ADAR1 protein degradation agent

Abstract

The invention discloses a compound shown as a general formula I, or a stereoisomer or pharmaceutically acceptable salt thereof, and a composition containing one or more compounds shown as the general formula I. The compound / composition disclosed by the invention is a novel RNA (Ribonucleic Acid) editing enzyme ADAR1 (ADAR1) regulator, and has the capability of degrading ADAR1 protein and is capable of remarkably inhibiting the proliferation of related cancer cells.

Description

TECHNICAL FIELD

[0001] The present patent relates to the field of medicine, and discloses a kind of degradable ADAR1 protein compound, or its stereoisomer or pharmaceutically acceptable salt and the pharmaceutical composition containing above-mentioned compound, and the application of using the compound in treating ADAR1 mediated disease.The present patent also discloses the method for preparing the compound. BACKGROUND

[0002] ADAR1 (Adenosine Deaminases Acting on RNA1) is an RNA editing enzyme, the most common function is to convert adenosine (A) on the RNA molecule into inosine (I), through A-to-I editing, ADAR1 can affect the splicing, transport, stability and translation of mRNA, and then regulate gene expression, and plays an important role in antiviral, tumor development, immune escape, etc. For example, in 2018, W. Nicholas Haining et al. in the paper published in Nature pointed out that most cancer patients do not respond to tumor immunotherapy or develop resistance to it, usually because acquired mutations impair antigen presentation, and the loss of RNA editing enzyme ADAR1 function in tumor cells makes the tumor extremely sensitive to immunotherapy and overcomes the resistance to tumor immunotherapy. Specifically, in the absence of ADAR1, the A-to-I editing of interferon-induced RNA is reduced, leading to the activation of downstream PKR and MDA5 pathways, further leading to growth inhibition and tumor immune response. The loss of ADAR1 overcomes the resistance to PD-1 checkpoint blockade caused by inactivation of tumor cell antigen presentation. Therefore, inducing sufficient immune response in interferon-sensitive tumors can enhance the effect of tumor immunotherapy and overcome the dependence of tumor immunotherapy on CD8+ T cell activation, which is a strategy to overcome immunotherapy resistance. The above more detailed molecular mechanism was further elucidated in 2022, three research groups published papers in Nature at the same time, the research showed that ADAR1 negatively regulates the activity of ZBP1, and in the absence of ADAR1 Zα domain function, ZBP1 can promote autoimmune diseases, on the contrary, overexpression of ADAR1 can inhibit the activation of ZBP1 through Z-RNA editing, thereby preventing ZBP1-mediated programmed necrosis and tumor immune activation. Knockout of ZBP1 rescues the phenotypic changes caused by loss of function of ADAR1 Zα domain. ADAR1 can inhibit ZBP1-mediated apoptosis, and the Zα domain of ADAR1 can inhibit ZBP1 by recognizing dsRNA, inhibiting caspase-8-dependent apoptosis and MLKL-mediated necrosis. In addition, ADAR1 can also reduce the expression level of dsRNA through A-to-I editing of endogenous Alu elements, thereby inhibiting the activation of ZBP1.

[0003] Further, ADAR1 has the function of regulating gene expression independently of A-to-I editing, ADAR1 can promote the interaction between HuR and SIRT1 mRNA, thereby increasing the stability of SIRT1 mRNA. The increase of SIRT1 expression in turn inhibits p16 INK4atranslation of mRNA, thereby inhibiting the occurrence of aging. The expression level of ADAR1 can be a potential biomarker of aging. These results deepen the understanding of the function of ADAR1, and by adjusting the expression level of ADAR1, new therapeutic means can be provided for the intervention and treatment of tumors, autoimmune diseases and aging.

[0004] However, there is currently a lack of specific small molecule modulators of ADAR1, which limits the development of drugs for the treatment of tumors, autoimmune diseases and aging, etc. by targeting ADAR1. The present application provides a novel class of ADAR1 modulator molecules. SUMMARY

[0005] In a first aspect, the present application provides a compound of general formula I, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0006]

[0007] In formula I, X is N or CH,

[0008] R 1 independently at each occurrence H, halogen, hydroxyl, thiol, nitro, amino, cyano, C1-6alkyl, C1-6alkyloxy, haloC1-6alkyl, or haloC1-6alkyloxy;

[0009] m is 0, 1, 2, 3, or 4;

[0010] R 2 independently at each occurrence H, halogen, hydroxyl, thiol, nitro, amino, cyano, C1-6alkyl, C1-6alkyloxy, haloC1-6alkyl, or haloC1-6alkyloxy;

[0011] n is 0, 1, 2, 3, or 4;

[0012] R 3 independently at each occurrence H, halogen, hydroxyl, thiol, nitro, amino, cyano, C1-6alkyl, C1-6alkyloxy, haloC1-6alkyl, or haloC1-6alkyloxy;

[0013] p is 0, 1, 2, 3, or 4;

[0014] linker is selected from formula II:

[0015] -Y-(CHR a ) a -(CHR b ) b -(CHR c ) c -Z-

[0016] (Formula II)

[0017] Y is CONH, NH, CH2or O, R a independently at each occurrence hydrogen, halogen, hydroxyl, thiol, nitro, amino, cyano, C 1-6 alkyl, C 1-6 alkyloxy, haloC 1-6 alkyl or haloC 1-6 alkyloxy, a is 0, 1, 2, 3 or 4, wherein one or more (e.g. 1, 2, 3 or 4) of the CH a ) a R groups are optionally replaced with O, NH or CO; or -Y-(CHR a ) a ) a together or -(CHR a ) a individually form a 4-6 membered (e.g. 4, 5, 6 membered) monocyclic saturated heterocycloalkyl or a 7 to 11 membered (e.g. 7, 8, 9, 10, 11 membered) spirocyclic saturated heterocycloalkyl, the ring heteroatoms of the heterocycloalkyl being 1 or 2 heteroatoms independently at each occurrence selected from nitrogen and oxygen, the heterocycloalkyl being optionally substituted with 1, 2, 3 or 4 substituents selected from halogen, hydroxyl, thiol, nitro, amino, cyano, C 1-6 alkyl, C 1-6 alkyloxy, haloC 1-6 alkyl or haloC 1-6 alkyloxy;

[0018] R b independently at each occurrence hydrogen, halogen, hydroxyl, thiol, nitro, amino, cyano, C 1-6 alkyl, C 1-6 alkyloxy, haloC 1-6 alkyl or haloC 1-6 alkyloxy, b is 0, 1, 2, 3 or 4, wherein one or more (e.g. 1, 2, 3 or 4) of the CH b ) b R groups are optionally replaced with O, NH or CO; or -(CHR b ) a ) a -(CHR b ) b together or -(CHR b ) b -(CHR c ) c together or (CHR b ) bIt is formed individually as a 4-6 member (e.g., 4, 5, 6) monocyclic saturated cycloalkyl or a 4-6 member (e.g., 4, 5, 6) monocyclic saturated heterocyclic alkyl or a 7-11 member (e.g., 7, 8, 9, 10, 11) spirocyclic saturated heterocyclic alkyl, wherein the heterocyclic heteroatom is one or two heteroatoms independently selected from nitrogen and oxygen each time it appears, and the cycloalkyl or heterocyclic alkyl is optionally substituted with one, two, three or four heteroatoms selected from halogen, hydroxyl, mercapto, nitro, amino, cyano, C 1-6 Alkyl, C 1-6 Alkyloxy group, halogenated C 1-6 Alkyl or halogenated C 1-6 Substituents of alkyloxy groups;

[0019] Z is CONH, NH, CH2 or O, R c Each time it appears, it is independently hydrogen, halogen, hydroxyl, mercapto, nitro, amino, cyano, C 1-6 Alkyl, C 1-6 Alkyloxy group, halogenated C 1-6 Alkyl or halogenated C 1-6 Alkyloxy group, where c is 0, 1, 2, 3, or 4, wherein (CHR) c ) c One or more (e.g., 1, 2, 3, or 4) CHR c You may choose to replace it with O, NH or CO; or -(CHR) b ) b -(CHR c ) c -Together or-(CHR) c ) c -Z- together or -(CHR) c ) c - Formed individually as a 4-6 member (e.g., 4, 5, 6 member) monocyclic saturated heterocyclic alkyl group or a 7-11 member (e.g., 7, 8, 9, 10, 11 member) spirocyclic saturated heterocyclic alkyl group, wherein the heterocyclic heteroatom is one or two heteroatoms independently selected from nitrogen and oxygen each time it appears, and the heterocyclic alkyl group is optionally substituted with one, two, three or four heteroatoms selected from halogen, hydroxyl, mercapto, nitro, amino, cyano, C 1-6 Alkyl, C 1-6 Alkyloxy group, halogenated C 1-6 Alkyl or halogenated C 1-6 Substituents of alkyloxy groups;

[0020] The E3 binder portion is selected from the structure described in Equation III:

[0021]

[0022] In Equation III, R 4 R5 , R 6 , R 7 each independently H, halogen, hydroxyl, thiol, nitro, amino, cyano, C 1-6 alkyl, C 1-6 alkyloxy, haloC 1-6 alkyl, or haloC 1-6 alkyloxy;

[0023] M is CH or N;

[0024] L is (CH2) d wherein d is 0, 1, 2, or 3, and one or more (e.g., 1, 2, or 3) CH2in (CH2) d is optionally replaced with NH or CO;

[0025] R 8 is, at each occurrence, independently halogen, hydroxyl, thiol, nitro, amino, cyano, C1-6alkyl, C1-6alkyloxy, C1-6alkylthio, haloC1-6alkyl, or haloC1-6alkylthio;

[0026] e is 0, 1, 2, 3, or 4.

[0027] In a second aspect, the present application provides a compound of the general formula IV, or a stereoisomer or a pharmaceutically acceptable salt thereof:

[0028]

[0029] In formula IV, X, R 1 , R 2 , R 3 , m, n, p, linker are as described in the first aspect with respect to formula I, and R 9 , R 10 are as described in the first aspect with respect to R 4 , R 5 in formula III, respectively.

[0030] In a third aspect, the present application provides a compound of the general formula V, or a stereoisomer, or a pharmaceutically acceptable salt thereof:

[0031]

[0032] In formula IV, X, R 1 , R 2 , R 3 , R 8 , linker, L, M, m, n, p, e are as described in the first aspect.

[0033] In a fourth aspect, the present application provides the compound of one of the first through third aspects, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein X is N.

[0034] In a fifth aspect, the present application provides the compound of one of the first through fourth aspects, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein R 1 independently at each occurrence is halogen or Ci-6alkyl; and / or m is 1 or 2.

[0035] In a sixth aspect, the present application provides the compound of one of the first through fifth aspects, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein R 2 independently at each occurrence is halogen; and / or n is 1.

[0036] In a seventh aspect, the present application provides the compound of one of the first through sixth aspects, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein R 3 independently at each occurrence is halogen; and / or p is 1.

[0037] In an eighth aspect, the present application provides the compound of one of the first through seventh aspects, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein linker is selected from:

[0038]

[0039]

[0040]

[0041] In a ninth aspect, the present application provides the compound of one of the first through eighth aspects, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein E3 binder is selected from:

[0042]

[0043]

[0044] In a tenth aspect, the present application provides the following compounds, or a stereoisomer or pharmaceutically acceptable salt thereof:

[0045]

[0046]

[0047]

[0048]

[0049]

[0050]

[0051]

[0052]

[0053]

[0054]

[0055]

[0056]

[0057]

[0058]

[0059]

[0060]

[0061]

[0062]

[0063]

[0064]

[0065]

[0066]

[0067]

[0068]

[0069]

[0070]

[0071] In an eleventh aspect, the present application provides a pharmaceutical composition comprising a compound of one of the first through tenth aspects, or a stereoisomer or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0072] In a twelfth aspect, the present application provides a method for degrading ADAR1 protein, the method comprising contacting the ADAR1 protein with a compound of one of the first through tenth aspects, or a stereoisomer or pharmaceutically acceptable salt thereof.

[0073] In a 13th aspect, the present application provides a method of treating a disease mediated by ADAR1 protein comprising administering to a patient having a disease mediated by ADAR1 protein a compound of one of the first through tenth aspects, or a stereoisomer or pharmaceutically acceptable salt thereof.

[0074] In a 14th aspect, the present application provides a compound of one of the first through tenth aspects, or a stereoisomer or pharmaceutically acceptable salt thereof, for use in degrading ADAR1 protein.

[0075] In a 15th aspect, the present application provides a compound of one of the first through tenth aspects, or a stereoisomer or pharmaceutically acceptable salt thereof, for use in treating a disease mediated by ADAR1 protein.

[0076] In a 16th aspect, the present application provides use of a compound of one of the first through tenth aspects, or a stereoisomer or pharmaceutically acceptable salt thereof, in the manufacture of a medicament for degrading ADAR1 protein.

[0077] In a 17th aspect, the present application provides use of a compound of one of the first through tenth aspects, or a stereoisomer or pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating a disease mediated by ADAR1 protein. BRIEF DESCRIPTION OF DRAWINGS

[0078] Figure 1 Inhibition of MDA-MB-468 cell proliferation by the compound of Example 1

[0079] Figure 2 Degradation of ADAR1 protein by the compound of Example 1

[0080] Figure 3 Degradation of ADAR1 protein by the compound of Example 1 DETAILED DESCRIPTION

[0081] THF: tetrahydrofuran

[0082] HOBT: 1-hydroxybenzotriazole

[0083] EDCI: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride

[0084] DIEA: N,N-diisopropylethylamine

[0085] DMF: N,N-dimethylformamide

[0086] TFA: trifluoroacetic acid

[0087] TCFH: N,N,N',N'-tetramethyluronium hexafluorophosphate

[0088] NMP: N-methyl pyrrolidone

[0089] SFC: supercritical fluid chromatography

[0090] TMSOTf: trimethylsilyl triflate

[0091] PE: petroleum ether

[0092] BrettPhos: 2-(dicyclohexylphosphino)-3,6-dimethoxy-2'-4'-6'-triisopropyl-1,1'- biphenyl

[0093] RuPhos Pd G3: (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)(2- amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate

[0094] HPLC: high pressure liquid chromatography

[0095] Intermediate 1: (S)-2-(5-chloro-1-(4-chlorobenzyl)-2-oxo-1,2-dihydropyridine-3- carboxamido)-3-(2-(6-chloro-4-methylpyridin-3-yl)phenyl)propanoic acid

[0096]

[0097]

[0098] Step 1: 5-chloro-2-oxo-1,2-dihydropyridine-3-carboxylic acid methyl ester

[0099] To a solution of 5-chloro-2-hydroxy nicotinic acid (14 g, 80.7 mmol) in MeOH (100 mL) was added SOCl2(20 mL) dropwise at 0 °C, then heated to 60 °C and stirred for 2 h. After the reaction was completed, the reaction was concentrated under reduced pressure to give a yellow solid, which was the target product (11 g) and was used directly in the next reaction. LCMS (ESI + ): m / z = 188 [M+H] + .

[0100] Step 2: 5-chloro-1-(4-chlorobenzyl)-2-oxo-1,2-dihydropyridine-3-carboxylic acid methyl ester

[0101] To a solution of 5-chloro-2-oxo-l,2-dihydropyridine-3-carboxylic acid methyl ester (11 g, 58.6 mmol) in DMF (60 mL) was added K2CO3(24.3 g, 175.8 mmol) and 4-chlorobenzyl bromide (16 g, 76.1 mmol), then heated to 100 °C and stirred for 12 h. After the reaction was completed, the reaction solution was diluted with water (30 mL). The aqueous phase was extracted with EtOAc (30 mL x 3), and the organic phase was collected. The ester phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash preparative liquid chromatography (CH2Cl2:MeOH (v / v) = 96:4) to give the product as a yellow solid (11 g, yield 58%). LCMS (ESI + ): m / z = 312 [M+H] + .

[0102] Step 3: 5-chloro-l-(4-chlorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxylic acid

[0103] To a solution of 5-chloro-l-(4-chlorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxylic acid methyl ester (11 g, 35.2 mmol) in a mixed solution (150 mL, H2O:THF:MeOH (v / v / v) = 1:1:1) was added NaOH (5.63 g, 140.8 mmol), and the reaction was stirred at room temperature for 2 h. After the reaction was completed, the pH was adjusted to 2 with 1 M dilute hydrochloric acid. The aqueous phase was extracted with CH2Cl2(30 mL x 3), and the organic phase was combined. The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the product as a white solid (8.5 g), which was used directly in the next reaction. LCMS (ESI + ): m / z = 298 [M+H] + .

[0104] Step 4: (S)-2-((tert-butoxycarbonyl)amino)-3-(2-(6-chloro-4-methylpyridin-3- yl)phenyl)propanoic acid

[0105] In a 250 mL round-bottom flask, was added (S)-3-(2-bromophenyl)-2-((tert- butoxycarbonyl)amino)propanoic acid (10 g, 29.1 mmol), 2-chloro-4-methylpyridine-5- boronic acid (12.47 g, 72.7 mmol), Pd(dppf)Cl2(2.13 g, 2.91 mmol), Cs2CO3(28.44 g, 87.3 mmol), 1,4-dioxane (120 mL) and H2O (30 mL), heated to 100 °C and stirred for 16 h. After the reaction was completed, the reaction was diluted with water (300 mL) and extracted with CH2Cl2(300 mL x 3). The aqueous phase was adjusted to pH = 4 with 4 M HCl and extracted again with CH2Cl2(300 mL x 3). The organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a light yellow solid (10 g), which was used directly in the next step. LCMS (ESI + ): m / z = 391 [M+H] + .

[0106] Step 5: Preparation of (S)-methyl 2-amino-3-(2-(6-chloro-4-methylpyridin-3- yl)phenyl)propanoate

[0107] To a solution of (S)-2-((tert-butoxycarbonyl)amino)-3-(2-(6-chloro-4- methylpyridin-3-yl)phenyl)propanoic acid in methanol (15 mL) was added SOCl2(6 mL) dropwise at 0 °C, then warmed to 25 °C and stirred for 2 h. After the reaction was completed, the reaction was concentrated under reduced pressure to give a yellow solid product (5.5 g), which was used directly in the next step. LCMS (ESI + ): m / z = 305 [M+H] + .

[0108] Step 6: Preparation of (S)-methyl 2-(5-chloro-1-(4-chlorobenzyl)-2-oxo-1,2- dihydropyridine-3-carboxamido)-3-(2-(6-chloro-4-methylpyridin-3-yl)phenyl)propanoate

[0109] In a 100 mL round-bottom flask, add 5-chloro-l-(4-chlorobenzyl)-2-oxo-l,2- dihydropyridine-3-carboxylic acid (4 g, 13.4 mmol), HOBT (2.72 g, 20.1 mmol), EDCI (3.85 g, 20.1 mmol), DIEA (5.20 g, 40.2 mmol) and DMF (30 mL), then add (S)-methyl 2-amino-3-(2-(6-chloro-4-methylpyridin-3-yl)phenyl)propanoate (4.08 g, 13.4 mmol), stir the reaction at room temperature for 16 h. After the reaction is completed, dilute with water (100 mL). Extract the aqueous phase with EtOAc (100 mL x 3), combine the organic phases. Dry the organic phase over anhydrous Na2SO4, filter, and concentrate under reduced pressure, then purify with flash preparation chromatography (CH2Cl2:MeOH (v / v) = 95:5) to obtain a yellow solid (5 g, yield 57%). LCMS (ESI + ): m / z = 584 [M+H] + .

[0110] Step 7: (S)-2-(5-chloro-l-(4-chlorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxamido)-3-(2-(6-chloro-4-methylpyridin-3-yl)phenyl)propanoic acid

[0111] Dissolve (S)-methyl 2-(5-chloro-l-(4-chlorobenzyl)-2-oxo-l,2-dihydropyridine-3- carboxamido)-3-(2-(6-chloro-4-methylpyridin-3-yl)phenyl)propanoate (4.5 g, 7.7 mmol) in MeOH (20 mL). Add H2O (4 mL), LiOH (1.84 g, 77 mmol) to the system, stir the reaction at room temperature for 2 h. After the reaction is completed, dilute with water (50 mL), adjust the pH to 5 with 2 M hydrochloric acid, filter, and dry to obtain a white solid (4 g), which is used directly in the next reaction. LCMS (ESI + ): m / z = 570 [M+H] + .

[0112] Intermediate 2: (S)-2-(5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3- carboxamido)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)propanoic acid

[0113]

[0114] Step 1: methyl 5-fluoro-2-oxo-l,2-dihydropyridine-3-carboxylate

[0115] To a solution of 5-fluoro-2-hydroxybenzoic acid (10 g, 63.7 mmol) in MeOH (100 mL) was added SOCl2(20 mL) dropwise at 0 °C, and the reaction was stirred at room temperature for 4 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain a white solid, which was the target product (10 g) and was used directly in the next reaction. LCMS (ESI + ): m / z = 172 [M+H] + .

[0116] Step 2: 5-Fluoro-l-(4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxylic acid methyl ester

[0117] To a solution of 5-fluoro-2-oxo-l,2-dihydropyridine-3-carboxylic acid methyl ester (10 g, 58.4 mmol) in DMF (100 mL) was added K2CO3(16.1 g, 116.8 mmol) and 4-fluorobenzyl bromide (9.9 g, 52.6 mmol), and then the reaction was stirred at 90 °C for 6 h. After the reaction was completed, the reaction solution was diluted with water (100 mL). The aqueous phase was extracted with EtOAc (100 mL x 3), and the organic phase was collected. The ester phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash preparative liquid chromatography (CH2Cl2:MeOH (v / v) = 90:10) to obtain a yellow oil (9.0 g, yield 52%). LCMS (ESI + ): m / z = 280 [M+H] + .

[0118] Step 3: 5-Fluoro-l-(4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxylic acid

[0119] To a solution of 5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxylic acid methyl ester (9.0 g, 32.2 mmol) in a mixed solvent (100 mL, H2O:MeOH (v / v) = 1:1) was added NaOH (2.58 g, 64.4 mmol), and the reaction was stirred at room temperature for 4 h. After the reaction was completed, the reaction solution was diluted with water (50 mL), the pH was adjusted to 6 with 1M dilute hydrochloric acid, filtered, and dried to obtain a white solid product (7.8 g), which was used directly in the next reaction. LCMS (ESI + ): m / z = 266 [M+H] + .

[0120] Step 4: (S)-2-((tert-Butoxycarbonyl)amino)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)propanoic acid

[0121] In a 500 mL round-bottom flask, was added (S)-3-(2-bromophenyl)-2-((tert- butoxycarbonyl)amino)propanoic acid (30 g, 87.2 mmol), 6-fluoro-4-methylpyridine-3- boronic acid (27.0 g, 174.4 mmol), Pd(dppf)Cl2(6.38 g, 8.72 mmol), Cs2CO3(56.8 g, 174.4 mmol), 1,4-dioxane (300 mL) and H2O (50 mL), heated to 100 °C and stirred for 16 h. After the reaction was completed, the reaction was diluted with water (500 mL), adjusted to pH = 6 with 4 M hydrochloric acid, extracted with EtOAc (300 mL x 3), the organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (CH2Cl2:MeOH (v / v) = 90:10) to obtain a yellow oil (18 g, yield 44%). LCMS (ESI + ): m / z = 375 [M+H] + .

[0122] Step 5: (S)-methyl 2-((tert-butoxycarbonyl)amino)-3-(2-(6-fluoro-4- methylpyridin-3-yl)phenyl)propanoate

[0123] In a 250 mL round-bottom flask, was added (S)-2-((tert-butoxycarbonyl)amino)-3-(2-(6- fluoro-4-methylpyridin-3-yl)phenyl)propanoic acid (18.0 g, 48.1 mmol), K2CO3(13.3 g, 96.2 mmol), DMF (100 mL) and iodomethane (20.5 g, 144.3 mmol), stirred at room temperature for 4 h. After the reaction was completed, diluted with water (500 mL), extracted with EtOAc (300 mL x 3), the organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (PE:EtOAc (v / v) = 70:30) to obtain a yellow oil product (13 g, yield 62%). LCMS (ESI + ): m / z = 389 [M+H] + .

[0124] Step 6: (S)-methyl 2-amino-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)propanoate

[0125] (S)-2-((tert-butoxycarbonyl)amino)-3-(2-(6-fluoro-4-methylpyridin-3- yl)phenyl)propanoic acid methyl ester (13.0 g, 33.5 mmol) was dissolved in CH2Cl2(200 mL), then TFA (19.1 g, 167.5 mmol) was added, the reaction was stirred at room temperature for 2 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain a yellow solid product (8.0 g), which was directly used in the next reaction. LCMS (ESI + ): m / z = 289 [M+H] + .

[0126] Step 7: (S)-2-(5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3- carboxamido)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)propanoic acid methyl ester

[0127] In a 100 mL round-bottom flask, 5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2- dihydropyridine-3-carboxylic acid (7.3 g, 27.7 mmol), TCFH (15.540 g, 55.4 mmol), N-methylimidazole (4.5 g, 55.4 mmol) and CH2Cl2(200 mL) were added, then (S)-2-amino-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)propanoic acid methyl ester (8.0 g, 27.7 mmol) was added, and the reaction was stirred at room temperature for 2 h. After the reaction was completed, water (200 mL) was added for dilution, and extraction was performed with CH2Cl2(200 mL x 3). The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (CH2Cl2:MeOH (v / v) = 90: 10) to obtain a yellow oily product (8.2 g, yield 49%). LCMS (ESI + ): m / z = 536 [M+H] + .

[0128] Step 8: (S)-2-(5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3- carboxamido)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)propanoic acid

[0129] (S)-2-(5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxamido)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)propanoic acid methyl ester (8.2 g, 15.3 mmol) was dissolved in MeOH (50 mL). To the system was added H2O (50 mL), NaOH (1.2 g, 30.6 mmol), and the reaction was stirred at room temperature for 4 h. After the reaction was completed, the reaction was diluted with water (50 mL), adjusted to pH = 6 with 1 M hydrochloric acid, filtered, and dried to give a white solid (6.1 g), which was used directly in the next reaction. LCMS (ESI + ): m / z = 522 [M+H] + .

[0130] Example 1: 5-chloro-N-((2S)-3-(2-(6-chloro-4-methylpyridin-3-yl)phenyl)-l-((4-((2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4-yl)amino)butyl)amino)-l-oxopropan-2-yl)-l-(4-chlorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxamide

[0131]

[0132] Step 1: 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-l,3-dione

[0133] 3-aminopiperidine-2,6-dione hydrochloride (1.0 g, 6.1 mmol), 3-fluorophthalic anhydride (1.01 mg, 6.1 mmol), and AcONa (1.0 g, 12.2 mmol) were dissolved in AcOH (10 mL), and the reaction was stirred at 120 °C for 6 h. After the reaction was completed, the reaction was diluted with water (100 mL), filtered, and dried to give a white solid product (1.0 g), which was used directly in the next reaction. LCMS (ESI + ): m / z = 277 [M+H] + .

[0134] Step 2: tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4-yl)amino)butyl)carbamate

[0135] 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (1.0 g, 3.6 mmol), DIEA (930 mg, 7.2 mmol) were dissolved in NMP (10 mL), then N-tert-butoxycarbonyl-1,4-butanediamine (680 mg, 3.6 mmol) was added, heated to 90 °C and stirred for 6 h. After the reaction was completed, it was diluted with water (100 mL) and extracted with EtOAc (100 mL x 3). The organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc (v / v) = 1:1) to give the product as a yellow oil (800 mg, yield 44%). LCMS (ESI + ): m / z = 467 [M+Na] + .

[0136] Step 3: 4-((4-aminobutyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0137] tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butyl)carbamate (700 mg, 1.6 mmol) was dissolved in CH2Cl2(10 mL), and a 1,4-dioxane solution of HCl (4 M, 2 mL) was added. The reaction was stirred at room temperature for 2 h. After the reaction was completed, the reaction was concentrated under reduced pressure to give the product as a yellow solid (600 mg), which was used directly in the next step. LCMS (ESI + ): m / z = 345 [M+H] + .

[0138] Step 4: 5-chloro-N-((2S)-3-(2-(6-chloro-4-methylpyridin-3-yl)phenyl)-1-((4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butyl)amino)-1-oxopropan-2-yl)-1-(4-chlorobenzyl)-2-oxo-1,2-dihydropyridine-3-carboxamide

[0139] In a 25 mL round-bottom flask, was added (S)-2-(5-chloro-l-(4-chlorobenzyl)-2- oxo-l,2-dihydropyridine-3-carboxamido)-3-(2-(6-chloro-4-methylpyridin-3- yl)phenyl)propanoic acid (200 mg, 0.35 mmol), HOBT (71 mg, 0.53 mmol), EDCI (101 mg, 0.53 mmol), DIEA (91 mg, 0.70 mmol) and CH2Cl2(10 mL), then added 4-((4-aminobutyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-l,3-dione (121 mg, 0.35 mmol), stirred at room temperature for 2 h. After the reaction was completed, diluted with water (50 mL). The aqueous phase was extracted with CH2Cl2(50 mL x 3), and the organic phases were combined. The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure, and purified by flash chromatography (CH2Cl2:MeOH (v / v) = 90: 10) to obtain a yellow solid (140 mg, yield 40%). LCMS (ESI + ): m / z = 896 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 11.10 (s, 1H), 9.72 - 9.65 (m, 1H), 8.61 - 8.55 (m, 1H), 8.16 - 8.06 (m, 3H), 7.58 - 7.50 (m, 1H), 7.46 - 7.41 (m, 3H), 7.40 - 7.34 (m, 3H), 7.34 - 7.21 (m, 2H), 7.10 - 7.02 (m, 2H), 6.99 (d, J = 7.0 Hz, 1H), 6.56 - 6.47 (m, 1H), 5.17 (s, 2H), 5.09 - 5.00 (m, 1H), 4.71 - 4.48 (m, 1H), 3.29 - 3.19 (m, 2H), 3.06 - 2.83 (m, 4H), 2.79 - 2.65 (m, 1H), 2.63 - 2.54 (m, 2H), 2.08 - 1.97 (m, 1H), 1.92 (d, J = 6.8 Hz, 3H), 1.54 - 1.31 (m, 4H).

[0140] Example Two

[0141] Apparatus: Thar prep 80; Column: CHIRALPAK AS-H 250mm x 20mm, 5pm; Modifier: 40% MeOH (FA 0.2%): 60% CO2; Total Flow: 40 g / min) and the first peak was collected and lyophilized to get the product as a yellow solid (44.6 mg, yield 32%, Rt: 14.56 min). LCMS (ESI + ): m / z = 896 [M+H] + . 1 H NMR (400 MHz, Chloroform-d) δ 10.33 - 9.60 (m, 1H), 8.53 - 7.97 (m, 3H), 7.66 - 7.29 (m, 8H), 7.24 - 7.17 (m, 1H), 7.13 - 6.73 (m, 3H), 6.54 - 5.98 (m, 2H), 5.10 (s, 2H), 4.98 - 4.35 (m, 2H), 3.38 - 3.04 (m, 4H), 3.04 - 2.59 (m, 4H), 2.21 - 1.92 (m, 5H), 1.77 - 1.35 (m, 4H).

[0142] Example Three

[0143] Apparatus: Thar prep 80; Column: CHIRALPAK AS-H 250mm x 20mm, 5pm; Modifier: 40% MeOH (FA 0.2%): 60% CO2; Total Flow: 40 g / min) and the second peak was collected and lyophilized to get the product as a yellow solid (47.3 mg, yield 34%, Rt: 27.16 min). LCMS (ESI + ): m / z = 896 [M+H] + . 1 H NMR (400 MHz, Chloroform-d) δ 10.33 - 9.60 (m, 1H), 8.53 - 7.97 (m, 3H), 7.66 - 7.29 (m, 8H), 7.24 - 7.17 (m, 1H), 7.13 - 6.73 (m, 3H), 6.54 - 5.98 (m, 2H), 5.10 (s, 2H), 4.98 - 4.35 (m, 2H), 3.38 - 3.04 (m, 4H), 3.04 - 2.59 (m, 4H), 2.21 - 1.92 (m, 5H), 1.77 - 1.35 (m, 4H).

[0144] Example Four and Example Five

[0145]

[0146] Step 1: N-((2S)-1-((4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4- yl)amino)butyl)amino)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)-1-oxopropan-2- yl)-5-fluoro-1-(4-fluorobenzyl)-2-oxo-1,2-dihydropyridine-3-carboxamide

[0147] Into a 25 mL round-bottom flask, was added (S)-2-(5-fluoro-1-(4-fluorobenzyl)-2-oxo-1,2- dihydropyridine-3-carboxamido)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)propanoic acid (200 mg, 0.38 mmol), HOBT (78 mg, 0.58 mmol), EDCI (110 mg, 0.58 mmol), DIEA (99 mg, 0.77 mmol) and CH2Cl2(10 mL), then 4-((4-aminobutyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (132 mg, 0.38 mmol) was added, and the reaction was stirred at room temperature for 2 h. After the reaction was completed, water (50 mL) was added for dilution. The aqueous phase was extracted with CH2Cl2(50 mL x 3), and the organic phases were combined. The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. Purification by flash preparative chromatography (CH2Cl2:MeOH (v / v) = 90:10) yielded a yellow solid (170 mg, 47% yield). LCMS (ESI): m / z = 848 [M+H] + ): m / z = 848 [M+H] + .

[0148] Step 2: Resolution

[0149] N-((2S)-1-((4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butyl)amino)- 3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)-1-oxopropan-2-yl)-5-fluoro-1-(4-fluorobenzyl)- 2-oxo-1,2-dihydropyridine-3-carboxamide was resolved by SFC (Apparatus: Thar prep 80; Column: CHIRALPAK AS-H 250 mm x 20 mm, 5 μm; Modifier: 40% MeOH (FA 0.2%): 60% CO2; Total Flow: 40 g / min).

[0150] Example Four: The first peak was collected and freeze-dried to get yellow solid product (62.6 mg, yield 37%, Rt: 6.87 min), LCMS (ESI + ): m / z = 848 [M+H] + . 1 H NMR (400 MHz, Chloroform-d) δ 10.35 - 9.82 (m, 1H), 8.54 - 7.85 (m, 3H), 7.57 - 7.28 (m, 6H), 7.13 - 6.97 (m, 4H), 6.95 - 6.73 (m, 2H), 6.55 - 6.01 (m, 2H), 5.25 - 5.01 (m, 2H), 4.98 - 4.78 (m, 1H), 4.73 - 4.41 (m, 1H), 3.39 - 3.07 (m, 4H), 3.04 - 2.62 (m, 4H), 2.22 - 2.01 (m, 4H), 1.70 - 1.36 (m, 5H).

[0151] Example Five: The second peak was collected and freeze-dried to get yellow solid product (51.3 mg, yield 30%, Rt: 12.73 min), LCMS (ESI + ): m / z = 848 [M+H] + . 1 H NMR (400 MHz, Chloroform-d) δ 10.34 - 9.87 (m, 1H), 8.43 - 7.85 (m, 3H), 7.54 - 7.27 (m, 6H), 7.12 - 7.02 (m, 4H), 6.91 - 6.78 (m, 2H), 6.48 - 6.18 (m, 2H), 5.18 - 5.02 (m, 2H), 4.95 - 4.88 (m, 1H), 4.65 - 4.47 (m, 1H), 3.31 - 3.11 (m, 4H), 3.02 - 2.64 (m, 4H), 2.20 - 2.03 (m, 4H), 1.64 - 1.42 (m, 5H).

[0152] Example Six and Example Seven

[0153]

[0154]

[0155] Step 1: tert-Butyl (2-(4-(2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4-yl)piperazin-l- yl)ethyl)carbamate

[0156] 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-l,3-dione (200 mg, 0.72 mmol), DIEA (281 mg, 2.17 mmol) were dissolved in NMP (2 mL), then 1-(N-Boc- aminoethyl)piperazine (200 mg, 0.87 mmol) was added, heated to 100 °C and stirred for 12 h. After the reaction was completed, diluted with water (20 mL), extracted with EtOAc (20 mL x 3), the organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure, the residue was separated and purified by silica gel column chromatography (CH2Cl2:MeOH (v / v) = 97:3) to give a yellow solid product (180 mg, yield 49%). LCMS (ESI + ): m / z = 486 [M+H] + .

[0157] Step 2: 4-(4-(2-aminoethyl)piperazin-l-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-l,3-dione

[0158] tert-butyl (2-(4-(2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4-yl)piperazin-l- yl)ethyl)carbamate (180 mg, 0.37 mmol) was dissolved in CH2Cl2(6 mL), added HCl solution in 1,4-dioxane (4 M, 3 mL), stirred at room temperature for 2 h. After the reaction was completed, the reaction was concentrated under reduced pressure to give a yellow solid product (165 mg), which was directly used in the next step. LCMS (ESI + ): m / z = 386 [M+H] + .

[0159] Step 3: N-((2S)-l-((2-(4-(2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4-yl)piperazin-l- yl)ethyl)amino)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)-l-oxopropan-2-yl)-5-fluoro-l- (4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxamide

[0160] In a 25 mL round-bottom flask, was added (S)-2-(5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2- dihydropyridine-3-carboxamido)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)propanoic acid (203 mg, 0.38 mmol), TCFH (164 mg, 0.58 mmol), N-methylimidazole (96 mg, 1.17 mmol) and CH2Cl2(5 mL), then added 4-(4-(2-aminoethyl)piperazin-l-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-l,3-dione (150 mg, 0.38 mmol), stirred at room temperature for 2 h. After the reaction was completed, diluted with water (20 mL), extracted with CH2Cl2(20 mL x 3), the organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure, the residue was separated and purified by silica gel column chromatography (CH2Cl2:MeOH (v / v) = 97:3) to give yellow solid product (140 mg, yield 41%). LCMS (ESI + ): m / z = 889 [M+H] + .

[0161] Step 4: Resolution

[0162] N-((2S)-l-((2-(4-(2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4-yl)piperazin-l- yl)ethyl)amino)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)-l-oxopropan-2-yl)-5-fluoro-l- (4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxamide was resolved by SFC (Apparatus: Thar prep 80; Column: CHIRALPAK AS-H 250 mm x 20 mm, 5 μm; Modifier: 40% MeOH (NH4OH 0.2%):60% CO2; Total Flow: 40 g / min).

[0163] Example Six: The first peak was collected and freeze-dried to give yellow solid product (51.5 mg, RT: 4.3 min), LCMS (ESI + ): m / z = 889 [M+H] + . 1H NMR (400 MHz, Chloroform-d) δ 12.13 (s, 1H), 10.45 - 10.04 (m, 1H), 8.48 - 7.86 (m, 3H), 7.71 - 7.60 (m, 1H), 7.56 - 7.42 (m, 2H), 7.40 - 7.27 (m, 5H), 7.18 - 7.03 (m, 4H), 6.90 - 6.80 (m, 1H), 5.14 (s, 2H), 5.02 - 4.87 (m, 1H), 4.68 - 4.50 (m, 1H), 3.89 - 3.56 (m, 7H), 3.37 - 3.04 (m, 5H), 2.99 - 2.47 (m, 5H), 2.22 - 2.04 (m, 4H).

[0164] Example Seven: The second peak was collected and lyophilized to give a yellow solid product (53.8 mg, RT: 5.1 min), LCMS (ESI + ): m / z = 889 [M+H] + . 1 H NMR (400 MHz, Chloroform-d) δ 12.13 (s, 1H), 10.45 - 10.04 (m, 1H), 8.48 - 7.86 (m, 3H), 7.71 - 7.60 (m, 1H), 7.56 - 7.42 (m, 2H), 7.40 - 7.27 (m, 5H), 7.18 - 7.03 (m, 4H), 6.90 - 6.80 (m, 1H), 5.14 (s, 2H), 5.02 - 4.87 (m, 1H), 4.68 - 4.50 (m, 1H), 3.89 - 3.56 (m, 7H), 3.37 - 3.04 (m, 5H), 2.99 - 2.47 (m, 5H), 2.22 - 2.04 (m, 4H).

[0165] Example Eight and Example Nine

[0166]

[0167]

[0168] Step 1: tert-Butyl ((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)azetidin-3-yl)methyl)carbamate

[0169] 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-l,3-dione (200 mg, 0.72 mmol), DIEA (281 mg, 2.17 mmol) were dissolved in NMP (2 mL), then 3-(Boc-aminomethyl)azetidine (162 mg, 0.87 mmol) was added, heated to 100 °C and stirred for 3 h. After the reaction was completed, it was diluted with water (20 mL) and extracted with EtOAc (20 mL x 3). The organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (CH2Cl2:MeOH (v / v) = 97:3) to give a yellow solid product (160 mg, yield 47%). LCMS (ESI): m / z = 465 [M+Na] + ): m / z = 465 [M+Na] + .

[0170] Step 2: 4-(3-(aminomethyl)azetidin-l-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-l,3-dione

[0171] tert-butyl ((l-(2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4-yl)azetidin-3- yl)methyl)carbamate (160 mg, 0.36 mmol) and pyridine (3 mL) were dissolved in CH2Cl2(6 mL), and TMSOTf (3 mL) was added. The reaction was stirred at room temperature for 5 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure to give a yellow solid product (165 mg), which was used directly in the next step. LCMS (ESI): m / z = 343 [M+H] + ): m / z = 343 [M+H] + .

[0172] Step 3: N-((2S)-l-(((l-(2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4-yl)azetidin-3- yl)methyl)amino)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)-l-oxopropan-2-yl)-5- fluoro-l-(4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxamide

[0173] In a 25 mL round-bottom flask, was added (S)-2-(5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2- dihydropyridine-3-carboxamido)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)propanoic acid (411 mg, 0.79 mmol), HOBT (160 mg, 1.18 mmol), EDCI (227 mg, 1.18 mmol), DIEA (305 mg, 2.36 mmol) and CH2Cl2(5 mL), then 4-(3-(aminomethyl)azetidin-l-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-l,3-dione (250 mg, 0.79 mmol) was added, and the reaction was stirred at room temperature for 2 h. After the reaction was completed, it was diluted with water (50 mL). The aqueous phase was extracted with CH2Cl2(50 mL x 3), and the organic phases were combined. The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. Purification by flash chromatography (CH2Cl2:MeOH (v / v) = 90: 10) gave a white solid (200 mg, yield 24%). LCMS (ESI + ): m / z = 846 [M+H] + .

[0174] Step 4: Resolution

[0175] N-((2S)-l-(((l-(2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4-yl)azetidin-3-yl)methyl)amino)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)-l-oxopropan-2-yl)-5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxamide was resolved by SFC (Apparatus: Thar prep 80; Column: CHIRALPAK AS-H 250 mm x 20 mm, 5 μm; Modifier: 40% MeOH (NH4OH 0.2%): 60% CO2; Total Flow: 40 g / min).

[0176] Example Eight: The first peak was collected and lyophilized to give a yellow solid product (53.1 mg, RT: 6.2 min). LCMS (ESI + ): m / z = 846 [M+H] + . 1H NMR (400 MHz, Chloroform-d) δ 10.38 - 9.83 (m, 1H), 8.48 - 8.20 (m, 1H), 8.18 - 8.06 (m, 1H), 8.05 - 7.90 (m, 1H), 7.41 (d, J = 7.2 Hz, 1H), 7.37 - 7.27 (m, 5H), 7.14 - 6.94 (m, 5H), 6.86 (d, J = 10.1 Hz, 2H), 6.53 - 6.34 (m, 1H), 5.21 - 5.08 (m, 1H), 4.99 - 4.81 (m, 2H), 4.70 - 4.44 (m, 1H), 4.28 - 4.00 (m, 2H), 3.94 - 3.76 (m, 2H), 3.55 - 3.32 (m, 2H), 3.03 - 2.92 (m, 1H), 2.92 - 2.65 (m, 5H), 2.20 - 2.02 (m, 4H).

[0177] Example Nine: The second peak was collected and lyophilized to yield a yellow solid product (50.0 mg, RT: 8.7 min), LCMS (ESI + ): m / z = 846 [M+H] + . 1 H NMR (400 MHz, Chloroform-d) δ 10.29 - 9.87 (m, 1H), 8.82 - 7.86 (m, 3H), 7.41 (d, J = 6.4 Hz, 1H), 7.37 - 7.26 (m, 5H), 7.16 - 6.93 (m, 5H), 6.88 - 6.81 (m, 2H), 6.53 - 6.32 (m, 1H), 5.21 - 5.06 (m, 1H), 5.02 - 4.87 (m, 2H), 4.64 - 4.45 (m, 1H), 4.24 - 4.00 (m, 2H), 3.93 - 3.75 (m, 2H), 3.55 - 3.20 (m, 2H), 2.99 - 2.93 (m, 1H), 2.92 - 2.66 (m, 5H), 2.18 - 1.99 (m, 4H).

[0178] Example Ten and Example Eleven

[0179]

[0180]

[0181] Step 1: tert-Butyl (2-((2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4- yl)amino)ethyl)carbamate

[0182] Under nitrogen protection, 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (200 mg, 0.72 mmol), DIEA (281 mg, 2.17 mmol) were dissolved in NMP (10 mL), then N-tert-butoxycarbonyl-1,2-ethanediamine (140 mg, 0.87 mmol) was added, heated to 100 °C and stirred for 16 h. After the reaction was completed, it was diluted with water (100 mL) and extracted with EtOAc (100 mL x 3). The organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (PE:EtOAc (v / v) = 1:1) to give the product as a yellow oil (250 mg, yield 78%). LCMS (ESI + ): m / z = 439 [M+Na] + .

[0183] Step 2: 4-((2-aminoethyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0184] tert-butyl (2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethyl)carbamate (250 mg, 0.60 mmol) was dissolved in CH2Cl2(5 mL), and a 1,4-dioxane solution of HCl (4 M, 15 mL) was added. The reaction was stirred at room temperature for 3 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure to give the product as a yellow solid (180 mg), which was used directly in the next step. LCMS (ESI + ): m / z = 317 [M+H] + .

[0185] Step 3: N-((2S)-1-((2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethyl)amino)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)-1-oxopropan-2-yl)-5-fluoro-1-(4-fluorobenzyl)-2-oxo-1,2-dihydropyridine-3-carboxamide

[0186] In a 25 mL round-bottom flask, was added (S)-2-(5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2- dihydropyridine-3-carboxamido)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)propanoic acid (247 mg, 0.47 mmol), HOBT (96 mg, 0.71 mmol), EDCI (136 mg, 0.71 mmol), DIEA (184 mg, 1.42 mmol) and CH2Cl2(10 mL), then added 4-((2-aminoethyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-l,3-dione (150 mg, 0.47 mmol), stirred at room temperature for 16 h. After the reaction was completed, diluted with water (50 mL). The aqueous phase was extracted with CH2Cl2(50 mL x 3), the organic phases were combined. The organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure, purified by flash preparative chromatography (CH2Cl2:MeOH (v / v) = 90: 10) to give a yellow solid (200 mg, yield 48%). LCMS (ESI + ): m / z = 820 [M+H] + .

[0187] Step 4: Resolution

[0188] N-((2S)-l-((2-((2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4-yl)amino)ethyl)amino)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)-l-oxopropan-2-yl)-5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxamide was resolved by SFC (Apparatus: Thar prep 80; Column: CHIRALPAK AS-H 250 mm x 20 mm, 5 μm; Modifier: 40% MeOH (FA 0.2%): 60% CO2; Total Flow: 40 g / min).

[0189] Example Ten: The first peak was collected and lyophilized to give a yellow solid product (80 mg, RT: 5.16 min), LCMS (ESI + ): m / z = 820 [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ 11.09 (s, 1H), 9.89 - 9.82 (m, 1H), 8.56 (t, J = 3.8 Hz, 1H), 8.39 - 8.26 (m, 1H), 8.18 - 8.11 (m, 1H), 8.01 - 7.73 (m, 1H), 7.57 - 7.40 (m, 3H), 7.39 - 7.33 (m, 1H), 7.29 - 7.16 (m, 4H), 7.16 - 7.08 (m, 1H), 7.07 - 6.97 (m, 3H), 6.73 - 6.64 (m, 1H), 5.14 (s, 2H), 5.09 - 5.00 (m, 1H), 4.69 - 4.45 (m, 1H), 3.30 - 3.10 (m, 4H), 2.99 - 2.81 (m, 2H), 2.79 - 2.73 (m, 1H), 2.62 - 2.53 (m, 2H), 2.05 - 1.90 (m, 4H).

[0190] Example Eleven: The second peak was collected and lyophilized to give a yellow solid product (70 mg, RT: 7.7 min), LCMS (ESI + ): m / z = 820 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 11.05 (s, 1H), 9.86 (d, J = 7.9 Hz, 1H), 8.56 (t, J = 3.8 Hz, 1H), 8.40 - 8.25 (m, 1H), 8.18 - 8.10 (m, 1H), 8.05 - 7.71 (m, 1H), 7.58 - 7.40 (m, 3H), 7.41 - 7.32 (m, 1H), 7.30 - 7.16 (m, 4H), 7.15 - 7.09 (m, 1H), 7.08 - 6.95 (m, 3H), 6.73 - 6.63 (m, 1H), 5.14 (s, 2H), 5.10 - 4.99 (m, 1H), 4.71 - 4.47 (m, 1H), 3.41 - 3.10 (m, 4H), 3.01 - 2.81 (m, 2H), 2.81 - 2.71 (m, 1H), 2.63 - 2.52 (m, 2H), 2.07 - 1.90 (m, 4H).

[0191] Example Twelve and Example Thirteen

[0192]

[0193] Step 1: tert-Butyl (2-((2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4-yl)oxy)ethyl)carbamate

[0194] In a 50 ml reaction flask, N-Boc-bromoethylamine (237 mg, 1.06 mmol), DMF (5 mL), 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (376 mg, 1.37 mmol) and Et3N (320 mg, 3.17 mmol) were added, and the reaction was stirred at 70°C for 3 hours. After the reaction was completed, the reaction solution was poured into water (80 mL), and the aqueous phase was extracted with EtOAc (50 mL x 3). The organic phase was washed with saturated NaCl aqueous solution, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was separated by silica gel column chromatography (DCM:MeOH (v / v)=20:1) to obtain the product (331 mg, yield: 75%). MS (ESI) m / z: 317

[0195] [M+H-100] + .

[0196] Step 2: 4-(2-Aminoethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0197] In a 50 ml reaction flask, tert-butyl (2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethyl)carbamate (331 mg, 0.79 mmol) was dissolved in DCM (10.0 mL), and then TFA (6.0 mL) was added. The reaction was stirred at room temperature for 1 hour. After the reaction was completed, it was concentrated under reduced pressure to obtain an oil (250 mg), which was directly used in the next reaction. MS (ESI) m / z: 317 [M+H] + .

[0198] Step 3: N-((2S)-1-((2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethyl)amino)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)-1-oxopropan-2-yl)-5-fluoro-1-(4-fluorobenzyl)-2-oxo-1,2-dihydropyridine-3-carboxamide

[0199] In a 25 mL round-bottom flask, was added (S)-2-(5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2- dihydropyridine-3-carboxamido)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)propanoic acid (411 mg, 0.79 mmol), HOBT (160 mg, 1.2 mmol), EDCI (227 mg, 1.2 mmol), DIEA (305 mg, 2.4 mmol) and CH2Cl2(10 mL), then 4-(2- aminoethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-l,3-dione (250 mg, 0.79 mmol) was added, and the reaction was stirred at room temperature for 2 h. After the reaction was completed, it was diluted with water (50 mL). The aqueous phase was extracted with CH2Cl2(50 mL x 3), and the organic phases were combined. The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. Purification by flash chromatography (CH2Cl2:MeOH (v / v) = 96:4) gave a white solid (200 mg, yield 31%). LCMS (ESI + ): m / z = 821 [M+H] + .

[0200] Step 4: Resolution

[0201] N-((2S)-l-((2-((2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4-yl)oxy)ethyl)amino)-3-(2-(6- fluoro-4-methylpyridin-3-yl)phenyl)-l-oxopropan-2-yl)-5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2- dihydropyridine-3-carboxamide was resolved by SFC (Apparatus: Thar prep 80; Column: CHIRALPAK AS-H 250 mm x 20 mm, 5 μm; Modifier: 40% MeOH (NH4OH 0.2%):60% CO2; Total Flow: 40 g / min).

[0202] Example Twelve: The first peak was collected and lyophilized to give a yellow solid product (64.6 mg, RT: 38.7 min). LCMS (ESI + ): m / z = 821 [M+H] + . 1H NMR (400 MHz, Chloroform-d) δ 10.21 - 9.95 (m, 1H), 8.44 - 7.90 (m, 3H), 7.71 - 7.61 (m, 1H), 7.51 - 7.44 (m, 1H), 7.44 - 7.28 (m, 4H), 7.24 - 7.12 (m, 3H), 7.11 - 6.98 (m, 3H), 6.96 - 6.77 (m, 2H), 5.10 (s, 2H), 4.99 - 4.88 (m, 1H), 4.67 - 4.50 (m, 1H), 4.24 - 4.04 (m, 2H), 3.71 - 3.54 (m, 2H), 3.26 - 2.66 (m, 5H), 2.14 - 2.04 (m, 4H).

[0203] Example Thirteen: The second peak was collected and lyophilized to give a yellow solid product (70.0 mg, RT: 45.5 min), LCMS (ESI + ): m / z = 821 [M+H] + . 1 H NMR (400 MHz, Chloroform-d) δ 10.28 - 9.99 (m, 1H), 8.59 - 7.89 (m, 3H), 7.71 - 7.61 (m, 1H), 7.51 - 7.44 (m, 1H), 7.44 - 7.28 (m, 4H), 7.25 - 7.13 (m, 3H), 7.12 - 6.99 (m, 3H), 6.97 - 6.79 (m, 2H), 5.16 - 4.89 (m, 3H), 4.62 - 4.50 (m, 1H), 4.24 - 4.05 (m, 2H), 3.69 - 3.51 (m, 2H), 3.22 - 2.69 (m, 5H), 2.18 - 2.03 (m, 4H).

[0204] Example Fourteen: 5-Chloro-N-((2S)-3-(2-(6-chloro-4-methylpyridin-3-yl)phenyl)-1-(((1r,4S)-4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)cyclohexyl)amino)-1-oxopropan-2-yl)-1-(4-chlorobenzyl)-2-oxo-1,2-dihydropyridine-3-carboxamide

[0205]

[0206] Step 1: tert-Butyl ((1r,4r)-4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4- yl)amino)cyclohexyl)carbamate

[0207] Under nitrogen protection, 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (0.2 g, 0.7 mmol), DIEA (281 mg, 2.1 mmol) were dissolved in NMP (3 mL), then trans-tert-butyl (4-aminocyclohexyl)carbamate (187 mg, 0.8 mmol) was added, heated to 100 °C and stirred for 6 h. After the reaction was completed, it was diluted with water (30 mL) and extracted with EtOAc (30 mL x 3). The organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (PE:EtOAc (v / v) = 1:1) to give the product (300 mg, yield 79%) as yellow oil. LCMS (ESI + ): m / z = 493 [M+Na] + .

[0208] Step 2: 4-(((1r,4r)-4-aminocyclohexyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0209] tert-Butyl ((1r,4r)-4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)cyclohexyl)carbamate (300 mg, 0.64 mmol) was dissolved in CH2Cl2(5 mL), and a 1,4-dioxane solution of HCl (4 M, 15 mL) was added. The reaction was stirred at room temperature for 3 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure to give the product (200 mg) as a yellow solid, which was used directly in the next step. LCMS (ESI + ): m / z = 371 [M+H] + .

[0210] Step 3: 5-chloro-N-((2S)-3-(2-(6-chloro-4-methylpyridin-3-yl)phenyl)-1-(((1r,4S)-4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)cyclohexyl)amino)-1-oxopropan-2-yl)-1-(4-chlorobenzyl)-2-oxo-1,2-dihydropyridine-3-carboxamide

[0211] In a 25 mL round-bottom flask, was added (S)-2-(5-chloro-l-(4-chlorobenzyl)-2- oxo-l,2-dihydropyridine-3-carboxamido)-3-(2-(6-chloro-4-methylpyridin-3-yl)phenyl) propanoic acid (385 mg, 0.67 mmol), HOBT (137 mg, 1.0 mmol), EDCI (194 mg, 1.0 mmol), DIEA (262 mg, 2.0 mmol) and CH2Cl2(10 mL), then added 4-(((lr,4r)-4-aminocyclohexyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-l,3-dione (250 mg, 0.68 mmol), stirred at room temperature for 16 h. After the reaction was completed, diluted with water (50 mL). The aqueous phase was extracted with CH2Cl2(50 mL x 3), the organic phases were combined. The organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure, purified by flash chromatography (CH2Cl2:MeOH (v / v) = 90: 10) to obtain a yellow solid (200 mg, yield 34%). LCMS (ESI + ): m / z = 922 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 9.80 - 9.62 (m, 1H), 8.69 - 8.49 (m, 1H), 8.23 - 7.83 (m, 3H), 7.58 (t, J = 8.1 Hz, 1H), 7.50 - 7.24 (m, 8H), 7.18 (dd, J = 8.9, 1.9 Hz, 1H), 7.09 - 7.00 (m, 2H), 6.22 - 6.14 (m, 1H), 5.18 (s, 2H), 5.10 - 5.01 (m, 1H), 4.80 - 4.48 (m, 1H), 3.67 - 3.39 (m, 2H), 2.99 - 2.71 (m, 3H), 2.63 - 2.51 (m, 2H), 2.13 - 1.87 (m, 6H), 1.83 - 1.54 (m, 2H), 1.44 - 1.12 (m, 4H).

[0212] Example Fifteen and Example Sixteen

[0213]

[0214] Step 1: tert-Butyl (3-((2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4-yl)amino)propyl) carbamate

[0215] 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (2.0 g, 7.2 mmol), DIEA (1.86 g, 14.4 mmol) were dissolved in NMP (20 mL), then N-tert-butoxycarbonyl-1,3-propanediamine (1.26 g, 7.2 mmol) was added, heated to 90 °C and stirred for 6 h. After the reaction was completed, it was diluted with water (100 mL) and extracted with EtOAc (100 mL x 3). The organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (PE:EtOAc (v / v) = 1:1) to give the product as a yellow oil (1.2 g, yield 34%). LCMS (ESI + ): m / z = 431 [M+H] + .

[0216] Step 2: 4-((3-aminopropyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0217] tert-butyl (3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)propyl)carbamate (1.2 g, 2.8 mmol) was dissolved in CH2Cl2(10 mL), and a 1,4-dioxane solution of HCl (4 M, 5 mL) was added. The reaction was stirred at room temperature for 2 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure to give the product as a yellow solid (1.0 g), which was used directly in the next reaction. LCMS (ESI + ): m / z = 331 [M+H] + .

[0218] Step 3: 5-chloro-N-((2S)-3-(2-(6-chloro-4-methylpyridin-3-yl)phenyl)-1-((3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)propyl)amino)-1-oxopropan-2-yl)-1-(4-chlorobenzyl)-2-oxo-1,2-dihydropyridine-3-carboxamide

[0219] In a 25 mL round-bottom flask, was added (S)-2-(5-chloro-l-(4-chlorobenzyl)-2-oxo-l,2- dihydropyridine-3-carboxamido)-3-(2-(6-chloro-4-methylpyridin-3-yl)phenyl)propanoic acid (200 mg, 0.35 mmol), HOBT (71 mg, 0.53 mmol), EDCI (101 mg, 0.53 mmol), DIEA (91 mg, 0.70 mmol) and CH2Cl2(10 mL), then added 4-((3- aminopropyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-l,3-dione (116 mg, 0.35 mmol), stirred at room temperature for 2 h. After the reaction was completed, diluted with water (50 mL). The aqueous phase was extracted with CH2Cl2(50 mL x 3), the organic phases were combined. The organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure, purified by flash preparative chromatography (CH2Cl2:MeOH (v / v) = 90: 10) to give a yellow solid (140 mg, yield 41%). LCMS (ESI + ): m / z = 882 [M+H] + .

[0220] Step 4: Resolution

[0221] 5-chloro-N-((2S)-3-(2-(6-chloro-4-methylpyridin-3-yl)phenyl)-l-((3-((2-(2,6- dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4-yl)amino)propyl)amino)-l-oxopropan-2-yl)-l- (4-chlorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxamide was resolved by SFC (Apparatus: SFC Thar prep 80; Column: CHIRALPAK OD-H 250 mm x 20 mm, 5 μm; Modifier: 40% MeOH (FA 0.2%): 60% CO2; Total Flow: 40 g / min).

[0222] Example Fifteen: The first peak was collected and freeze-dried to give a yellow solid product (52.1 mg, RT: 12.5 min), LCMS (ESI + ): m / z = 882 [M+H] + . 1H NMR (400 MHz, Chloroform-d) δ 10.35 - 9.53 (m, 1H), 8.93 - 8.01 (m, 3H), 7.73 - 7.29 (m, 8H), 7.16 - 6.03 (m, 6H), 5.14 (s, 2H), 5.00 - 4.39 (m, 2H), 3.50 - 3.06 (m, 4H), 3.03 - 2.42 (m, 5H), 2.29 - 1.87 (m, 4H), 1.90 - 1.55 (m, 2H).

[0223] Example Sixteen: The second peak was collected and lyophilized to give a yellow solid product (58.0 mg, RT: 21.4 min), LCMS (ESI + ): m / z = 882 [M+H] + . 1 H NMR (400 MHz, Chloroform-d) δ 10.13 - 9.75 (m, 1H), 8.48 - 8.01 (m, 3H), 7.66 - 7.28 (m, 8H), 7.12 - 6.70 (m, 4H), 6.67 - 6.11 (m, 2H), 5.11 (s, 2H), 4.99 - 4.82 (m, 1H), 4.70 - 4.41 (m, 1H), 3.39 - 3.10 (m, 4H), 3.02 - 2.61 (m, 5H), 2.22 - 1.97 (m, 4H), 1.89 - 1.62 (m, 2H).

[0224] Example Seventeen and Eighteen

[0225]

[0226] Step 1: N-((2S)-1-((3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)propyl)amino)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)-1-oxopropan-2-yl)-5-fluoro-1-(4-fluorobenzyl)-2-oxo-1,2-dihydropyridine-3-carboxamide.

[0227] In a 25 mL round-bottom flask, was added (S)-2-(5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2- dihydropyridine-3-carboxamido)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)propanoic acid (200 mg, 0.38 mmol), HOBT (78 mg, 0.58 mmol), EDCI (110 mg, 0.58 mmol), DIEA (99 mg, 0.77 mmol) and CH2Cl2(10 mL), then added 4-((3-aminopropyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-l,3-dione (127 mg, 0.38 mmol), stirred at room temperature for 2 h. After the reaction was completed, diluted with water (50 mL). The aqueous phase was extracted with CH2Cl2(50 mL x 3), the organic phases were combined. The organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure, purified by flash chromatography (CH2Cl2:MeOH (v / v) = 90: 10) to give a yellow solid (150 mg, yield 42%). LCMS (ESI + ): m / z = 834 [M+H] + .

[0228] Step 2: Resolution

[0229] N-((2S)-l-((3-((2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4-yl)amino)propyl)amino)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)-l-oxopropan-2-yl)-5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxamide was resolved by SFC (Apparatus: SFC Thar prep 80; Column: CHIRALPAK OD-H 250 mm x 20 mm, 5 μm; Modifier: 40% MeOH (NH4OH 0.2%):60% CO2; Total Flow: 40 g / min).

[0230] Example Seventeen: The first peak was collected and freeze-dried to give a yellow solid product (54.6 mg, RT: 5.24 min), LCMS (ESI + ): m / z = 834 [M+H] + . 1H NMR (400 MHz, Chloroform-d) δ 10.32 - 9.91 (m, 1H), 8.89 - 7.91 (m, 3H), 7.48 - 7.26 (m, 7H), 7.16 - 6.89 (m, 5H), 6.88 - 6.65 (m, 2H), 5.12 (s, 2H), 5.01 - 4.85 (m, 1H), 4.69 - 4.49 (m, 1H), 3.43 - 3.09 (m, 4H), 3.03 - 2.64 (m, 5H), 2.20 - 2.04 (m, 4H), 1.80 - 1.60 (m, 2H).

[0231] Example Eighteen: The second peak was collected and lyophilized to give a yellow solid product (66.7 mg, RT: 8.41 min), LCMS (ESI + ): m / z = 834 [M+H] + . 1 H NMR (400 MHz, Chloroform-d) δ 10.34 - 9.87 (m, 1H), 8.45 - 7.79 (m, 3H), 7.56 - 7.27 (m, 7H), 7.18 - 6.97 (m, 5H), 6.94 - 6.65 (m, 2H), 5.11 (s, 2H), 5.01 - 4.81 (m, 1H), 4.74 - 4.46 (m, 1H), 3.49 - 3.08 (m, 4H), 3.06 - 2.61 (m, 5H), 2.31 - 2.00 (m, 4H), 1.89 - 1.57 (m, 2H).

[0232] Example Nineteen and Example Twenty

[0233]

[0234]

[0235] Step 1: (S)-2-((tert-butoxycarbonyl)amino)-3-(2-(6-fluoropyridin-3-yl)phenyl)propanoic acid

[0236] In a 100 mL round-bottom flask, (S)-3-(2-bromophenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid (1.0 g, 2.9 mmol), 6-fluoropyridine-3-boronic acid (410 mg, 2.9 mmol), Pd(PPh3)4(340 g, 0.3 mmol), Cs2CO3(1.9 g, 5.8 mmol), 1,4-dioxane (20 mL) and H2O (5 mL) were added, and the reaction was stirred at 90 °C for 16 h under nitrogen. After the reaction was completed, the reaction solution was diluted with water (50 mL), adjusted to pH = 6 with 4 M hydrochloric acid, and extracted with EtOAc (100 mL x 3). The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (CH2Cl2:MeOH (v / v) = 90:10) to obtain a yellow oil (0.7 g, yield 55%). LCMS (ESI + ): m / z = 361 [M+H] + .

[0237] Step 2: (S)-methyl 2-((tert-butoxycarbonyl)amino)-3-(2-(6-fluoropyridin-3- yl)phenyl)propanoate

[0238] In a 250 mL round-bottom flask, (S)-2-((tert-butoxycarbonyl)amino)-3-(2-(6- fluoropyridin-3-yl)phenyl)propanoic acid (0.7 g, 1.9 mmol), K2CO3(537 mg, 3.9 mmol), DMF (5 mL) and iodomethane (827 mg, 5.83 mmol) were added, and the reaction was stirred at room temperature for 4 h. After the reaction was completed, the reaction solution was diluted with water (50 mL), extracted with EtOAc (100 mL x 3), and the organic phase was dried over anhydrous Na2SO4. After filtration, the residue was concentrated under reduced pressure and separated and purified by silica gel column chromatography (PE:EtOAc (v / v) = 70:30) to obtain a yellow oil product (0.3 g, yield 40%). LCMS (ESI + ): m / z = 375 [M+H] + .

[0239] Step 3: (S)-methyl 2-amino-3-(2-(6-fluoropyridin-3-yl)phenyl)propanoate

[0240] (S)-methyl 2-((tert-butoxycarbonyl)amino)-3-(2-(6-fluoropyridin-3-yl)phenyl)propanoate (0.3 g, 0.8 mmol) was dissolved in CH2Cl2(10 mL), and then TFA (953 mg, 8.0 mmol) was added. The reaction was stirred at room temperature for 2 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain a yellow solid product (0.2 g), which was directly used in the next reaction. LCMS (ESI + ): m / z = 275 [M+H]+ .

[0241] Step 4: (S)-methyl 2-(5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3- carboxamido)-3-(2-(6-fluoropyridin-3-yl)phenyl)propanoate

[0242] Into a 25 mL round-bottom flask, was placed 5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2- dihydropyridine-3-carboxylic acid (193 mg, 0.73 mmol), TCFH (409 mg, 1.46 mmol), N- methylimidazole (180 mg, 2.19 mmol) and CH2Cl2(5 mL), followed by (S)-methyl 2- amino-3-(2-(6-fluoropyridin-3-yl)phenyl)propanoate (200 mg, 0.730 mmol). The reaction mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was diluted with water (50 mL) and extracted with CH2Cl2(50 mL x 3). The organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (CH2Cl2:MeOH (v / v) = 90: 10) to give the product as a yellow oil (200 mg, 49% yield). LCMS (ESI + ): m / z = 522 [M+H] + .

[0243] Step 5: (S)-2-(5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxamido)-3- (2-(6-fluoropyridin-3-yl)phenyl)propanoic acid

[0244] (S)-methyl 2-(5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxamido)-3- (2-(6-fluoropyridin-3-yl)phenyl)propanoate (200 mg, 0.380 mmol) was dissolved in MeOH (3 mL). To the system was added H2O (3 mL) and NaOH (46 mg, 1.15 mmol) and the reaction mixture was stirred at room temperature for 4 h. After completion of the reaction, the reaction mixture was diluted with water (20 mL) and the pH was adjusted to 6 with 1 M HCl. The mixture was filtered and dried to give a yellow solid (190 mg), which was used directly in the next step. LCMS (ESI + ): m / z = 508 [M+H] + .

[0245] Step 6: N-((2S)-1-((3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4- yl)amino)propyl)amino)-3-(2-(6-fluoropyridin-3-yl)phenyl)-1-oxopropan-2-yl)-5- fluoro-1-(4-fluorobenzyl)-2-oxo-1,2-dihydropyridine-3-carboxamide.

[0246] In a 25 mL round-bottom flask, (S)-2-(5-fluoro-1-(4-fluorobenzyl)-2-oxo-1,2- dihydropyridine-3-carboxamido)-3-(2-(6-fluoropyridin-3-yl)phenyl)propanoic acid (190 mg, 0.37 mmol), HOBT (76 mg, 0.56 mmol), EDCI (108 mg, 0.56 mmol), DIEA (97 mg, 0.75) and CH2Cl2(10 mL) were added, then 4-((3- aminopropyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (124 mg, 0.37 mmol) was added, and the reaction was stirred at room temperature for 2 h. After the reaction was completed, water (50 mL) was added for dilution. The aqueous phase was extracted with CH2Cl2(50 mL x 3), and the organic phases were combined. The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. Purification by flash preparative chromatography (CH2Cl2:MeOH (v / v) = 90:10) yielded a yellow solid (160 mg, 47% yield). LCMS (ESI + ): m / z = 820 [M+H] + .

[0247] Step 7: Resolution

[0248] N-((2S)-1-((3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)propyl)amino)-3-(2-(6-fluoropyridin-3-yl)phenyl)-1-oxopropan-2-yl)-5-fluoro-1-(4- fluorobenzyl)-2-oxo-1,2-dihydropyridine-3-carboxamide was resolved by SFC (Apparatus: Thar prep 80; Column: CHIRALPAK AS-H 250 mm x 20 mm, 5 μm; Modifier: 40% MeOH (NH4OH 0.2%):60% CO2; Total Flow: 40 g / min).

[0249] Example Nineteen: The first peak was collected and freeze-dried to obtain a yellow solid product (61.1 mg, RT: 7.37 min). LCMS (ESI + ): m / z = 820 [M+H] + . 1H NMR (400 MHz, Chloroform-d) δ 10.13 (s, 1H), 9.05 - 8.80 (m, 1H), 8.41 - 8.08 (m, 2H), 7.92 - 7.71 (m, 1H), 7.53 - 7.27 (m, 7H), 7.21 - 6.90 (m, 6H), 6.82 - 6.54 (m, 2H), 5.12 (s, 2H), 5.01 - 4.83 (m, 1H), 4.74 - 4.49 (m, 1H), 3.50 - 3.24 (m, 2H), 3.21 - 2.98 (m, 4H), 2.97 - 2.62 (m, 3H), 2.20 - 1.99 (m, 1H), 1.79 - 1.48 (m, 2H).

[0250] Example Twenty: The second peak was collected and lyophilized to yield a yellow solid product (72.4 mg, RT: 12.19 min), LCMS (ESI + ): m / z = 820 [M+H] + . 1 H NMR (400 MHz, Chloroform-d) δ 10.23 - 9.96 (m, 1H), 8.60 - 8.41 (m, 1H), 8.36 - 8.05 (m, 2H), 7.79 (t, J = 8.0 Hz, 1H), 7.50 - 7.27 (m, 7H), 7.15 (d, J = 7.0 Hz, 1H), 7.12 - 7.02 (m, 3H), 6.98 (d, J = 8.2 Hz, 1H), 6.78 (d, J = 8.3 Hz, 1H), 6.62 - 6.46 (m, 1H), 6.33 - 6.20 (m, 1H), 5.11 (s, 2H), 5.00 - 4.87 (m, 1H), 4.69 - 4.56 (m, 1H), 3.42 - 2.99 (m, 6H), 2.93 - 2.67 (m, 3H), 2.20 - 2.03 (m, 1H), 1.80 - 1.62 (m, 2H).

[0251] Example Twenty-one: N-((2S)-1-(4-(((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4- yl)amino)methyl)piperidin-1-yl)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)-1-oxopropan- 2-yl)-5-fluoro-1-(4-fluorobenzyl)-2-oxo-1,2-dihydropyridine-3-carboxamide

[0252]

[0253] Step 1: tert-butyl 4-(((2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4- yl)amino)methyl)piperidine- 1 -carboxylate

[0254] 2-(2,6-dioxopiperidin-3-yl)-4-((piperidin-4-ylmethyl)amino)isoindoline-l,3-dione (0.7 g, 1.5 mmol) was dissolved in CH2Cl2(6 mL), and HCl solution in 1,4-dioxane (4 M, 3 mL) was added. The reaction was stirred at room temperature for 2 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain a yellow solid product (0.6 g), which was used directly in the next reaction. LCMS (ESI + ): m / z = 371 [M+H] + .

[0255] Step 2: 2-(2,6-dioxopiperidin-3-yl)-4-((piperidin-4-ylmethyl)amino)isoindoline-l,3-dione

[0256] tert-butyl 4-(((2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4- yl)amino)methyl)piperidine- 1 -carboxylate (0.7 g, 1.5 mmol) was dissolved in CH2Cl2(6 mL), and HCl solution in 1,4-dioxane (4 M, 3 mL) was added. The reaction was stirred at room temperature for 2 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain a yellow solid product (0.6 g), which was used directly in the next reaction. LCMS (ESI + ): m / z = 371 [M+H] + .

[0257] Step 3: N-((2S)-l-(4-(((2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4- yl)amino)methyl)piperidin-l-yl)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)-l- oxopropan-2-yl)-5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxamide

[0258] In a 25 mL round-bottom flask, was added (S)-2-(5-fluoro-l-(4-fluorobenzyl)-2- oxo-l,2-dihydropyridine-3-carboxamido)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl) propanoic acid (282 mg, 0.54 mmol), HOBT (109 mg, 0.81 mmol), EDCI (155 mg, 0.81 mmol), DIEA (209 mg, 1.62 mmol) and DMF (6 mL), then added 2-(2,6-dioxopiperidin-3-yl)-4- ((piperidin-4-ylmethyl)amino)isoindoline-l,3-dione (200 mg, 0.54 mmol), stirred at room temperature for 3 h. After the reaction was completed, diluted with water (100 mL). The aqueous phase was extracted with CH2Cl2(100 mL x 3), the organic phases were combined. The organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure, and purified by flash preparation chromatography (CH2Cl2:MeOH (v / v) = 95:5) to obtain a yellow solid (130 mg, yield 25%). LCMS (ESI + ): m / z = 874 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 11.10 (s, 1H), 10.31 - 9.79 (m, 1H), 8.74 - 8.33 (m, 1H), 8.16 - 8.06 (m, 1H), 8.05 - 7.91 (m, 1H), 7.67 - 7.38 (m, 3H), 7.36 - 6.95 (m, 9H), 6.70 - 6.55 (m, 1H), 5.17 (s, 2H), 5.11 - 4.74 (m, 2H), 4.35 - 4.13 (m, 1H), 3.26 - 3.06 (m, 2H), 2.93 - 2.78 (m, 1H), 2.73 - 2.39 (m, 7H), 2.12 - 1.95 (m, 4H), 1.85 - 1.40 (m, 3H), 1.08 - 0.75 (m, 2H).

[0259] Example Twenty-two: 5-Chloro-N-((2S)-3-(2-(6-chloro-4-methylpyridin-3-yl)phenyl)-l-(4-(((2-(2,6-dioxopiperidin-3-yl)-l,3-dioxoisoindolin-4-yl)amino)methyl)piperidin-l-yl)-l- oxopropan-2-yl)-l-(4-chlorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxamide

[0260]

[0261]

[0262] In a 25 mL round-bottom flask, was added (S)-2-(5-chloro-l-(4-chlorobenzyl)-2- oxo-l,2-dihydropyridine-3-carboxamido)-3-(2-(6-chloro-4-methylpyridin-3-yl)phenyl) propanoic acid (308 mg, 0.54 mmol), HOBT (109 mg, 0.81 mmol), EDCI (155 mg, 0.81 mmol), DIEA (209 mg, 1.62 mmol) and DMF (6 mL), then added 2-(2,6-dioxopiperidin-3-yl)-4- ((piperidin-4-ylmethyl)amino)isoindoline-l,3-dione (200 mg, 0.54 mmol), stirred at room temperature for 3 h. After the reaction was completed, diluted with water (100 mL). The aqueous phase was extracted with CH2Cl2(100 mL x 3), the organic phases were combined. The organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure, and purified by flash preparation chromatography (CH2Cl2:MeOH (v / v) = 95:5) to obtain a yellow solid (150 mg, yield 27%). LCMS (ESI + ): m / z = 922 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 10.03 - 9.74 (m, 1H), 8.64 - 8.49 (m, 1H), 8.25 - 7.91 (m, 2H), 7.73 - 7.35 (m, 6H), 7.35 - 6.98 (m, 6H), 6.76 - 6.48 (m, 1H), 5.20 (s, 2H), 5.13 - 4.73 (m, 2H), 4.26 (s, 1H), 3.25 - 2.99 (m, 3H), 2.94 - 2.77 (m, 2H), 2.67 - 2.52 (m, 5H), 2.15 - 1.92 (m, 4H), 1.85 - 1.43 (m, 3H), 1.08 - 0.81 (m, 2H).

[0263] Example Twenty-Three: (S)-N-(l-((3-((4-(2,4-dioxotetrahydropyrimidin-l(2H)-yl)-3- fluorophenyl)amino)propyl)amino)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)-l- oxopropan-2-yl)-5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxamide

[0264]

[0265]

[0266] Step 1: 3-((4-bromo-2-fluorophenyl)amino)propanoic acid

[0267] In a 100 mL round-bottom flask, was placed 4-bromo-2-fluoroaniline (5.0 g, 26.0 mmol), acrylic acid (2.08 g, 28.0 mmol), H2O (40 mL) and AcOH (10 mL), heated to 100 °C and stirred for 6 h. After the reaction was completed, the reaction was diluted with water (50 mL) and extracted with CH2Cl2(50 mL x 3), and the organic phase was collected. The organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a white solid (6.0 g), which was used directly in the next step. LCMS (ESI + ): m / z = 262, 264 [M+H] + .

[0268] Step 2: 1-(4-bromo-2-fluorophenyl)dihydropyrimidine-2,4(lH,3H)-dione

[0269] 3-((4-bromo-2-fluorophenyl)amino)propanoic acid (6.0 g, 22.9 mmol) and urea (3.44 g, 57.3 mmol) were dissolved in AcOH (30 ml), heated to 100 °C and stirred for 6 h. After the reaction was completed, the reaction was concentrated under reduced pressure, the residue was diluted with water (50 mL), the pH was adjusted to 10 with a saturated NaHCO3solution, and then extracted with EtOAc (30 mL x 3), and the organic phase was collected. The organic phase was dried over Na2SO4, filtered and concentrated under reduced pressure, and the residue was purified by flash column chromatography (PE:EtOAc (v / v) = 1:1) to give a white solid (2.3 g, yield 33%) as the target product. LCMS (ESI + ): m / z = 287, 289 [M+H] + .

[0270] Step 3: tert-butyl (3-((4-(2,4-dioxotetrahydropyrimidin-l(2H)-yl)-3-fluorophenyl)amino)propyl)carbamate

[0271] In a 100 mL round-bottom flask, was added 1-(4-bromo-2-fluorophenyl)dihydropyrimidine- 2,4(lH,3H)-dione (1.5 g, 5.24 mmol), N-tert-butoxycarbonyl-l,3-propanediamine (912 mg, 5.24 mmol), Pd2(dba)3(48 mg, 0.05 mmol), BrettPhos (55 mg, 0.10 mmol), Cs2CO3(1.7 g, 5.24 mmol) and dioxane (20 mL) and heated to 90 °C with stirring for 16 h. After completion of the reaction, diluted with water (50 mL) and extracted with EtOAc (50 mL x 3) and the combined organic phase was dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography (CH2Cl2:MeOH (v / v) = 98:2) to afford white solid (160 mg, yield 8%). LCMS (ESI + ): m / z = 403 [M+Na] + .

[0272] Step 4: l-(4-((3-aminopropyl)amino)-2-fluorophenyl)dihydropyrimidine-2,4(lH,3H)- dione

[0273] tert-Butyl (3-((4-(2,4-dioxotetrahydropyrimidin-l(2H)-yl)-3-fluorophenyl)amino)propyl) carbamate (160 mg, 0.42 mmol) was dissolved in HCl in 1,4-dioxane solution (2 M, 5 mL) at 0 °C and stirred at room temperature for 3 h. After completion of the reaction, the reaction was concentrated under reduced pressure to afford white solid (140 mg) which was used directly for the next step. LCMS (ESI + ): m / z = 281 [M+H] + .

[0274] Step 5: (S)-N-(l-((3-((4-(2,4-dioxotetrahydropyrimidin-l(2H)-yl)-3-fluorophenyl)amino)propyl) amino)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)-l-oxopropan-2-yl)-5-fluoro-l-(4- fluorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxamide

[0275] In a 25 mL round-bottom flask, was added (S)-2-(5-fluoro-l-(4-fluorobenzyl)-2- oxo-l,2-dihydropyridine-3-carboxamido)-3-(2-(6-fluoro-4-methylpyridin-3- yl)phenyl)propanoic acid (219 mg, 0.421 mmol), HOBT (56 mg, 0.42 mmol), EDCI (80 mg, 0.42 mmol), DIEA (108 mg, 0.84 mmol) and DMF (10 mL), then added l-(4-((3-aminopropyl)amino)-2-fluorophenyl)dihydropyrimidine-2,4(lH,3H)-dione (140 mg, 0.42 mmol), stirred at room temperature for 5 h. After the reaction was completed, diluted with water (100 mL). The aqueous phase was extracted with CH2Cl2(100 mL x 3), and the organic phases were combined. The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by SFC (Apparatus: SFC Thar prep 80; Column: CHIRALPAK AS-H 250 mm x 20 mm, 5 μm; Modifier: 40% MeOH (FA 0.2%): 60% CO2; Total Flow: 40 g / min) to give a yellow solid (50 mg, 15% yield). LCMS (ESI + ): m / z = 784 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 10.33 (s, 1H), 10.00 - 9.73 (m, 1H), 8.56 (t, J = 3.8 Hz, 1H), 8.21 - 8.06 (m, 2H), 8.07 - 7.70 (m, 1H), 7.55 - 7.35 (m, 3H), 7.33 - 7.14 (m, 4H), 7.11 - 6.95 (m, 3H), 6.41 - 6.25 (m, 2H), 6.05 - 5.88 (m, 1H), 5.15 (s, 2H), 4.77 - 4.44 (m, 1H), 3.57 (t, J = 6.7 Hz, 2H), 3.15 - 2.99 (m, 2H), 2.98 - 2.84 (m, 2H), 2.82 - 2.71 (m, 1H), 2.67 (t, J = 6.7 Hz, 2H), 2.60 - 2.52 (m, 1H), 1.98 (s, 3H), 1.67 - 1.45 (m, 2H).

[0276] Example Twenty-Four: (S)-N-(1-((2-((4-(2,4-dioxotetrahydropyrimidin-1 (2H)-yl)-3- fluorophenyl)amino)ethyl)amino)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)-1- oxopropan-2-yl)-5-fluoro-1-(4-fluorobenzyl)-2-oxo-1,2-dihydropyridine-3-carboxamide

[0277]

[0278]

[0279] Step 1: tert-Butyl (2-((4-(2,4-dioxotetrahydropyrimidin-1 (2H)-yl)-3-fluorophenyl)amino)ethyl) carbamate

[0280] In a 100 mL round-bottom flask, 1-(4-bromo-2-fluorophenyl)dihydropyrimidine-2,4(1H,3H)- dione (1.5 g, 5.24 mmol), N-tert-butoxycarbonyl-1,2-ethanediamine (839 mg, 5.24 mmol), Pd2(dba)3 (48 mg, 0.05 mmol), BrettPhos (55 mg, 0.10 mmol), Cs2CO3 (1.7 g, 5.24 mmol) and dioxane (20 mL) were added and heated to 90 °C with stirring for 16 h. After the reaction was completed, water (50 mL) was added for dilution, and the mixture was extracted with EtOAc (50 mL x 3), and the organic phases were combined. The organic phase was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (CH2Cl2:MeOH (v / v) = 98:2) to give a white solid (220 mg, yield 11%). LCMS (ESI + ): m / z = 389 [M+Na] + .

[0281] Step 2: 1-(4-((2-aminoethyl)amino)-2-fluorophenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0282] Tert-butyl (2-((4-(2,4-dioxotetrahydropyrimidin-1 (2H)-yl)-3-fluorophenyl)amino)ethyl) carbamate (220 mg, 0.60 mmol) was dissolved in a 1,4-dioxane solution of HCl (2 M, 5 mL) at 0 °C, and the reaction was stirred at room temperature for 3 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure to give a white solid (186 mg), which was used directly in the next step. LCMS (ESI + ): m / z = 267 [M+H] + .

[0283] Step 3: (S)-N-(l-((2-((4-(2,4-dioxotetrahydropyrimidin-l(2H)-yl)-3-fluorophenyl)amino)ethyl)amino)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)-l-oxopropan-2-yl)-5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxamide

[0284] In a 25 mL round-bottom flask, was added (S)-2-(5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxamido)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)propanoic acid (313 mg, 0.60 mmol), HOBT (81 mg, 0.60 mmol), EDCI (115 mg, 0.60 mmol), DIEA (155 mg, 1.2 mmol) and DMF (10 mL), then 1-(4-((2-aminoethyl)amino)-2-fluorophenyl)dihydropyrimidine-2,4(lH,3H)-dione (186 mg, 0.70 mmol) was added, and the reaction was stirred at room temperature for 5 h. After the reaction was completed, water (100 mL) was added for dilution. The aqueous phase was extracted with CH2Cl2(100 mL x 3), and the organic phases were combined. The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. Purification by TLC (CH2Cl2:MeOH (v / v) = 95:5) gave a white solid (55 mg, yield 12%). LCMS (ESI + ): m / z = 770 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 10.33 (s, 1H), 9.89 (d, J = 7.9 Hz, 1H), 8.56 (t, J = 3.8 Hz, 1H), 8.24 - 8.12 (m, 2H), 8.06 - 7.72 (m, 1H), 7.53 - 7.36 (m, 3H), 7.33 - 7.14 (m, 4H), 7.09 - 6.96 (m, 3H), 6.42 - 6.33 (m, 2H), 6.11 - 5.92 (m, 1H), 5.16 (s, 2H), 4.73 - 4.47 (m, 1H), 3.56 (t, J = 6.7 Hz, 2H), 3.18 - 3.06 (m, 2H), 3.02 - 2.90 (m, 2H), 2.83 - 2.73 (m, 1H), 2.66 (t, J = 6.7 Hz, 2H), 2.61 - 2.52 (m, 1H), 1.98 (d, J = 3.8 Hz, 3H).

[0285] Example Twenty-Five: (S)-5-chloro-N-(3-(2-(6-chloro-4-methylpyridin-3-yl)phenyl)-1- ((4-((4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3-fluorophenyl)amino)butyl)amino)-1- oxopropan-2-yl)-1-(4-chlorobenzyl)-2-oxo-1,2-dihydropyridine-3-carboxamide

[0286]

[0287] Step 1: 1-(4-bromo-2-fluorophenyl)-3-((2-(trimethylsilyl)ethoxy)methyl)dihydropyrimidine- 2,4(1H,3H)-dione

[0288] In a 250 mL round-bottom flask, 1-(4-bromo-2-fluorophenyl)dihydropyrimidine-2,4(1H,3H)- dione (5.0 g, 17.4 mmol) was dissolved in DMF (100 mL) at 0 °C. Then NaH (1.04 g, 60%, 26.1 mmol) was added carefully to maintain the temperature. The reaction was stirred for 10 min, and then 2-(trimethylsilyl)ethoxymethyl chloride (3.87 g, 20.9 mmol) was added. The reaction was stirred at room temperature for 2 h. After the reaction was completed, the reaction solution was slowly poured into ice water (100 mL), and the aqueous phase was extracted with EtOAc (50 mL x 3). The organic phase was combined and dried over Na2SO4. After filtration, the filtrate was concentrated under reduced pressure, and the residue was separated and purified by column chromatography on silica gel (PE:EtOAc (v / v) = 1:1) to obtain a yellow solid (2.60 g, yield 36%). LCMS (ESI): m / z = 439 [M+Na] + + .

[0289] Step 2: tert-butyl (4-((4-(2,4-dioxo-3-((2-(trimethylsilyl)ethoxy)methyl)tetrahydropyrimidin-1(2H)- yl)-3-fluorophenyl)amino)butyl)carbamate

[0290] ​In a 100 mL round-bottom flask, 1-(4-bromo-2-fluorophenyl)-3-((2- (trimethylsilyl)ethoxy)methyl)dihydropyrimidine-2,4(lH,3H)-dione (2.50 g, 5.99 mmol), N-tert-butoxycarbonyl-l,4-butanediamine (1.28 g, 6.59 mmol), Cs2CO3(5.97 g, 18.0 mmol), RuPhos Pd G3 (511 mg, 0.60 mmol) and 1,4-dioxane solution (35 mL) were added, heated to 110 °C and stirred for 3 h. After the reaction was completed, water (150 mL) was added for dilution, extracted with EtOAc (150 mL x 3), and the organic phase was combined. The organic phase was dried over Na2SO4, filtered and concentrated under reduced pressure, and the residue was purified by flash column chromatography (PE:EtOAc (v / v) = 1:1) to obtain a white solid (2.0 g, yield 64%). LCMS (ESI + ): m / z = 425 [M-100+H] + .

[0291] Step 3: l-(4-((4-aminobutyl)amino)-2-fluorophenyl)dihydropyrimidine-2,4(lH,3H)- dione

[0292] Tert-butyl (4-((4-(2,4-dioxo-3-((2-(trimethylsilyl)ethoxy)methyl)tetrahydropyrimidin- l(2H)-yl)-3-fluorophenyl)amino)butyl)carbamate (1.90 g, 3.62 mmol) was dissolved in CH2Cl2(60 mL) at 0 °C, TFA (19 mL) was added, and the reaction was stirred at room temperature for 16 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the white solid was obtained by reverse phase preparative separation (950 mg, yield 89%). LCMS (ESI + ): m / z = 295 [M+H] + .

[0293] Step 4: (S)-5-chloro-N-(3-(2-(6-chloro-4-methylpyridin-3-yl)phenyl)-l-((4-((4-(2,4- dioxotetrahydropyrimidin-l(2H)-yl)-3-fluorophenyl)amino)butyl)amino)-l- oxopropan-2-yl)-l-(4-chlorobenzyl)-2-oxo-l,2-dihydropyridine-3-carboxamide

[0294] In a 25 mL round-bottom flask, (S)-2-(5-chloro-l-(4-chlorobenzyl)-2-oxo-l,2- dihydropyridine-3-carboxamido)-3-(2-(6-chloro-4-methylpyridin-3-yl)phenyl)propanoic acid (100 mg, 0.17 mmol), HOBT (26 mg, 0.19 mmol), EDCI (41 mg, 0.21 mmol), DIEA (92 mg, 0.7 mmol) and DMF (2 mL) were added at 0 °C, then l-(4-((2-aminoethyl)amino)-2-fluorophenyl)dihydropyrimidine-2,4(lH,3H)-dione (62 mg, 0.21 mmol) was added, and the reaction was stirred at room temperature for 5 h. After the reaction was completed, water (100 mL) was added for dilution. The aqueous phase was extracted with CH2Cl2(100 mL x 3), and the organic phases were combined. The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. Purification by preparative TLC (CH2Cl2:MeOH (v / v) = 95:5) gave a white solid (50 mg, 34% yield). LCMS (ESI + ): m / z = 846 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 10.31 (s, 1H), 9.72 - 9.65 (m, 1H), 8.61 - 8.54 (m, 1H), 8.19 - 7.89 (m, 3H), 7.47 - 7.34 (m, 6H), 7.34 - 7.22 (m, 2H), 7.08 - 6.98 (m, 2H), 6.35 (d, J = 10.7 Hz, 2H), 5.99 - 5.94 (m, 1H), 5.17 (s, 2H), 4.78 - 4.49 (m, 1H), 3.56 (t, J = 6.7 Hz, 2H), 3.07 - 2.88 (m, 4H), 2.79 - 2.73 (m, 1H), 2.66 (t, J = 6.7 Hz, 2H), 2.61 - 2.51 (m, 1H), 1.93 (d, J = 8.6 Hz, 3H), 1.48 - 1.32 (m, 4H).

[0295] Example Twenty-Six: (S)-N-(l-(4-(((4-(2,4-dioxotetrahydropyrimidin-l(2H)-yl)-3- fluorophenyl)amino)amino)methyl)piperidin-l-yl)-3-(2-(6-fluoro-4-methylpyridin-3- yl)phenyl)-l-oxopropan-2-yl)-5-fluoro-l-(4-fluorobenzyl)-2-oxo-l,2-dihydropyridine-3- carboxamide

[0296]

[0297]

[0298] Step 1: tert-butyl 4-(((4-(2,4-dioxotetrahydropyrimidin-1 (2H)-yl)-3- fluorophenyl)amino)methyl)piperidine-1-carboxylate

[0299] In a 100 mL round-bottom flask, was added 1-(4-bromo-2-fluorophenyl)dihydropyrimidine- 2,4(1H,3H)-dione (2.6 g, 9.1 mmol), 1-tert-butoxycarbonyl-4-aminomethylpiperidine (1.9 g, 9.1 mmol), Pd2(dba)3(83 mg, 0.09 mmol), BrettPhos (97 mg, 0.18 mmol), Cs2CO3(2.9 g, 9.1 mmol) and dioxane (30 mL), heated to 90 °C and stirred for 16 h. After the reaction was completed, diluted with water (50 mL) and extracted with EtOAc (50 mL x 3), combined the organic phase. The organic phase was dried over Na2SO4, filtered and concentrated under reduced pressure, the residue was purified by flash column chromatography (CH2Cl2:MeOH (v / v) = 98:2) to give a white solid (300 mg, yield 8%). LCMS (ESI + ): m / z = 443 [M+Na] + .

[0300] Step 2: 1-(2-fluoro-4-((piperidin-4-ylmethyl)amino)phenyl)dihydropyrimidine-2,4(1H,3H)- dione

[0301] Tert-butyl 4-(((4-(2,4-dioxotetrahydropyrimidin-1 (2H)-yl)-3-fluorophenyl)amino)methyl)piperidine-1-carboxylate (300 mg, 0.71 mmol) was dissolved in HCl in 1,4-dioxane solution (2 M, 10 mL) at 0 °C, and stirred at room temperature for 3 h. After the reaction was completed, the reaction was concentrated under reduced pressure to give a white solid (261 mg), which was used directly in the next step. LCMS (ESI + ): m / z = 321 [M+H] + .

[0302] Step 3: (S)-N-(1-(4-(((4-(2,4-dioxotetrahydropyrimidin-1 (2H)-yl)-3-fluorophenyl)amino)amino)methyl)piperidin-1-yl)-3-(2-(6-fluoro-4-methylpyridin-3-yl)phenyl)-1-oxopropan-2-yl)-5-fluoro-1-(4-fluorobenzyl)-2-oxo-1,2-dihydropyridine-3-carboxamide

[0303] In a 25 mL round-bottom flask, was added (S)-2-(5-fluoro-l-(4-fluorobenzyl)-2- oxo-l,2-dihydropyridine-3-carboxamido)-3-(2-(6-fluoro-4-methylpyridin-3- yl)phenyl)propanoic acid (371 mg, 0.71 mmol), HOBT (96 mg, 0.71 mmol), EDCI (136 mg, 0.71 mmol), DIEA (183 mg, 1.4 mmol) and CH2Cl2(10 mL) at 0 °C, then added l-(4-((2-aminoethyl)amino)-2-fluorophenyl)dihydropyrimidine-2,4(lH,3H)-dione (261 mg, 0.71 mmol), and stirred at room temperature for 5 h. After the reaction was completed, diluted with water (100 mL). The aqueous phase was extracted with CH2Cl2(100 mL x 3), and the organic phases were combined. The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give a white solid (40 mg, yield 7%). LCMS (ESI + ): m / z = 824 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 10.33 (s, 1H), 10.18 - 9.95 (m, 1H), 8.59 - 8.52 (m, 1H), 8.18 - 7.92 (m, 2H), 7.52 - 7.42 (m, 2H), 7.36 - 6.97 (m, 8H), 6.46 - 6.34 (m, 2H), 6.18 - 6.05 (m, 1H), 5.17 (s, 2H), 5.07 - 4.76 (m, 1H), 4.37 - 4.17 (m, 1H), 3.65 - 3.51 (m, 2H), 3.46 - 3.20 (m, 1H), 3.15 - 2.75 (m, 4H), 2.71 - 2.53 (m, 4H), 2.13 - 1.92 (m, 3H), 1.81 - 1.43 (m, 3H), 1.02 - 0.77 (m, 2H).

[0304] Example Twenty-Seven, CellTiter-Glo 2.0 assay to detect the effect of compounds on tumor cell proliferation

[0305] 1) Culture MDA-MB-468 cells (Shanghai Cell Bank of Chinese Academy of Sciences, ADAR1 high expression cell strain), and plate them when they are in the logarithmic phase of vigorous growth. On the first afternoon, inoculate 150 μL of 8000 cells per well in a 96-well cell culture plate (no cells are added to the outer circle and PBS is added) while reserving wells without cells and only adding culture medium. Incubate at 37°C, 5% CO2, and 90% humidity overnight.

[0306] 2) The next day, prepare compound solutions with different concentration gradients (100 μM, 30 μM, 10 μM, 3 μM, 1 μM, 0.3 μM, 0.1 μM, and 0.03 μM). Remove the culture medium from the 96-well plate, add 150 uL of compound solution to each well at each concentration, and incubate at 37°C, 5% CO2, and 90% humidity for 72 h.

[0307] After 72 h of cell culture, thaw the CellTiter-Glo 2.0 reagent in a 22°C water bath, remove the 96-well cell culture plate, and add 150 uL of CellTiter-Glo 2.0 reagent to each well. Mix the contents on an orbital shaker for 2 min, and then incubate the plate at room temperature for 10 min.

[0308] 3) After the incubation is complete, transfer all the solutions in the 96-well cell culture plate to a white 96-well plate, and use a BioTek enzyme marker to read the luminescence values and save the analysis data.

[0309] Table 1 Proliferation inhibition ability of example compounds on breast cancer cell strain MDA-MB-468

[0310]

[0311] From the above data, it can be seen that the IC50 of the compound of Example 1 for inhibiting the proliferation of MDA-MB-468 cells is 101.3 nM. Figure 1 50 = 101.3 nM.

[0312] Example Twenty-Eight, Western Blot Detection of the Degradation Ability of Compounds on ADAR1 Protein

[0313] 1) Culture MDA-MB-468 cells (Shanghai Cell Bank of Chinese Academy of Sciences), and when the coverage is 60-70%, add 10 uM of the compound and incubate for 0-72 h. Centrifuge at 1000 rpm for 3 min to collect the cells, add 100 μL of ice-cold RIPA lysis buffer, mix well, and then stand on ice for 30 min. Every 10 min, vortex and invert to ensure complete lysis, and then centrifuge and place on ice.

[0314] ​2) After quantification with protein standard, loading and electrophoresis; after electrophoresis, the gel was removed, and the PVDF membrane soaked in methanol for 30s was clamped with sponge, filter paper, etc., and inserted into the electrophoresis tank, and electrophoretic transfer was carried out at a constant voltage of 100V for 1.5h.

[0315] 3) After the completion of the electric transfer, the PVDF membrane was placed in a blocking solution of 5% skim milk for 1h, washed slowly in TBST for 3 times, 10min each time, and the washed PVDF membrane was placed in a box containing the first antibody and incubated on a 4℃ shaker overnight; after taking out the PVDF membrane, washing it slowly in TBST for 3 times, the second antibody was added, and incubated at room temperature for 1h; after the end, the PVDF membrane was washed with TBST for 3 times; the color developing solution was mixed and uniformly added to the surface of the membrane, and then exposed and developed in the iBright imager, and the picture was taken and saved.

[0316] Table 2 Degradation ability of example compounds on ADAR1 protein

[0317]

[0318] From the above table, it can be seen that after the MDA-MB-468 cells were treated with 10uM of the compound of Example 1, the degradation of ADAR1 protein began to appear at 8h, and the degradation was more obvious after 48h. Figure 2

[0319] Example Twenty-Nine, Detection of the Influence of Compounds on ADAR1 Protein Degradation by HiBiT Reporter Assay (in A549 HiBiT-ADAR1 cell)

[0320] 1) A549 HiBiT-ADAR1 cells (Kanglong Huazheng (Beijing) New Drug Technology Co., Ltd.) were cultured, and the cells were plated when they were in the logarithmic phase of vigorous growth. On the first afternoon, 30uL of 1250 cells per well were inoculated in a 384-well cell culture plate, and incubated at 37℃, 5% CO2, 90% humidity for overnight.

[0321] 2) The next day, compound solutions with different concentration gradients (10uM, 3uM, 1uM, 0.3uM, 0.1uM, 0.03uM, 0.01uM, 0.003uM, 0.001uM) were prepared and added to the 384-well plate, and incubated at 37℃, 5% CO2, 90% humidity for 48h.

[0322] 3) After 48h of cell culture, HiBiT lysis detection reagent was added to the cell culture well, and incubated at room temperature for 30min.

[0323] After incubation, PerkinElemer EnVision microplate reader read the luminescence value, and saved the analysis data. ​

[0324] By Figure 3 It is known that the compound of Example 1 treated A549 HiBiT-ADAR1 cells for 48h, the DC of ADAR1 protein degradation 50 = 849 nM, wherein the DC 50 refers to the concentration of the compound that causes half of the ADAR1 protein to be degraded.

Claims

1. A compound of Formula I, or a stereoisomer, or a pharmaceutically acceptable salt thereof: Formula I wherein X is N or CH, m is 0, 1, 2, 3, or 4; R 1 independently at each occurrence H, halogen, hydroxyl, thiol, nitro, amino, cyano, Ci-6 alkyl, Ci-6 alkyloxy, haloCi-6 alkyl, or haloCi-6 alkyloxy; n is 0, 1, 2, 3, or 4; R 2 independently at each occurrence H, halogen, hydroxyl, thiol, nitro, amino, cyano, Ci-6 alkyl, Ci-6 alkyloxy, haloCi-6 alkyl, or haloCi-6 alkyloxy; p is 0, 1, 2, 3, or 4; R 3 independently at each occurrence H, halogen, hydroxyl, thiol, nitro, amino, cyano, Ci-6 alkyl, Ci-6 alkyloxy, haloCi-6 alkyl, or haloCi-6 alkyloxy; linker is selected from Formula II: Formula II -Y-(CHR a ) a -(CHR b ) b -(CHR c ) c -Z- E3 binder moiety is selected from structures as described in Formula III: Y is CONH, NH, CH2or O, R a independently at each occurrence hydrogen, halogen, hydroxyl, thiol, nitro, amino, cyano, C 1-6 alkyl, C 1-6 alkyloxy, haloC 1-6 alkyl or haloC 1-6 alkyloxy, a is 0, 1, 2, 3 or 4, wherein (CHR a ) a one or more (e.g., 1, 2, 3 or 4) of the CH a optionally replaced with O, NH or CO; or -Y-(CHR a ) a - together or -(CHR a ) a individually form a 4-6 membered (e.g., 4, 5, 6 membered) monocyclic saturated heterocycloalkyl or a 7 to 11 membered (e.g., 7, 8, 9, 10, 11 membered) spirocyclic saturated heterocycloalkyl, the ring heteroatoms of the heterocycloalkyl being 1 or 2 heteroatoms independently selected at each occurrence from nitrogen and oxygen, the heterocycloalkyl being optionally substituted with 1, 2, 3 or 4 substituents selected from halogen, hydroxyl, thiol, nitro, amino, cyano, C 1-6 alkyl, C 1-6 alkyloxy, haloC 1-6 alkyl or haloC 1-6 alkyloxy; R b independently at each occurrence is hydrogen, halogen, hydroxyl, thiol, nitro, amino, cyano, C 1-6 alkyl, C 1-6 alkyloxy, haloC 1-6 alkyl, or haloC 1-6 alkyloxy, b is 0, 1, 2, 3, or 4, wherein one or more (such as 1, 2, 3, or 4) of the CH b ) b R b optionally replaced with O, NH, or CO; or -(CHR a ) a -(CHR b ) b together or -(CHR b ) b -(CHR c ) c together or (CHR b ) b independently at each occurrence is hydrogen, halogen, hydroxyl, thiol, nitro, amino, cyano, C 1-6 alkyl, C 1-6 alkyloxy, haloC 1-6 alkyl, or haloC 1-6 alkyloxy; Z is CONH, NH, CH2or O, R c independently at each occurrence hydrogen, halogen, hydroxyl, thiol, nitro, amino, cyano, C 1-6 alkyl, C 1-6 alkyloxy, haloC 1-6 alkyl or haloC 1-6 alkyloxy, c is 0, 1, 2, 3 or 4, wherein (CHR c ) c one or more (e.g., 1, 2, 3 or 4) of the CH c optionally replaced with O, NH or CO; or -(CHR b ) b -(CHR c ) c - together or -(CHR c ) c -Z- together or -(CHR c ) c - individually form a 4-6 membered (e.g., 4, 5, 6 membered) monocyclic saturated heterocycloalkyl or a 7 to 11 membered (e.g., 7, 8, 9, 10, 11 membered) spirocyclic saturated heterocycloalkyl, the ring heteroatoms of the heterocycloalkyl being 1 or 2 heteroatoms independently selected at each occurrence from nitrogen and oxygen, the heterocycloalkyl being optionally substituted with 1, 2, 3 or 4 substituents selected from halogen, hydroxyl, thiol, nitro, amino, cyano, C 1-6 alkyl, C 1-6 alkyloxy, haloC 1-6 alkyl or haloC 1-6 alkyloxy; M is CH or N; R 4 , R 5 , R 6 , R 7 are each independently H, halogen, hydroxyl, thiol, nitro, amino, cyano, C 1-6 alkyl, C 1-6 alkyl, C 1-6 alkyl, or haloC 1-6 alkyloxy; e is 0, 1, 2, 3, or 4. L is (CH2) d wherein d is 0, 1, 2, or 3, and one or more (e.g., 1, 2, or 3) CH2in (CH2) d is optionally replaced with NH or CO; R 8 halo, hydroxy, mercapto, nitro, amino, cyano, Ci-6 alkyl, Ci-6 alkyloxy, Ci-6 alkylthio, haloCi-6 alkyl, or haloCi-6 alkylthio; 2. A compound of Formula IV, or a stereoisomer, or a pharmaceutically acceptable salt thereof:

3. A compound of Formula V, or a stereoisomer, or a pharmaceutically acceptable salt thereof: In formula IV, X, R 1 , R 2 , R 3 , linker, m, n, p have the meanings indicated for formula I in claim 1. R 9 R 10 Its references are respectively the same as R in Formula III of claim 1. 4 R 5 As stated above.

4. The structure according to claim 1 is selected from the following structures: In formula IV, X, R 1 , R 2 , R 3 , m, n, p, R 8 , linker, L, M, e refer to the same as in claim 1.

5. A pharmaceutical composition comprising a compound according to any one of claims 1-4, or a stereoisomer, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

6. A method for degrading ADAR1 protein, the method comprising contacting ADAR1 protein with a compound according to any one of claims 1-4, or a stereoisomer, or a pharmaceutically acceptable salt thereof.

7. A method of treating a disease mediated by ADAR1 protein, comprising administering to a patient having a disease mediated by ADAR1 a compound according to any one of claims 1-4, or a stereoisomer, or a pharmaceutically acceptable salt thereof. ​

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