-dione (5)

[0451] The title compound (3.5 g, crude, 2HCl) was obtained as a white solid by a similar method to step 4 of Example 9. MS (M+H) + = 292.1.

[0452] Step 5. Synthesis of N-(4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-3-fluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 19)

[0453] The title compound (130.61 mg, 103.76 μmol, 76.00% yield, 97% purity, 3TFA) was obtained as a white solid by a similar method to step 4 of Example 6. MS (M+H) + = 879.3.

[0454] 1H NMR (400 MHz, DMSO-d6) δ = 10.84 (s, 1H), 10.43 (s, 1H), 9.73 (s, 1H), 9.55 (br s, 1H), 9.48 - 9.26 (m, 1H), 8.18 (d,J= 3.5 Hz, 1H), 7.81 (br d,J= 14.1 Hz, 1H), 7.76 - 7.72 (m, 4H), 7.71 - 7.66 (m, 1H), 7.62 - 7.56 (m, 1H), 7.48 - 7.41 (m, 2H), 7.39 - 7.32 (m, 2H), 7.16 (br t,J= 8.6 Hz, 1H), 6.87 (br d,J= 13.6 Hz, 1H), 6.81 (br d,J= 7.9 Hz, 1H), 3.98 - 3.80 (m, 3H), 3.62 - 3.48 (m, 2H), 3.26 - 2.94 (m, 8H), 2.80 - 2.64 (m, 3H), 2.21 - 2.12 (m, 1H), 1.99 - 1.90 (m, 1H), 1.81 - 1.70 (m, 2H), 1.69 - 1.61 (m, 2H), 1.60 - 1.45 (m, 1H), 1.43 - 1.29 (m, 3H).

[0455]

[0456] Example 20. Synthesis of N-(4-((2-((4-((4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-3-fluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 20)

[0457]

[0458] The title compound (66.63 mg, 57.52 μmol, 13.97% yield, 94% purity, 2TFA) was obtained as a pink solid by a similar method to step 4 of Example 6. MS (M+H) + = 861.3.

[0459] 1H NMR (400 MHz, DMSO-d6) δ = 10.83 (s, 1H), 10.42 (s, 1H), 9.57 - 9.48 (m, 3H), 8.15 (d,J= 3.6 Hz, 1H), 7.80 - 7.75 (m, 4H), 7.75 - 7.65 (m, 5H), 7.62 - 7.56 (m, 1H), 7.40 - 7.32 (m, 2H), 7.17 (t,J= 8.8 Hz, 1H), 6.88 (dd,J= 1.8, 14.1 Hz, 1H), 6.83 - 6.79 (m, 1H), 3.94 - 3.88 (m, 3H), 3.63 - 3.56 (m, 2H), 3.22 - 3.08 (m, 6H), 3.05 - 2.95 (m, 2H), 2.85 - 2.76 (m, 2H), 2.75 - 2.68 (m, 1H), 2.20 - 2.11 (m, 1H), 1.98 - 1.92 (m, 1H), 1.81 - 1.52 (m, 5H), 1.42 - 1.30 (m, 3H).

[0460]

[0461] Example 21. Synthesis of N-(4-((2-((4-((4-(2-(4-(4-((2,6-dioxopiperidin-3-yl)amino)-3-fluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)picolinamide (Compound 21)

[0462]

[0463] The title compound (88.8 mg, 70.24 μmol, 28.38% yield, 95% purity, 3TFA salt) was obtained as a gray solid by a similar method to step 9 of Example 12. MS (M+H) + = 859.4.

[0464] 1H NMR (400 MHz, DMSO-d6) δ = 10.81 (s, 1H), 10.68 (s, 1H), 9.84 - 9.71 (m, 1H), 9.66 - 9.42 (m, 3H), 8.76 (d,J= 4.5 Hz, 1H), 8.22 - 8.15 (m, 2H), 8.12 - 8.05 (m, 1H), 7.92 (d,J= 8.9 Hz, 2H), 7.85 - 7.75 (m, 4H), 7.73 - 7.68 (m, 3H), 6.92 - 6.84 (m, 1H), 6.81 - 6.75 (m, 1H), 6.69 - 6.63 (m, 1H), 4.31 (br dd,J= 4.8, 11.7 Hz, 1H), 3.69 - 3.51 (m, 4H), 3.29 - 3.07 (m, 7H), 2.93 - 2.81 (m, 4H), 2.80 - 2.66 (m, 1H), 2.14 - 1.90 (m, 2H), 1.83 - 1.71 (m, 2H), 1.70 - 1.56 (m, 2H), 1.49 - 1.30 (m, 3H).

[0465]

[0466] Example 22. Synthesis of N-(4-((2-((4-((4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-2-fluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 22)

[0467]

[0468] The title compound (52.50 mg, 44.35 μmol, 10.77% yield, 92% purity, 2TFA) was obtained as a pink solid by a similar method to step 4 of Example 6. MS (M+H) + = 861.3.

[0469] 1H NMR (400 MHz, DMSO-d6) δ = 10.84 (s, 1H), 10.43 (s, 1H), 9.60 - 9.50 (m, 2H), 8.16 (d,J= 3.8 Hz, 1H), 7.80 - 7.76 (m, 4H), 7.75 - 7.65 (m, 6H), 7.62 - 7.56 (m, 1H), 7.40 - 7.32 (m, 2H), 7.13 - 7.01 (m, 3H), 3.84 (dd,J= 4.9, 11.9 Hz, 1H), 3.61 (br d,J= 10.6 Hz, 2H), 3.52 (br d,J= 12.4 Hz, 2H), 3.26 - 3.11 (m, 6H), 3.07 - 2.97 (m, 2H), 2.88 - 2.76 (m, 2H), 2.69 - 2.62 (m, 1H), 2.26 - 2.18 (m, 1H), 2.03 - 1.96 (m, 1H), 1.86 - 1.51 (m, 5H), 1.45 - 1.30 (m, 3H).

[0470]

[0471] Example 23. Synthesis of N-(4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-2,6-difluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 23)

[0472]

[0473] Step 1. Synthesis of tert-butyl 4-(4-bromo-2,6-difluorophenyl)piperazine-1-carboxylate (2)

[0474] The title compound (6.12 g, 15.57 mmol, 49% yield, 96% purity) was obtained as a light yellow solid by a similar method to step 1 of Example 9.

[0475] Step 2. Synthesis of tert-butyl 4-(4-(2,6-bis(benzyloxy)pyridin-3-yl)-2,6-difluorophenyl)piperazine-1-carboxylate (4)

[0476] The title compound (8.48 g, 13.85 mmol, 85% yield, 96% purity) as a green oil was obtained by a similar method as step 2 of Example 9. MS(M+H) + = 588.3.

[0477] Step 3. Synthesis of tert-butyl 4-(4-(2,6-dioxopiperidin-3-yl)-2,6-difluorophenyl)piperazine-1-carboxylate (5)

[0478] The title compound (1.42 g, 3.47 mmol, 24% yield, 100% purity) was obtained as a gray solid by a similar method to step 3 of Example 9. MS (M+H) + = 410.1.

[0479] Step 4. Synthesis of 3-(3,5-difluoro-4-(piperazin-1-yl)phenyl)piperidine-2,6-dione (6)

[0480] The title compound (1.2 g, 3.47 mmol, HCl salt) was obtained as a gray solid by a similar method to step 4 of Example 9. MS (M+H) + = 310.2.

[0481] Step 5. Synthesis of N-(4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-2,6-difluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 23)

[0482] The title compound (83.25 mg, 70.30 μmol, 21% yield, 95% purity, 2TFA) was obtained as a white solid by a similar method to step 4 of Example 6. MS (M+H) + = 897.4.

[0483] 1H NMR (400 MHz, DMSO-d6) δ = 10.88 (s, 1H), 10.43 (s, 1H), 9.71 (s, 1H), 9.58 - 9.49 (m, 2H), 9.20 (br s, 1H), 8.18 (d,J= 3.6 Hz, 1H), 7.83 - 7.67 (m, 5H), 7.62 - 7.56 (m, 1H), 7.44 (br d,J= 4.0 Hz, 2H), 7.38 - 7.32 (m, 2H), 7.03 (br d,J= 10.4 Hz, 2H), 3.87 (br dd,J= 12.4, 4.8 Hz, 1H), 3.58 - 3.47 (m, 2H), 3.43 - 3.26 (m, 4H), 3.20 - 2.86 (m, 6H), 2.71 - 2.54 (m, 4H), 2.33 - 2.20 (m, 1H), 2.00 - 1.96 (m, 1H), 1.71 - 1.62 (m, 4H), 1.40 - 1.30 (m, 3H).

[0484]

[0485] Example 24. Synthesis of N-(4-((2-((4-((4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-2,6-difluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 24)

[0486]

[0487] The title compound (85.09 mg, 76.10 μmol, 26% yield, 99% purity, 2TFA) was obtained as a white solid by a similar method to step 4 of Example 6. MS (M+H) + = 879.4.

[0488] 1H NMR (400 MHz, DMSO-d6) δ = 10.88 (s, 1H), 10.44 (s, 1H), 9.77 (br s, 1H), 9.64 - 9.61 (m, 2H), 9.54 (s, 1H), 8.17 (d,J= 4.0 Hz, 1H), 7.79 - 7.68 (m, 8H), 7.62 - 7.57 (m, 1H), 7.39 - 7.33 (m, 2H), 7.03 (br d,J= 10.4 Hz, 2H), 3.88 (br dd,J= 12.4, 4.8 Hz, 1H), 3.56 (br d,J=10.4 Hz, 2H), 3.43 - 3.37 (m, 4H), 3.20 - 3.14 (m, 6H), 2.95 - 2.85 (m, 2H), 2.71 - 2.62 (m, 1H), 2.55 - 2.53 (m, 1H), 2.33 - 2.20 (m, 1H), 2.00 - 1.96 (m, 1H), 1.85 - 1.75 (m, 2H), 1.70 - 1.55 (m, 2H), 1.45 - 1.30 (m, 3H).

[0489]

[0490] Example 25. Synthesis of N-(4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-3,5-difluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 25)

[0491]

[0492] Step 1. Synthesis of tert-butyl 4-(4-bromo-3,5-difluorophenyl)piperazine-1-carboxylate (2)

[0493] The title compound (12.5 g, 25.85 mmol, 82.42% yield, 78% purity) was obtained as a yellow solid by a similar method to step 1 of Example 9.

[0494] Step 2. Synthesis of tert-butyl 4-(4-(2,6-bis(benzyloxy)pyridin-3-yl)-3,5-difluorophenyl)piperazine-1-carboxylate (4)

[0495] The title compound (14.45 g, 19.67 mmol, 76.11% yield, 80% purity) was obtained as a yellow solid by a similar method to step 2 of Example 9.

[0496] Step 3. Synthesis of tert-butyl 4-(4-(2,6-dioxopiperidin-3-yl)-3,5-difluorophenyl)piperazine-1-carboxylate (5)

[0497] The title compound (6 g, 11.58 mmol, 58.85% yield, 79% purity) was obtained as a gray solid by a similar method to step 3 of Example 9. MS (M+H) + = 410.1.

[0498] Step 4. Synthesis of 3-(2,6-difluoro-4-(piperazin-1-yl)phenyl)piperidine-2,6-dione (6)

[0499] The title compound (5 g, crude, HCl) was obtained as a gray solid by a similar method to step 4 of Example 9. MS (M+H) + = 310.1.

[0500] Step 5. Synthesis of N-(4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-3,5-difluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 25)

[0501] The title compound (33.91 mg, 25.94 μmol, 7.83% yield, 94.78% purity, 3TFA) was obtained as a white solid by a similar method to step 4 of Example 6. MS (M+H) + = 897.3.

[0502] 1H NMR (400 MHz, DMSO-d6) δ = 10.90 (s, 1H), 10.42 (s, 1H), 9.67 (s, 1H), 9.51 (s, 1H), 9.47 - 9.39 (m, 1H), 9.05 (br s, 1H), 8.17 (d,J= 3.5 Hz, 1H), 7.80 (br d,J= 14.6 Hz, 1H), 7.77 - 7.72 (m, 3H), 7.71 - 7.66 (m, 1H), 7.62 - 7.55 (m, 1H), 7.46 - 7.39 (m, 2H), 7.38 - 7.31 (m, 2H), 6.79 (br d,J= 12.5 Hz, 2H), 4.09 (br d,J= 6.9 Hz, 1H), 3.96 (br d,J= 10.9 Hz, 2H), 3.61 - 3.53 (m, 2H), 3.24 - 3.15 (m, 2H), 3.14 - 2.98 (m, 7H), 2.66 - 2.57 (m, 3H), 2.15 - 2.04 (m, 1H), 2.00 - 1.91 (m, 1H), 1.75 - 1.55 (m, 4H), 1.43 - 1.21 (m, 3H).

[0503]

[0504] Example 26. Synthesis of N-(4-((2-((4-((4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-3,5-difluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 26)

[0505]

[0506] Step 1. Synthesis of tert-butyl 4-(4-bromo-3,5-difluorophenyl)piperazine-1-carboxylate (2)

[0507] The title compound (12.5 g, 25.85 mmol, 82.42% yield, 78% purity) was obtained as a yellow solid by a similar method to step 1 of Example 9. MS (M+H) + = 377.0.

[0508] Step 2. Synthesis of tert-butyl 4-(4-(2,6-bis(benzyloxy)pyridin-3-yl)-3,5-difluorophenyl)piperazine-1-carboxylate (4)

[0509] The title compound (14.45 g, 19.67 mmol, 76.11% yield, 80% purity) was obtained as a yellow solid by a similar method to step 2 of Example 9. MS (M+H) + = 588.2.

[0510] Step 3. Synthesis of tert-butyl 4-(4-(2,6-dioxopiperidin-3-yl)-3,5-difluorophenyl)piperazine-1-carboxylate (5)

[0511] The title compound (6 g, 11.58 mmol, 58.85% yield, 79% purity) was obtained as a gray solid by a similar method to step 3 of Example 9. MS (M+H) + = 410.1.

[0512] Step 4. Synthesis of 3-(2,6-difluoro-4-(piperazin-1-yl)phenyl)piperidine-2,6-dione (6)

[0513] The title compound (5 g, crude, HCl salt) was obtained as a gray solid by a similar method to step 4 of Example 9. MS (M+H) + = 310.2.

[0514] Step 5. Synthesis of N-(4-((2-((4-((4-(2-(4-(4-(2,6-dioxopiperidin-3-yl)-3,5-difluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 26)

[0515] The title compound (84.75 mg, 74.26 μmol, 25.58% yield, 97% purity, 2TFA) was obtained as a white solid by a similar method to step 4 of Example 6. MS (M+H) + = 879.3.

[0516] 1 H NMR (400 MHz, DMSO-d6) δ = 10.90 (s, 1H), 10.43 (s, 1H), 9.66 (br d,J= 4.4 Hz, 1H), 9.57 (s, 1H), 9.51 (s, 1H), 8.16 (d,J= 3.6 Hz, 1H), 7.79 - 7.67 (m, 9H), 7.62 - 7.56 (m, 1H), 7.39 - 7.33 (m, 2H), 6.79 (br d,J= 12.4 Hz, 2H), 4.09 (dd,J= 12.4, 4.8 Hz, 1H), 3.96 (br d,J= 9.2 Hz, 2H), 3.59 - 3.57 (m, 2H), 3.18 - 3.06 (m, 8H), 2.83 - 2.73 (m, 3H), 2.54 - 2.52 (m, 1H), 2.16 - 2.03 (m, 1H), 1.97 - 1.95 (m, 1H), 1.75 - 1.72 (m, 2H), 1.66 - 1.61 (m, 2H), 1.41 - 1.32 (m, 3H).

[0517]

[0518] Example 27. Synthesis of N-(4-((2-((4-((4-(2-(4-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 27)

[0519]

[0520] The title compound (64.59 mg, 53.24 μmol, 20.79% yield, 91% purity, 2TFA salt) was obtained as a yellowish-white solid by a similar method to step 4 of Example 6. MS (M+H) + = 876.5.

[0521] 1H NMR (400 MHz, DMSO-d6) δ = 10.79 (s, 1H), 10.44 (s, 1H), 9.79 - 9.37 (m, 4H), 8.17 (d,J= 3.6 Hz, 1H), 7.84 - 7.66 (m, 9H), 7.64 - 7.56 (m, 1H), 7.40 - 7.32 (m, 2H), 6.90 (t,J= 9.4 Hz, 1H), 6.55 (dd,J= 2.4, 14.9 Hz, 1H), 6.46 (dd,J= 2.1, 8.7 Hz, 1H), 4.29 (br dd,J= 4.8, 11.6 Hz, 1H), 3.57 (br d,J= 10.8 Hz, 2H), 3.34 - 3.12 (m, 8H), 3.01 - 2.91 (m, 2H), 2.89 - 2.79 (m, 2H), 2.79 - 2.68 (m, 1H), 2.63 - 2.54 (m, 1H), 2.12 - 2.03 (m, 1H), 1.94 - 1.81 (m, 1H), 1.79 - 1.71 (m, 2H), 1.65 - 1.61 (m, 2H), 1.47 - 1.31 (m, 3H).

[0522]

[0523] Example 28. Synthesis of N-(4-((2-((4-((4-(2-(4-(4-((2,6-dioxopiperidin-3-yl)amino)-3-fluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 28)

[0524]

[0525] Step 1. Synthesis of 3-((4-(4-(2-(1-aminopiperidin-4-yl)ethyl)piperazin-1-yl)-2-fluorophenyl)amino) piperidine-2,6-dione (2)

[0526] The title compound (600 mg, crude, 2TFA) as a blue oil was obtained by a similar method to step 1 of Example 16 and was used directly in the following reaction. MS(M+H) += 432.4.

[0527] Step 2. Synthesis of N-(4-((2-((4-((4-(2-(4-(4-((2,6-dioxopiperidin-3-yl)amino)-3-fluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 28)

[0528] The title compound (37.75 mg, 27.89 μmol, 7.15% yield, 90% purity, 3TFA) was obtained as a brown solid by a similar method to step 2 of Example 16. MS (M+H) + = 876.4.

[0529] 1 H NMR (400 MHz, DMSO-d6) δ = 10.81 (s, 1H), 10.44 (s, 1H), 9.75 (br s, 1H), 9.61 (s, 1H), 9.55 - 9.49 (m, 2H), 8.16 (d,J= 3.6 Hz, 1H), 7.79 - 7.68 (m, 9H), 7.62 - 7.57 (m, 1H), 7.39 - 7.33 (m, 2H), 6.87 (dd,J= 14.4, 2.4 Hz, 1H), 6.81 - 6.76 (m, 1H), 6.66 (dd,J= 8.4, 1.6 Hz, 1H), 4.31 (dd,J= 12.0, 4.8 Hz, 1H), 3.71 - 3.58 (m, 4H), 3.19 - 3.12 (m, 6H), 2.89 - 2.86 (m, 4H), 2.80 - 2.71 (m, 1H), 2.59 - 2.54 (m, 1H), 2.09 - 2.04 (m, 1H), 2.01 - 1.95 (m, 1H), 1.76 - 1.74 (m, 2H), 1.68 - 1.62 (m, 2H), 1.45 - 1.37 (m, 3H).

[0530]

[0531] Example 29. Synthesis of N-(4-(2-(4-(4-((2,6-dioxopiperidin-3-yl)amino)-3-fluorophenyl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 29)

[0532]

[0533] The title compound (70.5 mg, 60.32 μmol, 16.07% yield, 96% purity, 2TFA) was obtained as a light brown solid by a similar method to step 2 of Example 16. MS (M+H) + = 894.3.

[0534] 1 H NMR (400 MHz, DMSO-d6) δ = 10.82 (s, 1H), 10.45 (s, 1H), 9.73 - 9.63 (m, 1H), 9.56 (s, 1H), 9.48 (br s, 1H), 9.25 - 8.91 (m, 1H), 8.19 (d,J= 4.0 Hz, 1H), 7.82 (br d,J= 14.0 Hz, 1H), 7.76 - 7.73 (m, 4H), 7.73 - 7.67 (m, 1H), 7.63 - 7.57 (m, 1H), 7.47 - 7.44 (m, 2H), 7.39 - 7.33 (m, 2H), 6.87 (dd,J= 14.4, 2.4 Hz, 1H), 6.81 - 6.77 (m, 1H), 6.67 (dd,J= 8.4, 1.2 Hz, 1H), 4.32 (br dd,J= 11.6, 4.8 Hz, 1H), 3.76 - 3.62 (m, 4H), 3.19 - 3.08 (m, 6H), 2.89 - 2.75 (m, 3H), 2.73 - 2.67 (m, 2H), 2.60 - 2.54 (m, 1H), 2.10 - 2.05 (m, 1H), 2.02 - 1.94 (m, 1H), 1.76 - 1.71 (m, 2H), 1.68 - 1.61 (m, 2H), 1.38 - 1.30 (m, 3H).

[0535]

[0536] Example 30. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-((2,6-dioxopiperidin-3-yl)amino)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 30)

[0537]

[0538] Step 1. Synthesis of tert-butyl 4-(4-fluoropyridin-2-yl)piperazine-1-carboxylate (2)

[0539] To a solution of 2-bromo-4-fluoro-pyridine (5 g, 28.41 mmol), tert-butyl piperazine-1-carboxylate (5.29 g, 28.41 mmol), and Cs2CO3 (18.51 g, 56.82 mmol) in dioxane (80 mL) was added RuPhos Pd G4 (1.21 g, 1.42 mmol) under N2 at 20 °C. The resulting mixture was stirred under N2 at 90 °C for 2 h. LCMS confirmed complete consumption of the starting material and the appearance of a peak of the target mass. The reaction mixture was filtered and washed with EtOAc (50 mL). The filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by flash silica gel chromatography (40 g silica gel column, eluent 0-10% EtOAc / HEX) to give the title compound (2.5 g, 8.53 mmol, 30.03% yield, 96% purity) as a light yellow solid.

[0540] Step 2. Synthesis of tert-butyl 4-(4-fluoro-5-iodopyridin-2-yl)piperazine-1-carboxylate (3)

[0541] To a solution of tert-butyl 4-(4-fluoropyridin-2-yl)piperazine-1-carboxylate (4.8 g, 17.06 mmol) in DMF (60 mL) was added NIS (7.68 g, 34.12 mmol) at 20 °C, and the resulting mixture was stirred at 20 °C for 2 h. The major peak of the desired mass was identified by LCMS. Water (300 mL) was added to the reaction mixture, and it was extracted with EtOAc (100 mL x 3). The combined organic layer was washed with brine (100 mL), dried over Na2SO4, and concentrated. The crude product was purified by flash silica gel chromatography (40 g silica gel column, eluent 0-7% EtOAc / petroleum ether) to give the title compound (6 g, 14.14 mmol, 82.90% yield, 96% purity) as a yellow solid. MS(M+H) + = 408.1.

[0542] Step 3. Synthesis of tert-butyl 4-(5-((2,6-bis(benzyloxy)pyridin-3-yl)amino)-4-fluoropyridin-2-yl)piperazine-1-carboxylate (5)

[0543] To a solution of tert-butyl 4-(4-fluoro-5-iodopyridin-2-yl)piperazine-1-carboxylate (5 g, 12.28 mmol), 2,6-dibenzyloxypyridin-3-amine (3.39 g, 11.05 mmol), and Cs2CO3 (10.00 g, 30.70 mmol) in dioxane (80 mL) was added RuPhos Pd G4 (626.48 mg, 736.70 μmol) under N2 at 20 °C. The resulting mixture was stirred at 100 °C for 14 h. LCMS confirmed the presence of a portion of the starting material and the presence of a peak of the desired mass. The reaction mixture was filtered and washed with EtOAc (50 mL). The filtrate was concentrated to obtain the residue. The residue was purified by flash silica gel chromatography (40 g silica gel column, eluent 0-15% EtOAc / petroleum ether) to give the title compound (1.5 g, 3.50 mmol, 28.50% yield, 95% purity) as a dark brown oil.

[0544] Step 4. Synthesis of tert-butyl 4-(5-((2,6-dioxopiperidin-3-yl)amino)-4-fluoropyridin-2-yl)piperazine-1-carboxylate (6)

[0545] To a solution of Pd(OH)2 (400 mg, 20% purity) in THF (20 mL) was added a solution of tert-butyl 4-(5-((2,6-bis(benzyloxy)pyridin-3-yl)amino)-4-fluoropyridin-2-yl)piperazine-1-carboxylate (1.1 g, 1.88 mmol) in MeOH (20 mL) under N2 at 20 °C. The resulting mixture was stirred under H2 (50 psi) at 30 °C for 36 h. LCMS confirmed complete consumption of the starting material and a peak of the desired mass. The mixture was filtered and washed with THF (150 mL) and MeOH (150 mL). The filtrate was concentrated to give the crude product. The crude product was triturated with MTBE (10 mL). The mixture was filtered and washed with petroleum ether (10 mL). The filter cake was collected and dried to obtain the title compound as a black solid (500 mg, 1.10 mmol, 58.80% yield, 90% purity).

[0546] Step 5. Synthesis of 3-((4-fluoro-6-(piperazin-1-yl)pyridin-3-yl)amino)piperidine-2,6-dione (7)

[0547] The title compound (500 mg, crude, 2HCl) was obtained as a black solid by a similar method to step 2 of Example 18. MS (M+H) + = 308.1.

[0548] Step 6. Synthesis of tert-butyl (4-(2-(4-(5-((2,6-dioxopiperidin-3-yl)amino)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamate (9)

[0549] The title compound (300 mg, 460.99 μmol, 38.95% yield, 82% purity) was obtained as a black solid by a similar method to step 3 of Example 18. MS (M+H) + = 534.3.

[0550] Step 7. Synthesis of 3-((6-(4-(2-(1-aminopiperidin-4-yl)ethyl)piperazin-1-yl)-4-fluoropyridin-3-yl)amino)piperidine-2,6-dione (10)

[0551] The title compound (400 mg, crude, 2TFA) as a black oil was obtained by a similar method to step 1 of Example 16. MS (M+H) + = 434.3.

[0552] Step 8. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-((2,6-dioxopiperidin-3-yl)amino)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 30)

[0553] The title compound (15.2 mg, 13.34 μmol, 4.41% yield, 97% purity, 2TFA) was obtained as a white solid by a similar method to step 2 of Example 16. MS (M+H) + = 877.5.

[0554] 1 H NMR (400 MHz, DMSO-d6) δ = 10.80 (s, 1H), 10.43 (s, 1H), 9.59 - 9.44 (m, 3H), 8.16 (d,J= 3.7 Hz, 1H), 7.83 (d,J= 11.6 Hz, 1H), 7.80 - 7.66 (m, 9H), 7.63 - 7.56 (m, 1H), 7.41 - 7.22 (m, 3H), 6.92 (br d,J= 14.1 Hz, 1H), 4.34 (br d,J= 6.8 Hz, 1H), 4.21 - 4.12 (m, 2H), 3.63 - 3.51 (m, 2H), 3.25 - 2.94 (m, 8H), 2.85 - 2.60 (m, 3H), 2.58 - 2.52 (m, 1H), 2.10 - 1.95 (m, 2H), 1.77 - 1.56 (m, 4H), 1.56 - 1.23 (m, 3H).

[0555]

[0556] Example 31. Synthesis of N-(4-(2-(4-(5-((2,6-dioxopiperidin-3-yl)amino)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 31)

[0557]

[0558] The title compound (17.8 mg, 11.07 μmol, 3.66% yield, 84% purity, 4TFA) was obtained as a white solid by a similar method to step 2 of Example 16. MS (M+H) + = 895.6.

[0559] 1 H NMR (400 MHz, DMSO-d6) δ = 10.80 (s, 1H), 10.42 (s, 1H), 9.70 (s, 1H), 9.58 - 9.42 (m, 2H), 9.23 - 9.11 (m, 1H), 8.18 (d,J= 3.5 Hz, 1H), 7.85 - 7.78 (m, 2H), 7.77 - 7.71 (m, 4H), 7.68 (br t,J= 7.4 Hz, 1H), 7.62 - 7.56 (m, 1H), 7.47 - 7.30 (m, 4H), 6.92 (br d,J= 14.3 Hz, 1H), 4.34 (br dd,J= 4.9, 11.6 Hz, 1H), 4.23 - 4.11 (m, 2H), 3.62 - 3.46 (m, 2H), 3.21 - 3.11 (m, 2H), 3.10 - 2.92 (m, 6H), 2.78 - 2.55 (m, 3H), 2.44 - 2.37 (m, 1H), 2.10 - 1.97 (m, 2H), 1.77 - 1.56 (m, 4H), 1.50 - 1.23 (m, 3H).

[0560]

[0561] Example 32. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-((2,6-dioxopiperidin-3-yl)amino)-3-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 32)

[0562]

[0563] The title compound (7.5 mg, 5.91 μmol, 2.31% yield, 87% purity, 2TFA salt) was obtained as a yellow solid by a similar method to step 4 of Example 6. MS (M+H) + = 877.5.

[0564] 1 H NMR (400 MHz, DMSO-d6) δ = 10.83 (s, 1H), 10.44 (s, 1H), 9.90 - 9.44 (m, 3H), 8.16 (d,J= 3.6 Hz, 1H), 7.86 - 7.63 (m, 10H), 7.62 - 7.53 (m, 2H), 7.41 - 7.31 (m, 2H), 7.05 (dd,J= 2.1, 14.4 Hz, 1H), 4.32 (br dd,J= 4.8, 11.9 Hz, 1H), 3.73 - 3.62 (m, 2H), 3.57 (br d,J= 9.8 Hz, 2H), 3.26 - 2.99 (m, 8H), 2.92 - 2.79 (m, 2H), 2.77 - 2.66 (m, 1H), 2.63 - 2.54 (m, 1H), 2.15 - 2.03 (m, 1H), 1.98 - 1.83 (m, 1H), 1.82 - 1.70 (m, 2H), 1.65 - 1.61 (m, 2H), 1.50 - 1.31 (m, 3H).

[0565]

[0566] Example 33. Synthesis of N-(4-(2-(4-(5-((2,6-dioxopiperidin-3-yl)amino)-3-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 33)

[0567]

[0568] Step 1. Synthesis of tert-butyl 4-(5-((2,6-bis(benzyloxy)pyridin-3-yl)amino)-3-fluoropyridin-2-yl)piperazine-1-carboxylate (2)

[0569] The title compound (2.1 g, 3.48 mmol, 62.64% yield, 97% purity) was obtained as a black-brown oil by a similar method to step 3 of Example 30. MS (M+H) + = 586.3.

[0570] Step 2. Synthesis of tert-butyl 4-(5-((2,6-dioxopiperidin-3-yl)amino)-3-fluoropyridin-2-yl)piperazine-1-carboxylate (3)

[0571] The title compound (1.4 g, crude) was obtained as a blackish brown solid by a similar method to step 4 of Example 30. MS (M+H) + = 408.1.

[0572] Step 3. Synthesis of 3-((5-fluoro-6-(piperazin-1-yl)pyridin-3-yl)amino)piperidine-2,6-dione (4)

[0573] The title compound (1 g, crude, HCl salt) was obtained as a gray solid by a similar method to step 2 of Example 18. MS (M+H) + = 308.2.

[0574] Step 4. Synthesis of N-(4-(2-(4-(5-((2,6-dioxopiperidin-3-yl)amino)-3-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 33)

[0575] The title compound (63.06 mg, 53.91 μmol, 21.69% yield, 96% purity, 2TFA salt) was obtained as a yellow solid by a similar method to step 4 of Example 6. MS (M+H) + = 895.4.

[0576] 1 H NMR (400 MHz, DMSO-d6) δ = 10.83 (s, 1H), 10.44 (s, 1H), 9.72 (br d,J= 6.8 Hz, 1H), 9.65 - 9.43 (m, 1H), 9.31 - 9.12 (m, 1H), 8.18 (d,J= 3.5 Hz, 1H), 7.85 - 7.71 (m, 6H), 7.71 - 7.65 (m, 1H), 7.63 - 7.55 (m, 2H), 7.44 (br s, 2H), 7.39 - 7.31 (m, 2H), 7.06 (dd,J= 2.1, 14.5 Hz, 1H), 4.38 - 4.28 (m, 1H), 3.72 - 3.46 (m, 4H), 3.24 - 2.98 (m, 9H), 2.78 - 2.55 (m, 3H), 2.14 - 2.03 (m, 1H), 1.97 - 1.83 (m, 1H), 1.77 - 1.55 (m, 4H), 1.40 - 1.25 (m, 3H).

[0577]

[0578] Example 34. Synthesis of N-(4-(2-(4-(6-(2,6-dioxopiperidin-3-yl)pyridin-3-yl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 34)

[0579]

[0580] Step 1. Synthesis of tert-butyl (4-(2-(4-(6-(2,6-dioxopiperidin-3-yl)pyridin-3-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamate (3)

[0581] To a solution of 3-(5-(piperazin-1-yl)pyridin-2-yl)piperidine-2,6-dione (800 mg, 2.06 mmol, TFA) in DCM (20 mL) were added TEA (521.13 mg, 5.15 mmol) and tert-butyl (4-(2-oxoethyl)piperidin-1-yl)carbamate (383.98 mg, 1.03 mmol) at 15 °C. The mixture was stirred at 15 °C for 30 min, and NaBH(OAc)3 (654.91 mg, 3.09 mmol) was slowly added. The resulting mixture was stirred at 15 °C for 16 h. The peak 64% of the target mass was identified by LCMS. The mixture was diluted with water (30 mL) and extracted with DCM (30 mL x 2). The mixed organic layer was washed with NaHCO3 solution (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (Biotage, 4 g silica gel column, eluent 25% MeOH / EtOAc) to obtain the title compound (450 mg, 808.98 μmol, 78.54% yield, 90% purity) as a yellow solid. MS (M+H) + = 501.1.

[0582] Step 2. Synthesis of 3-(5-(4-(2-(1-aminopiperidin-4-yl)ethyl)piperazin-1-yl)pyridin-2-yl)piperidine-2,6-dione (4)

[0583] The title compound (360 mg, crude, 2TFA) was obtained as a yellow oil by a similar method to step 8 of Example 12. The product was used directly in the following reaction. MS(M+H) + = 401.3.

[0584] Step 3. Synthesis of N-(4-(2-(4-(6-(2,6-dioxopiperidin-3-yl)pyridin-3-yl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 34)

[0585] The title compound (17.38 mg, 13.57 μmol, 14.80% yield, 94% purity, 3TFA) was obtained as a white solid by a similar method to step 9 of Example 12. MS (M+H) + = 862.3.

[0586] 1 H NMR (400 MHz, DMSO-d6) δ = 10.85 (s, 1H), 10.42 (s, 1H), 9.69 (s, 1H), 9.62 - 9.49 (m, 2H), 9.13 (br s, 1H), 8.32 (d,J= 2.4 Hz, 1H), 8.18 (d,J= 3.6 Hz, 1H), 7.80 (br d,J= 14.4 Hz, 1H), 7.76 - 7.68 (m, 4H), 7.62 - 7.56 (m, 1H), 7.53 (br dd,J= 8.4, 1.6 Hz, 1H), 7.44 (br s, 2H), 7.38 - 7.32 (m, 3H), 4.03 - 3.88 (m, 3H), 3.62 (br d,J= 11.6 Hz, 2H), 3.27 - 3.01 (m, 8H), 2.70 - 2.63 (m, 2H), 2.62 - 2.55 (m, 2H), 2.29 - 2.17 (m, 1H), 2.15 - 2.05 (m, 1H), 1.79 - 1.56 (m, 4H), 1.36 - 1.26 (m, 3H).

[0587]

[0588] Example 35. Synthesis of N-(4-((2-((4-((4-(2-(4-(6-(2,6-dioxopiperidin-3-yl)pyridin-3-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 35)

[0589]

[0590] Step 1. Synthesis of tert-butyl 4-(2',6'-bis(benzyloxy)-[2,3'-bipyridin]-5-yl)piperazine-1-carboxylate (2)

[0591] The title compound (9 g, crude) was obtained as a light yellow solid by a similar method to step 2 of Example 9. MS (M+H) + = 553.4.

[0592] Step 2. Synthesis of tert-butyl 4-(6-(2,6-dioxopiperidin-3-yl)pyridin-3-yl)piperazine-1-carboxylate (3)

[0593] To a solution of Pd / C (778.32 mg, 731.37 μmol, 10% purity, wet) and Pd(OH)2 / C (500 mg, 731.37 μmol, 20% purity, wet) in THF (50 mL) were added AcOH (439.19 mg, 7.31 mmol) and tert-butyl 4-(2',6'-bis(benzyloxy)-[2,3'-bipyridin]-5-yl)piperazine-1-carboxylate (4.3 g, 7.31 mmol) under Ar. The suspension was degassed and purged three times with H2. The mixture was stirred at 30 °C under H2 (50 psi) for 48 h. The target mass peak was identified as ~62% by LCMS. The reaction mixture was filtered and washed with MeOH (200 mL). The filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography (Biotage, 40 g silica gel column, 100% EtOAc) to obtain the title compound (1.5 g, 3.53 mmol, 48.20% yield, 88% purity) as a brown solid. MS (M+H) + = 375.2.

[0594] Step 3. Synthesis of 3-(5-(piperazin-1-yl)pyridin-2-yl)piperidine-2,6-dione (4)

[0595] To a solution of tert-butyl 4-(6-(2,6-dioxopiperidin-3-yl)pyridin-3-yl)piperazine-1-carboxylate (500 mg, 1.18 mmol) in DCM (10 mL) was added TFA (6 mL, 80.77 mmol). The mixture was stirred at 15 °C for 2 h. The peak of the desired mass was identified by LCMS. The mixture was concentrated under reduced pressure to obtain the title compound (1.4 g, crude, TFA) as a brown oil. MS (M+H) + = 275.1.

[0596] Step 4. Synthesis of N-(4-((2-((4-((4-(2-(4-(6-(2,6-dioxopiperidin-3-yl)pyridin-3-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 35)

[0597] The title compound (95.58 mg, 78.17 μmol, 19.07% yield, 97% purity, 3TFA) was obtained as a white solid by a similar method to step 4 of Example 6. MS (M+H) + = 844.3.

[0598] 1 H NMR (400 MHz, DMSO-d6) δ = 10.91 (s, 1H), 10.44 (s, 1H), 10.05 - 9.98 (m, 1H), 9.96 - 9.86 (m, 1H), 9.68 (s, 1H), 9.59 (s, 1H), 8.35 (d,J= 2.0 Hz, 1H), 8.18 (d,J= 3.6 Hz, 1H), 7.80 - 7.57 (m, 10H), 7.41 (br d,J= 8.8 Hz, 1H), 7.39 - 7.33 (m, 2H), 4.04 (dd,J= 10.0, 5.2 Hz, 1H), 3.98 (br d,J= 9.8 Hz, 2H), 3.65 - 3.55 (m, 2H), 3.26 - 3.08 (m, 8H), 2.93 - 2.91 (m, 2H), 2.68 - 2.52 (m, 2H), 2.31 - 2.21 (m, 1H), 2.13 - 2.07 (m, 1H), 1.86 - 1.76 (m, 2H), 1.70 - 1.52 (m, 2H), 1.48 - 1.32 (m, 3H).

[0599]

[0600] Example 36. Synthesis of N-(4-((2-((4-((4-(2-(4-(6-(2,6-dioxopiperidin-3-yl)-5-fluoropyridin-3-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 36)

[0601]

[0602] Step 1. Synthesis of 2-bromo-3-fluoro-5-iodopyridine (2)

[0603] AmylONO (1.48 g, 12.63 mmol, 1.7 mL) was added to a solution of 3-fluoro-5-iodopyridin-2-amine (2 g, 8.40 mmol) and CuBr2 (2.25 g, 10.07 mmol, 471.71 μL) in ACN (20 mL) at 20 °C, and the mixture was stirred at 50 °C for 3 h. TLC (petroleum ether:EtOAc=3:1) confirmed complete consumption of the starting material and the formation of a new spot. The mixture was diluted with water (10 mL) and extracted with EtOAc (10 mL x 3). The combined organic layer was washed with brine (10 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (12 g SepaFlash® silica flash column, eluent 20% EtOAc / petroleum ether, gradient @ 100 mL / min) to give the title compound (0.6 g, crude) as a yellow solid.

[0604] Step 2. Synthesis of tert-butyl 4-(6-bromo-5-fluoropyridin-3-yl)piperazine-1-carboxylate (3)

[0605] The title compound (420 mg, 1.06 mmol, 53.38% yield, 91% purity) was obtained as a yellow oil by a similar method as step 1 of Example 9. MS (M+H) + =360.1

[0606] Step 3. Synthesis of tert-butyl 4-(2',6'-bis(benzyloxy)-3-fluoro-[2,3'-bipyridin]-5-yl)piperazine-1-carboxylate (4)

[0607] The title compound (270 mg, 468.41 μmol, 84.37% yield, 99% purity) was obtained as a yellow solid by a similar method to step 2 of Example 10. MS (M+H) + = 571.3.

[0608] Step 4. Synthesis of tert-butyl 4-(6-(2,6-dioxopiperidin-3-yl)-5-fluoropyridin-3-yl)piperazine-1-carboxylate (5)

[0609] The title compound (230 mg, crude) as a yellow oil was obtained by synthesis in a similar manner to step 4 of Example 13. MS (M+H) + = 393.2.

[0610] Step 5. Synthesis of 3-(3-fluoro-5-(piperazin-1-yl)pyridin-2-yl)piperidine-2,6-dione (6)

[0611] To a solution of tert-butyl 4-(6-(2,6-dioxopiperidin-3-yl)-5-fluoropyridin-3-yl)piperazine-1-carboxylate (230 mg, 586.10 μmol) in DCM (5 mL) was added HCl / dioxane (2 M, 9.58 mL), and the mixture was stirred at 20 °C for 4 h. LCMS confirmed complete consumption of the starting material. The mixture was concentrated under reduced pressure to give the title compound (0.2 g, crude, HCl salt) as a yellow solid. MS (M+H) + = 293.2.

[0612] Step 6. Synthesis of N-(4-((2-((4-((4-(2-(4-(6-(2,6-dioxopiperidin-3-yl)-5-fluoropyridin-3-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzenamide (Compound 36)

[0613] The title compound (32.28 mg, 29.32 μmol, 11.45% yield, 99% purity, 2TFA salt) was obtained as a yellow solid by a similar method to step 4 of Example 6. MS (M+H) + = 862.5.

[0614] 1 H NMR (400 MHz, MeOD) δ = 8.15 (d,J= 2.0 Hz, 1H), 8.07 (dd,J= 2.1, 4.5 Hz, 1H), 7.82 - 7.70 (m, 5H), 7.68 - 7.56 (m, 5H), 7.38 - 7.25 (m, 3H), 4.22 (dd,J= 5.1, 10.6 Hz, 1H), 4.10 - 3.91 (m, 1H), 3.79 - 3.57 (m, 2H), 3.27 - 3.10 (m, 9H), 2.80 - 2.62 (m, 4H), 2.42 - 2.28 (m, 1H), 2.24 - 2.13 (m, 1H), 1.80 - 1.69 (m, 4H), 1.61 - 1.48 (m, 2H), 1.42 - 1.28 (m, 1H).

[0615]

[0616] Example 37. Synthesis of 5-((2-((4-((4-(2-(4-(6-(2,6-dioxopiperidin-3-yl)-5-fluoropyridin-3-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)-N-(2-fluorophenyl)picolinamide (Compound 37)

[0617]

[0618] Step 1. Synthesis of tert-butyl (4-(2-(4-(6-(2,6-dioxopiperidin-3-yl)-5-fluoropyridin-3-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamate (3)

[0619] To a solution of 3-(3-fluoro-5-(piperazin-1-yl)pyridin-2-yl)piperidine-2,6-dione (0.1 g, 304.16 μmol, HCl salt) and tert-butyl (4-(2-oxoethyl)piperidin-1-yl)carbamate (0.1 g, 412.69 μmol) in DCM (2 mL) were added TEA (101.78 mg, 1.01 mmol, 140 μL) and MgSO4 (109.84 mg, 912.49 μmol), and the mixture was stirred at 20 °C for 15 min. NaBH(OAc)3 (193.39 mg, 912.49 μmol) was added, and the mixture was stirred at 20 °C for 14 h. The peak of the target mass (73%) was confirmed by LCMS. The mixture was diluted with water (10 mL) and extracted with DCM (10 mL x 2). The combined organic layer was washed with water (10 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (5 g SepaFlash® silica flash column, eluent 0–10% MeOH / EtOAc, gradient @ 50 mL / min) to give the title compound (70 mg, 134.97 μmol, 44.37% yield) as a yellow solid. MS (M+H) + = 519.4.

[0620] Step 2. Synthesis of 3-(5-(4-(2-(1-aminopiperidin-4-yl)ethyl)piperazin-1-yl)-3-fluoropyridin-2-yl)piperidine-2,6-dione (4)

[0621] The title compound (70 mg, crude, TFA salt) as a yellow oil was obtained by a similar method to step 8 of Example 12. MS (M+H) + = 419.4.

[0622] Step 3. Synthesis of 5-((2-((4-((4-(2-(4-(6-(2,6-dioxopiperidin-3-yl)-5-fluoropyridin-3-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)-N-(2-fluorophenyl)picolinamide (Compound 37)

[0623] The title compound (7.9 mg, 7.17 μmol, 5.45% yield, 99% purity, 2TFA salt) was obtained as a white solid by a similar method to step 9 of Example 12. MS (M+H) + = 863.6.

[0624] 1 H NMR (400 MHz, DMSO-d6) δ = 10.88 (s, 1H), 10.27 (s, 1H), 10.09 (s, 1H), 9.76 (s, 1H), 9.72 - 9.61 (m, 1H), 9.60 - 9.51 (m, 1H), 9.26 (d,J= 2.3 Hz, 1H), 8.57 (dd,J= 2.4, 8.5 Hz, 1H), 8.32 (d,J= 3.4 Hz, 1H), 8.24 - 8.12 (m, 2H), 7.83 - 7.68 (m, 4H), 7.47 - 7.39 (m, 1H), 7.38 - 7.30 (m, 1H), 7.29 - 7.18 (m, 2H), 4.15 (br dd,J= 5.3, 10.9 Hz, 2H), 4.02 - 3.97 (m, 1H), 3.65 - 3.54 (m, 2H), 3.28 - 3.04 (m, 8H), 2.85 - 2.68 (m, 2H), 2.58 - 2.52 (m, 1H), 2.45 - 2.38 (m, 1H), 2.30 - 2.17 (m, 1H), 2.10 - 2.01 (m, 1H), 1.79 - 1.59 (m, 4H), 1.40 - 1.30 (m, 3H).

[0625]

[0626] Example 38. Synthesis of 5-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)-N-(2-fluorophenyl)picolinamide (Compound 38)

[0627]

[0628] Step 1. Synthesis of N-(2-fluorophenyl)-5-nitropicolinamide (3)

[0629] The title compound (10 g, 38.28 mmol, 64.36% yield) was obtained as a brown solid by a similar method to step 1 of Example 7.

[0630] Step 2. Synthesis of 5-amino-N-(2-fluorophenyl)picolinamide (4)

[0631] N-(2-fluorophenyl)-5-nitropicolinamide (9 g, 34.46 mmol) was slowly added to a solution of Pd / C (1 g, 10% purity) in EtOH (200 mL) under Ar. The resulting mixture was purged and degassed three times with Ar, and then purged and degassed three times with H2. The mixture was stirred at 20 °C under H2 (50 psi) for 14 h. The peak of the target mass was confirmed by LCMS. The reaction mixture was filtered through Celite and washed with EtOH (300 mL). The filtrate was concentrated to obtain the title compound (4 g, 16.61 mmol, 48.20% yield, 96% purity) as a brown solid.

[0632] Step 3. Synthesis of 5-((2-chloro-5-fluoropyrimidin-4-yl)amino)-N-(2-fluorophenyl)picolinamide (6)

[0633] The title compound (2.7 g, 7.46 mmol, 43.15% yield) was obtained as a yellow solid by a similar method to step 3 of Example 7, and was used directly in the next reaction. MS(M+H)+ = 362.0.

[0634] Step 4. Synthesis of methyl 4-((5-fluoro-4-((6-((2-fluorophenyl)carbamoyl)pyridin-3-yl)amino)pyrimidin-2-yl)amino)benzoate (8)

[0635] To a solution of 5-((2-chloro-5-fluoropyrimidin-4-yl)amino)-N-(2-fluorophenyl)picolinamide (2.2 g, 6.08 mmol) and methyl 4-aminobenzoate (919.34 mg, 6.08 mmol) in dioxane (30 mL) was added TsOH (2.09 g, 12.16 mmol) at 20 °C, and the resulting mixture was stirred at 100 °C for 14 h. The main peak of the desired mass was identified by LCMS. After cooling to 20 °C, the mixture was filtered. The filter cake was collected and dried to give the title compound (4 g, crude) as a yellow solid, which was used directly in the next reaction. MS(M+H) + = 477.2.

[0636] Step 5. Synthesis of 4-((5-fluoro-4-((6-((2-fluorophenyl)carbamoyl)pyridin-3-yl)amino)pyrimidin-2-yl)amino)benzoic acid (9)

[0637] The title compound (3.5 g, 7.21 mmol, 98.15% yield, Na salt) was obtained as a white solid by a similar method to step 5 of Example 7, and the product was used directly in the next reaction. MS(M+H) + = 463.0.

[0638] Step 6. Synthesis of 5-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)-N-(2-fluorophenyl)picolinamide (Compound 38)

[0639] The title compound (71.7 mg, 64.41 μmol, 10.42% yield, 98% purity, 2TFA) was obtained as a white solid by a similar method to step 9 of Example 12. MS (M+H) + = 863.7.

[0640] 1 H NMR (400 MHz, DMSO-d6) δ = 10.90 (s, 1H), 10.27 (s, 1H), 10.09 (s, 1H), 9.76 (s, 1H), 9.59 - 9.52 (m, 1H), 9.26 (d,J= 2.3 Hz, 1H), 8.57 (dd,J= 2.3, 8.6 Hz, 1H), 8.32 (d,J= 3.3 Hz, 1H), 8.24 - 8.18 (m, 1H), 8.16 (d,J= 8.6 Hz, 1H), 8.08 (d,J= 10.9 Hz, 1H), 7.81 - 7.75 (m, 2H), 7.75 - 7.69 (m, 2H), 7.39 - 7.32 (m, 1H), 7.29 - 7.20 (m, 2H), 6.90 (d,J= 13.8 Hz, 1H), 4.41 (br d,J= 13.8 Hz, 2H), 3.95 (br dd,J= 4.8, 13.3 Hz, 1H), 3.63 - 3.54 (m, 2H), 3.25 - 2.95 (m, 8H), 2.89 - 2.66 (m, 3H), 2.58 - 2.52 (m, 1H), 2.23 - 2.13 (m, 1H), 2.02 - 1.94 (m, 1H), 1.78 - 1.59 (m, 4H), 1.45 - 1.28 (m, 3H).

[0641]

[0642] Example 39. Synthesis of N-(2-chlorophenyl)-5-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)picolinamide (Compound 39)

[0643]

[0644] Step 1. Synthesis of 2,6-bis(benzyloxy)-6'-chloro-4'-fluoro-3,3'-bipyridine (2)

[0645] The title compound (7.85 g, 17.91 mmol, 54.23% yield, 96% purity) was obtained as a yellow solid by a similar method to step 1 of Example 10. MS (M+H) + = 421.2.

[0646] Step 2. Synthesis of tert-butyl 4-(2',6'-bis(benzyloxy)-4-fluoro-[3,3'-bipyridin]-6-yl)piperazine- 1-carboxylate (3)

[0647] The title compound (7.4 g, 12.97 mmol, 66.56% yield, 100% purity) was obtained as a white solid by a similar method to step 1 of Example 30. MS (M+H) + = 571.3.

[0648] Step 3. tert-Butyl 4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazine-1-carboxylate (4)

[0649] To a solution of Pd(OH)2 / C (2 g, 20% purity) in MeOH (80 mL) was slowly added a solution of tert-butyl 4-(2',6'-bis(benzyloxy-4-fluoro-[3,3'-bipyridin]-6-yl)piperazine-1-carboxylate (7.4 g, 12.97 mmol) in THF (80 mL) under N2, and the mixture was stirred at 30 °C under H2 (45 Psi) for 40 h. After completion of the reaction, the peak of the target mass was confirmed by LCMS. The mixture was filtered and washed with THF (800 mL). The filtrate was concentrated under reduced pressure to obtain the title compound (3.94 g, crude) as a white solid. MS(M+H) + = 393.2.

[0650] Step 4. Synthesis of 3-(4-fluoro-6-(piperazin-1-yl)pyridin-3-yl)piperidine-2,6-dione (5)

[0651] To a solution of tert-butyl 4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazine-1-carboxylate (3.94 g, 10.04 mmol) in DCM (20 mL) was added HCl / dioxane (2 M, 100 mL), and the mixture was stirred at 15 °C for 14 h. The peak of the desired mass was confirmed by LCMS. The mixture was concentrated under reduced pressure to obtain the title compound (4.2 g, crude, HCl salt) as a white solid.

[0652] Step 5. Synthesis of tert-butyl (4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamate (6)

[0653] The title compound (2 g, 3.86 mmol, 60.37% yield) was obtained as a yellow solid by a similar method to step 1 of Example 37. MS (M+H) + = 519.4.

[0654] Step 6. Synthesis of 3-(6-(4-(2-(1-aminopiperidin-4-yl)ethyl)piperazin-1-yl)-4-fluoropyridin-3-yl)piperidine-2,6-dione (7)

[0655] The title compound (2 g, crude, TFA salt) as a yellow oil was obtained by synthesis in a similar manner to step 8 of Example 12.

[0656] Step 7. Synthesis of N-(2-chlorophenyl)-5-nitropicolinamide (9)

[0657] The title compound (10 g, crude) was obtained as a yellow solid by a similar method to step 1 of Example 7. MS (M+H) + = 278.1.

[0658] Step 8. Synthesis of 5-amino-N-(2-chlorophenyl)picolinamide (10)

[0659] To a solution of N-(2-chlorophenyl)-5-nitropicolinamide (10 g, 36.01 mmol) and NH4Cl (5.78 g, 108.04 mmol) in EtOH (100 mL) and water (30 mL) was added Fe (6.03 g, 108.04 mmol), and the mixture was stirred at 80 °C for 1 h. The peak of the target mass was identified by LCMS. The mixture was filtered and filtered with MeOH (100 mL) and THF (50 mL). The filtrate was concentrated under reduced pressure. The product was diluted with water (50 mL) and adjusted to pH = 8 with NaHCO3. The mixture was extracted with EtOAc (20 mL x 3). The combined organic layer was washed with water (30 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain the title compound (1 g, crude) as a yellow solid. The filter cake was diluted with DMF (100 mL) and stirred at 20 °C for 3 hours. The mixture was filtered and washed with DMF (50 mL). The filtrate was diluted with EtOAc (100 mL) and water (100 mL), and the mixture was adjusted to pH = 8 with NaHCO3. The mixture was extracted with EtOAc (50 mL x 2). The combined organic layer was washed with brine (50 mL x 3), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain the title compound (4.2 g, crude) as a yellow solid. MS (M+H) + = 248.1.

[0660] Step 9. Synthesis of 5-((2-chloro-5-fluoropyrimidin-4-yl)amino)-N-(2-chlorophenyl)picolinamide (11)

[0661] To a solution of 2,4-dichloro-5-fluoropyrimidine (1 g, 4.04 mmol) and 5-amino-N-(2-chlorophenyl)picolinamide (670.00 mg, 4.01 mmol) in NMP (10 mL) was added DIPEA (1.57 g, 12.11 mmol, 2.11 mL), and the mixture was stirred in a microwave at 100 °C for 1 h. LCMS confirmed the remaining starting material and a small amount of the target mass. The mixture was stirred in a microwave at 120 °C for 2 h. LCMS confirmed the peak of the target mass (28%). Water (30 mL) was slowly added to the mixture, and the suspension was filtered. The filter cake was washed with water (10 mL), and the filter cake was concentrated under reduced pressure to obtain the title compound (1.1 g, crude) as a yellow solid. MS (M+H) + = 378.1.

[0662] Step 10. Synthesis of methyl 4-((4-((6-((2-chlorophenyl)carbamoyl)pyridin-3-yl)amino)-5-fluoropyrimidin-2-yl)amino)benzoate (12)

[0663] The title compound (920 mg, crude) was obtained as a yellow solid by a similar method to step 4 of Example 7. MS (M+H) + = 493.2.

[0664] Step 11. Synthesis of 4-((4-((6-((2-chlorophenyl)carbamoyl)pyridin-3-yl)amino)-5-fluoropyrimidin-2-yl)amino)benzoic acid (13)

[0665] The title compound (80 mg, 163.72 μmol, 8.77% yield, 98% purity) was obtained as a yellow solid by a similar method to step 5 of Example 7. MS (M+H) + = 479.1.

[0666] Step 12. Synthesis of N-(2-chlorophenyl)-5-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)picolinamide (Compound 39)

[0667] The title compound (19.82 mg, 19.55 μmol, 11.70% yield, 98% purity, TFA salt) was obtained as a white solid by a similar method to step 6 of Example 1. MS (M+H) + = 879.4.

[0668] 1 H NMR (400 MHz, DMSO-d6) δ = 10.89 (s, 1H), 10.52 (s, 1H), 10.09 (s, 1H), 9.76 (s, 1H), 9.70 - 9.55 (m, 1H), 9.53 - 9.38 (m, 1H), 9.27 (d,J= 2.1 Hz, 1H), 8.57 (dd,J= 2.4, 8.6 Hz, 1H), 8.44 (d,J= 7.3 Hz, 1H), 8.32 (d,J= 3.4 Hz, 1H), 8.18 (d,J= 8.6 Hz, 1H), 8.08 (d,J=11.0 Hz, 1H), 7.85 - 7.68 (m, 3H), 7.59 (d,J= 8.0 Hz, 1H), 7.43 (t,J= 7.4 Hz, 1H), 7.27 - 7.17 (m, 1H), 6.90 (d,J= 13.6 Hz, 1H), 4.48 - 4.36 (m, 2H), 3.95 (br dd,J= 4.8, 13.2 Hz, 1H), 3.59 (br d,J= 11.4 Hz, 2H), 3.29 - 2.96 (m, 8H), 2.84 - 2.69 (m, 2H), 2.57 (br d,J= 3.1 Hz, 1H), 2.47 - 2.43 (m, 1H), 2.26 - 2.11 (m, 1H), 2.03 - 1.92 (m, 1H), 1.80 - 1.58 (m, 4H), 1.45 - 1.32 (m, 3H).

[0669]

[0670] Example 40. N-(2-chlorophenyl)-4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)-2-fluorobenzamide (Compound 40)

[0671]

[0672] Step 1. Synthesis of N-(2-chlorophenyl)-2-fluoro-4-nitrobenzamide (3)

[0673] The title compound (16 g, crude) was obtained as a yellow solid by a similar method to step 1 of Example 7. MS (M+H) + = 294.9.

[0674] Step 2. Synthesis of 4-amino-N-(2-chlorophenyl)-2-fluorobenzamide (4)

[0675] The title compound (9.4 g, 35.51 mmol, 65.41% yield) was obtained as a yellow solid by a similar method to step 8 of Example 39. MS (M+H) + = 265.1.

[0676] Step 3. Synthesis of 4-((2-chloro-5-fluoropyrimidin-4-yl)amino)-N-(2-chlorophenyl)-2-fluorobenzamide (6)

[0677] The title compound (2.8 g, 7.09 mmol, 46.88% yield) was obtained as a yellow solid by a similar method to step 3 of Example 7. MS (M+H) + = 395.0.

[0678] Step 4. Synthesis of methyl 4-((4-((4-((2-chlorophenyl)carbamoyl)-3-fluorophenyl)amino)-5-fluoropyrimidin-2-yl)amino)benzoate (8)

[0679] To a solution of 4-((2-chloro-5-fluoropyrimidin-4-yl)amino)-N-(2-chlorophenyl)-2-fluorobenzamide (1 g, 2.53 mmol) and methyl 4-aminobenzoate (382.51 mg, 2.53 mmol) in dioxane (20 mL) was added TsOH·H2O (962.66 mg, 5.06 mmol) at 20 °C, and the resulting mixture was stirred at 100 °C for 12 h. LCMS confirmed complete consumption of the starting material and a peak of the target mass (81%). The mixture was cooled to 20 °C and filtered. The filter cake was washed with MTBE (40 mL). The filter cake was dried to give the title compound (1.2 g, 2.35 mmol, 93.01% yield) as a yellow solid. MS(M+H) + = 510.0.

[0680] Step 5. Synthesis of 4-((4-((4-((2-chlorophenyl)carbamoyl)-3-fluorophenyl)amino)-5-fluoropyrimidin-2-yl)amino)benzoic acid (9)

[0681] The title compound (1 g, 2.02 mmol, 85.69% yield) was obtained as a white solid by a similar method to step 5 of Example 7. MS (M+H) + = 496.1.

[0682] Step 6. Synthesis of N-(2-chlorophenyl)-4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)-2-fluorobenzamide (Compound 40)

[0683] The title compound (49.02 mg, 50.31 μmol, 24.95% yield, 92% purity) was obtained as a white solid by a similar method to step 6 of Example 1. MS (M+H) + = 896.4.

[0684] 1H NMR (400 MHz, DMSO-d6) δ = 10.93 - 10.87 (m, 1H), 9.94 (s, 1H), 9.77 - 9.65 (m, 2H), 9.64 - 9.53 (m, 1H), 9.52 - 9.41 (m, 1H), 8.32 - 8.26 (m, 1H), 8.16 - 8.02 (m, 2H), 8.00 - 7.93 (m, 1H), 7.87 - 7.66 (m, 5H), 7.57 (dd,J= 1.2, 8.1 Hz, 1H), 7.44 - 7.36 (m, 1H), 7.30 - 7.22 (m, 1H), 6.90 (d,J= 13.6 Hz, 1H), 4.41 (br d,J= 13.2 Hz, 2H), 3.95 (br dd,J= 4.8, 12.8 Hz, 1H), 3.63 - 3.53 (m, 2H), 3.24 - 3.12 (m, 4H), 3.12 - 2.96 (m, 4H), 2.90 - 2.67 (m, 3H), 2.58 - 2.26 (m, 1H), 2.24 - 2.13 (m, 1H), 2.01 - 1.92 (m, 1H), 1.79 - 1.68 (m, 2H), 1.67 - 1.58 (m, 2H), 1.43 - 1.21 (m, 3H).

[0685]

[0686] Example 41. Synthesis of N-(4-(2-(4-(6-((2,6-dioxopiperidin-3-yl)amino)-5-fluoropyridin-3-yl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 41)

[0687]

[0688] The title compound (63.44 mg, 55.36 μmol, 22.28% yield, 98% purity, 2TFA salt) was obtained as a yellowish-white solid by a similar method to step 4 of Example 6. MS (M+H) + = 895.5.

[0689] 1 H NMR (400 MHz, DMSO-d6) δ = 10.78 (s, 1H), 10.44 (s, 1H), 9.81 - 9.66 (m, 1H), 9.63 - 9.36 (m, 2H), 8.19 (d,J= 3.6 Hz, 1H), 7.86 - 7.65 (m, 6H), 7.63 - 7.54 (m, 2H), 7.50 - 7.31 (m, 5H), 6.64 - 6.44 (m, 1H), 4.78 - 4.71 (m, 1H), 3.71 - 3.54 (m, 4H), 3.25 - 3.05 (m, 6H), 2.91 (br t,J= 11.9 Hz, 2H), 2.76 - 2.65 (m, 3H), 2.60 - 2.55 (m, 1H), 2.27 - 2.13 (m, 1H), 2.06 - 1.96 (m, 1H), 1.77 - 1.57 (m, 4H), 1.43 - 1.32 (m, 3H).

[0690]

[0691] Example 42. Synthesis of N-(4-((2-((4-((4-(2-(4-(6-((2,6-dioxopiperidin-3-yl)amino)-5-fluoropyridin-3-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 42)

[0692]

[0693] Step 1. Synthesis of tert-butyl 4-(5-fluoropyridin-3-yl)piperazine-1-carboxylate (2)

[0694] The title compound (15.5 g, 52.34 mmol, 92.11% yield, 95% purity) was obtained as a yellow solid by a similar method to step 1 of Example 30. MS (M+H) + = 282.2.

[0695] Step 2. Synthesis of tert-butyl 4-(5-fluoro-6-iodopyridin-3-yl)piperazine-1-carboxylate (3)

[0696] The title compound (8.8 g, 20.53 mmol, 37.26% yield, 95% purity) was obtained as a blackish brown solid by a similar method to step 2 of Example 30. MS (M+H) + = 408.1.

[0697] Step 3. Synthesis of tert-butyl 4-(6-((2,6-bis(benzyloxy)pyridin-3-yl)amino)-5-fluoropyridin-3-yl)piperazine-1-carboxylate (4)

[0698] The title compound (3.6 g, 6.15 mmol, 83.44% yield) was obtained as a blackish brown solid by a similar method to step 3 of Example 30. MS (M+H) + = 586.4.

[0699] Step 4. Synthesis of tert-butyl 4-(6-((2,6-dioxopiperidin-3-yl)amino)-5-fluoropyridin-3-yl)piperazine-1-carboxylate (5)

[0700] To a solution of Pd(OH)2 / C (0.8 g, 20% purity) in THF (50 mL) was added a solution of tert-butyl 4-(6-((2,6-bis(benzyloxy)pyridin-3-yl)amino)-5-fluoropyridin-3-yl)piperazine-1-carboxylate (2 g, 3.41 mmol) in MeOH (50 mL) under N2, and the mixture was stirred under H2 (45 Psi) at 30 °C for 30 h. The peak of the target mass (82%) was identified by LCMS. The mixture was filtered and washed with THF (800 mL). The filtrate was dried under reduced pressure to obtain the title compound (1.35 g, crude) as a yellow solid. MS (M+H) + = 408.4.

[0701] Step 5. Synthesis of 3-((3-fluoro-5-(piperazin-1-yl)pyridin-2-yl)amino)piperidine-2,6-dione (6)

[0702] The title compound (1.6 g, crude, HCl salt) was obtained as a yellow solid by a similar method to step 2 of Example 18. MS (M+H) + = 308.2.

[0703] Step 6. Synthesis of N-(4-((2-((4-((4-(2-(4-(6-((2,6-dioxopiperidin-3-yl)amino)-5-fluoropyridin-3-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 42)

[0704] The title compound (36.46 mg, 31.68 μmol, 12.37% yield, 96% purity, 2TFA salt) was obtained as a yellowish-white solid by a similar method to Example 6-4. MS (M+H) + = 877.5.

[0705] 1 H NMR (400 MHz, DMSO-d6) δ = 10.77 (s, 1H), 10.44 (s, 1H), 9.70 - 9.21 (m, 3H), 8.22 - 8.13 (m, 1H), 7.81 - 7.65 (m, 9H), 7.63 - 7.55 (m, 2H), 7.45 - 7.31 (m, 3H), 6.61 - 6.50 (m, 1H), 4.80 - 4.70 (m, 1H), 3.70 - 3.51 (m, 4H), 3.24 - 3.05 (m, 6H), 2.96 - 2.75 (m, 6H), 2.26 - 2.13 (m, 1H), 2.05 - 1.96 (m, 1H), 1.78 - 1.58 (m, 4H), 1.44 - 1.29 (m, 3H).

[0706]

[0707] Example 43. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 43)

[0708]

[0709] The title compound (67.88 mg, 59.52 μmol, 20.50% yield, 94% purity, 2TFA) was obtained as a white solid by a similar method to step 4 of Example 6. MS (M+H) + = 844.3.

[0710] 1 H NMR (400 MHz, DMSO-d6) δ = 10.84 (s, 1H), 10.43 (s, 1H), 9.57 (br s, 2H), 9.51 (br s, 1H), 8.16 (d,J= 3.6 Hz, 1H), 8.03 (d,J= 2.4 Hz, 1H), 7.79 - 7.56 (m, 10H), 7.53 (dd,J= 9.2, 2.4 Hz, 1H), 7.39 - 7.31 (m, 2H), 6.97 (d,J= 8.8 Hz, 1H), 4.39 (d,J= 10.8 Hz, 1H), 3.80 (dd,J=12.8, 5.2 Hz, 1H), 3.61 - 3.59 (m, 2H), 3.21 - 3.05 (m, 9H), 2.84 - 2.79 (m, 2H), 2.74 - 2.65 (m, 1H), 2.55 - 2.53 (m, 1H), 2.27 - 2.16 (m, 1H), 1.99 - 1.94 (m, 1H), 1.76 - 1.71 (bm, 2H), 1.65 - 1.62 (m, 2H), 1.42 - 1.33 (m, 3H).

[0711]

[0712] Example 44. Synthesis of N-(4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 44)

[0713]

[0714] Step 1. Synthesis of tert-butyl (4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamate (3)

[0715] The title compound (220 mg, 404.29 μmol, 56.15% yield, 92% purity) was obtained as a light yellow solid by a similar method to step 4 of Example 1. MS (M+H) + = 501.4.

[0716] Step 2. Synthesis of 3-(6-(4-(2-(1-aminopiperidin-4-yl)ethyl)piperazin-1-yl)pyridin-3-yl)piperidine-2,6-dione (4)

[0717] The title compound (470 mg, crude, 2TFA) as a yellow oil was obtained by a similar method to step 5 of Example 1. MS (M+H) + = 401.1.

[0718] Step 3, Synthesis of N-(4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 44)

[0719] The title compound (75.5 mg, 52.13 μmol, 18.02% yield, 91% purity, 4TFA) was obtained as a yellow solid by a similar method to step 6 of Example 1. MS (M+H) + = 862.3.

[0720] 1 H NMR (400 MHz, DMSO-d6) δ = 10.85 (s, 1H), 10.43 (s, 1H), 9.75 - 9.64 (m, 2H), 9.56 (s, 1H), 9.33 (br s, 1H), 8.18 (d,J= 4.0 Hz, 1H), 8.03 (d,J= 2.0 Hz, 1H), 7.76 - 7.72 (m, 4H), 7.71 - 7.66 (m, 1H), 7.62 - 7.52 (m, 2H), 7.48 - 7.42 (m, 2H), 7.39 - 7.32 (m, 2H), 6.99 (br d,J=8.8 Hz, 1H), 4.38 (br d,J= 12.0 Hz, 2H), 3.80 (dd,J= 12.4, 4.8 Hz, 1H), 3.63 - 3.50 (m, 2H), 3.24 - 2.99 (m, 8H), 2.79 - 2.61 (m, 3H), 2.57 - 2.51 (m, 1H), 2.25 - 2.15 (m, 1H), 2.01 - 1.92 (m, 1H), 1.78 - 1.70 (m, 2H), 1.67 - 1.58 (m, 2H), 1.38 - 1.26 (m, 3H).

[0721]

[0722] Example 45. N-(4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-3-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 45)

[0723]

[0724] Step 1. Synthesis of tert-butyl 4-(5-bromo-3-fluoropyridin-2-yl)piperazine-1-carboxylate (2)

[0725] To a solution of 5-bromo-2,3-difluoropyridine (10 g, 51.55 mmol) and tert-butyl piperazine-1-carboxylate (9.60 g, 51.55 mmol) in DMF (100 mL) was added DIPEA (20.03 g, 155.01 mmol, 27 mL), and the mixture was stirred at 80 °C for 14 h. TLC (petroleum ether:EtOAc=10:1) confirmed complete consumption of the starting material and the formation of a new spot. The mixture was diluted with water (100 mL) and extracted with EtOAc (50 mL x 3), the combined organic layers were washed with brine (100 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (120 gSepaFlash® silica flash column, eluent 10–15% EtOAc / petroleum ether, gradient @200 mL / min) to give the title compound (16.4 g, 44.62 mmol, 86.55% yield, 98% purity) as a yellow oil. MS(M-Boc+H) + = 260.1.

[0726] Step 2. Synthesis of tert-butyl 4-(2',6'-bis(benzyloxy)-5-fluoro-[3,3'-bipyridin]-6-yl)piperazine-1-carboxylate (3)

[0727] To a solution of tert-butyl 4-(5-bromo-3-fluoropyridin-2-yl)piperazine-1-carboxylate (2.5 g, 6.94 mmol), 2,6-bis(benzyloxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (3.19 g, 7.63 mmol), and Cs2CO3 (6.78 g, 20.82 mmol) in dioxane (60 mL) and water (10 mL) was added Pd(dtbpf)Cl2 (226.25 mg, 347.15 μmol), and the mixture was stirred at 100 °C under N2 for 14 h. The target mass was confirmed by LCMS. The mixture was diluted with water (100 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (12 g SepaFlash® silica flash column, eluent 10–15% EtOAc / petroleum ether, gradient @ 200 mL / min) to give the title compound (1.52 g, 2.45 mmol, 35.31% yield, 92% purity) as a yellow solid. MS (M+H) + = 571.4.

[0728] Step 3. Synthesis of tert-butyl 4-(5-(2,6-dioxopiperidin-3-yl)-3-fluoropyridin-2-yl)piperazine-1-carboxylate (4)

[0729] To a solution of Pd(OH)2 / C (0.5 g, 20% purity) in MeOH (10 mL) was slowly added a solution of tert-butyl 4-(2',6'-bis(benzyloxy)-5-fluoro-[3,3'-bipyridin]-6-yl)piperazine-1-carboxylate (1.52 g, 2.66 mmol) in THF (10 mL) under N2, and the mixture was stirred under H2 (45 psi) at 30 °C for 30 h. The peak of the target mass (89%) was identified by LCMS. The mixture was filtered and washed with THF (700 mL). The filtrate was concentrated under reduced pressure to give the title compound (1 g, crude) as a white solid. MS (M+H) + = 393.2.

[0730] Step 4. Synthesis of 3-(5-fluoro-6-(piperazin-1-yl)pyridin-3-yl)piperidine-2,6-dione (5)

[0731] To a solution of tert-butyl 4-(5-(2,6-dioxopiperidin-3-yl)-3-fluoropyridin-2-yl)piperazine-1-carboxylate (1 g, 2.55 mmol) in DCM (10 mL) was added HCl / dioxane (2 M, 80 mL), and the mixture was stirred at 20 °C for 14 h. The peak of the target mass (67%) was confirmed by LCMS. The mixture was concentrated under reduced pressure to obtain the title compound (1 g, crude, HCl salt) as a white solid. MS (M+H) + = 293.3.

[0732] Step 5. Synthesis of N-(4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-3-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 45)

[0733] To a solution of 3-(5-fluoro-6-(piperazin-1-yl)pyridin-3-yl)piperidine-2,6-dione (160 mg, 486.66 μmol, HCl salt) and 2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)-N-(4-(2-oxoethyl)piperidin-1-yl)benzamide (150 mg, 248.51 μmol) in DMSO (5 mL) was added NaOAc (61.16 mg, 745.54 μmol), and the mixture was stirred at 30 °C for 0.5 h. NaBH(OAc)3 (158.01 mg, 745.54 μmol) was added, and the mixture was stirred at 30 °C for 1 h. The target mass peak (61%) was confirmed by LCMS. After pouring water (20 mL) into the mixture, the mixture was filtered. The filtrate was washed with water (5 mL) and concentrated under reduced pressure. The product was dried by prep-HPLC (column: Phenomenex Luna C18 150*25 mm*10 μm; mobile phase: [H2O (0.1% TFA) -ACN]; gradient: 3%-33% B over 15 min) and lyophilized to obtain the title compound (35.44 mg, 32.09 μmol, 12.91% yield, 90% purity, TFA salt) as a white solid. MS (M+H) + = 880.4.

[0734] 1H NMR (400 MHz, DMSO-d6) δ = 10.89 (s, 1H), 10.42 (s, 1H), 9.73 - 9.42 (m, 2H), 9.09 - 8.85 (m, 1H), 8.17 (d,J= 3.6 Hz, 1H), 7.99 - 7.91 (m, 1H), 7.83 - 7.65 (m, 6H), 7.62 - 7.52 (m, 2H), 7.46 - 7.28 (m, 4H), 4.06 (br d,J= 11.6 Hz, 2H), 3.91 (br dd,J= 4.8, 12.4 Hz, 1H), 3.65 - 3.48 (m, 2H), 3.26 - 2.99 (m, 8H), 2.71 - 2.54 (m, 3H), 2.46 - 2.36 (m, 1H), 2.30 - 2.21 (m, 1H), 2.03 - 1.94 (m, 1H), 1.75 - 1.52 (m, 4H), 1.36 - 1.22 (m, 3H).

[0735]

[0736] Example 46. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-3-fluoropyridin-2-yl) piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 46)

[0737]

[0738] The title compound (30.67 mg, 25.61 μmol, 10.00% yield, 91% purity, 2TFA salt) was obtained as a white solid by a similar method to step 4 of Example 6. MS (M+H) + = 862.8.

[0739] 1H NMR (400 MHz, DMSO-d6) δ = 10.90 (s, 1H), 10.44 (s, 1H), 10.00 - 9.43 (m, 3H), 8.16 (br d,J= 3.1 Hz, 1H), 7.96 (s, 1H), 7.83 - 7.66 (m, 9H), 7.64 - 7.55 (m, 2H), 7.41 - 7.30 (m, 2H), 4.14 - 4.02 (m, 2H), 3.91 (br dd,J= 4.4, 12.6 Hz, 1H), 3.60 (br d,J= 10.0 Hz, 2H), 3.20 (br d,J= 10.8 Hz, 7H), 2.94 - 2.76 (m, 2H), 2.75 - 2.64 (m, 1H), 2.57 (br d,J= 3.8 Hz, 1H), 2.34 - 2.20 (m, 1H), 2.04 - 1.93 (m, 1H), 1.82 - 1.60 (m, 4H), 1.49 - 1.28 (m, 3H).

[0740]

[0741] Example 47. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 47)

[0742]

[0743] Step 1. Synthesis of tert-butyl (4-(2-fluorobenzamido)phenyl)carbamate (2)

[0744] To a solution of 2-fluorobenzoic acid (3 g, 21.41 mmol), DIPEA (8.30 g, 64.23 mmol), and HATU (9.77 g, 25.69 mmol) in DMF (30 mL) was added tert-butyl N-(4-aminophenyl)carbamate (4.46 g, 21.41 mmol) at 20 °C, and the resulting mixture was stirred at 20 °C for 20 min. The main peak of the desired mass was identified by LCMS. Water (200 mL) was poured into the mixture, and the mixture was stirred slowly. The mixture was filtered and washed with water (100 mL). The filter cake was collected and dried to give the title compound (7 g, crude) as a gray solid, which was used directly in the next reaction. MS (M+H) + = 331.1.

[0745] Step 2. Synthesis of N-(4-aminophenyl)-2-fluorobenzamide (3)

[0746] The title compound (5.6 g, crude, HCl) was obtained as a gray solid by a similar method to step 2 of Example 7. MS (M+H) + = 231.0.

[0747] Step 3. Synthesis of N-(4-((2-chloro-5-(trifluoromethyl)pyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (4)

[0748] The title compound (5 g, crude) was obtained as a dark brown oil by a similar method to step 3 of Example 7. The product was used directly in the following reaction. MS (M+H) + = 411.0.

[0749] Step 4. Synthesis of methyl 4-((4-((4-(2-fluorobenzamido)phenyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)benzoate (5)

[0750] To a solution of N-(4-((2-chloro-5-(trifluoromethyl)pyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (2.5 g, 6.09 mmol) and methyl 4-aminobenzoate (920.03 mg, 6.09 mmol) in dioxane (30 mL) was added TsOH (2.10 g, 12.17 mmol) at 20 °C, and the resulting mixture was stirred at 100 °C for 2 h. The peak of the desired mass was confirmed by LCMS. The mixture was concentrated under reduced pressure to obtain the title compound (3.5 g, crude) as a gray solid. The product was used directly in the next reaction. MS(M+H) + = 526.1.

[0751] Step 5. Synthesis of 4-((4-((4-(2-fluorobenzamido)phenyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)benzoic acid (6)

[0752] The title compound (350 mg, crude) was obtained as a yellow solid by a similar method to step 5 of Example 7, and was used directly in the following reaction. MS(M+H) + = 512.1.

[0753] Step 6. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 47)

[0754] The title compound (21.8 mg, 16.64 μmol, 5.67% yield, 87% purity, 2TFA) was obtained as a yellow solid by a similar method to step 9 of Example 12. MS (M+H) + = 912.7.

[0755] 1H NMR (400 MHz, DMSO-d6) δ = 10.90 (s, 1H), 10.53 (s, 1H), 9.99 - 9.86 (m, 1H), 9.72 - 9.24 (m, 1H), 8.97 - 8.83 (m, 1H), 8.40 (s, 1H), 8.08 (d,J= 10.9 Hz, 1H), 7.80 - 7.72 (m, 3H), 7.68 - 7.53 (m, 5H), 7.45 (br d,J= 8.5 Hz, 2H), 7.39 - 7.32 (m, 2H), 6.91 (br d,J= 13.8 Hz, 1H), 4.42 (br d,J= 12.6 Hz, 2H), 3.95 (br dd,J= 4.8, 12.9 Hz, 1H), 3.63 - 3.54 (m, 2H), 3.21 - 3.00 (m, 8H), 2.79 - 2.65 (m, 3H), 2.56 - 2.54 (m, 1H), 2.25 - 2.14 (m, 1H), 2.02 - 1.93 (m, 1H), 1.73 - 1.55 (m, 4H), 1.38 - 1.20 (m, 3H).

[0756]

[0757] Example 48. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluorobenzamide (Compound 48)

[0758]

[0759] The title compound (105.4 mg, 81.41 μmol, 21.68% yield, 93% purity, 3TFA salt) was obtained as a yellow solid by a similar method to step 9 of Example 12. MS (M+H) + = 862.5.

[0760] 1H NMR (400 MHz, DMSO-d6) δ = 10.89 (s, 1H), 10.43 (s, 1H), 9.80 - 9.47 (m, 3H), 8.19 - 8.13 (m, 1H), 8.08 (d,J= 10.9 Hz, 1H), 7.82 - 7.65 (m, 9H), 7.63 - 7.56 (m, 1H), 7.40 - 7.25 (m, 2H), 6.90 (br d,J= 13.6 Hz, 1H), 4.41 (br d,J= 13.4 Hz, 2H), 4.03 - 3.90 (m, 1H), 3.58 (br d,J=10.9 Hz, 2H), 3.27 - 2.68 (m, 10H), 2.59 - 2.52 (m, 2H), 2.26 - 2.11 (m, 1H), 2.02 - 1.90 (m, 1H), 1.80 - 1.54 (m, 4H), 1.47 - 1.29 (m, 3H).

[0761]

[0762] Example 49. Synthesis of N-(4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 49)

[0763]

[0764] A solution of 2-fluoro-4-((5-fluoro-4-((4-(2-fluorobenzamido)phenyl)amino)pyrimidin-2-yl)amino)-N-(4-(2-oxoethyl)piperidin-1-yl)benzamide (150 mg, 248.51 μmol), 3-(4-fluoro-6-(piperazin-1-yl)pyridin-3-yl)piperidine-2,6-dione (98.04 mg, 298.21 μmol, HCl) and NaOAc (61.16 mg, 745.54 μmol) in DMSO (3 mL) was stirred at 20 °C for 30 min. NaBH(OAc)3 (79.00 mg, 372.77 μmol) was added and the resulting mixture was stirred at 20 °C for 1 h. The main peak of the target mass was identified by LCMS. Aqueous NaHCO3 solution (20 mL) was poured into the mixture. The mixture was filtered and washed with water (10 mL). The filter cake was collected to obtain the crude product. The crude product was purified by prep-HPLC (column: Phenomenex Luna C18 150*25 mm*10 μm; mobile phase: [H2O (0.1% TFA) -ACN]; gradient: 3%-33% B over 15 min) and lyophilized to obtain the title compound (25.9 mg, 21.74 μmol, 8.75% yield, 93% purity, 2TFA) as a white solid. MS (M+H) + = 880.4.

[0765] 1H NMR (400 MHz, DMSO-d6) δ = 10.90 (s, 1H), 10.43 (s, 1H), 9.77 - 9.61 (m, 1H), 9.55 (br s, 1H), 9.34 - 9.18 (m, 1H), 8.18 (d,J= 3.5 Hz, 1H), 8.08 (d,J= 11.0 Hz, 1H), 7.81 (br d,J= 13.9 Hz, 1H), 7.74 (s, 4H), 7.68 (br t,J= 6.8 Hz, 1H), 7.63 - 7.56 (m, 1H), 7.49 - 7.40 (m, 2H), 7.39 - 7.31 (m, 2H), 6.90 (br d,J= 13.7 Hz, 1H), 4.47 - 4.33 (m, 2H), 3.95 (br dd,J= 4.7, 12.5 Hz, 1H), 3.64 - 3.48 (m, 2H), 3.18 - 2.83 (m, 7H), 2.79 - 2.64 (m, 3H), 2.59 - 2.54 (m, 1H), 2.49 - 2.41 (m, 1H), 2.25 - 2.12 (m, 1H), 2.02 - 1.92 (m, 1H), 1.81 - 1.56 (m, 4H), 1.55 - 1.25 (m, 3H).

[0766]

[0767] Example 50. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-3-fluoropicolinamide (Compound 50)

[0768]

[0769] Step 1. Synthesis of tert-butyl (4-((2-chloro-5-fluoropyrimidin-4-yl)amino)phenyl)carbamate (2)

[0770] The title compound (25 g, crude) as a blackish brown solid was obtained by a similar method to step 3 of Example 7, and was used directly in the following reaction. MS(M+H) + = 339.1.

[0771] Step 2. Synthesis of N-(2-chloro-5-fluoropyrimidin-4-yl)benzene-1,4-diamine (3)

[0772] The title compound (22 g, crude, HCl) was obtained as a gray solid by a similar method to step 2 of Example 7. MS (M+H) + = 239.1.

[0773] Step 3. Synthesis of N-(4-((2-chloro-5-fluoropyrimidin-4-yl)amino)phenyl)-3-fluoropicolinamide (4)

[0774] N in DMF (15 mL) 1 -(2-Chloro-5-fluoropyrimidin-4-yl)benzene-1,4-diamine (1.5 g, 5.45 mmol), 3-fluoropyridine-2-carboxylic acid (846.26 mg, 6.00 mmol), and DIPEA (2.82 g, 21.81 mmol) were added HATU (2.70 g, 7.09 mmol) to a solution at 20 °C, and the resulting mixture was stirred at 20 °C for 1 h. The main peak of the desired mass was identified by LCMS. Water (120 mL) was slowly poured into the mixture. The mixture was filtered and washed with water (30 mL). The filter cake was collected and dried to give the title compound (1.4 g, crude) as a brown solid, which was used directly in the next reaction. MS(M+H) + = 362.0.

[0775] Step 4. Synthesis of methyl 4-((5-fluoro-4-((4-(3-fluoropicolinamido)phenyl)amino)pyrimidin-2-yl)amino)benzoate (5)

[0776] To a solution of N-(4-((2-chloro-5-fluoropyrimidin-4-yl)amino)phenyl)-3-fluoropicolinamide (600 mg, 1.66 mmol) and methyl 4-aminobenzoate (250.73 mg, 1.66 mmol) in dioxane (8 mL) was added TsOH (571.25 mg, 3.32 mmol) at 20 °C, and the resulting mixture was stirred at 100 °C for 14 h. The main peak of the desired mass was confirmed by LCMS. The mixture was concentrated to give the title compound (800 mg, crude) as a yellow solid, which was used directly in the next reaction. MS(M+H) + = 477.2.

[0777] Step 5. Synthesis of 4-((5-fluoro-4-((4-(3-fluoropicolinamido)phenyl)amino)pyrimidin-2-yl)amino)benzoic acid (6)

[0778] The title compound (800 mg, crude) was obtained as a yellow solid by a similar method to step 5 of Example 7. MS (M+H) + = 463.1.

[0779] Step 6. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-3-fluoropicolinamide (Compound 50)

[0780] The title compound (29.3 mg, 25.51 μmol, 4.95% yield, 95% purity, 2TFA) was obtained as a yellow solid by a similar method to step 9 of Example 12. MS (M+H) + = 863.5.

[0781] 1H NMR (400 MHz, DMSO-d6) δ = 10.91 (s, 1H), 10.65 (s, 1H), 9.69 - 9.60 (m, 1H), 9.57 - 9.53 (m, 1H), 8.58 (br d,J= 4.6 Hz, 1H), 8.17 (d,J= 3.5 Hz, 1H), 8.08 (d,J= 11.0 Hz, 1H), 7.98 - 7.92 (m, 1H), 7.83 - 7.73 (m, 8H), 7.71 - 7.67 (m, 2H), 6.91 (br d,J= 14.2 Hz, 1H), 4.47 - 4.36 (m, 2H), 3.95 (br dd,J= 4.7, 12.7 Hz, 1H), 3.63 - 3.57 (m, 2H), 3.23 - 3.11 (m, 6H), 3.09 - 3.00 (m, 2H), 2.93 - 2.65 (m, 3H), 2.59 - 2.53 (m, 1H), 2.22 - 2.14 (m, 1H), 2.00 - 1.93 (m, 1H), 1.79 - 1.71 (m, 2H), 1.67 - 1.59 (m, 2H), 1.44 - 1.33 (m, 3H).

[0782]

[0783] Example 51. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)-3-fluorophenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-3-fluoropicolinamide (Compound 51)

[0784]

[0785] Step 1. Synthesis of methyl 2-fluoro-4-((5-fluoro-4-((4-(3-fluoropicolinamido)phenyl)amino)pyrimidin-2-yl)amino)benzoate (2)

[0786] To a solution of N-(4-((2-chloro-5-fluoropyrimidin-4-yl)amino)phenyl)-3-fluoropicolinamide (600 mg, 1.66 mmol) and methyl 4-amino-2-fluoro-benzoate (280.57 mg, 1.66 mmol) in dioxane (8 mL) was added TsOH (571.25 mg, 3.32 mmol) at 20 °C, and the resulting mixture was stirred at 100 °C for 14 h. The main peak of the desired mass was confirmed by LCMS. The reaction mixture was concentrated to give the title compound (800 mg, crude) as a yellow solid, which was used directly in the next reaction. MS(M+H) + = 495.2.

[0787] Step 2. Synthesis of 2-fluoro-4-((5-fluoro-4-((4-(3-fluoropicolinamido)phenyl)amino)pyrimidin-2-yl)amino)benzoic acid (3)

[0788] The title compound (750 mg, crude) was obtained as a yellow solid by a similar method to step 5 of Example 7. MS (M+H) + = 481.1.

[0789] Step 3. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)-3-fluorophenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-3-fluoropicolinamide (Compound 51)

[0790] The title compound (17 mg, 12.37 μmol, 6.23% yield, 89% purity, 3TFA) was obtained as a yellow solid by a similar method to step 9 of Example 12. MS (M+H) + = 881.6.

[0791] 1H NMR (400 MHz, DMSO-d6) δ = 10.91 (s, 1H), 10.64 (s, 1H), 9.72 (s, 1H), 9.54 (br s, 2H), 9.23 - 9.10 (m, 1H), 8.58 (d,J= 4.5 Hz, 1H), 8.18 (d,J= 3.7 Hz, 1H), 8.08 (d,J= 11.0 Hz, 1H), 7.98 - 7.92 (m, 1H), 7.84 - 7.73 (m, 6H), 7.44 (s, 1H), 6.90 (d,J= 13.2 Hz, 1H), 4.41 (br dd,J= 0.9, 13.2 Hz, 2H), 3.98 - 3.92 (m, 1H), 3.63 - 3.53 (m, 2H), 3.21 - 2.99 (m, 8H), 2.72 - 2.64 (m, 4H), 2.22 - 2.14 (m, 1H), 2.00 - 1.93 (m, 1H), 1.74 - 1.52 (m, 4H), 1.51 - 1.25 (m, 3H).

[0792]

[0793] Example 52. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-3-fluoroisonicotinamide (Compound 52)

[0794]

[0795] Step 1. Synthesis of N-(4-((2-chloro-5-fluoropyrimidin-4-yl)amino)phenyl)-3-fluoroisonicotinamide (2)

[0796] The title compound (2.5 g, crude) was obtained as a gray solid by a similar method to step 1 of Example 47. MS (M+H) + = 362.0.

[0797] Step 2. Synthesis of 4-((5-fluoro-4-((4-(3-fluoroisonicotinamido)phenyl)amino)pyrimidin-2-yl)amino)benzoic acid (3)

[0798] To a solution of N-(4-((2-chloro-5-fluoropyrimidin-4-yl)amino)phenyl)-3-fluoroisonicotinamide (400 mg, 1.11 mmol) and 4-aminobenzoic acid (151.64 mg, 1.11 mmol) in i-PrOH (6 mL) was added HCl (12 M, 184.30 μL) at 20 °C, and the resulting mixture was stirred at 80 °C for 14 h. LCMS confirmed that 30% of the target mass peak and the starting material remained. The mixture was further stirred at 100 °C for 4 h. LCMS confirmed that 50% of the target mass peak was present. After cooling to 20 °C, the reaction mixture was filtered and washed with MeCN (5 mL). The filtrate was discarded. The filter cake was collected and dried to obtain the title compound (250 mg, crude) as a yellow solid, which was used directly in the next reaction. MS(M+H) + = 463.2.

[0799] Step 3. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-3-fluoroisonicotinamide (Compound 52)

[0800] The title compound (50.5 mg, 43.05 μmol, 9.95% yield, 93% purity, 2TFA) was obtained as a yellow solid by a similar method to step 9 of Example 12. MS (M+H) + = 863.6.

[0801] 1H NMR (400 MHz, DMSO-d6) δ = 10.91 (s, 1H), 10.73 (s, 1H), 10.02 - 9.70 (m, 2H), 9.65 - 9.55 (m, 1H), 8.78 (d,J= 1.1 Hz, 1H), 8.61 (dd,J= 1.0, 4.8 Hz, 1H), 8.19 (br s, 1H), 8.08 (d,J= 10.9 Hz, 1H), 7.83 - 7.77 (m, 4H), 7.74 - 7.69 (m, 5H), 6.91 (d,J= 13.6 Hz, 1H), 4.42 (br d,J= 12.3 Hz, 2H), 3.95 (dd,J= 4.6, 12.7 Hz, 1H), 3.63 - 3.54 (m, 2H), 3.33 - 3.14 (m, 6H), 3.10 - 3.00 (m, 2H), 2.98 - 2.87 (m, 2H), 2.79 - 2.67 (m, 1H), 2.60 - 2.53 (m, 1H), 2.24 - 2.11 (m, 1H), 2.02 - 1.93 (m, 1H), 1.82 - 1.73 (m, 2H), 1.70 - 1.60 (m, 2H), 1.50 - 1.37 (m, 3H).

[0802]

[0803] Example 53. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)-3-fluorophenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-3-fluoroisonicotinamide (Compound 53)

[0804]

[0805] Step 1. Synthesis of methyl 2-fluoro-4-((5-fluoro-4-((4-(3-fluoroisonicotinamido)phenyl)amino)pyrimidin-2-yl)amino)benzoate (2)

[0806] To a solution of N-(4-((2-chloro-5-fluoropyrimidin-4-yl)amino)phenyl)-3-fluoroisonicotinamide (500 mg, 1.38 mmol) and methyl 4-amino-2-fluoro-benzoate (233.81 mg, 1.38 mmol) in dioxane (10 mL) was added TsOH (476.04 mg, 2.76 mmol) at 20 °C, and the resulting mixture was stirred at 100 °C for 14 h. LCMS showed that most of the starting material remained. The mixture was further stirred at 100 °C for 16 h. LCMS showed that most of the starting material was consumed. After cooling to 20 °C, the reaction mixture was filtered. The filtrate was discarded. The filter cake was collected and dried to give the title compound (700 mg, crude) as a yellow solid. MS(M+H) + = 495.2.

[0807] Step 2. Synthesis of 2-fluoro-4-((5-fluoro-4-((4-(3-fluoroisonicotinamido)phenyl)amino)pyrimidin-2-yl)amino)benzoic acid (3)

[0808] To a solution of methyl 2-fluoro-4-((5-fluoro-4-((4-(3-fluoroisonicotinamido)phenyl)amino)pyrimidin-2-yl)amino)benzoate (400 mg, 809.02 μmol) in THF (5 mL) was added a solution of NaOH (161.80 mg, 4.05 mmol) in water (2 mL) at 20 °C, and the resulting mixture was stirred at 20 °C for 2 h. LCMS showed that most of the starting material was consumed. The mixture was further stirred at 20 °C for 14 h. LCMS confirmed that some of the starting material remained and the target mass was determined. The reaction mixture was neutralized with 1 N HCl aqueous solution. The mixture was concentrated to remove the organic solvent. The residue was filtered and washed with water (3 mL). The filtrate was discarded. The filter cake was lyophilized to obtain the title compound (300 mg, crude) as a yellow solid, which was used directly in the following reaction. MS(M+H) + = 481.1.

[0809] Step 3. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)-3-fluorophenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-3-fluoroisonicotinamide (Compound 53)

[0810] The title compound (19 mg, 16.28 μmol, 4.10% yield, 95% purity, 2TFA) was obtained as a yellow solid by a similar method to step 8 of Example 12. MS (M+H) + = 881.5.

[0811] 1H NMR (400 MHz, DMSO-d6) δ = 10.91 (s, 1H), 10.71 (s, 1H), 9.72 (br d,J= 1.1 Hz, 1H), 9.62 - 9.49 (m, 2H), 9.31 - 9.11 (m, 1H), 8.77 (s, 1H), 8.60 (d,J= 4.9 Hz, 1H), 8.19 (d,J= 3.4 Hz, 1H), 8.08 (d,J= 10.9 Hz, 1H), 7.80 - 7.69 (m, 5H), 7.44 - 7.41 (m, 2H), 6.90 (d,J=13.4 Hz, 1H), 4.44 - 4.37 (m, 2H), 3.95 (br dd,J= 4.6, 12.9 Hz, 1H), 3.63 - 3.50 (m, 2H), 3.23 - 2.97 (m, 8H), 2.78 - 2.63 (m, 3H), 2.53 - 2.52 (m, 1H), 2.22 - 2.12 (m, 1H), 2.02 - 1.93 (m, 1H), 1.76 - 1.55 (m, 4H), 1.45 - 1.18 (m, 3H).

[0812]

[0813] Example 54. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2,6-difluorobenzamide (Compound 54)

[0814]

[0815] Step 1. Synthesis of N-(4-((2-chloro-5-fluoropyrimidin-4-yl)amino)phenyl)-2,6-difluorobenzamide (2)

[0816] The title compound (2.5 g, crude) was obtained as a brown solid by a similar method to step 1 of Example 47, and the product was used directly in the following reaction. MS(M+H) + = 379.1.

[0817] Step 2. Synthesis of methyl 4-((4-((4-(2,6-difluorobenzamido)phenyl)amino)-5-fluoropyrimidin-2-yl)amino)benzoate (3)

[0818] The title compound (1.2 g, crude) was obtained as a white solid by a similar method to step 1 of Example 53. MS (M+H) + = 494.2.

[0819] Step 3. Synthesis of 4-((4-((4-(2,6-difluorobenzamido)phenyl)amino)-5-fluoropyrimidin-2-yl)amino)benzoic acid (4)

[0820] To a solution of methyl 4-((4-((4-(2,6-difluorobenzamido)phenyl)amino)-5-fluoropyrimidin-2-yl)amino)benzoate (1.2 g, 2.43 mmol) in THF (10 mL) and MeOH (5 mL) was added NaOH (291.81 mg, 7.30 mmol) in water (10 mL) at 20 °C. The mixture was stirred at 50 °C for 1 h. LCMS confirmed complete consumption of the starting material and the main peak of the desired mass. The mixture was concentrated to remove the organic solvent. The residue was diluted with water (20 mL), and the mixture was filtered. The filter cake was collected and lyophilized to give the title compound (1 g, crude, Na) as a yellow solid. MS (M+H) + = 480.2.

[0821] Step 4. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2,6-difluorobenzamide (Compound 54)

[0822] The title compound (17.6 mg, 13.97 μmol, 7.02% yield, 97% purity, 3TFA) was obtained as a white solid by a similar method to step 9 of Example 12. MS (M+H)+ = 880.3.

[0823] 1 H NMR (400 MHz, DMSO-d6) δ = 10.91 (s, 1H), 10.81 (s, 1H), 9.67 - 9.54 (m, 3H), 8.17 (d,J= 3.5 Hz, 1H), 8.08 (d,J= 11.0 Hz, 1H), 7.82 - 7.77 (m, 4H), 7.69 (br d,J= 9.0 Hz, 4H), 7.63 - 7.57 (m, 1H), 7.27 (t,J= 8.0 Hz, 2H), 6.91 (d,J= 13.6 Hz, 1H), 4.42 (br d,J= 12.0 Hz, 2H), 3.95 (dd,J= 4.8, 12.8 Hz, 1H), 3.63 - 3.55 (m, 2H), 3.22 - 3.09 (m, 6H), 3.09 - 2.99 (m, 2H), 2.87 - 2.69 (m, 2H), 2.52 - 2.51 (m, 1H), 2.45 - 2.38 (m, 1H), 2.23 - 2.14 (m, 1H), 2.00 - 1.93 (m, 1H), 1.79 - 1.60 (m, 4H), 1.45 - 1.31 (m, 3H).

[0824]

[0825] Example 55. Synthesis of 4-((4-((4-(2,6-difluorobenzamido)phenyl)amino)-5-fluoropyrimidin-2-yl)amino)-N-(4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluorobenzamide (Compound 55)

[0826]

[0827] Step 1. Synthesis of methyl 4-((4-((4-(2,6-difluorobenzamido)phenyl)amino)-5-fluoropyrimidin-2-yl)amino)-2-fluorobenzoate (2)

[0828] The title compound (1.5 g, crude) was obtained as a brown solid by a similar method to step 1 of Example 53. The product was used directly in the following reaction. MS (M+H) + = 512.1.

[0829] Step 2. Synthesis of 4-((4-((4-(2,6-difluorobenzamido)phenyl)amino)-5-fluoropyrimidin-2-yl)amino)-2-fluorobenzoic acid (3)

[0830] The title compound (1.5 g, 2.56 mmol, 87.40% yield, 85% purity) was obtained as a gray solid by a similar method to step 5 of Example 7.

[0831] Step 3. Synthesis of 4-((4-((4-(2,6-difluorobenzamido)phenyl)amino)-5-fluoropyrimidin-2-yl)amino)-N-(4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluorobenzamide (Compound 55)

[0832] The title compound (18.6 mg, 13.80 μmol, 7.18% yield, 92% purity, 3TFA) was obtained as a white solid by a similar method to step 9 of Example 12. MS (M+H) + = 898.6.

[0833] 1H NMR (400 MHz, DMSO-d6) δ = 10.91 (s, 1H), 10.81 (s, 1H), 9.70 (s, 1H), 9.62 - 9.50 (m, 1H), 9.20 - 9.12 (m, 1H), 8.18 (d,J= 3.7 Hz, 1H), 8.08 (d,J= 10.9 Hz, 1H), 7.82 (br d,J= 14.3 Hz, 1H), 7.79 - 7.75 (m, 2H), 7.73 - 7.68 (m, 2H), 7.64 - 7.57 (m, 1H), 7.43 (br d,J= 4.4 Hz, 2H), 7.30 - 7.23 (m, 2H), 6.90 (d,J= 13.6 Hz, 1H), 4.47 - 4.35 (m, 2H), 4.01 - 3.89 (m, 1H), 3.58 (br d,J= 11.9 Hz, 2H), 3.22 - 2.94 (m, 8H), 2.73 - 2.63 (m, 2H), 2.60 - 2.51 (m, 2H), 2.22 - 2.13 (m, 1H), 2.01 - 1.92 (m, 1H), 1.75 - 1.53 (m, 4H), 1.50 - 1.25 (m, 3H).

[0834]

[0835] Example 56. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2,4-difluorobenzamide (Compound 56)

[0836]

[0837] Step 1. Synthesis of N-(4-((2-chloro-5-fluoropyrimidin-4-yl)amino)phenyl)-2,4-difluorobenzamide (2)

[0838] The title compound (2.5 g, 5.94 mmol, 81.72% yield, 90% purity) was obtained as a brown solid by a similar method to step 1 of Example 7. MS (M+H)+ = 379.0.

[0839] Step 2. Synthesis of methyl 4-((4-((4-(2,4-difluorobenzamido)phenyl)amino)-5-fluoropyrimidin-2-yl)amino)benzoate (3)

[0840] The title compound (700 mg, crude) was obtained as a white solid by a similar method to step 1 of Example 53, and the product was used directly in the following reaction. MS(M+H) + = 494.2.

[0841] Step 3. Synthesis of 4-((4-((4-(2,4-difluorobenzamido)phenyl)amino)-5-fluoropyrimidin-2-yl)amino)benzoic acid (4)

[0842] The title compound (400 mg, crude, Na) was obtained as a yellow solid by a similar method to step 2 of Example 53. MS (M+H) + = 480.0.

[0843] Step 4. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl) piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2,4-difluorobenzamide (Compound 56)

[0844] The title compound (65.9 mg, 54.01 μmol, 13.57% yield, 90.8% purity, 2TFA) was obtained as a white solid by a similar method to step 9 of Example 12. MS (M+H) + = 880.6.

[0845] 1H NMR (400 MHz, DMSO-d6) δ = 10.91 (s, 1H), 10.44 (s, 1H), 9.93 - 9.60 (m, 2H), 9.59 - 9.53 (m, 1H), 8.17 (d,J= 2.9 Hz, 1H), 8.08 (d,J= 11.0 Hz, 1H), 7.82 - 7.75 (m, 5H), 7.73 - 7.67 (m, 4H), 7.44 (dt,J= 2.3, 9.9 Hz, 1H), 7.24 (dt,J= 2.2, 8.4 Hz, 1H), 6.91 (d,J= 13.7 Hz, 1H), 4.42 (br d,J= 13.1 Hz, 2H), 3.95 (dd,J= 4.5, 13.0 Hz, 1H), 3.64 - 3.53 (m, 2H), 3.24 - 3.00 (m, 8H), 2.96 - 2.80 (m, 2H), 2.79 - 2.68 (m, 1H), 2.59 - 2.52 (m, 1H), 2.25 - 2.13 (m, 1H), 2.02 - 1.92 (m, 1H), 1.81 - 1.71 (m, 2H), 1.69 - 1.60 (m, 2H), 1.47 - 1.35 (m, 3H).

[0846]

[0847] Example 57. Synthesis of 4-((4-((4-(2,4-difluorobenzamido)phenyl)amino)-5-fluoropyrimidin-2-yl)amino)-N-(4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridinyl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluorobenzamide (Compound 57)

[0848]

[0849] Step 1. Synthesis of methyl 4-((4-((4-(2,4-difluorobenzamido)phenyl)amino)-5-fluoropyrimidin-2-yl)amino)-2-fluorobenzoate (2)

[0850] The title compound (750 mg, crude) was obtained as a white solid by a similar method to step 1 of Example 53, and was used directly in the following reaction. MS(M+H) + = 512.1.

[0851] Step 2. Synthesis of 4-((4-((4-(2,4-difluorobenzamido)phenyl)amino)-5-fluoropyrimidin-2-yl)amino)-2-fluorobenzoic acid (3)

[0852] The title compound (400 mg, crude, Na) was obtained as a yellow solid by a similar method to step 2 of Example 53. MS (M+H) + = 498.2.

[0853] Step 3. Synthesis of 4-((4-((4-(2,4-difluorobenzamido)phenyl)amino)-5-fluoropyrimidin-2-yl)amino)-N-(4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluorobenzamide (Compound 57)

[0854] The title compound (48.5 mg, 40.92 μmol, 10.65% yield, 95% purity, 2TFA) was obtained as a white solid by a similar method to step 9 of Example 12. MS (M+H) + = 898.6.

[0855] 1H NMR (400 MHz, DMSO-d6) δ = 10.90 (s, 1H), 10.43 (s, 1H), 9.72 (br s, 1H), 9.66 - 9.52 (m, 1H), 9.33 - 9.18 (m, 1H), 8.18 (d,J= 3.5 Hz, 1H), 8.08 (d,J= 11.0 Hz, 1H), 7.83 - 7.69 (m, 6H), 7.47 - 7.40 (m, 3H), 7.24 (dt,J= 2.3, 8.3 Hz, 1H), 6.90 (d,J= 13.6 Hz, 1H), 4.46 - 4.36 (m, 2H), 3.95 (dd,J= 5.0, 12.8 Hz, 1H), 3.58 (br d,J= 11.1 Hz, 2H), 3.23 - 3.01 (m, 8H), 2.79 - 2.65 (m, 3H), 2.59 - 2.53 (m, 1H), 2.24 - 2.13 (m, 1H), 2.02 - 1.92 (m, 1H), 1.76 - 1.56 (m, 4H), 1.42 - 1.25 (m, 3H).

[0856]

[0857] Example 58. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-methoxybenzamide (Compound 58)

[0858]

[0859] Step 1. Synthesis of N-(4-((2-chloro-5-fluoropyrimidin-4-yl)amino)phenyl)-2-methoxybenzamide (2)

[0860] The title compound (2.8 g, crude) was obtained as a yellow solid by a similar method to step 1 of Example 47. MS (M+H) + = 373.0.

[0861] Step 2. Synthesis of methyl 4-((5-fluoro-4-((4-(2-methoxybenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzoate (3)

[0862] The title compound (1.2 g, crude) was obtained as a white solid by a similar method to step 1 of Example 53. MS (M+H) + = 488.3.

[0863] Step 3. Synthesis of 4-((5-fluoro-4-((4-(2-methoxybenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzoic acid (4)

[0864] The title compound (1.4 g, crude, Na salt) was obtained as a yellow solid by a similar method to step 5 of Example 7. MS (M+H) + = 474.2.

[0865] Step 4. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-methoxybenzamide (Compound 58)

[0866] The title compound (9.8 mg, 8.80 μmol, 5.69% yield, 99% purity, 2TFA salt) was obtained as a white solid by a similar method to step 9 of Example 12. MS (M+H) + = 874.6.

[0867] 1H NMR (400 MHz, DMSO-d6) δ = 11.00 - 10.83 (m, 1H), 10.14 (s, 1H), 9.64 - 9.40 (m, 3H), 8.20 - 8.01 (m, 2H), 7.83 - 7.62 (m, 9H), 7.45 (s, 1H), 7.23 - 7.16 (m, 1H), 7.11 - 7.03 (m, 1H), 6.94 - 6.86 (m, 1H), 4.46 - 4.37 (m, 2H), 3.98 - 3.88 (m, 4H), 3.63 - 3.53 (m, 2H), 3.23 - 2.97 (m, 7H), 2.84 - 2.70 (m, 2H), 2.57 - 2.57 (m, 3H), 2.24 - 2.10 (m, 1H), 2.03 - 1.91 (m, 1H), 1.78 - 1.58 (m, 4H), 1.43 - 1.28 (m, 3H).

[0868]

[0869] Example 59. Synthesis of N-(4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-methoxybenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 59)

[0870]

[0871] Step 1. Synthesis of methyl 2-fluoro-4-((5-fluoro-4-((4-(2-methoxybenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzoate (2)

[0872] The title compound (1.4 g, crude) was obtained as a yellow solid by a similar method to step 1 of Example 53. MS (M+H) + = 506.3.

[0873] Step 2. Synthesis of 2-fluoro-4-((5-fluoro-4-((4-(2-methoxybenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzoic acid (3)

[0874] The title compound (1.5 g, crude, Na salt) was obtained as a yellow solid by a similar method to step 5 of Example 7. MS (M+H) + = 492.2.

[0875] Step 3. Synthesis of N-(4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-ylethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-methoxybenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 59)

[0876] The title compound (21.8 mg, 19.08 μmol, 10.16% yield, 98% purity, 2TFA salt) was obtained as a white solid by a similar method to step 9 of Example 12. MS (M+H) + = 892.6.

[0877] 1 H NMR (400 MHz, DMSO-d6) δ = 10.90 (s, 1H), 10.15 - 10.07 (m, 1H), 9.74 - 9.65 (m, 1H), 9.56 - 9.41 (m, 1H), 9.24 - 8.96 (m, 1H), 8.17 (br d,J= 2.8 Hz, 1H), 8.11 - 8.00 (m, 1H), 7.83 - 7.62 (m, 6H), 7.54 - 7.46 (m, 1H), 7.44 - 7.36 (m, 2H), 7.19 (br d,J= 8.4 Hz, 1H), 7.10 - 7.03 (m, 1H), 6.94 - 6.84 (m, 1H), 4.46 - 4.32 (m, 2H), 3.99 - 3.84 (m, 4H), 3.63 - 3.48 (m, 2H), 3.24 - 2.92 (m, 7H), 2.78 - 2.63 (m, 2H), 2.46 - 2.35 (m, 3H), 2.25 - 2.08 (m, 1H), 2.02 - 1.90 (m, 1H), 1.75 - 1.54 (m, 4H), 1.38 - 1.21 (m, 3H).

[0878]

[0879] Example 60. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluoro-4-methoxybenzamide (Compound 60)

[0880]

[0881] Step 1. Synthesis of N-(4-((2-chloro-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluoro-4-methoxybenzamide (2)

[0882] The title compound (2.54 g, 6.11 mmol, 56.03% yield, 94% purity) was obtained as a yellow solid by a similar method to step 1 of Example 47. MS (M+H) + = 391.2.

[0883] Step 2. Synthesis of methyl 4-((5-fluoro-4-((4-(2-fluoro-4-methoxybenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzoate (3)

[0884] The title compound (1.1 g, crude) was obtained as a white solid by a similar method to step 1 of Example 53. MS (M+H) + = 506.1.

[0885] Step 3. Synthesis of 4-((5-fluoro-4-((4-(2-fluoro-4-methoxybenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzoic acid (4)

[0886] The title compound (360 mg, crude, Li salt) was obtained as a white solid by a similar method to step 5 of Example 7. MS (M+H) + = 492.1.

[0887] Step 4. Synthesis of N-(4-((2-((4-((4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)carbamoyl)phenyl)amino)-5-fluoropyrimidin-4-yl)amino)phenyl)-2-fluoro-4-methoxybenzamide (Compound 60)

[0888] The title compound (55.6 mg, 47.16 μmol, 23.18% yield, 95% purity, 2TFA salt) was obtained as a white solid by similar synthesis to step 9 of Example 12. MS (M+H) + = 892.5.

[0889] 1 H NMR (400 MHz, DMSO-d6) δ = 10.90 (s, 1H), 10.20 (s, 1H), 9.67 - 9.45 (m, 3H), 8.16 (t,J= 3.4 Hz, 1H), 8.08 (d,J= 11.1 Hz, 1H), 7.82 - 7.63 (m, 9H), 6.98 (dd,J= 2.4, 12.5 Hz, 1H), 6.93 - 6.87 (m, 2H), 4.42 (br d,J= 13.9 Hz, 1H), 3.95 (br dd,J= 4.7, 12.6 Hz, 1H), 3.85 (s, 3H), 3.60 - 3.59 (m, 2H), 3.23 - 3.11 (m, 6H), 3.09 - 2.97 (m, 2H), 2.90 - 2.68 (m, 2H), 2.59 - 2.53 (m, 3H), 2.25 - 2.12 (m, 1H), 2.02 - 1.91 (m, 1H), 1.80 - 1.58 (m, 4H), 1.45 - 1.31 (m, 3H).

[0890]

[0891] Example 61. Synthesis of N-(4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluoro-4-methoxybenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 61)

[0892]

[0893] Step 1. Synthesis of methyl 2-fluoro-4-((5-fluoro-4-((4-(2-fluoro-4-methoxybenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzoate (2)

[0894] The title compound (1.3 g, crude) was obtained as a white solid by a similar method to step 1 of Example 53. MS (M+H) + = 524.1.

[0895] Step 2. Synthesis of 2-fluoro-4-((5-fluoro-4-((4-(2-fluoro-4-methoxybenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzoic acid (3)

[0896] The title compound (360 mg, crude) was obtained as a white solid by a similar method to step 5 of Example 7. MS (M+H) + = 510.1.

[0897] Step 3. Synthesis of N-(4-(2-(4-(5-(2,6-dioxopiperidin-3-yl)-4-fluoropyridin-2-yl)piperazin-1-yl)ethyl)piperidin-1-yl)-2-fluoro-4-((5-fluoro-4-((4-(2-fluoro-4-methoxybenzamido)phenyl)amino)pyrimidin-2-yl)amino)benzamide (Compound 61)

[0898] The title compound (26 mg, 18.90 μmol, 9.63% yield, 91% purity, 3TFA salt) was obtained as a white solid by a similar method to step 9 of Example 12. MS (M+H) + = 910.5.

[0899] 1 H NMR (400 MHz, MeOD) δ = 8.13 - 8.05 (m, 2H), 7.77 (td,J= 4.4, 8.5 Hz, 3H), 7.71 - 7.57 (m, 4H), 7.32 (dd,J= 2.0, 8.6 Hz, 1H), 6.94 - 6.82 (m, 2H), 6.78 (d,J= 13.2 Hz, 1H), 4.60 - 4.39 (m, 1H), 3.95 (dd,J= 4.7, 12.7 Hz, 1H), 3.90 (s, 3H), 3.74 - 3.57 (m, 2H), 3.28 - 3.12 (m, 9H), 2.84 - 2.58 (m, 4H), 2.35 - 2.22 (m, 1H), 2.18 - 2.08 (m, 1H), 1.82 - 1.66 (m, 4H), 1.63 - 1.48 (m, 2H), 1.45 - 1.32 (m, 1H).

[0900]

[0901] Experimental Example 1. Measurement of AURKA protein degradation capacity using a luciferase assay.

[0902] Production and culture of HeLa LgBit (AURKA-HiBit KI) cell line

[0903] A HeLa cell line was transfected with the LgBit vector to create a stable overexpressing cell line. Next, a DNA donor was designed and manufactured to express the HiBit amino acid sequence at the C-terminal end of the Aurora kinase A (AURKA) gene endogenously present in the cells, and a guide RNA was also manufactured to enable its insertion. This was then inserted into the cells together with a vector capable of expressing clustered regularly interspaced palindromic repeats (CRISPR) / Cas9. Only single cells with complete insertion of the HiBit gene were selected and passaged for use.

[0904] For cell culture, DMEM (Gibco, Cat. No. 11995-065; Lot. No. 2994601), FBS (Gibco, Cat. No. 16000-044; Lot. No. U3109271P), penicillin / streptomycin (PS) (Gibco, Cat. No. 15140-122; Lot. No. 227766), 90 ㎟ cell culture dish (SPL, Cat. No. 20100), 96-well white plate (SPL, Cat. No. 30196), PBS pH7.4 (Gibco, Cat. No. 10010-023; Lot. No. 3034813), TrypLE TM Express (Gibco, Cat. No. 12605-010; Lot. No. 2807766), counting chamber (Hematocytometer) (Hirschmann, Cat. No. 8100204; Lot. No. 8021087), and 0.4% trypan blue solution (DYNEBIO, Cat. No. CBT3710; Lot. No. 20240731) were used. Culture conditions were DMEM medium supplemented with 10% FBS and 1% PS, and cultured in a CO2 incubator (Thermo, Cat. No. 4111) at 5% CO2 and 37°C.

[0905] Compound treatment and luciferase assay experimental method of the present invention

[0906] 90mm for HeLa LgBit (AURKA-HiBit KI) 2 After culturing in cell culture dishes, 3 x 10 in each well of a 96-well white plate 4Distribute to the dogs. The total medium volume was 150 ㎕ and cultured in a CO2 incubator for 24 hours. After culture, Eudurazine (Promega, Cat. No. N257B; Lot. No. 495218) was added to each well of a 96-well white plate to make 1% of the total volume, and cultured in a CO2 incubator for another 2 hours.

[0907] The compound of the present invention was completely dissolved in DMSO (Sigma-Aldrich Cat. No. D2438, Lot. No. RNBM8168) and used in the experiment, and the final concentration of each compound was 10 nM or 100 nM when treating the compound. After treating the compound in each well of a 96-well white plate, it was placed in a plate reader (BMG Labtech, CLARIOstar Plus) and analyzed in real time. The analysis wavelength was set to 470-480 nM, and the luminescence value was measured after 12 hours. The luminescence value was converted into an Excel program, and the ratio of the decrease compared to the DMSO treatment group when treating with 10 nM and 100 nM of the compound was expressed as the activity.

[0908] The activity values ​​according to the compound 10 nM treatment group are as described in Table 3 below (++++: 0.3 or less; +++: 0.3 to 0.5; ++: 0.5 to 0.7; +: 0.7 to 0.8).

[0909] [Table 3]

[0910]

[0911]

[0912] The activity values ​​according to the compound 100 nM treatment group are as shown in Table 4 below (++++: 0.2 or less; +++: 0.2 to 0.4; ++: 0.4 to 0.6; +: 0.6 to 0.8).

[0913] [Table 4]

[0914]

[0915]

[0916] 2. Measurement of cell line killing effect

[0917] NCI-H446, HEL, SET-2, and K-562 cell line culture

[0918] NCI-H446 (hereinafter H446), HEL92.1.7 (hereinafter HEL), and SET-2 cell lines were provided by ATCC, and K-562 cell line was provided by Korea Cell Line Bank. For cell culture, RPMI medium 1640 HEPES (Gibco, Cat. No. 22400-089; Lot. No. 2867221), FBS (Gibco, Cat. No. 16000-044; Lot. No. U2781540P), penicillin / streptomycin (PS) (Gibco, Cat. No. 15140-122; Lot. No. 227766), 75T cell culture flask (SPL, Cat. No. 70075), 96-well cell culture plate (SPL, Cat. No. 30096), PBS pH7.4 (Gibco, Cat. No. 10010-023; Lot. No. 2561466), TrypLE TMExpress (Gibco, Cat. No. 12605-010; Lot. No. 2492811), a counting chamber (Hematocytometer) (Hirschmann, Cat. No. 8100204), and 0.4% trypan blue solution (DYNEBIO, Cat. No. CBT3710; Lot. No. 20231017) were used. The three cell lines, H446, SET-2, and K-562, were cultured in RPMI1640 medium containing 10% FBS and 1% PS, and the HEL cell line was cultured in RPMI1640 medium containing 20% ​​FBS and 1% PS. Cells were cultured in a CO2 incubator (ThermoScientific, Cat. No. 4111) at 37°C and 5% CO2 conditions.

[0919] Compound treatment and cell viability assay experimental method of the present invention

[0920] Comparative and example compounds were completely dissolved in DMSO (Sigma-Aldrich Cat. No. D2438, Lot. No. RNBL7287) and used in the experiment.

[0921] H446, HEL, SET-2, K-562 cell lines per well in a 96-well cell culture plate

[0922] 6 x 10 3 Dog, 5 x 10 3 Dog, 2 x 10 4 Dog, 2 x 10 3 The cells were seeded so that they would be 150 μl, and the medium volume of each well was adjusted to 150 μl. H446 cells were seeded in a 96-well cell culture plate and compound treatment was performed 24 hours later, and HEL, SET-2, and K562 cells were treated with compounds 5 hours after seeding in a 96-well cell culture plate.

[0923] The intracellular treatment capacity of the compound of the present invention was set to a maximum concentration of 1000 nM, and this was diluted by 1 / 3 and treated at each concentration, with a minimum concentration of 0.15 nM. After treating the compound so that the total solution volume in each well was 200 μl, the cells were cultured in a CO2 incubator (Thermo Fisher Science, Cat. No. 4111) for 5 days.

[0924] Afterwards, 20 ㎕ of EZ-Cytox (DOGEN, Cat. NO. EZ-3000, Lot. No. DLS2403) was treated to each well and cultured for 4 hours in a CO2 incubator under the same conditions. After the culture was completed, the absorbance of the sample was measured by setting the wavelength of the plate reader (BMG Labtech, CLARIOstar Plus) to 450 nM, and the measurement was performed after shaking for 2 minutes in the plate reader. The final measurement values ​​were organized in an Excel file and displayed as a graph using the Prism-GraphPad program, and the IC 50 The value was measured.

[0925] IC of this compound against human bone marrow-derived carcinoma cell lines HEL, SET-2, and K562 50 The values ​​are as shown in Table 5 below (+++++: less than 50 nM, ++++: 50 to 100 nM, +++: 100 to 150 nM, ++: 150 to 200 nM, +: 200 to 250 nM).

[0926] [Table 5]

[0927]

[0928]

[0929] IC of this compound against human small cell lung cancer cell line H446 50The values ​​are as shown in Table 6 below (++++: less than 10 nM, +++: 10 to 20 nM, ++: 20 to 30 nM, +: 30 to 50 nM). This shows an overall improved effect compared to the cell killing effect after treatment with the comparative compound (see International Patent Publication No. WO2024 / 155112 A1).

[0930] [Table 6]

[0931]

Claims

1. A compound represented by the following chemical formula I: [Chemical Formula I] In the above chemical formula I, PTM is an Aurora kinase A (AURKA) protein binding moiety represented by the following chemical formula II, [Chemical Formula II] {In the above chemical formula II, R1 is hydrogen or halo; R2 is hydrogen, halo or CF3; R3 is CH, N or CF; R4 is -CONH- or -NHCO-; is phenyl, pyridyl or pyrimidinyl, which may be substituted with 1 to 2 halo or OCH3. ULM is an E3 ubiquitin ligase binding moiety represented by the following chemical formula III, [Chemical Formula III] {In the above chemical formula III, U is a covalent bond or -NH-; Silver has 1 to 3 halos, C 1-3 Alkyl or OC 1-3 phenyl, pyridyl or pyrimidinyl which may be substituted with alkyl} Linker is a group that connects ULM and PTM, represented by the following chemical formula IV. [Chemical Formula IV] {In the above chemical formula IV, X1 to X4 are each independently CH or N; X5 is hydrogen or CH3; L M is -CH2CH2-, -CH2NH-, -CH2N(CH3)-, -CH2CHOH- or -CH2CH2CH2-; L P is -NHCO-, -CO- or -NHCH2-; X1 and L P are each {Covalently linked to ULM and PTM via 2. A compound in which the moiety represented by chemical formula II in paragraph 1 is a moiety represented by chemical formulas II-1 to II-4 below. [Chemical Formula II-1] {In the above chemical formula II-1, R2 and R5 are each independently hydrogen, F or Cl; R3 is CH, N or CF} [Chemical Formula II-2] {In the above chemical formula II-2, R1 is hydrogen or F; R2 is hydrogen, F or CF3; R5 is hydrogen, F or OCH3} [Chemical Formula II-3] {In the above chemical formula II-3, R1 is hydrogen or F; R5 is hydrogen or F; R 6A and R 6B are each independently CH or N} [Chemical Formula II-4] {In the above chemical formula II-4, R1 is hydrogen or F; R 5A and R 5B are each independently hydrogen, F or OCH3} 3. A compound in which the moiety represented by chemical formula III in paragraph 1 is a moiety represented by chemical formulas III-1 to III-4 below. [Chemical Formula III-1] {In the above chemical formula III-1, U2 and U3 are each independently hydrogen, F or Cl. [Chemical Formula III-2] {In the above chemical formula III-2, U2 is hydrogen, F or Cl respectively; U 4A and U 4B {One of them is CH and the other is N} [Chemical Formula III-3] {In the above chemical formula III-3, U2 and U3 are each independently hydrogen, F or OCH3} [Chemical Formula III-4] {In the above chemical formula III-4, U2 is hydrogen, F or Cl; U 4A and U 4B {One of them is CH and the other is N} 4. In the first paragraph, the group represented by the chemical formula IV is a compound represented by the following chemical formulas IV-1 to IV-4 {wherein, the group and {Covalently linked to ULM and PTM, respectively}. [Chemical Formula IV-1] {In the above chemical formula IV-1, X5 is hydrogen or CH3; X6 is hydrogen or OH} [Chemical Formula IV-2] {In the above chemical formula IV-2, X7 is -CH2- or -CO-; [Chemical Formula IV-3] {In the above chemical formula IV-3, X4 is CH or N, X6 is hydrogen or CH3} [Chemical Formula IV-4] {In the above chemical formula IV-4, X5 is hydrogen or CH3} 5. A compound according to claim 1, wherein the compound is selected from compounds 1 to 61 below.

6. A compound according to any one of claims 1 to 5, which induces degradation of aurora kinase A protein.

7. A pharmaceutical composition comprising a compound according to any one of claims 1 to 5 and at least one pharmaceutically acceptable carrier.

8. A pharmaceutical composition for preventing or treating a disease associated with aurora kinase A protein, comprising a compound according to any one of claims 1 to 5.

Images