Compound for targeted degradation of DHHC family proteins and preparation method and application thereof

Abstract

The invention discloses a compound as shown in a formula (I) for targeted degradation of DHHC family proteins and application of the compound. Through rational design and chemical synthesis, a protein degradation agent (PROTAC) compound targeting DHHC family proteins is constructed, and activity and selectivity evaluation is carried out at the cellular level. According to the invention, the feasibility of selectively degrading specific DHHC family proteins by using the PROTAC strategy is explored for the first time, and the application of the PROTAC technology in the field of protein post-translational modification is expanded; meanwhile, the obtained selective protein degradation agent can be used as a favorable probe tool for researching new functions of DHHC family protein biology and developing a therapeutic drug with an anti-tumor effect. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the field of biomedicine, and particularly relates to a compound for targeted degradation of DHHC and its use. Background Art

[0002] Aspartate-Histidine-Histidine-Cysteine (DHHC) palmitoyltransferases (PATs) are a class of acyltransferases that contain two catalytic Zn 2+ ions and a conserved DHHC tetrapeptide sequence, also known as zinc finger DHHC (zDHHC). zDHHC is mainly responsible for the S-palmitoylation modification of proteins, which regulates protein function, stability, localization, and protein-protein interactions by covalently linking 16-carbon palmitic acid to the Cys residues of proteins. DHHC family proteins are widely present in the plasma membranes of organelles in eukaryotic cells including yeast, animals, and plants, and 23 zDHHCs have been identified to be expressed in mammalian cells. Notably, S-palmitoylation is reversible in enzyme kinetics, with lipid addition catalyzed by PATs and lipid removal catalyzed by a series of serine hydrolase family acyl protein thioesterases (APTs), including APT1 / 2, ABHD10, and ABHD17A / B / C. The activities of these PATs and APTs proteins are strictly regulated by cells and organisms, jointly and dynamically regulating the S-palmitoylation modification state of proteins.

[0003] Studies have clearly shown that abnormal functions of DHHC family proteins are associated with various diseases. For example, elevated levels of zDHHC7 promote the STAT3-HIF1α-zDHHC7 positive feedback loop, thus promoting the development of hepatocellular carcinoma, etc.

[0004] Existing inhibitors of DHHC family proteins generally have disadvantages such as low in vitro and in vivo activities, poor selectivity, and lack of drugability. In addition, in addition to the well-defined acyltransferase enzyme activity function, zDHHC has also been reported to be able to affect the localization, stability, and activity regulation of cellular proteins through other domains of the protein. Therefore, there is an urgent need to develop probe molecules with high selectivity, strong activity, and comprehensive intervention in the functions of DHHC proteins. Summary of the Invention

[0005] The present invention aims to overcome the deficiencies of the prior art and provides a compound for targeted degradation of DHHC family proteins, its preparation method, and the use of the compound in the preparation of drugs for treating diseases related to DHHC family proteins.

[0006] Specifically, the present invention provides a compound of formula (I):

[0007]

[0008] Among them,

[0009] A is CH or N; R is a halogen or -CH2CN;

[0010] is a single bond or a double bond;

[0011] n is a positive integer from 3 to 20;

[0012] B is S, O or CH, CH2;

[0013] L is a linking fragment;

[0014] D is Among them, M1 is absent, CH2, N or O; M2 is C=O or CH2. When M1 is absent, L is directly connected to the benzene ring carbon atom on D.

[0015] As an option, when A is CH, B is S and is a single bond, R is Cl, Br, I,

[0016] When A is N, B is CH and is a double bond with E configuration, R is -CH2CN.

[0017] As an option, the said L is selected from

[0018]

[0019] As an option, in L, x is an integer from 1 to 6; y is an integer from 1 to 10; n1, n2, n3 and n4 are each independently selected from 1, 2, 3.

[0020] Preferably, it has the structure shown in the general formula (II) or (III):

[0021]

[0022] The definitions of L and D are the same as those in the general formula (I) or other alternative schemes.

[0023] As an option, D is When (wherein, M1 is absent, CH2, N, O; M2 is C=O or CH2;), L is selected from

[0024] Furthermore, L is x is selected from integers from 1 to 4; y is selected from 6, 7, 8.

[0025] x is selected from integers from 1 to 4; y is selected from 6, 7, 8.

[0026] As an option, when D is L is selected from x is an integer selected from 1 to 4.

[0027] As an option, D is selected from

[0028] Preferably, L is x is an integer selected from 1 to 4; D is

[0029] In some embodiments of the present invention, the compound is:

[0030]

[0031] S-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethoxy)acetamido)ethyl) 2-bromopalmitate TM1

[0032]

[0033] S-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxo)ethoxy)ethoxy)acetamido)ethyl) 2-bromopalmitate TM2

[0034]

[0035] S-(14-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-4-oxo-6,9,12-trioxa-3-azatetradecyl) 2-bromopalmitate TM3

[0036]

[0037] S-(17-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-4-oxo-6,9,12,15-tetraoxa-3-azapentadecyl) 2-bromopalmitate TM4

[0038] (E)-N-(cyanomethyl)-4-(2-(2-((2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethoxy)acetamido)-N-tetradecylbut-2-enamide TM5

[0039]

[0040] (E)-N-(Cyanomethyl)-4-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethoxy)acetamido)-N-tetradecylbut-2-enamide TM6

[0041]

[0042] (E)-N-(Cyanomethyl)-4-(2-(2-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethoxy)ethoxy)ethoxy)acetamido)-N-tetradecylbut-2-enamide TM7

[0043]

[0044] (E)-N-(4-((Cyanomethyl)(tetradecyl)amino)-4-oxobutan-2-en-1-yl)-14-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-3,6,9,12-tetraoxatetradecanamide TM8

[0045]

[0046] S-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)acetamido)ethyl) 2-bromohexadecanoate thioester TM9

[0047]

[0048] S-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxo)ethoxy)ethoxy)acetamido)ethyl) 2-bromohexadecanoate thioester TM10

[0049]

[0050] S-(14-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-4-oxo-6,9,12-trioxa-3-azatetradecyl) 2-bromohexadecanoate thioester TM11

[0051]

[0052] S-(17-((2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-4-oxo-6,9,12,15-tetraoxa-3-azapentadecyl) 2-bromohexadecanoate Thioester TM12

[0053]

[0054] S-(2-(7-((2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)heptanamido)ethyl) 2-bromohexadecanoate Thioester TM13

[0055]

[0056] S-(2-(9-((2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)nonanamido)ethyl) 2-bromohexadecanoate Thioester TM14

[0057]

[0058] S-(2-(3-(3-(((S)-1-((2S,4R)-4-Hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)propylamino)ethyl) 2-bromohexadecanoate Thioester TM15

[0059]

[0060] S-((S)-15-((2S,4R)-4-Hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-16,16-dimethyl-4,13-dioxo-7,10-dioxa-3,14-diazapentadecyl) 2-bromohexadecanoate Thioester TM16

[0061]

[0062] S-((S)-18-((2S,4R)-4-Hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-19,19-dimethyl-4,16-dioxo-7,10,13-trioxa-3,17-diazadocosyl) 2-bromohexadecanoate Thioester TM17

[0063]

[0064] S-((S)-21-((2S,4R)-4-Hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-22,22-dimethyl-4,19-dioxo-7,10,13,16-tetraoxa-3,20-diazatricosane) 2-bromopalmitate TM18

[0065]

[0066] S-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)oxy)ethoxy)ethoxy)acetamido)ethyl) 2-bromopalmitate TM19

[0067]

[0068] S-(14-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)oxy)-4-oxo-6,9,12-trioxa-3-azatetradecyl) 2-bromopalmitate TM20

[0069]

[0070] S-(2-(2-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-2,7-diazaspiro[3.5]nonan-7-yl)acetamido)ethyl) 2-bromopalmitate TM21

[0071]

[0072] S-(2-(1-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)azetidin-3-yl)piperidine-4-carboxamido)ethyl) 2-bromopalmitate TM22

[0073]

[0074] S-(2-(1'-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-[1,4'-bipiperidine]-4-carboxamido)ethyl) 2-bromopalmitate TM23

[0075]

[0076] S-(2-(2-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)acetamido)ethyl) 2-bromohexadecanoate thioester TM24

[0077]

[0078] S-(2-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine-4-carboxamido)ethyl) 2-bromohexadecanoate thioester TM25

[0079]

[0080] S-(2-(4-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)benzamido)ethyl) 2-bromohexadecanoate thioester TM26

[0081] A pharmaceutical preparation comprising a therapeutically effective amount of the compound or a pharmaceutically acceptable salt or configurational isomer thereof according to any one of the technical solutions, and necessary excipients.

[0082] Use of the compound or a pharmaceutically acceptable salt or configurational isomer thereof according to any one of the above in the preparation of a drug for treating diseases alleviated, improved or cured by degrading DHHC family proteins.

[0083] Through rational design and chemical synthesis, the present invention constructs a series of PROTAC compounds targeting DHHC family proteins, and conducts activity and selectivity evaluations at the cellular level, with the expectation of discovering protein degraders selective for specific DHHC family members. On this basis, using the techniques and means of chemical biology, the pathways and mechanisms by which active PROTAC molecules degrade DHHC family proteins are studied. The present invention explores for the first time the feasibility of selectively degrading specific DHHC family proteins using the PROTAC strategy, expanding the application of PROTAC technology in the field of protein post-translational modification; at the same time, the obtained selective protein degraders can be used as advantageous probe tools for studying the new biological functions of DHHC family proteins, and can also be used to develop drugs for treating diseases caused by abnormalities of DHHC family proteins.

[0084] In some embodiments of the present invention, a pharmaceutical composition is provided, wherein the pharmaceutical composition comprises a therapeutically effective amount of the compound or a pharmaceutically acceptable salt thereof described above, and one or more pharmaceutically acceptable carriers, diluents or excipients.

[0085] In some embodiments of the present invention, there is provided the use of the compound or its pharmaceutically acceptable salt or pharmaceutical composition described in any one of the above in the preparation of a drug for preventing and / or treating DHHC1 abnormality diseases.

[0086] In some embodiments of the present invention, there is provided the use of the compound or its pharmaceutically acceptable salt or pharmaceutical composition described in any one of the above in preventing and / or treating DHHC1 abnormality diseases.

[0087] In some embodiments of the present invention, the use of the compound or its pharmaceutically acceptable salt or pharmaceutical composition described in any one of the above, wherein the diseases are selected from cancer, blood diseases, viral infectious diseases, neurodegenerative diseases, autoimmune diseases, metabolic diseases, cardiovascular diseases.

[0088] In some embodiments of the present invention, the use of the compound or its pharmaceutically acceptable salt or pharmaceutical composition described in any one of the above, wherein the blood diseases include leukemia, anemia, platelet dysfunction, hemophilia, multiple myeloma, etc.

[0089] Definitions and Explanations

[0090] Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered indeterminate or unclear without a specific definition, but should be understood in its ordinary meaning. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0091] It should be understood that the substitutions and combinations of substitutions described herein, whether explicitly stated or not, refer to substitutions that conform to the valence of the substituted member. For example, substitution applied to a carbon member refers to the tetravalence of C; when applied to a nitrogen member, it refers to the trivalence of N; when generally indicating a positive charge, it refers to the four bonds of a nitrogen member. Options allowed by valence are part of the art.

[0092] Unless otherwise specified, a solid wedge bond and a dashed wedge bond represent the absolute configuration of a stereocenter, a straight solid bond and a straight dashed bond represent the relative configuration of a stereocenter, and a wavy line represents a solid wedge bond or a dashed wedge bond or a wavy line represents a straight solid bond and a straight dashed bond

[0093] When it is not specified which atom of the listed substituent is connected to the substituted group, such a substituent can be bonded through any of its atoms. For example, a phenyl group as a substituent can be connected to the substituted group through any carbon atom on the benzene ring; a ring system formed by a substituent drawing a bond to the central ring (as shown in ) represents alternative substitution at all substitutable positions on the ring system.

[0094] The compounds provided by the present invention have good targeting effects on the DHHC1 family. BRIEF DESCRIPTION OF THE DRAWINGS

[0095] Figure 1 To evaluate the degradation activity of mzDHHC(1-9, 11-13) on TM1-TM4 by Western-blot (the PCCL group is not administered and is only used to judge the transfection efficiency, the same below).

[0096] Figure 2 To evaluate the degradation activity of mzDHHC(14-24) on TM1-TM4 by Western-blot.

[0097] Figure 3 To evaluate the degradation activity of mzDHHC1 on TM1-TM20 by Western-blot.

[0098] Figure 4 To evaluate the degradation activity of mzDHHC1 on TM21-TM26 by Western-blot. DETAILED DESCRIPTION OF THE INVENTION

[0099] The present invention will be described in detail below by way of examples, but this does not mean any adverse limitation to the present invention. The present invention has been described in detail herein, and its specific embodiments have also been disclosed. It will be obvious to those skilled in the art that various changes and improvements can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

[0100] In the examples, the experimental methods without specific conditions are conventional methods and conventional conditions well known in the art, or are operated according to the conditions recommended by the instrument manufacturer.

[0101] Example 1. S-(2-(tert-Butylcarbonyl)amino)ethyl) 2-Bromothiohexadecanoate A1

[0102]

[0103] To commercially available 2-bromohexadecanoic acid (1 g, 2.98 mmol, 1.1 eq) in 50 mL of anhydrous dichloromethane (DCM), commercially available 2-(Boc-amino)ethanethiol (480.4 mg, 2.71 mmol, 1 eq), EDCI (621.4 mg, 3.25 mmol, 1.2 eq) and DMAP (330.6 mg, 2.71 mmol, 1 eq) were added successively. After stirring the reaction system at room temperature for 12 hours, the mixture was washed successively with 80 mL of water, 80 mL of 0.5 M aqueous hydrochloric acid and 80 mL of saturated brine. After drying over anhydrous Na2SO4 and concentrating thoroughly, the crude product was subjected to column chromatography (EtOAc / hexanes = 0%-3%) to obtain 976 mg of a white pasty solid.

[0104] Yield 73.1%; LC-MS (ESI, m / z): calcd for C 18 H 37 BrNOS + , [M - Boc + 2H] + 394.2, found 394.1; 1 H NMR (400 MHz, CDCl3) δ 4.80 (s, 1H), 4.36 (dd, J = 8.0, 6.3 Hz, 1H), 3.33 (q, J = 6.7 Hz, 2H), 3.07 (t, J = 6.5 Hz, 2H), 2.09 - 1.95 (m, 2H), 1.44 (s, 9H), 1.25 (s, 24H), 0.87 (d, J = 7.1 Hz, 3H).

[0105] Example 2. S-(2-Aminoethyl) 2-bromothiohexadecanoate hydrochloride A

[0106]

[0107] At room temperature, 2 M HCl-EtOAc solution (9.89 mL, 19.79 mmol, 10 eq) was slowly added to A1 (976 mg, 1.979 mmol, 1 eq). After stirring the reaction system for 8 h, it was concentrated under reduced pressure. 848 mg of a white solid was obtained.

[0108] Yield 100%; LC-MS (ESI, m / z): calcd for C 18 H 37 BrNOS + , [M + H] + 394.2, found 394.1.

[0109] Example 3. (E)-4-((tert-Butoxycarbonyl)amino)but-2-enoic acid B1

[0110]

[0111] At 0 °C, commercially available 2-(diethoxyphosphoryl)acetic acid (259 mg, 1.32 mmol, 1.05 eq) was dissolved in 4 mL of anhydrous THF. A commercially available 1 M solution of lithium bis(trimethylsilyl)amide in THF (2.638 mL, 2.638 mmol, 2.1 eq) was slowly added dropwise thereto under an ice bath. After the reaction system was stirred for 1 h, the reaction system was cooled to -20 °C. Commercially available tert-butyl (2-oxoethyl)carbamate (200 mg, 1.256 mmol, 1 eq) was dissolved in 2 mL of anhydrous THF and slowly added dropwise to the reaction system at -20 °C, and then stirred for 2 h. The temperature was raised to room temperature and stirring was continued for 18 h. After monitoring by TLC and LC-MS that the raw materials had completely reacted, 0.4 mL of AcOH was added dropwise to quench the reaction. The mixture was diluted with 8 mL of water and extracted with EtOAc (6 mL × 3). The organic phase was dried over anhydrous Na2SO4 and concentrated thoroughly. The crude product was purified by column chromatography (EtOAc / hexanes = 16% - 18%, 0.1% AcOH) to obtain 134.5 mg of a grayish-yellow solid.

[0112] Yield: 50.6%; LC-MS (ESI, m / z): calcd for C9H 14 NO4 – , [M-H] – 200.1, found 200.1; 1 1H NMR (400 MHz, CDCl3) δ 6.99 (m, 1H), 5.93 (m, 1H), 4.77 (s, 1H), 3.94 (s, 2H), 1.44 (s, 9H).

[0113] Example 4. 2-(Tetradecylamino)acetonitrile B2

[0114]

[0115] At room temperature, commercially available tetradecan-1-amine (700 mg, 3.29 mmol, 1 eq) and K2CO3 (908.04 mg, 6.58 mmol, 2 eq) were dissolved in 5 mL of anhydrous DMF. 2-Bromoacetonitrile (174.6 μL, 2.46 mmol, 0.75 eq) was added to the mixture. The reaction system was heated to 85 °C and stirred for 8 h. After monitoring by TLC that the raw materials had completely reacted, the reaction system was cooled to room temperature, diluted with 30 mL of saturated NaHCO3 solution, and extracted with EtOAc (20 mL × 3). The organic phases were combined, washed with 100 mL of saturated brine, and concentrated under reduced pressure. The crude product was purified by column chromatography (EtOAc / hexanes = 0% - 3%) to obtain 497 mg of a pale yellow solid.

[0116] Yield 59.9%; LC-MS (ESI, m / z): calcd for C 16 H 33 N2 + , [M + H] + 253.3, found 253.3.

[0117] Example 5. tert-Butyl (E)-(4-((cyanomethyl)(tetradecyl)amino)-4-oxobut-2-en-1-yl)carbamate B3

[0118]

[0119] At 0 °C, B1 (70 mg, 0.348 mmol, 1 eq), HATU (198.4 mg, 0.522 mmol, 1.5 eq) were dissolved in 3 mL of anhydrous DCM. DIPEA (151.3 μL, 0.87 mmol, 2.5 eq) was slowly added dropwise under an ice bath. After stirring the reaction for 10 min, the reaction system was warmed to room temperature. A 3 mL anhydrous DCM solution of B2 (87.8 mg, 0.348 mmol, 1 eq) was added to the reaction system, and stirring was continued for 8 h. After monitoring by TLC and LC-MS that the raw materials had completely reacted, the reaction solution was diluted with 10 mL of water, extracted with DCM (8 mL × 3), concentrated under reduced pressure, and the crude product was purified by column chromatography (EtOAc / hexanes = 15% - 20%) to obtain 35.8 mg of a colorless crystalline solid.

[0120] Yield 23.6%; LC-MS (ESI, m / z): calcd for C 25 H 46 N3O3 + , [M + H] + 436.4, found 436.3; 1 1H NMR (400 MHz, CDCl3) δ 6.91 (d, J = 14.4 Hz, 1H), 6.31 - 6.26 (m, 1H), 4.74 (s, 1H), 4.32 (s, 2H), 3.93 - 3.89 (m, 2H), 3.43 (t, J = 7.7 Hz, 2H), 1.65 (d, J = 6.7 Hz, 2H), 1.44 (s, 9H), 1.24 (s, 22H), 0.86 (s, 3H).

[0121] Example 6. (E)-4-Amino-N-(cyanomethyl)-N-tetradecylbut-2-enamide trifluoroacetate B

[0122]

[0123] At room temperature, B3 (35.8 mg, 0.082 mmol, 1 eq) was dissolved in a 15% v / v TFA / DCM solution (635 μL, 1.211 mmol, 15 eq), and the mixture was stirred for 12 h. After monitoring the complete reaction of the starting materials by TLC and LC-MS, it was diluted with 2 mL of DCM, concentrated under reduced pressure, and the above operation was repeated 3 times to remove the remaining TFA. It was dried under vacuum to obtain 36.8 mg of a black liquid.

[0124] Yield 100%; LC-MS (ESI, m / z): calcd for C 20 H 40 N3O2 + , [M + H2O + H] + 354.3, found 354.3.

[0125] Example 7. tert-Butyl 2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethoxy)acetate i-1

[0126]

[0127] 2-(2,6-Dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (120 mg, 0.44 mmol, 1 eq) synthesized by the mature route and Na2CO3 (232.1 mg, 2.19 mmol, 5 eq) were dissolved in 3 mL of DMF. Under N2 protection, the mixture was heated to 100 °C and stirred for 15 min. tert-Butyl 2-(2-(p-toluenesulfonyloxy)ethoxy)acetate (158.4 mg, 0.48 mmol, 1.1 eq) synthesized by the mature route was dissolved in 3 mL of DMF and slowly added dropwise to the reaction system. The reaction system was stirred for another 8 h. After monitoring the complete reaction of the starting materials by TLC, the reaction system was cooled to room temperature, diluted with 40 mL of water, and extracted with EtOAc (30 mL × 3). The organic phases were combined, washed with 150 mL of saturated brine, concentrated under reduced pressure, and the crude product was purified by column chromatography to obtain 137 mg of a yellowish-green oily solid.

[0128] Yield: 72.0%; LC-MS (ESI, m / z): calcd for C 17 H 17 N2O8 + , [M - t Bu + 2H] + 377.2, found 377.1.

[0129] Example 8. tert-Butyl 2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethoxy)ethoxy)acetate i-2

[0130]

[0131] The preparation method was the same as that of compound i-1. tert-Butyl 2-(2-(2-(tosyloxy)ethoxy)ethoxy)acetate synthesized by a mature route was used instead of tert-Butyl 2-(tosyloxy)ethoxy)acetate to obtain 101 mg of a yellowish green oily solid.

[0132] Yield: 48.2%; LC-MS (ESI, m / z): calcd for C 19 H 21 N2O9 + , [M - t Bu + 2H] + 421.2, found 421.1.

[0133] Example 9. tert-Butyl 2-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethoxy)ethoxy)ethoxy)acetate i-3

[0134]

[0135] The preparation method was the same as that of compound i-1. tert-Butyl 2-(2-(2-(2-(tosyloxy)ethoxy)ethoxy)ethoxy)acetate synthesized by a mature route was used instead of tert-Butyl 2-(tosyloxy)ethoxy)acetate to obtain 141 mg of a yellowish green oily solid.

[0136] Yield: 61.6%; LC-MS (ESI, m / z): calcd for C 21 H 25 N2O 10 + , [M - t Bu + 2H] + 465.2, found 465.1.

[0137] Example 10. tert-Butyl 4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-3,6,9,12-tetraoxatetradecanoate i-4

[0138]

[0139] The preparation method was the same as that of compound i-1. 14-(p-Toluenesulfonyloxy)-3,6,9,12-tetraoxatetradecanoic acid tert-butyl ester synthesized by a mature route was used to replace 2-(2-(p-Toluenesulfonyloxy)ethoxy)acetic acid tert-butyl ester, and 131 mg of a yellowish-green oily solid was obtained.

[0140] Yield: 52.7%; LC-MS (ESI, m / z): calcd for C 23 H 29 N2O 11 + , [M - t Bu + 2H] + 509.2, found 509.2.

[0141] Example 11. 2-(2-((2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethoxy)acetic acid trifluoroacetate i-1-a

[0142]

[0143] At room temperature, i-1 (137 mg, 0.317 mmol, 1 eq) was dissolved in a 50% v / v TFA / DCM solution (968 μL, 6.34 mmol, 20 eq), and stirred for 12 h. After monitoring by TLC that the raw materials had completely reacted, it was diluted with 1 mL of DCM, concentrated under reduced pressure, and the above operation was repeated 3 times, and then dried in vacuo to remove the remaining TFA, obtaining 155.3 mg of a yellowish-green liquid.

[0144] Yield: 100%; LC-MS (ESI, m / z): calcd for C 17 H 15 N2O8 - , [M - H] - 375.1, found 375.1.

[0145] Example 12. 2-(2-(2-((2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethoxy)ethoxy)acetic acid trifluoroacetate i-2-a

[0146]

[0147] The preparation method was the same as that of compound i-1-a. Compound i-2 was used to replace compound i-1, and 113.3 mg of a yellowish-green liquid was obtained.

[0148] Yield: 100%; LC-MS (ESI, m / z): calcd for C 19 H 19N2O9 - ,[M-H] - 419.1, found 418.9。

[0149] Example 13. 2-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethoxy)ethoxy)acetic acid trifluoroacetate i-3-a

[0150]

[0151] Prepared in the same manner as compound i-1-a, using compound i-3 instead of compound i-1, to obtain 156.7 mg of a yellowish green liquid.

[0152] Yield: 100%; LC-MS (ESI, m / z): calcd for C 21 H 23 N2O 10 - ,[M-H] - 463.1, found 462.9。

[0153] Example 14. 14-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-3,6,9,12-tetraoxatetradecanoic acid trifluoroacetate i-4-a

[0154]

[0155] Prepared in the same manner as compound i-1-a, using compound i-4 instead of compound i-1, to obtain 144.4 mg of a yellowish green liquid.

[0156] Yield: 100%; LC-MS (ESI, m / z): calcd for C 23 H 27 N2O 11 - ,[M-H] - 507.2, found 507.1。

[0157] Example 15. 2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)acetic acid tert-butyl ester i-5

[0158]

[0159] The preparation method is the same as that of compound i-1, using 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione to replace 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione. The crude product was not purified at all, and the obtained 216 mg of pale yellow liquid was directly subjected to the next reaction.

[0160] LC-MS(ESI,m / z): calcd for C 17 H 17 N2O8 + , [M- t Bu+2H] + 377.2, found 376.9.

[0161] Example 16. tert-Butyl 2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)acetate i-6

[0162]

[0163] The preparation method is the same as that of compound i-2, using 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione to replace 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione. The crude product was not purified at all, and the obtained 421 mg of pale yellow liquid was directly subjected to the next reaction.

[0164] LC-MS(ESI,m / z): calcd for C 19 H 21 N2O9 + , [M- t Bu+2H] + 421.2, found 420.9.

[0165] Example 17. tert-Butyl 2-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethoxy)acetate i-7

[0166]

[0167] The preparation method is the same as that of compound i-3, using 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione to replace 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione. The crude product was not purified at all, and the obtained 231 mg of yellowish green liquid was directly subjected to the next reaction.

[0168] LC-MS (ESI, m / z): calculated for C 21 H 25 N2O 10 + , [M - t Bu + 2H] + 465.2, found 465.0.

[0169] Example 18. 14 - ((2-(2,6 - dioxopiperidin - 3 - yl)-1,3 - dioxoisoindolin - 5 - yl)oxy)-3,6,9,12 - tetraoxatetradecanoic acid tert - butyl ester i - 8

[0170]

[0171] The preparation method is the same as that of compound i - 4, using 2-(2,6 - dioxopiperidin - 3 - yl)-5 - hydroxyisoindoline - 1,3 - dione instead of 2-(2,6 - dioxopiperidin - 3 - yl)-4 - hydroxyisoindoline - 1,3 - dione. The crude product was purified by column chromatography to obtain 50 mg of a yellow - green oily solid.

[0172] Yield: 44.2%; LC - MS (ESI, m / z): calculated for C 23 H 29 N2O 11 + , [M - t Bu + 2H] + 509.2, found 509.0.

[0173] Example 19. 2 - (2 - ((2-(2,6 - dioxopiperidin - 3 - yl)-1,3 - dioxoisoindolin - 5 - yl)oxy)ethoxy)acetic acid trifluoroacetate i - 5 - a

[0174]

[0175] The preparation method is the same as that of compound i - 1 - a, using unpurified compound i - 5 instead of compound i - 1 to obtain 159.2 mg of a yellow - green liquid.

[0176] LC - MS (ESI, m / z): calculated for C 17 H 15 N2O8 - , [M - H] - 375.1, found 374.9.

[0177] Example 20. 2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)acetic acid trifluoroacetate i-6-a

[0178]

[0179] The preparation method was the same as that of compound i-1-a. Using unpurified compound i-6 instead of compound i-1, 313 mg of yellowish green liquid was obtained.

[0180] LC-MS(ESI,m / z): calcd for C 19 H 19 N2O9 - , [M-H] - 419.1, found 418.9.

[0181] Example 21. 2-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethoxy)ethoxy)acetic acid trifluoroacetate i-7-a

[0182]

[0183] The preparation method was the same as that of compound i-1-a. Using unpurified compound i-7 instead of compound i-1, 163 mg of yellowish green liquid was obtained.

[0184] LC-MS(ESI,m / z): calcd for C 21 H 23 N2O 10 - , [M-H] - 463.1, found 462.9.

[0185] Example 22. 14-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-3,6,9,12-tetraoxatetradecanoic acid trifluoroacetate i-8-a

[0186]

[0187] The preparation method was the same as that of compound i-1-a. Using compound i-8 instead of compound i-1, 153.6 mg of yellowish green liquid was obtained.

[0188] LC-MS(ESI,m / z): calcd for C 23 H 27 N2O 11 -, [M-H] - 507.2, found 506.9.

[0189] Example 23. 7 - ((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)tert-butyl heptanoate i-9

[0190]

[0191] The preparation method is the same as that of compound i-1. Using commercially available tert-butyl 7-bromoheptanoate to replace compound tert-butyl 2-(2-(p-toluenesulfonyloxy)ethoxy)acetate, 222 mg of green oily liquid was obtained.

[0192] LC-MS(ESI, m / z): calcd for C 20 H 23 N2O7 + , [M- t Bu+2H] + 403.2, found 403.0.

[0193] Example 24. 9 - ((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)tert-butyl nonanoate i-10

[0194]

[0195] The preparation method is the same as that of compound i-1. Using commercially available tert-butyl 9-bromononanoate to replace compound tert-butyl 2-(2-(p-toluenesulfonyloxy)ethoxy)acetate, 269 mg of green oily liquid was obtained.

[0196] LC-MS(ESI, m / z): calcd for C 22 H 27 N2O7 + , [M- t Bu+2H] + 431.2, found 431.0.

[0197] Example 25. 7 - ((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)heptanoic acid trifluoroacetate i-9-a

[0198]

[0199] The preparation method is the same as that of compound i-1-a. Using compound i-9 to replace compound i-1, the crude product was purified by preparative thin layer chromatography (MeOH / DCM = 2%), and 57 mg of white oily solid was obtained.

[0200] LC-MS (ESI, m / z): calculated for C 20 H 23 N2O7 + , [M+H] + 403.1, found 403.0; 1 1H NMR (400 MHz, CDCl3) δ 8.55 (s, 1H), 7.65 (dd, J = 8.6, 7.2 Hz, 1H), 7.43 (dd, J = 7.3 Hz, 1H), 7.19 (dd, J = 8.5 Hz, 1H), 4.95 (m, 1H), 4.16 (t, J = 6.4 Hz, 2H), 2.90 - 2.73 (m, 3H), 2.35 (t, J = 7.4 Hz, 2H), 2.14 - 2.08 (m, 1H), 1.87 (t, J = 7.5 Hz, 2H), 1.66 (q, J = 7.5 Hz, 2H), 1.53 (m, 2H), 1.43 (m, 2H).

[0201] Example 26. 9-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)nonanoic acid trifluoroacetate i-10-a

[0202]

[0203] Prepared in the same way as compound i-1-a, using compound i-10 instead of compound i-1. The crude product was purified by preparative thin layer chromatography (MeOH / DCM = 2.5%) to obtain 13.8 mg of a white oily solid.

[0204] LC-MS (ESI, m / z): calculated for C 22 H 27 N2O7 + , [M+H] + 431.2, found 431.0; 1 1H NMR (400 MHz, CDCl3) δ 8.60 (s, 1H), 7.65 (dd, J = 7.9, 7.9 Hz, 1H), 7.43 (dd, J = 7.3 Hz, 1H), 7.19 (dd, J = 8.5 Hz, 1H), 4.94 (dd, J = 11.8, 5.2 Hz, 1H), 4.16 (t, J = 6.3 Hz, 2H), 2.89 - 2.72 (m, 3H), 2.32 (s, 2H), 2.13 - 2.09 (m, 1H), 2.00 (m, 2H), 1.85 (t, J = 7.7 Hz, 2H), 1.62 (s, 4H), 1.45 (s, 4H).

[0205] tert-Butyl 3-(3-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)propionate i-11

[0206]

[0207] At 0 °C, the VHL ligand (101.7 mg, 0.229 mmol, 1 eq) synthesized via the mature route, commercially available 3-(3-(tert-butoxy)-3-oxopropoxy)propanoic acid (50 mg, 0.229 mmol, 1 eq), EDCI (48.5 mg, 0.252 mmol, 1.1 eq) and HOBt (35.22 mg, 0.229 mmol, 1 eq) were dissolved in 1 mL of anhydrous DCM. DIPEA (199.14 μL, 1.145 mmol, 5 eq) was slowly added dropwise under an ice bath, and then the reaction system was raised to room temperature and stirred for another 8 h. After monitoring by TLC and LC-MS that the raw materials had completely reacted, the reaction solution was diluted with 3 mL of water and extracted with DCM (2 mL × 3), and concentrated under reduced pressure. The crude product was purified by preparative thin layer chromatography (MeOH / DCM = 5%) to obtain 47.6 mg of a white oily solid.

[0208] Yield: 32.3%; LC-MS (ESI, m / z): calcd for C 33 H 49 N4O7S + , [M+H] + 645.3, found 645.1.

[0209] tert-Butyl 3-(2-(3-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethoxy)propionate i-12

[0210]

[0211] The preparation method was the same as that of compound i-11, using commercially available 3-(2-(3-(tert-butoxy)-3-oxopropoxy)ethoxy)propanoic acid instead of 3-(3-(tert-butoxy)-3-oxopropoxy)propanoic acid to obtain 70.8 mg of a white oily solid.

[0212] Yield: 54.2%; LC-MS (ESI, m / z): calcd for C 35 H 53 N4O8S + , [M+H] + 689.4, found 689.1.

[0213] Example 29. (S)-15-((2S,4R)-4-Hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carboxamido)-16,16-dimethyl-13-oxo-4,7,10-trioxa-14-azapentadecanoic acid tert-butyl ester i-13

[0214]

[0215] The preparation method was the same as that of compound i-11. Using commercially available compound 2,2-dimethyl-4-oxo-3,7,10,13-tetraoxahexadecanoic acid to replace compound 3-(3-(tert-butoxy)-3-oxopropoxy)propanoic acid, 75.6 mg of white oily solid was obtained.

[0216] Yield: 64.5%; LC-MS (ESI, m / z): calcd for C 37 H 57 N4O9S + , [M+H] + 733.4, found 733.1; 1 1H NMR (400 MHz, CDCl3) δ 8.66 (s, 1H), 7.54 (d, J = 7.8 Hz, 1H), 7.37 (m, 4H), 7.06 (d, J = 8.1 Hz, 1H), 5.09 - 5.03 (m, 1H), 4.73 (t, J = 7.9 Hz, 1H), 4.49 (s, 1H), 4.47 (s, 1H), 4.11 (m, 1H), 3.73 - 3.71 (m, 1H), 3.68 (t, J = 6.5 Hz, 4H), 3.60 (m, 8H), 3.57 - 3.54 (m, 1H), 2.51 (s, 3H), 2.48 (m, 4H), 2.06 - 2.03 (m, 2H), 1.45 (d, J = 7.0 Hz, 3H), 1.42 (s, 9H), 1.03 (s, 9H).

[0217] Example 30. (S)-18-((2S,4R)-4-Hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-19,19-dimethyl-16-oxo-4,7,10,13-tetraoxa-17-azaoctanoic acid tert-butyl ester i-14

[0218]

[0219] The preparation method was the same as that of compound i-11. Using commercially available compound 2,2-dimethyl-4-oxo-3,7,10,13,16-pentaoxanonadecane-19-oic acid to replace compound 3-(3-(tert-butoxy)-3-oxopropoxy)propanoic acid, 72.5 mg of white oily solid was obtained.

[0220] Yield: 66.7%; LC-MS (ESI, m / z): calcd for C 39 H 61 N4O 10 S + , [M+H] + 777.4, found 777.0.

[0221] Example 31. 3-(3-(((S)-1-((2S,4R)-4-Hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)propanoic acid trifluoroacetate i-11-a

[0222]

[0223] The preparation method was the same as that of compound i-1-a. Using compound i-11 to replace compound i-1, the crude product was purified by preparative thin-layer chromatography (MeOH / DCM = 8%), and 35.6 mg of yellowish-green liquid was obtained.

[0224] Yield: 79.5%; LC-MS (ESI, m / z): calcd for C 29 H 41 N4O7S + , [M+H] + 589.3, found 589.0.

[0225] Example 32. 3-(2-(3-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethoxy)propanoic acid trifluoroacetate i-12-a

[0226]

[0227] The preparation method was the same as that of compound i-1-a, using compound i-12 instead of compound i-1, to obtain 38.8 mg of a white oily solid.

[0228] Yield: 74.4%; LC-MS (ESI, m / z): calcd for C 31 H 45 N4O8S + , [M+H] + 633.3, found 633.0.

[0229] Example 33. (S)-15-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-16,16-dimethyl-13-oxo-4,7,10-trioxa-14-azapentadecanoic acid trifluoroacetate i-13-a

[0230]

[0231] The preparation method was the same as that of compound i-1-a, using compound i-13 instead of compound i-1, to obtain 31.6 mg of a white oily solid.

[0232] Yield: 57.3%; LC-MS (ESI, m / z): calcd for C 33 H 49 N4O9S + , [M+H] + 677.3, found 677.0.

[0233] Example 34. (S)-18-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-19,19-dimethyl-16-oxo-4,7,10,13-tetraoxa-17-azaicosanoic acid trifluoroacetate i-14-a

[0234]

[0235] The preparation method was the same as that of compound i-1-a, using compound i-14 to replace compound i-1, and 37.9 mg of white oily solid was obtained.

[0236] Yield: 65.1%; LC-MS (ESI, m / z): calcd for C 35 H 53 N4O 10 S + , [M+H] + 721.3, found 721.0.

[0237] Example 35. tert-Butyl 2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oxy)ethoxy)ethoxy)acetate i-15

[0238]

[0239] The preparation method was the same as that of compound i-2, using 3-(4-hydroxy-1-oxoisoindolin-2-yl)piperidine-2,6-dione synthesized by the mature route to replace 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione. The crude product was purified by column chromatography (EtOAc / hexanes = 40%-60%), and 84.1 mg of white oily solid was obtained.

[0240] Yield: 39.6%; LC-MS (ESI, m / z): calcd for C 19 H 23 N2O8 + , [M- t Bu+2H] + 407.2, found 407.0.

[0241] Example 36. tert-Butyl 2-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oxy)ethoxy)ethoxy)ethoxy)acetate i-16

[0242]

[0243] The preparation method was the same as that of compound i-3, using 3-(4-hydroxy-1-oxoisoindolin-2-yl)piperidine-2,6-dione to replace 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione. The crude product was purified by column chromatography (EtOAc / hexanes = 50%-70%), and 146 mg of white oily solid was obtained.

[0244] Yield: 25.0%; LC-MS (ESI, m / z): calcd for C 21 H 27 N2O9 + , [M - t Bu + 2H] + 451.2, found 451.0.

[0245] Example 37. 2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oxy)ethoxy)ethoxy)acetic acid trifluoroacetate i-15-a

[0246]

[0247] Prepared in the same manner as compound i-1-a, using compound i-15 instead of compound i-1, to obtain 94.7 mg of a black liquid.

[0248] Yield: 100%; LC-MS (ESI, m / z): calcd for C 19 H 23 N2O8 + , [M + H] + 407.1, found 407.0.

[0249] Example 38. 2-(2-(2-(2-((2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oxy)ethoxy)ethoxy)acetic acid trifluoroacetate i-16-a

[0250]

[0251] Prepared in the same manner as compound i-1-a, using compound i-16 instead of compound i-1, to obtain 36 mg of a brown liquid.

[0252] Yield: 100%; LC-MS (ESI, m / z): calcd for C 21 H 27 N2O9 + , [M + H] + 451.2, found 451.0.

[0253] Example 39. tert-Butyl 2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,7-diazaspiro[3.5]nonane-7-carboxylate i-17-1

[0254]

[0255] Dissolve 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (1 g, 3.62 mmol, 1 eq) and DIPEA (1.89 mL, 10.86 mmol, 3 eq) in 8 mL of DMSO, and then add commercially available tert-butyl 2,7-diazaspiro[3.5]nonane-7-carboxylate hydrochloride (1.05 g, 3.98 mmol, 1.1 eq). Heat the reaction system to 120 °C and stir for 1 h. After monitoring the reaction of the raw materials to completion by TLC, cool the reaction system to room temperature, quench the reaction with 30 mL of water, and then extract with EtOAc (20 mL × 3). Combine the organic phases, wash with 100 mL of saturated brine, and concentrate under reduced pressure. The crude product was purified by column chromatography to obtain 1.23 g of a yellowish-green oily solid.

[0256] Yield: 70.4%; LC-MS (ESI, m / z): calcd for C 20 H 23 N4O4 + , [M - Boc + 2H] + 383.2, found 382.9.

[0257] Example 40. 5-(2,7-Diazaspiro[3.5]non-2-yl)-2-(2,6-dioxo-3-piperidinyl)isoindoline-1,3-dione i-17-2

[0258]

[0259] The preparation method is the same as that of Compound A. Using Compound i-17-1 (593.8 mg, 1.23 eq) instead of Compound A1, 514.3 mg of a yellow oily solid was obtained.

[0260] Yield: 100%; LC-MS (ESI, m / z): calcd for C 20 H 23 N4O4 + , [M + H] + 383.2, found 382.9.

[0261] Example 41. tert-Butyl 2-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,7-diazaspiro[3.5]non-7-yl)acetate i-17-3

[0262]

[0263] Dissolve i-17-2 (514.3 mg, 1.23 mmol, 1 eq) in 5 mL of DMF. At room temperature, successively add commercially available tert-butyl bromoacetate (286.7 mg, 1.47 mmol, 1.2 eq) and DIPEA (322 μL, 1.5 mmol, 1.5 eq), and stir at room temperature for 3 h. After monitoring the complete reaction of the starting materials by TLC, pour the reaction solution into 40 mL of saturated aqueous sodium bicarbonate. Extract the aqueous phase with EtOAc (25 mL × 3). Combine the organic phases, wash the organic phases with 150 mL of saturated brine, concentrate under reduced pressure, and purify the crude product by column chromatography (EtOAc / hexanes = 20% - 40%) to obtain 288 mg of a yellowish-green oily solid.

[0264] Yield: 47.2%; LC-MS (ESI, m / z): calcd for C 26 H 33 N4O6 + , [M + H] + 497.2, found 496.9.

[0265] Example 42. 2-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,7-diazaspiro[3.5]non-7-yl)acetic acid trifluoroacetate i-17-4

[0266]

[0267] The preparation method is the same as that of compound i-1-a, using compound i-17-3 instead of compound i-1, to obtain 297.5 mg of a yellow oily solid.

[0268] Yield: 92.5%; LC-MS (ESI, m / z): calcd for C 22 H 23 N4O6 - , [M - H] - 439.2, found 439.0.

[0269] Example 43. 1-((Benzyloxy)carbonyl)azetidin-3-yl)piperidine-4-carboxylic acid tert-butyl ester i-18-1

[0270]

[0271] Dissolve the commercially available compound tert-butyl piperidine-4-carboxylate (1.85 g, 10 mmol, 1 eq) and the commercially available compound benzyl 3-oxoazetidine-1-carboxylate (2.05 g, 10 mmol, 1 eq) in 20 mL of anhydrous MeOH. Dropwise add AcOH (114 μL, 2 mmol, 0.2 eq) to the reaction solution at room temperature, stir for 1 h, and then add sodium cyanoborohydride (2.52 g, 40 mmol, 4 eq) to the reaction solution in portions under an ice bath. Raise the reaction system to room temperature and continue the reaction for 8 h. After monitoring by TLC and LC-MS that the raw materials have completely reacted, concentrate the reaction solution under reduced pressure, adjust the pH to approximately 10 with saturated aqueous sodium bicarbonate, extract 3 times with DCM, and purify the crude product by column chromatography (EtOAc / hexanes = 0%-5%) to obtain 1.3 g of a colorless liquid.

[0272] Yield: 34.8%; LC-MS (ESI, m / z): calcd for C 21 H 31 N2O4 + , [M + H] + 375.2, found 375.0; 1 1H NMR (400 MHz, CDCl3) δ 7.30 - 7.26 (m, 3H), 7.26 - 7.18 (m, 2H), 5.00 (s, 2H), 3.97 (t, J = 6.7 Hz, 1H), 3.92 (dd, J = 8.8, 7.1 Hz, 2H), 3.80 (dd, J = 8.8, 5.5 Hz, 2H), 2.63 (m, 1H), 1.84 - 1.77 (m, 4H), 1.69 - 1.53 (m, 4H), 1.35 (s, 9H); 13 13C NMR (101 MHz, CDCl3) δ 174.17, 171.38, 156.51, 136.73, 128.55, 128.11, 128.08, 80.34, 66.72, 64.46, 54.46, 53.92, 49.43, 41.83, 30.72, 28.14, 27.86, 21.12, 19.21, 13.80.

[0273] Example 44. 1-(Azetidin-3-yl)piperidine-4-carboxylic acid tert-butyl ester i-18-2

[0274]

[0275] Dissolve i-18-1 (679.9 mg, 1.82 mmol, 1 eq) and 10% palladium on carbon (136 mg) in 10 mL of anhydrous MeOH. Bubble in hydrogen, then displace the hydrogen with nitrogen, repeat 3 times, and finally bubble in hydrogen again. Under a hydrogen atmosphere, react the reaction solution at 50 °C for 2 h. After monitoring by TLC that the raw materials have completely reacted, cool the reaction solution to room temperature, filter, and concentrate the obtained filtrate under reduced pressure to obtain 436.8 mg of a colorless liquid.

[0276] Yield: 100%; LC-MS (ESI, m / z): calcd for C 13 H 25 N2O2 + , [M+H] + 241.2, found 241.0.

[0277] Example 45. 1-(1-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)azetidin-3-yl)piperidine-4-carboxylic acid tert-butyl ester i-18-3

[0278]

[0279] The preparation method is the same as that of compound i-17-1, using compound i-18-2 instead of tert-butyl 2,7-diazaspiro[3.5]nonane-7-carboxylate hydrochloride, changing the reaction time to 12 h, and purifying the crude product by column chromatography (EtOAc / DCM = 0%-15%) to obtain 567.8 mg of a yellow oily solid.

[0280] Yield: 68%; LC-MS (ESI, m / z): calcd for C 26 H 33 N4O6 + , [M+H] + 497.2, found 496.9; 11H NMR (400 MHz, DMSO-D6) δ 11.08 (s, 1H), 7.64 (dd, J = 8.3 Hz, 1H), 6.78 (dd, J = 2.1 Hz, 1H), 6.64 (dd, J = 8.4, 2.1 Hz, 1H), 5.05 (dd, J = 12.9, 5.4 Hz, 1H), 4.08 (m, 2H), 3.80 (dd, J = 8.7, 5.2 Hz, 2H), 3.27 (p, J = 5.7 Hz, 1H), 2.91 - 2.83 (m, 1H), 2.73 (d, J = 10.8 Hz, 2H), 2.62 - 2.51 (m, 2H), 2.24 - 2.17 (m, 1H), 2.03 - 1.97 (m, 1H), 1.92 (m, 2H), 1.78 (m, 2H), 1.56 - 1.50 (m, 2H), 1.39 (s, 9H); 13 13C NMR (101 MHz, DMSO-D6) δ 174.27, 173.36, 170.66, 168.00, 167.70, 155.47, 134.34, 125.38, 117.36, 114.72, 105.00, 80.02, 55.86, 54.81, 49.22, 49.09, 41.57, 34.73, 31.50, 29.01, 28.24, 28.19, 23.03, 22.73。

[0281] Example 46. 1-(1-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)azetidin-3-yl)piperidine-4-carboxylic acid trifluoroacetate i-18-4

[0282]

[0283] Prepared in the same way as compound i-1-a, using compound i-18-3 instead of compound i-1, 339.8 mg of yellow oily solid was obtained.

[0284] Yield: 100%; LC-MS (ESI, m / z): calcd for C 22 H 25 N4O6 + , [M + H] + 441.2, found 440.8.

[0285] Example 47. 1'-Benzyl 4-(tert-butyl) [1,4'-bipiperidine]-1',4-dicarboxylate i-19-1

[0286]

[0287] The preparation method was the same as that of compound i-18-1. The commercially available benzyl 4-oxopiperidine-1-carboxylate was used instead of benzyl 3-oxoazetidine-1-carboxylate. The crude product was purified by column chromatography (EtOAc / hexanes = 2%-10%) to obtain 827 mg of a yellow oily solid.

[0288] Yield: 32.1%; LC-MS (ESI, m / z): calcd for C 23 H 35 N2O4 + , [M+H] + 403.3, found 403.0.

[0289] Example 48. tert-Butyl 1,4'-bipiperidine-4-carboxylate i-19-2

[0290]

[0291] Dissolve i-19-1 (827 mg, 2.05 mmol, 1 eq) in 20 mL of anhydrous ACN. Under an ice bath and in the dark, slowly add the commercially available trimethylsilyl iodide (1.23 g, 6.15 mmol, 3 eq). After the addition, evacuate and refill with nitrogen three times. Raise the reaction system to room temperature and stir in the dark for 1.5 h. After monitoring by TLC that the raw materials have completely reacted, quench the reaction with MeOH. Concentrate the reaction solution under reduced pressure. The crude product was purified by column chromatography to obtain 549 mg of a colorless liquid.

[0292] Yield: 99%; LC-MS (ESI, m / z): calcd for C 15 H 29 N2O2 + , [M+H] + 269.2, found 269.1.

[0293] Example 49. tert-Butyl 1'-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-[1,4'-bipiperidine]-4-carboxylate i-19-3

[0294]

[0295] The preparation method was the same as that of compound i-17-1. Use compound i-19-2 instead of tert-butyl 2,7-diazaspiro[3.5]nonane-7-carboxylate hydrochloride, and change the reaction time to 12 h. The crude product was purified by column chromatography (EtOAc / DCM = 0%-20%) to obtain 203.4 mg of a yellow oily solid.

[0296] Yield: 20.9%; LC-MS (ESI, m / z): calcd for C 24 H 29 N4O6 + ,[M - t Bu + 2H] + 469.3, found 469.1.

[0297] Example 50. 1'-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-[1,4'-bipiperidine]-4-carboxylic acid trifluoroacetate i-19-4

[0298]

[0299] Prepared in the same way as compound i-1-a, using compound i-19-3 instead of compound i-1, 181.6 mg of yellow oily solid was obtained.

[0300] Yield: 100%; LC-MS (ESI, m / z): calcd for C 24 H 29 N4O6 + ,[M + H] + 469.2, found 469.1.

[0301] Example 51. 2-(4-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)acetic acid tert-butyl ester i-20-1

[0302]

[0303] Prepared in the same way as compound i-17-1, using commercially available 2-(piperazin-1-yl)acetic acid tert-butyl ester hydrochloride instead of 2,7-diazaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester hydrochloride, and changing the reaction time to 40 h. The crude product was purified by trituration to obtain 823.4 mg of yellow oily solid.

[0304] Yield: 100%; LC-MS (ESI, m / z): calcd for C 23 H 29 N4O6 + ,[M + H] + 457.2, found 456.9; 11H NMR (400 MHz, DMSO-D6) δ 11.09 (s, 1H), 7.67 (dd, J = 8.6 Hz, 1H), 7.34 (dd, J = 2.4 Hz, 1H), 7.25 (dd, J = 8.7, 2.4 Hz, 1H), 5.07 (dd, J = 12.9, 5.4 Hz, 1H), 3.44 (t, J = 5.1 Hz, 4H), 3.18 (s, 2H), 2.93 - 2.83 (m, 1H), 2.64 (t, J = 5.2 Hz, 4H), 2.60 - 2.50 (m, 2H), 2.06 - 1.97 (m, 1H), 1.41 (s, 9H).

[0305] Example 52. 2-(4-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)acetic acid i-20-2

[0306]

[0307] The preparation method is the same as that of compound i-1-a. Using compound i-20-1 to replace compound i-1, 351.7 mg of yellow oily solid was obtained.

[0308] Yield: 100%; LC-MS (ESI, m / z): calcd for C 19 H 21 N4O6 + , [M + H] + 401.1, found 400.8.

[0309] Example 53. 1-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine-4-carboxylic acid tert-butyl ester i-21-1

[0310]

[0311] The preparation method is the same as that of compound i-17-1. Using commercially available piperidine-4-carboxylic acid tert-butyl ester to replace compound 2,7-diazaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester hydrochloride, changing the reaction temperature to 120 °C and the reaction time to 12 h. The crude product was purified by trituration to obtain 475.4 mg of yellow oily solid.

[0312] Yield: 59.9%; LC-MS (ESI, m / z): calcd for C 23 H 28 N3O6 + , [M + H] + 442.2, found 441.9; 11H NMR (400 MHz, DMSO-D6) δ 11.09 (s, 1H), 7.66 (dd, J = 8.6 Hz, 1H), 7.32 (dd, J = 2.4 Hz, 1H), 7.24 (dd, J = 8.7, 2.4 Hz, 1H), 5.07 (dd, J = 12.9, 5.4 Hz, 1H), 3.97 (m, 2H), 3.07 (m, 2H), 2.91 - 2.84 (m, 1H), 2.62 - 2.57 (m, 1H), 2.57 - 2.51 (m, 2H), 2.03 - 1.98 (m, 1H), 1.86 (dd, J = 13.0, 3.3 Hz, 2H), 1.56 (m, 2H), 1.40 (s, 9H).

[0313] Example 54. 1-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine-4-carboxylic acid i-21-2

[0314]

[0315] The preparation method was the same as that of compound i-1-a, using compound i-21-1 instead of compound i-1, to obtain 217.7 mg of a yellow oily solid.

[0316] Yield: 99.2%; LC-MS (ESI, m / z): calcd for C 19 H 20 N3O6 + , [M + H] + 386.1, found 385.9.

[0317] Example 55. tert-Butyl 4-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)benzoate i-22-1

[0318]

[0319] The preparation method was the same as that of compound i-17-1, using commercially available tert-butyl 4-(piperazin-1-yl)benzoate instead of tert-butyl 2,7-diazaspiro[3.5]nonane-7-carboxylate hydrochloride. The crude product was purified by column chromatography, and the reaction time was changed to 12 h to obtain 471 mg of a yellow oily solid.

[0320] Yield: 100%; LC-MS (ESI, m / z): calcd for C 28 H 31 N4O6 + , [M + H] + 519.2, found 518.8;1 1H NMR (400 MHz, DMSO-D6) δ 11.10 (s, 1H), 7.75 (dd, J = 4.9 Hz, 1H), 7.72 (dd, 2H), 7.38 (dd, J = 2.4 Hz, 1H), 7.28 (dd, J = 8.6, 2.4 Hz, 1H), 6.98 (m, 2H), 5.08 (dd, J = 12.9, 5.4 Hz, 1H), 3.57 (m, 8H), 2.92 - 2.84 (m, 1H), 2.66 - 2.52 (m, 2H), 2.04 - 1.99 (m, 1H), 1.51 (s, 9H).

[0321] Example 56. 4-(4-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)benzoic acid i-22-2

[0322]

[0323] The preparation method was the same as that of compound i-1-a, using compound i-22-1 instead of compound i-1, and 269 mg of yellow oily solid was obtained.

[0324] Yield: 100%; LC-MS (ESI, m / z): calcd for C 24 H 23 N4O6 + , [M + H] + 463.1, found 462.8.

[0325] Example 57. S-(2-(2-(2-((2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethoxy)acetamido)ethyl) 2-bromohexadecanoic acid thioester TM1

[0326]

[0327] The preparation method was the same as that of compound B, using compound i-1-a instead of compound B1 and using compound A instead of compound B2. The crude product was purified by preparative thin-layer chromatography (MeOH / DCM = 1%), and 14.7 mg of white oily solid was obtained.

[0328] Yield: 30.8%; LC-MS (ESI, m / z): calcd for C 35 H 51 BrN3O8S + , [M + H] + 752.3, found 752.3; 1H NMR (400MHz, CDCl3) δ8.10 (s, 1H), 7.71 (m, 1H), 7.66 (s, 1H), 7.49 (dd, J = 7.3Hz, 1H), 7.21(dd,J=8.3Hz,1H),5.20(m,1H),4.33(m,1H),4.30(t,J=1.9Hz,2H),4.09(s,2H) ,3.91(t,J=3.5Hz,2H),3.61(td,J=8.8,4.5Hz,2H),3.13(m,2H),2.90-2.79(m,3H), 2.20-2.14(m,1H),1.99-1.87(m,2H),1.45-1.40(m,2H),1.24(s,22H),0.86(s,3H).

[0329] Example 58. S-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethoxy)ethoxy)acetylamino)ethyl)2-bromohexadecanoic acid thioester TM2

[0330]

[0331] The preparation method is the same as compound B, using compound i-2-a instead of compound B1, and compound A instead of compound B2. The crude product is purified by preparative thin layer chromatography (MeOH / DCM=1%) to obtain 15.6 mg of white oily solid.

[0332] Yield: 28.1%; LC-MS (ESI, m / z): calculated for C 37 H 55 BrN3O9S + , [M+H] + 796.3, found796.3; 1H NMR (400MHz, CDCl3) δ8.30(s,1H),7.68(dd,J=8.5,7.3Hz,1H),7.47(dd,J=7.1Hz,1H),7.31(m,1H ),7.24(s,1H),4.93(m,1H),4.37(m,2H),4.32(m,1H),3.98(s,2H),3.97(t,J=4.5Hz,2H),3.83(td ,J=3.8,1.8Hz,2H),3.69(t,J=3.7Hz,2H),3.42(td,J=6.1,3.6Hz,2H),3.05(t,J=2.2Hz,2H),2.87 -2.73(m,3H),2.14-2.11(m,1H),2.01-1.94(m,2H),1.45-1.42(m,2H),1.24(s,22H),0.86(s,3H).

[0333] Example 59. S-(14-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-4-oxo-6,9,12-trioxa-3-azatetradecyl)2-bromohexadecanoic acid thioester TM3

[0334]

[0335] The preparation method is the same as compound B, using compound i-3-a instead of compound B1, and compound A instead of compound B2. The crude product is purified by preparative thin layer chromatography (MeOH / DCM=1%) to obtain 13.9 mg of white oily solid.

[0336] Yield: 23.7%; LC-MS (ESI, m / z): calculated for C 39 H 59 BrN3O 10 S + , [M+H] + 840.3, found840.3; 1H NMR (400MHz, CDCl3) δ8.34(s,1H),7.67(dd,J=7.9,7.9Hz,1H),7.46(dd,J=7.3Hz,1H),7.37(s,1H),7.24(s ,1H),4.94(m,1H),4.35(dd,J=4.7Hz,1H),4.33(t,J=5.6Hz,2H),3.96(s,2H),3.94(t,J=4.6Hz,2H),3.82( t,J=3.1Hz,2H),3.69(t,J=4.5Hz,2H),3.65(s,2H),3.47(s,2H),3.44(d,J=6.2Hz,2H),3.08(t,J=7.1Hz,2 H),2.88-2.72(m,3H),2.14-2.08(m,1H),2.04-1.94(m,2H),1.46-1.40(m,2H),1.24(s,22H),0.86(s,3H).

[0337] Example 60. S-(17-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-4-oxo-6,9,12,15-tetraoxa-3-azaheptadecanyl)2-bromohexadecanoic acid thioester TM4

[0338]

[0339] The preparation method is the same as compound B, using compound i-4-a instead of compound B1, and compound A instead of compound B2. The crude product is purified by preparative thin layer chromatography (MeOH / DCM=1%) to obtain 22.6 mg of white oily solid.

[0340] Yield: 36.6%; LC-MS (ESI, m / z): calculated for C 41 H 63 BrN3O 11 S + , [M+H] + 884.3, found884.3; 1H NMR (400MHz, DMSO-D6) δ11.07(s,1H),7.85(t,J=5.7Hz,1H),7.77(dd,J=7.8,7.8Hz,1H),7.50(dd,J=8.7Hz,1H) ,7.42(dd,J=7.4Hz,1H),5.05(dd,J=12.8,5.3Hz,1H),4.75(dd,J=7.0,7.0Hz,1H),4.30(t,J=4.3Hz,2H),3.82(s ,2H),3.77(m,2H),3.61(m,2H),3.50(m,10H),3.45(td,J=11.5Hz,2H),2.99(t,J=6.3Hz,2H),2.89-2.80(m,1H), 2.66-2.50(m,2H),2.00-1.91(m,2H),1.87-1.78(m,1H),1.36-1.29(m,2H),1.19(s,22H),0.80(d,J=6.5Hz,3H).

[0341] Example 61. (E)-N-(Cyanomethyl)-4-(2-(2-((2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethoxy)acetamido)-N-tetradecylbut-2-enamide TM5

[0342]

[0343] The preparation method is the same as compound B, using compound i-1-a instead of compound B1, and using compound B instead of compound B2. The crude product is purified by preparative thin layer chromatography (MeOH / DCM=4%) to obtain 18 mg of white oily solid.

[0344] Yield: 35.1%; LC-MS (ESI, m / z): calculated for C 37 H 54 N5O9 + , [M+H2O+H] + 712.4, found712.4; 1H NMR (400MHz, CDCl3) δ10.39(s,1H),7.73(m,1H),7.70(m,1H),7.49(m,1H),7. 19(d,J=8.5Hz,1H),6.96-6.86(m,1H),6.53(m,1H),4.85(dd,J=6.7,6.6Hz,1H ),4.30(s,2H),4.21(d,2H),4.03(s,2H),3.99(m,2H),3.88(t,J=10.5Hz,2H), 2.75(m,2H),2.54-1.80(m,4H),1.46-1.40(m,2H),1.23(s,22H),0.86(s,3H).

[0345] Example 62. (E)-N-(Cyanomethyl)-4-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethoxy)ethoxy)acetylamino)-N-tetradecylbut-2-enamide TM6

[0346]

[0347] The preparation method is the same as compound B, using compound i-2-a instead of compound B1, and using compound B instead of compound B2. The crude product is purified by preparative thin layer chromatography (MeOH / DCM=5%) to obtain 11.8 mg of white oily solid.

[0348] Yield: 21.6%; LC-MS (ESI, m / z): calculated for C 39 H 58 N5O 10 + , [M+H2O+H] + 756.4, found756.4; 1H NMR (400MHz, CDCl3) δ9.49 (s, 1H), 7.67 (dd, J=7.8, 7.8Hz, 1H), 7.51 (m, 1H), 7.46 (dd, J= 7.2Hz,1H),7.21(dd,J=8.5Hz,1H),6.89-6.71(m,1H),6.49(m,1H),4.93(dd,J=10.2Hz,1 H),4.32(s,2H),4.02(s,2H),3.93(t,J=4.2Hz,4H),3.80(t,J=7.8Hz,2H),3.71(s,4H), 2.80(t,J=14.5Hz,2H),2.33-1.92(m,4H),1.46-1.40(m,2H),1.23(s,22H),0.86(s,3H).

[0349] Example 63. (E)-N-(Cyanomethyl)-4-(2-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethoxy)ethoxy)ethoxy)acetamido)-N-tetradecylbut-2-enamide TM7

[0350]

[0351] The preparation method is the same as compound B, using compound i-3-a instead of compound B1, and using compound B instead of compound B2. The crude product is purified by preparative thin layer chromatography (MeOH / DCM=6%) to obtain 12 mg of white oily solid.

[0352] Yield: 20.8%; LC-MS (ESI, m / z): calculated for C 41 H 62 N5O 11 + , [M+H2O+H] + 800.4, found800.5; 11H NMR (400 MHz, CDCl3) δ 9.32 (s, 1H), 7.66 (dd, J = 7.9 Hz, 1H), 7.51 (m, 1H), 7.45 (dd, J = 7.1 Hz, 1H), 7.23 (dd, J = 8.5 Hz, 1H), 6.92 - 6.68 (m, 1H), 6.50 (m, 1H), 4.94 (s, 1H), 4.31 (s, 2H), 4.05 (s, 2H), 3.99 (s, 2H), 3.91 (s, 2H), 3.78 (s, 2H), 3.65 (s, 8H), 2.77 (t, J = 13.5 Hz, 2H), 2.33 - 1.91 (m, 4H), 1.46 - 1.40 (m, 2H), 1.23 (s, 22H), 0.86 (s, 3H).

[0353] Example 64. (E)-N-(4-((Cyanomethyl)(tetradecyl)amino)-4-oxobut-2-en-1-yl)-14-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-3,6,9,12-tetraoxatetradecanamide TM8

[0354]

[0355] The preparation method was the same as that of Compound B, using Compound i-4-a instead of Compound B1 and Compound B instead of Compound B2. The crude product was purified by preparative thin-layer chromatography (MeOH / DCM = 7%) to obtain 10 mg of a white oily solid.

[0356] Yield: 16.4%; LC-MS (ESI, m / z): calcd for C 43 H 66 N5O 12 + , [M + H2O + H] + 844.5, found 844.5; 1 1H NMR (400 MHz, CDCl3) δ 9.35 (s, 1H), 7.67 (m, 1H), 7.58 (m, 1H), 7.45 (dd, J = 7.2 Hz, 1H), 7.23 (s, 1H), 6.94 - 6.75 (m, 1H), 6.47 (m, 1H) 4.93 (m, 1H), 4.32 (s, 2H), 4.06 (t, J = 5.3 Hz, 2H), 3.99 (s, 2H), 3.92 (m, 2H), 3.78 (m, 2H), 3.64 (s, 12H), 2.76 (m, 2H), 2.03 - 1.90 (m, 4H), 1.46 - 1.40 (m, 2H), 1.23 (s, 22H), 0.86 (s, 3H).

[0357] Example 65. S-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)acetamido)ethyl) 2-bromohexadecanoate TM9

[0358]

[0359] The preparation method is the same as that of compound TM1, using compound i-5-a instead of compound i-1-a. The crude product was purified by preparative thin-layer chromatography (MeOH / DCM = 1%), and 14.4 mg of white oily solid was obtained.

[0360] Yield: 27.5%; LC-MS (ESI, m / z): calcd for C 35 H 51 BrN3O8S + , [M+H] + 752.3, found 751.8; 1 H NMR (400 MHz, CDCl3) δ 8.15 (s, 1H), 7.81 (dd, J = 8.2 Hz, 1H), 7.38 (dd, J = 2.3 Hz, 1H), 7.27 (m, 1H), 6.90 (t, J = 5.8 Hz, 1H), 4.95 (dd, J = 12.5, 5.4 Hz, 1H), 4.34 (dd, J = 8.1, 6.3 Hz, 1H), 4.28 (m, 2H), 4.07 (s, 2H), 3.93 (m, 2H), 3.51 (q, J = 6.5 Hz, 2H), 3.10 (m, 2H), 2.92 - 2.72 (m, 3H), 2.18 - 2.12 (m, 1H), 2.03 - 1.94 (m, 2H), 1.47 - 1.40 (m, 2H), 1.24 (s, 22H), 0.86 (s, 3H); 13 C NMR (101 MHz, CDCl3) δ 196.22, 170.92, 169.64, 168.09, 167.09, 166.92, 163.93, 134.47, 125.79, 124.16, 121.19, 108.80, 77.31, 70.76, 69.81, 68.16, 53.98, 49.44, 38.58, 35.21, 32.01, 31.51, 29.78, 29.74, 29.68, 29.58, 29.45, 29.39, 29.24, 28.92, 27.28, 22.78, 22.76, 14.23, 1.11.

[0361] Example 66. S-(2-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)acetamido)ethyl) 2-bromohexadecanoate TM10

[0362]

[0363] The preparation method was the same as that of compound TM2, using compound i-6-a instead of compound i-2-a. The crude product was purified by preparative thin-layer chromatography (MeOH / DCM = 1%), and 28 mg of white oily solid was obtained.

[0364] Yield: 50.4%; LC-MS (ESI, m / z): calcd for C 37 H 55 BrN3O9S + ,[M + H] + 796.3, found 795.9; 1 1H NMR (400 MHz, CDCl3) δ 8.15 (s, 1H), 7.78 (d, J = 8.2 Hz, 1H), 7.37 (d, J = 2.3 Hz, 1H), 7.23 (dd, J = 8.3, 2.3 Hz, 1H), 7.19 (m, 1H), 4.95 (dd, J = 12.5, 5.4 Hz, 1H), 4.34 (dd, J = 8.1, 6.2 Hz, 1H), 4.30 (t, J = 3.8 Hz, 2H), 3.99 (s, 2H), 3.92 (m, 2H), 3.72 (ddd, J = 14.5, 3.6, 1.7 Hz, 4H), 3.44 (m, 2H), 3.05 (td, J = 6.8, 4.7 Hz, 2H), 2.91 - 2.71 (m, 3H), 2.16 - 2.11 (m, 1H), 2.05 - 1.94 (m, 2H), 1.51 - 1.37 (m, 2H), 1.23 (s, 22H), 0.85 (t, J = 7.0 Hz, 3H); 13 13C NMR (101 MHz, CDCl3) δ 196.12, 170.99, 170.27, 168.15, 167.15, 166.98, 164.15, 134.41, 125.71, 123.94, 121.03, 109.07, 77.32, 70.97, 70.72, 70.44, 69.53, 68.44, 54.08, 53.54, 49.40, 38.32, 35.23, 32.01, 31.51, 29.77, 29.74, 29.68, 29.59, 29.45, 29.40, 29.17, 28.92, 27.28, 22.78, 22.76, 14.23.

[0365] Example 67. S-(14-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-4-oxo-6,9,12-trioxa-3-azatetradecyl) 2-bromohexadecanoate TM11

[0366]

[0367] The preparation method was the same as that of compound TM3, using compound i-7-a instead of compound i-3-a. The crude product was purified by preparative thin-layer chromatography (MeOH / DCM = 1%), and 32.2 mg of a white oily solid was obtained.

[0368] Yield: 54.8%; LC-MS (ESI, m / z): calcd for C 39 H 59 BrN3O 10 S + , [M+H] + 840.3, found 839.8; 1 1H NMR (400 MHz, CDCl3) δ 8.15 (s, 1H), 7.78 (dd, J = 8.3 Hz, 1H), 7.37 (dd, J = 2.3 Hz, 1H), 7.31 (t, J = 5.5 Hz, 1H), 7.22 (dd, J = 8.3, 2.3 Hz, 1H), 4.96 (dd, J = 12.4, 5.3 Hz, 1H), 4.36 (dd, J = 8.1, 6.2 Hz, 1H), 4.26 (t, J = 4.6 Hz, 2H), 3.99 (s, 2H), 3.91 (m, 2H), 3.74 (ddd, 4H), 3.78 - 3.75 (m, 2H), 3.72 - 3.70 (m, 2H), 3.48 (q, J = 6.7 Hz, 2H), 3.10 (td, J = 6.8, 2.5 Hz, 2H), 2.92 - 2.73 (m, 3H), 2.16 - 2.12 (m, 1H), 2.05 - 1.95 (m, 2H), 1.52 - 1.38 (m, 2H), 1.24 (s, 22H), 0.88 (d, J = 6.7 Hz, 3H); 1313C NMR (101 MHz, CDCl3) δ 196.06, 170.96, 170.51, 168.15, 167.19, 167.01, 164.31, 134.36, 125.64, 123.80, 121.04, 109.13, 77.32, 71.08, 71.01, 70.66, 70.43, 69.49, 68.52, 54.07, 49.39, 38.32, 35.23, 32.01, 31.51, 29.77, 29.74, 29.68, 29.59, 29.45, 29.40, 29.20, 28.93, 27.29, 22.78, 14.23。

[0369] Example 68. S-(17-((2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-4-oxo-6,9,12,15-tetraoxa-3-azapentadecyl) 2-bromohexadecanoate TM12

[0370]

[0371] The preparation method was the same as that of compound TM4, using compound i-8-a instead of compound i-4-a. The crude product was purified by preparative thin-layer chromatography (MeOH / DCM = 1%), and 42.9 mg of a white oily solid was obtained.

[0372] Yield: 69.4%; LC-MS (ESI, m / z): calcd for C 41 H 63 BrN3O 11 S + , [M+H] + 884.3, found 883.9; 11H NMR (400 MHz, CDCl3) δ 8.24 (s, 1H), 7.78 (dd, J = 8.3 Hz, 1H), 7.36 (dd, J = 2.3 Hz, 1H), 7.24 (s, 1H), 7.22 (dd, J = 2.4 Hz, 1H), 4.95 (dd, J = 12.5, 5.4 Hz, 1H), 4.36 (dd, J = 8.2, 6.1 Hz, 1H), 4.27 (m, 2H), 3.97 (s, 2H), 3.91 (m, 2H), 3.74 (dd, J = 6.2, 3.1 Hz, 2H), 3.68 (s, 4H), 3.66 (s, 4H), 3.46 (q, J = 6.4 Hz, 2H), 3.09 (td, J = 6.8, 3.3 Hz, 2H), 2.90 - 2.73 (m, 3H), 2.17 - 2.13 (m, 1H), 2.06 - 1.95 (m, 2H), 1.51 - 1.37 (m, 2H), 1.24 (s, 22H), 0.86 (d, J = 7.0 Hz, 3H); 13 13C NMR (101 MHz, CDCl3) δ 196.00, 171.03, 170.77, 168.20, 167.20, 167.03, 164.34, 134.34, 125.64, 123.76, 120.97, 109.19, 77.32, 71.07, 71.04, 70.64, 70.33, 70.29, 69.45, 68.52, 54.08, 49.38, 38.35, 35.22, 32.01, 31.50, 29.77, 29.74, 29.68, 29.59, 29.45, 29.40, 29.18, 28.94, 27.29, 22.78, 14.23。

[0373] Example 69. S-(2-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)heptanamido)ethyl) 2-bromopalmitate TM13

[0374]

[0375] The preparation method was the same as that of compound TM1, using compound i-9-a instead of compound i-1-a. The crude product was purified by preparative thin layer chromatography (MeOH / DCM = 2.5%) to obtain 28.5 mg of a white oily solid.

[0376] Yield: 54.0%; 11H NMR (400 MHz, CDCl3) δ 8.31 (s, 1H), 7.66 (dd, J = 8.5, 7.3 Hz, 1H), 7.43 (dd, J = 6.6 Hz, 1H), 7.20 (dd, J = 7.9 Hz, 1H), 6.12 (t, J = 6.0 Hz, 1H), 4.97 (dd, J = 10.8, 5.4 Hz, 1H), 4.34 (dd, J = 8.1, 6.2 Hz, 1H), 4.16 (m, 2H), 3.42 (m, 2H), 3.06 (qd, J = 6.8, 3.1 Hz, 2H), 2.85 - 2.75 (m, 3H), 2.18 (t, J = 7.4 Hz, 2H), 2.00 - 1.93 (m, 1H), 1.84 (t, J = 6.5 Hz, 2H), 1.66 (p, J = 7.3 Hz, 4H), 1.57 - 1.51 (m, 2H), 1.43 - 1.39 (m, 2H), 1.23 (s, 24H), 0.86 (m, 3H); 13 13C NMR (101 MHz, CDCl3) δ 196.81, 173.48, 171.07, 168.37, 167.14, 165.98, 156.74, 136.69, 133.84, 118.98, 117.15, 115.85, 77.32, 69.30, 53.98, 49.20, 39.05, 38.70, 35.99, 35.19, 32.01, 31.50, 29.77, 29.74, 29.67, 29.58, 29.45, 29.39, 29.32, 28.92, 28.43, 28.25, 27.30, 25.23, 25.03, 22.78, 22.76, 14.23。

[0377] Example 70. S-(2-(9-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)nonanamido)ethyl) 2-bromopalmitate TM14

[0378]

[0379] The preparation method was the same as that of compound TM1, using compound i-10-a instead of compound i-1-a. The crude product was purified by preparative thin-layer chromatography (MeOH / DCM = 2%) to obtain 2.7 mg of a white oily solid.

[0380] Yield: 10.5%; 11H NMR (400 MHz, CDCl3) δ 8.31 (s, 1H), 7.66 (dd, J = 8.5, 7.3 Hz, 1H), 7.43 (dd, J = 6.6 Hz, 1H), 7.20 (dd, J = 7.9 Hz, 1H), 6.12 (t, J = 6.0 Hz, 1H), 4.97 (ddd, J = 12.5, 5.4, 1.6 Hz, 1H), 4.34 (dd, J = 8.1, 6.2 Hz, 1H), 4.16 (t, J = 1.4 Hz, 2H), 3.42 (m, 2H), 3.05 (td, J = 6.9, 3.7 Hz, 2H), 2.86 - 2.76 (m, 3H), 2.18 (t, J = 7.4 Hz, 2H), 2.02 - 1.96 (m, 1H), 1.86 (m, 2H), 1.66 (p, J = 7.3 Hz, 4H), 1.54 (m, 2H), 1.48 - 1.35 (m, 4H), 1.23 (s, 24H), 0.86 (m, 3H).

[0381] Example 71. S-(2-(3-(3-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)propanamido)ethyl) 2-bromohexadecanoate TM15

[0382]

[0383] The preparation method was the same as that of compound TM1, using compound i-11-a instead of compound i-1-a. The crude product was purified by preparative thin-layer chromatography (MeOH / DCM = 4% - 5%) to obtain 10.6 mg of a white oily solid.

[0384] Yield: 39.8%; LC-MS (ESI, m / z): calcd for C 47 H 76 BrN5O7S 2+ , [M + 2H] 2+ 482.7, found 482.6; 11H NMR (400 MHz, CDCl3) δ 8.66 (s, 1H), 7.40 - 7.34 (m, 5H), 7.21 (dd, J = 11.9, 5.8 Hz, 1H), 7.14 (t, J = 9.5 Hz, 1H), 5.13 - 5.08 (m, 1H), 4.72 (m, 1H), 4.64 (dd, J = 9.0, 5.6 Hz, 1H), 4.50 (s, 1H), 4.36 (dd, J = 8.2, 6.1 Hz, 1H), 4.07 (dt, J = 11.7 Hz, 1H), 3.77 - 3.73 (m, 1H), 3.69 (m, 4H), 3.65 - 3.61 (m, 1H), 3.44 - 3.34 (m, 2H), 3.07 (m, 2H), 2.52 (s, 3H), 2.47 (m, 4H), 2.43 - 2.37 (m, 2H), 2.05 - 1.98 (m, 2H), 1.47 (d, J = 7.0 Hz, 3H), 1.24 (s, 24H), 1.04 (s, 9H), 0.86 (s, 3H); 13 13C NMR (101 MHz, CDCl3) δ 197.49, 171.92, 171.79, 170.02, 150.41, 148.56, 143.29, 131.66, 130.95, 129.64, 126.47, 77.31, 70.19, 67.39, 66.71, 58.71, 57.41, 57.06, 54.02, 53.93, 53.52, 48.86, 38.88, 37.51, 36.53, 36.19, 35.63, 35.24, 32.01, 31.59, 30.21, 29.79, 29.74, 29.68, 29.59, 29.45, 29.40, 29.12, 28.94, 27.31, 26.56, 22.78, 22.37, 16.20, 14.23.

[0385] Example 72. S - ((S) - 15 - ((2S,4R) - 4 - hydroxy - 2 - ((((S) - 1 - (4 - (4 - methylthiazol - 5 - yl)phenyl)ethyl)carbamoyl)pyrrolidine - 1 - carbonyl) - 16,16 - dimethyl - 4,13 - dioxo - 7,10 - dioxo - 3,14 - diazaheptadecyl)) 2 - bromohexadecanoate TM16

[0386]

[0387] The preparation method was the same as that of compound TM1, using compound i - 12 - a instead of compound i - 1 - a. The crude product was purified by preparative thin - layer chromatography (MeOH / DCM = 4% - 5%) to obtain 12.2 mg of a white oily solid.

[0388] Yield: 25.4%; LC-MS (ESI, m / z): calcd for C 49 H 80 BrN5O8S2 2+ , [M + 2H] 2+ 504.7, found 504.6; 1 1H NMR (400 MHz, CDCl3) δ 8.66 (s, 1H), 7.43 (m, 1H), 7.37 (dd, J = 6.4 Hz, 4H), 7.05 (m, 1H), 7.01 (d, J = 5.3 Hz, 1H), 5.08 (p, J = 7.3 Hz, 1H), 4.71 (t, J = 7.3 Hz, 1H), 4.56 (d, J = 8.6 Hz, 1H), 4.48 (s, 1H), 4.35 (t, J = 6.1 Hz, 1H), 4.12 (m, 1H), 3.78 - 3.75 (m, 1H), 3.70 (m, 4H), 3.62 (m, 4H), 3.59 - 3.57 (m, 1H), 3.44 - 3.35 (m, 2H), 3.06 (m, 2H), 2.52 (s, 3H), 2.49 (m, 4H), 2.45 - 2.40 (m, 2H), 2.00 - 1.93 (m, 2H), 1.47 (d, J = 6.9 Hz, 3H), 1.24 (s, 24H), 1.04 (s, 9H), 0.86 (s, 3H); 13 13C NMR (101 MHz, CDCl3) δ 196.91, 172.16, 172.10, 171.87, 169.97, 150.39, 148.56, 143.29, 131.68, 130.93, 129.64, 126.50, 77.31, 70.42, 70.17, 67.31, 67.06, 58.56, 57.85, 56.91, 54.12, 48.89, 38.91, 36.88, 36.54, 35.96, 35.20, 32.01, 30.21, 29.79, 29.74, 29.68, 29.59, 29.45, 29.40, 29.25, 28.93, 27.31, 26.61, 22.79, 22.36, 16.20, 14.23。

[0389] Example 73. S - ((S) - 18 - ((2S,4R) - 4 - hydroxy - 2 - ((((S) - 1 - (4 - (4 - methylthiazol - 5 - yl)phenyl)ethyl)carbamoyl)pyrrolidine - 1 - carbonyl) - 19,19 - dimethyl - 4,16 - dioxo - 7,10,13 - trioxa - 3,17 - docosanyl) 2 - bromohexadecanoate TM17

[0390]

[0391] The preparation method is the same as that of compound TM1, using compound i-13-a instead of compound i-1-a. The crude product was purified by preparative thin-layer chromatography (MeOH / DCM = 4%-5%) to obtain 10.9 mg of a white oily solid.

[0392] Yield: 37.8%; LC-MS (ESI, m / z): calcd for C 51 H 84 BrN5O9S2 2+ , [M + 2H] 2+ 526.7, found 526.6; 1 H NMR (400 MHz, CDCl3) δ 8.66 (s, 1H), 7.45 (d, J = 7.8 Hz, 1H), 7.37 (dd, J = 6.6 Hz, 4H), 6.96 (d, J = 7.2 Hz, 1H), 6.91 (m, 1H), 5.09 - 5.05 (m, 1H), 4.71 (t, J = 8.1 Hz, 1H), 4.53 (d, J = 8.5 Hz, 1H), 4.48 (s, 1H), 4.35 (dd, J = 8.2, 6.1 Hz, 1H), 4.12 (dt, J = 11.4 Hz, 1H), 3.76 - 3.74 (m, 1H), 3.70 (t, J = 5.8 Hz, 4H), 3.63 (d, J = 7.4 Hz, 8H), 3.59 - 3.56 (m, 1H), 3.44 - 3.39 (m, 2H), 3.06 (m, 2H), 2.52 (s, 3H), 2.49 (m, 4H), 2.45 (s, 2H), 2.01 - 1.96 (m, 2H), 1.46 (d, J = 7.0 Hz, 3H), 1.24 (s, 24H), 1.03 (s, 9H), 0.86 (s, 3H); 13CNMR (101 MHz, CDCl3) δ 196.42, 172.10, 172.06, 171.99, 169.89, 150.39, 148.56, 143.30, 131.68, 130.93, 129.64, 126.52, 77.32, 70.57, 70.51, 70.42, 70.22, 70.13, 67.25, 67.17, 58.47, 57.88, 56.83, 54.11, 48.91, 38.86, 36.89, 36.67, 35.72, 35.21, 35.09, 32.01, 31.59, 30.21, 29.79, 29.74, 29.68, 29.59, 29.45, 29.40, 29.34, 28.94, 27.31, 26.60, 22.78, 22.35, 16.20, 14.23。

[0393] Example 74. S-((S)-21-((2S,4R)-4-Hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-22,22-dimethyl-4,19-dioxo-7,10,13,16-tetraoxa-3,20-diazatriacontan-1-yl) 2-bromopalmitate TM18

[0394]

[0395] The preparation method is the same as that of compound TM1, using compound i-14-a instead of compound i-1-a. The crude product was purified by preparative thin layer chromatography (MeOH / DCM = 4%-5%) to obtain 23.3 mg of a white oily solid.

[0396] Yield: 45.3%; LC-MS (ESI, m / z): calcd for C 53 H 88 BrN5O 10 S2 + , [M + 2H] 2+ 548.7, found 548.6; 11H NMR (400 MHz, CDCl3) δ 8.66 (s, 1H), 7.50 (d, J = 7.8 Hz, 1H), 7.37 (dd, J = 5.7 Hz, 4H), 7.05 (d, J = 9.9 Hz, 1H), 6.98 (t, J = 6.5 Hz, 1H), 5.07 (p, J = 7.1 Hz, 1H), 4.71 (t, J = 8.0 Hz, 1H), 4.52 (d, J = 8.5 Hz, 1H), 4.47 (s, 1H), 4.34 (m, 1H), 4.09 (d, J = 11.4 Hz, 1H), 3.75 - 3.72 (m, 1H), 3.70 (td, J = 6.2, 3.5 Hz, 4H), 3.61 (d, J = 3.1 Hz, 12H), 3.58 - 3.54 (m, 1H), 3.44 - 3.38 (m, 2H), 3.05 (m, 2H), 2.51 (s, 3H), 2.47 (t, J = 11.1 Hz, 4H), 2.44 (d, J = 5.7 Hz, 2H), 2.02 - 1.83 (m, 2H), 1.46 (d, J = 7.0 Hz, 3H), 1.23 (s, 24H), 1.03 (s, 9H), 0.85 (s, 3H); 13 13C NMR (101 MHz, CDCl3) δ 196.31, 172.15, 172.08, 171.94, 169.98, 150.42, 148.53, 143.35, 131.69, 130.88, 129.62, 126.52, 77.33, 70.55, 70.49, 70.44, 70.36, 70.24, 70.08, 67.24, 67.18, 58.51, 57.87, 56.80, 54.11, 48.89, 38.82, 36.86, 36.72, 35.76, 35.21, 35.14, 32.00, 29.77, 29.73, 29.68, 29.59, 29.44, 29.40, 29.35, 28.93, 27.30, 26.59, 22.78, 22.34, 16.19, 14.22。

[0397] Example 75. S-(2-(2-(2-((2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oxy)ethoxy)ethoxy)acetamido)ethyl) 2-bromohexadecanoate TM19

[0398]

[0399] The preparation method was the same as that of compound TM1, using compound i-15-a instead of compound i-1-a. The crude product was purified by preparative thin-layer chromatography to obtain 24 mg of a white oily solid.

[0400] Yield: 43.9%; LC-MS (ESI, m / z): calcd for C 37 H 57 BrN3O8S + , [M + H] + 782.3, found 781.9; 1 H NMR (400 MHz, CDCl3) δ 8.30 (s, 1H), 7.50 (dd, J = 7.5 Hz, 1H), 7.44 (dd, J = 7.8, 7.8 Hz, 1H), 7.31 (s, 1H), 7.06 (dd, J = 7.9 Hz, 1H), 5.21 (m, 1H), 4.45 (d, J = 16.8 Hz, 1H), 4.34 (ddd, J = 11.8, 6.0, 3.3 Hz, 2H), 4.28 (t, 2H), 4.05 (s, 2H), 3.89 (m, 2H), 3.72 (td, J = 2.3 Hz, 4H), 3.41 (q, J = 7.3 Hz, 2H), 3.03 (td, J = 6.7, 1.9 Hz, 2H), 2.93 - 2.79 (m, 2H), 2.49 - 2.38 (m, 1H), 2.23 - 2.17 (m, 1H), 2.05 - 1.93 (m, 2H), 1.50 - 1.35 (m, 2H), 1.25 (s, 22H), 0.88 (d, J = 6.5 Hz, 3H); 13 C NMR (101 MHz, CDCl3) δ 196.17, 171.30, 170.79, 169.69, 169.44, 153.76, 133.22, 130.10, 130.04, 116.67, 114.36, 77.31, 70.94, 70.84, 70.33, 69.70, 67.73, 54.07, 51.98, 45.25, 38.42, 35.20, 32.01, 31.65, 29.77, 29.74, 29.68, 29.59, 29.45, 29.40, 29.05, 28.93, 27.29, 23.45, 22.79, 14.23。

[0401] Example 76. S-(14-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oxy)-4-oxo-6,9,12-trioxa-3-azatetradecyl) 2-bromohexadecanoate TM20

[0402]

[0403] The preparation method was the same as that of compound TM1, using compound i-16-a instead of compound i-1-a. The crude product was purified by preparative thin-layer chromatography to obtain 42.4 mg of a white oily solid.

[0404] Yield: 73.4%; LC-MS (ESI, m / z): calcd for C 39 H 62 BrN3O9S 2+ , [M + 2H] 2+ 413.7, found 413.4; 1 H NMR (400 MHz, CDCl3) δ 8.33 (s, 1H), 7.49 (dd, J = 6.7 Hz, 1H), 7.43 (t, J = 7.7 Hz, 1H), 7.36 (s, 1H), 7.05 (dd, J = 9.0 Hz, 1H), 5.20 (dd, J = 13.3, 5.1 Hz, 1H), 4.44 (d, J = 16.7 Hz, 1H), 4.37 - 4.30 (m, 2H), 4.24 (m, 2H), 4.00 (s, 2H), 3.87 (m, 2H), 3.76 - 3.73 (m, 2H), 3.66 (s, 4H), 3.45 (t, J = 6.7 Hz, 2H), 3.10 - 3.04 (m, 2H), 2.91 - 2.76 (m, 2H), 2.45 - 2.34 (m, 1H), 2.23 - 2.17 (m, 1H), 2.05 - 1.91 (m, 2H), 1.51 - 1.36 (m, 2H), 1.25 (s, 22H), 0.86 (d, J = 7.0 Hz, 3H); 13 C NMR (101 MHz, CDCl3) δ 196.09, 171.34, 170.93, 169.71, 169.47, 153.88, 133.14, 130.00, 116.52, 114.34, 77.32, 71.11, 70.90, 70.65, 70.42, 70.29, 69.71, 67.82, 54.08, 51.95, 45.22, 38.40, 35.21, 32.01, 31.65, 29.77, 29.74, 29.68, 29.59, 29.45, 29.40, 29.14, 28.93, 27.29, 23.53, 22.79, 14.23.

[0405] Example 77. S-(2-(2-(2-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,7-diazaspiro[3.5]nonan-7-yl)acetamido)ethyl) 2-bromohexadecanoate TM21

[0406]

[0407] The preparation method was the same as that of compound TM1, using compound i-17-4 instead of compound i-1-a. The crude product was purified by preparative thin-layer chromatography (MeOH / DCM = 5%), and 10.7 mg of a yellowish-green oily solid was obtained.

[0408] Yield: 39.8%; LC-MS (ESI, m / z): calcd for C 40 H 59 BrN5O6S + , [M+H] + 816.3, found 816.1; 1 H NMR (400 MHz, CDCl3) δ 8.42 (s, 1H), 7.57 (dd, J = 8.3, 3.1 Hz, 1H), 7.41 (m, 1H), 7.04 (s, 1H), 6.82 (dd, J = 8.4, 2.2 Hz, 1H), 4.92 (dd, J = 12.1, 5.3 Hz, 1H), 4.37 (dd, J = 8.0, 6.2 Hz, 1H), 3.59 (s, 2H), 3.50 (q, J = 6.4 Hz, 2H), 3.31 (d, J = 6.4 Hz, 2H), 3.17 - 3.08 (m, 2H), 3.01 (s, 2H), 2.87 - 2.74 (m, 3H), 2.58 (t, J = 8.6 Hz, 2H), 2.44 (m, 2H), 2.23 - 2.15 (m, 1H), 2.13 - 1.99 (m, 4H), 1.99 - 1.90 (m, 2H), 1.46 - 1.43 (m, 2H), 1.24 (s, 22H), 0.86 (s, 3H); 13 C NMR (101 MHz, CDCl3) δ 196.35, 170.66, 125.68, 106.50, 77.32, 61.58, 54.07, 49.62, 49.17, 38.54, 37.35, 35.25, 32.01, 31.53, 30.26, 29.78, 29.74, 29.58, 29.45, 29.40, 28.94, 27.31, 22.78, 14.23.

[0409] Example 78. S-(2-(1-(1-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)azetidin-3-yl)piperidine-4-carboxamido)ethyl) 2-bromohexadecanoate TM22

[0410]

[0411] At 0 °C, compound i-18-4 (32.35 mg, 0.0734 mmol, 1 eq), A (31.53 mg, 0.0734 mmol, 1 eq), PyBop (38.17 mg, 0.0734 mmol, 1 eq) and HOBt (11.23 mg, 0.0881 mmol, 1.2 eq) were dissolved in 1.2 mL of DMF. DIPEA (33.17 μL, 0.2202 mmol, 3 eq) was slowly added dropwise under an ice bath. The reaction system was warmed to room temperature and stirred for 8 h. After TLC and LC-MS monitored that the raw materials had completely reacted, the reaction solution was diluted with 12 mL of water and extracted with DCM (6 mL × 3). The organic phases were combined, washed with saturated brine, concentrated under reduced pressure, and the crude product was purified by preparative thin-layer chromatography (MeOH / DCM = 2%), to obtain 20 mg of a yellow-green oily solid.

[0412] Yield: 33.4%; LC-MS (ESI, m / z): calcd for C 40 H 59 BrN5O6S + , [M+H] + 816.3, found 816.1; 1 H NMR (400 MHz, CDCl3) δ 8.12 (s, 1H), 7.63 (dd, J = 8.2 Hz, 1H), 6.77 (dd, J = 2.2 Hz, 1H), 6.66 (t, J = 5.6 Hz, 1H), 6.51 (dd, J = 8.3, 2.2 Hz, 1H), 4.91 (dd, J = 12.3, 5.4 Hz, 1H), 4.28 (dd, J = 13.3, 4.6 Hz, 1H), 4.08 (t, J = 7.6 Hz, 2H), 3.89 - 3.83 (m, 2H), 3.48 - 3.43 (m, 2H), 3.34 (m, 1H), 3.05 (t, J = 6.4 Hz, 2H), 2.90 - 2.86 (m, 2H), 2.86 - 2.66 (m, 3H), 2.60 - 2.50 (m, 1H), 2.16 - 2.10 (m, 1H), 2.09 - 1.99 (m, 2H), 1.99 - 1.96 (m, 2H), 1.96 - 1.91 (m, 2H), 1.86 - 1.78 (m, 2H), 1.46 - 1.39 (m, 2H), 1.24 (s, 22H), 0.85 (d, J = 7.1 Hz, 3H); 13CNMR(101MHz,CDCl3)δ201.95,171.11,169.18,168.39,167.93,167.52,154.99,134.32,125.37,118.17,114.27,105.17,55.55,54.86,52.11,50.09,49.41,49.14,40.16,36.01,32.01,31.53,29.78,29.75,29.70,29.62,29.46,28.89,28.37,27.89,27.28,22.85,22.79,14.23。

[0413] Example 79. S-(2-(1'-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-[1,4'-bipiperidine]-4-carboxamido)ethyl) 2-bromohexadecanoic acid thioester TM23

[0414]

[0415] The preparation method is the same as that of compound TM1, using compound i-19-4 instead of compound i-1-a. The crude product was purified by preparative thin-layer chromatography (MeOH / DCM = 5%), and 6.9 mg of yellowish-green oily solid was obtained.

[0416] Yield: 17.6%; LC-MS (ESI, m / z): calcd for C 42 H 63 BrN5O6S + , [M+H] + 844.3, found 844.3; 1 H NMR(400MHz,CDCl3)δ8.01(s,1H),7.66(dd,J = 8.5Hz,1H),7.26(dd,J = 2.1Hz,1H),7.03(dd,J = 8.7,2.3Hz,1H),5.87(s,1H),4.93(dd,J = 12.2,5.3Hz,1H),4.36(dd,J = 8.0,6.3Hz,1H),3.98(d,J = 12.4Hz,2H),3.49(s,2H),3.48 - 3.44(m,2H),3.01 - 2.98(m,2H),2.84 - 2.70(m,3H),2.61(s,1H),2.28(s,4H),2.11(d,J = 6.9Hz,2H),2.05 - 1.92(m,8H),1.92 - 1.87(m,2H),1.43 - 1.41(m,2H),1.24(s,22H),0.87(t,3H); 13CNMR (101 MHz, CDCl3) δ 171.01, 125.57, 118.00, 77.31, 53.86, 49.20, 47.61, 39.27, 32.02, 31.54, 29.79, 29.75, 29.59, 29.46, 29.39, 28.93, 27.36, 22.79, 14.23。

[0417] Example 80. S-(2-(2-(4-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)acetamido)ethyl) 2-bromopalmitate TM24

[0418]

[0419] The preparation method was the same as that of compound TM1, using compound i-20-2 instead of compound i-1-a. The crude product was purified by preparative thin-layer chromatography (MeOH / DCM = 4%), and 23.9 mg of a yellowish-green oily solid was obtained.

[0420] Yield: 66.2%; LC-MS (ESI, m / z): calcd for C 37 H 55 BrN5O6S + , [M + H] + 776.3, found 776.3; 1 H NMR (400 MHz, CDCl3) δ 8.25 (s, 1H), 7.69 (dd, J = 8.5 Hz, 1H), 7.40 (t, J = 6.1 Hz, 1H), 7.28 (dd, J = 2.3 Hz, 1H), 7.06 (dd, J = 8.5, 2.3 Hz, 1H), 4.93 (dd, J = 12.3, 5.3 Hz, 1H), 4.36 (dd, J = 8.1, 6.3 Hz, 1H), 3.53 (q, J = 6.2 Hz, 2H), 3.45 (t, J = 5.1 Hz, 4H), 3.21 - 3.09 (m, 2H), 3.07 (s, 2H), 2.90 - 2.74 (m, 3H), 2.68 (t, J = 5.1 Hz, 4H), 2.12 - 2.08 (m, 1H), 2.03 - 1.92 (m, 2H), 1.48 - 1.42 (m, 2H), 1.23 (s, 22H), 0.86 (s, 3H); 1313C NMR (101 MHz, CDCl3) δ 196.22, 171.16, 170.16, 168.39, 167.90, 167.25, 155.39, 134.34, 125.50, 120.07, 118.27, 108.96, 77.33, 61.41, 53.96, 53.06, 49.25, 47.72, 38.61, 35.22, 32.00, 31.52, 29.76, 29.73, 29.67, 29.56, 29.44, 29.39, 29.32, 28.93, 28.23, 27.30, 22.81, 22.78, 14.23。

[0421] Example 81. S-(2-(1-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine-4-carboxamido)ethyl) 2-bromohexadecanoate TM25

[0422]

[0423] The preparation method is the same as that of compound TM1, using compound i-21-2 instead of compound i-1-a. The crude product was purified by preparative thin-layer chromatography (MeOH / DCM = 4%), and 21.5 mg of a yellowish-green oily solid was obtained.

[0424] Yield: 60.7%; 1 1H NMR (400 MHz, CDCl3) δ 8.21 (s, 1H), 7.66 (dd, J = 8.5 Hz, 1H), 7.26 (dd, J = 2.2 Hz, 1H), 7.03 (dd, J = 8.6, 2.4 Hz, 1H), 5.97 (t, J = 5.7 Hz, 1H), 4.93 (dd, J = 12.2, 5.3 Hz, 1H), 4.37 (dd, J = 8.0, 6.3 Hz, 1H), 3.94 (dt, J = 13.2, 3.7 Hz, 2H), 3.49 (q, J = 6.2 Hz, 2H), 3.16 - 3.05 (m, 2H), 2.98 (t, J = 11.8 Hz, 2H), 2.87 - 2.72 (m, 3H), 2.32 (p, J = 3.3 Hz, 1H), 2.13 - 2.09 (m, 1H), 2.08 - 1.99 (m, 2H), 1.96 - 1.92 (m, 2H), 1.81 - 1.76 (m, 2H), 1.49 - 1.43 (m, 2H), 1.23 (s, 22H), 0.86 (s, 3H); 1313C NMR (101 MHz, CDCl3) δ 197.02, 174.28, 171.19, 168.47, 168.02, 167.31, 155.29, 134.42, 125.55, 119.17, 118.17, 108.88, 77.32, 53.82, 49.21, 47.54, 42.67, 39.46, 35.15, 32.01, 31.53, 29.78, 29.73, 29.67, 29.57, 29.45, 29.39, 29.21, 28.92, 28.02, 27.98, 27.31, 22.82, 22.78, 14.23。

[0425] Example 82. S-(2-(4-(4-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)benzamido)ethyl) 2-bromohexadecanoate TM26

[0426]

[0427] The preparation method was the same as that of compound TM1, using compound i-22-2 instead of compound i-1-a. The crude product was purified by preparative thin-layer chromatography (MeOH / DCM = 5%) to obtain 21.5 mg of a yellowish-green oily solid.

[0428] Yield: 60.7%; 1 1H NMR (400 MHz, CDCl3) δ 8.73 (dd, J = 4.5, 1.4 Hz, 1H), 8.45 (dd, J = 8.4, 1.4 Hz, 1H), 8.19 (dd, J = 9.1 Hz, 2H), 8.00 (s, 1H), 7.75 (dd, J = 8.5 Hz, 1H), 7.45 (dd, J = 8.3, 4.5 Hz, 1H), 7.35 (t, J = 6.5 Hz, 1H), 7.07 (m, 1H), 4.96 (m, 1H), 4.37 (m, 1H), 3.69 (s, 8H), 3.39 (m, 2H), 3.06 (m, 2H), 2.20 - 2.14 (m, 1H), 1.42 - 1.40 (m, 2H), 1.24 (s, 22H), 0.87 (s, 3H).

[0429] Example 83. Detection of the degradation effect of compounds TM1 - TM4 on the entire DHHC protein family by Western blot (pan-DHHC inhibitor (2-bromopalmitate, 2-BP) as a negative control).

[0430] Experimental method:

[0431] Ⅰ. Cell culture

[0432] HEK-293T cells were subcultured in a carbon dioxide cell incubator. The culture conditions were set as follows: in DMEM medium containing 10% fetal bovine serum, at 37 °C, and 5% CO2.

[0433] Ⅱ. Seeding cells, plasmid transfection, and drug administration

[0434] One day in advance, seed 24-well plates at a density of 200,000 cells per well. The next day, discard the original culture medium and replace it with 0.5 mL of fresh culture medium. Use JetPRIME reagent for plasmid transfection. The transfection system for 24-well plates is 0.5 mL, with 0.25 μg of plasmid, 0.5 μL of transfection reagent, and 50 μL of buffer. The specific operation is as follows: Calculate the required volume according to the plasmid concentration, add it to the buffer, vortex and centrifuge the mixture. Finally, add 0.5 μL of transfection reagent, vortex and centrifuge again, and let it stand at room temperature for 10 min. After standing, evenly drip the mixed system containing the plasmid and transfection reagent into the cells and gently mix. After 4 - 6 h, replace the original culture medium in the culture dish with fresh serum-containing culture medium. Discard the culture medium 24 h after transfection and perform drug treatment for 48 h.

[0435] Ⅲ. Western blot

[0436] (1) Preparation of protein samples: Discard the culture medium, rinse once with phosphate-buffered saline, collect cells with 100 μL of RIPA lysis buffer into a 1.5 mL microcentrifuge tube, flick it three times, lyse on ice for 10 min, repeat 3 times until the cells are completely lysed, centrifuge at 13,400 rpm at 4 °C for 30 min, transfer the supernatant to a new 1.5 mL microcentrifuge tube. Take an appropriate amount of the sample and add 5× protein loading buffer according to the ratio, then place it on a metal bath and heat at 95 °C for 5 min, briefly centrifuge to gather the water droplets on the tube wall to the bottom of the tube, and vortex to mix evenly.

[0437] (2) Gel preparation: a) Align the clean and water-free thick and thin glass plates and clamp them in a gel-making rack. Select the appropriate separation gel concentration according to the molecular weight of the target protein in this experiment, add the raw materials to the tube according to the corresponding formula, vortex to mix evenly, then add the separation gel between the thick and thin glass plates. Subsequently, add pure water above the separation gel to make its upper edge flat and wait for it to solidify. b) Stacking gel: Discard the water above the separation gel. Add the raw materials to the tube according to the stacking gel formula, vortex to mix evenly, pour it above the separation gel, then insert a clean and water-free gel comb and wait for it to solidify.

[0438] (3) Electrophoresis: a) Preparation: After the stacking gel solidifies, remove the prepared gel from the gel-making rack, clamp it and place it in the electrophoresis tank. Pour an appropriate amount of electrophoresis buffer, check for leakage, and then pull out the comb teeth to prepare for loading. b) Loading: Vortex and mix the samples prepared in the previous step, and sequentially add them into the loading wells in order. The loading volume is generally about 10 μL. After loading, turn on the electrophoresis instrument switch, select the constant voltage mode, set the voltage to 70 V for pre-electrophoresis for about 15 min, confirm whether the current is normal. The normal current for two gels is about 40 - 50 mA. Then increase the voltage to 130 - 200 V to continue electrophoresis, and stop electrophoresis when the bromophenol blue line reaches the bottom.

[0439] (4) Membrane transfer and blocking: a) Preparation: Prepare 1× transfer buffer: Take 70 mL of 10× transfer buffer, initially dilute it with pure water and then add 140 mL of absolute ethanol, and continue to make up to 700 mL with pure water to obtain 1× transfer buffer, and pre-cool it in a 4℃ refrigerator; Immerse the polyvinylidene fluoride membrane (PVDF membrane) in methanol for 1 min for activation, and then transfer it to 1× transfer buffer to terminate the activation. b) Membrane transfer: Use the wet transfer method. First, open the transfer cassette, face the black side down and immerse it in 1× transfer buffer, and place it layer by layer in the order of one layer of sponge - three layers of filter paper - gel - PVDF membrane - three layers of filter paper - one layer of sponge. During the process, the actions should be gentle to avoid generating bubbles. Finally, close the transfer cassette and clamp it, and place it in the electrotransfer tank. Put ice packs inside and outside the electrotransfer tank for subsequent cooling. Turn the electrophoresis instrument to the constant current mode, set the current to 330 mA to start membrane transfer, check whether the voltage is normal, and select an appropriate membrane transfer time according to the molecular weight of the target protein. c) Blocking: After completion, take out the transfer cassette, open it with the black side up, trim the band of the required protein according to the protein molecular weight standard indication on the PVDF membrane, and place the band in the pre-prepared 5% skim milk for blocking at room temperature for 1 h.

[0440] (5) Antibody incubation and exposure: a) Antibody incubation: After blocking, remove the milk, wash the remaining milk on the PVDF membrane with phosphate buffered saline with Tween (T-PBS) buffer, add the buffer containing the primary antibody corresponding to the target protein, and incubate overnight on a horizontal shaker in a 4℃ refrigerator. The next day, recover the primary antibody, rinse the PVDF membrane three times with T-PBS buffer, and then place it in 5% skim milk. Add the secondary antibody corresponding to the resistance at a ratio of 1:5000, and incubate on a horizontal shaker at room temperature for 1 h. b) Exposure: Remove the milk, rinse the PVDF membrane three times with T-PBS buffer and then prepare for exposure. Prepare an appropriate volume of ECL developing solution according to the ratio of Solution A: Solution B = 1:1. Place the PVDF membrane in the middle of the developing plate for development, quickly cover the ECL developing solution on the membrane, let it stand for 1 min, put it into the AI800 exposure machine, and set the exposure time for exposure.

[0441] Ⅳ. Cell Proliferation Inhibition Experiment

[0442] After digesting the HL60 cells in the logarithmic growth phase, they were blown into a single-cell suspension and inoculated into a 96-well culture plate (about 5×10 3 cells / well); 10% serum-containing complete medium was added to each well and incubated overnight in an incubator containing 5% carbon dioxide; test compounds with gradient concentrations (the pan-DHHC inhibitor 2-BP was used as a control compound) were added, and the cells were continuously cultured for 3 days in a normoxic incubator environment. The cell viability kit CCK-8 (from MCE, USA) was used to evaluate the inhibitory effect of the target compound on cell proliferation. The OD value of each well was detected by an enzyme-linked immunosorbent assay (detection wavelength: 450 nm), and the inhibition rate (%) was calculated as follows: Inhibition rate (%) = (OD control - OD drug administration) ÷ OD control × 100%.

[0443] The degradation rate calculation method is as follows: Quantitative analysis of the exposure results was performed using Image J software. First, the gray value of the exposure band was calculated using ImageJ software, and then the gray value of the target protein was divided by the gray value of the internal reference protein for normalization. The calculation formula is R 样本 = A 目的蛋白 / A 内参蛋白 × 100% (A is the gray value, and R is the degradation rate). Table 1 Results of the degradation of mzDHHC (1-9, 11-13) protein by representative compounds TM1-TM4 in HEK293 cells (unit: %)

[0444]

[0445]

[0446] Table 2 Results of the degradation of mzDHHC (14-24) protein by representative compounds TM1-TM4 in HEK293 cells (unit: %)

[0447]

[0448] Table 3 Results of the degradation of mzDHHC1 protein by compounds in HEK293 cells (unit: %)

[0449]

[0450] Table 4 The proliferation inhibition rates of representative compounds TM3 and TM4 on human acute promyelocytic leukemia cells (HL60 cells) are superior to those of the existing pan-DHHC inhibitor 2-BP (unit: %, 3 days)

[0451]

Claims

1. A compound represented by formula (I), or a pharmaceutically acceptable salt or configurational isomer thereof: A is CH or N; R is halogen or -CH2CN; is a single bond or a double bond; n is a positive integer from 3 to 20; B is S, O or CH, CH2; L is the connecting fragment; D is in, M1 is absent, CH2, N or O; M2 is C=O or CH2. When M1 is absent, L is directly connected to the benzene ring carbon atom on D.

2. The compound according to claim 1, characterized in that The L is selected from x is an integer of 1 to 6; y is an integer of 1 to 10; n1, n2, n3 and n4 are each independently selected from 1, 2 and 3.

3. The compound according to claim 2, characterized in that The D is selected from:

4. The compound according to claim 1, characterized in that: When A is CH, B is S and is a single bond, R is Cl, Br, I; When A is N, B is CH and It is a double bond of E configuration, R is -CH2CN; L is selected from x is selected from an integer of 1-4; y is selected from 6, 7, 8.

5. The compound according to any one of claims 1 to 4, characterized in that: Having the structure shown in formula (II) or (III) The definitions of L and D are the same as those in claim 1.

6. The compound according to claim 1, characterized in that L is x is an integer selected from 1 to 4; P is 7. The compound according to claim 1, characterized in that The structure is shown in any of the following formulas:

8. The compound according to any one of claims 1 to 7, characterized in that When there is a double bond, the configuration is E type.

9. A pharmaceutical preparation, characterized in that: Contains a therapeutically effective amount of the compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt or configurational isomer thereof, and necessary auxiliary materials.

10. Use of the compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt or configurational isomer thereof in the preparation of a drug for alleviating, improving or curing related diseases by degrading DHHC family proteins.

Images