PROTACs compound for targeted degradation of PRMT5 and application of PROTACs compound

Abstract

The invention discloses a PROTACs compound for targeted degradation of PRMT5 and application of the PROTACs compound. The PROTACs compound comprises a compound as shown in the following formula, and related analogues or pharmaceutically acceptable salts, metabolites or prodrugs of the compound: in the # imgabs0 # formula, X represents a ligand bound to PRMT5 protein, Z represents a ligand of E3 ubiquitin ligase, and Y represents a linking group for linking X and Z; according to the invention, CRBN is used as E3 ligase, and a series of PRMT5-targeted PROTAC degradation compounds are designed and synthesized. The compound can effectively induce PRMT5 degradation, significantly inhibit proliferation, growth, migration, infiltration, clone formation and metastasis of tumor cells, promote apoptosis of cancer cells, promote autophagy of the tumor cells, prevent or overcome drug resistance of BTK inhibitors and / or prolong the lifetime of tumor patients, and has wide application prospects.

Description

Field of the Invention

[0001] The present invention belongs to the technical field of anti-tumor drugs, and in particular relates to PROTACs compounds for targeted degradation of PRMT5 and applications thereof. Background of the Invention

[0003] Protein arginine methyltransferase (PRMT5) is a member of the protein arginine methyltransferase family responsible for epigenetic regulation and post-transcriptional modification. It catalyzes the transfer of the methyl group of S-adenosylmethionine (SAM) to the guanidinium nitrogen atom of the substrate arginine to achieve symmetrical dimethylation of arginine on the substrate protein. Studies have shown that PRMT5, as an epigenetic regulator, can be responsible for the methylation modification of histones such as H4R3, H3R8, and H3R2, thereby regulating gene expression. PRMT5 can also perform post-transcriptional modification functions, by catalyzing the methylation of non-histone proteins to regulate mRNA splicing, DNA replication and damage repair, and cell cycle and other biological processes.

[0004] PRMT5 is significantly overexpressed in many tumors: in breast cancer cells, the expression of PRMT5 is significantly higher than that in normal breast cells. PRMT5 promotes the occurrence and development of breast cancer by regulating the methylation of proteins such as AKT1, KLF5 and KLF4; in lung cancer cells, PRMT5 is also significantly overexpressed; the survival of acute myeloid leukemia cells is related to the catalytic activity of PRMT5; the expression of PRMT5 in prostate cancer cells is significantly higher than that in benign prostatic hyperplasia tissues, and it interacts with Sp1 and forms a complex with Brg1 to regulate the expression of AR, thereby affecting the growth of prostate cancer cells (Oncogene. 2017Mar 2; 36(9): 1223-1231.). In summary, PRMT5 is closely related to the occurrence and development of many cancers. Therefore, PRMT5 has attracted much attention as an emerging tumor treatment target.

[0005] Protein-targeted chimera (PROTAC) technology is a new means of inducing protein degradation. PROTAC molecules are usually composed of three parts: a target protein ligand, an E3 ligase ligand, and a linker connecting the two ligands. Under the action of such small molecules, the target protein and the E3 ligase are spatially close to each other to achieve ubiquitination of the target protein, and finally protein degradation dependent on the ubiquitin proteasome occurs. Compared with traditional inhibitors, PROTAC also shows great potential in selectivity, overcoming drug resistance, and inhibiting the non-enzyme functions of multiple target proteins (Nat Rev Cancer. 2021Oct; 21(10): 638-654.). Taking the human homolog of the Drosophila Zeste gene enhancer 2 (EZH2) as an example, the currently developed targeted EZH2 inhibitors can only achieve anti-tumor activity by inhibiting its enzymatic activity and inducing cancer-related gene silencing. However, the non-enzyme activity of EZH2 is also crucial for the activation of genes related to cancer progression, which leads to incomplete inhibition of its carcinogenic activity by inhibitors, while the development of PROTAC molecules targeting EZH2 achieves complete blocking of its carcinogenic function. In addition, compared with inhibitors, PROTAC degraders targeting EZH2 also showed significant antiproliferative activity in cell lines resistant to inhibitors. In summary, the use of PROTAC technology to induce the degradation of the target protein PRMT5 may also achieve complete blocking of the oncogenic function, which is a potential strategy for targeting PRMT5 to treat cancer. Summary of the invention

[0006] The present invention provides a class of PROTACs compounds targeting degradation of PRMT5, which can effectively induce PRMT5 degradation and significantly inhibit the proliferation of tumor cells. The present invention uses CRBN as an E3 ligase to design and synthesize a series of PROTAC degradation compounds targeting PRMT5. Compared with the recently reported MS4322 (which takes 6-8 days to exert the maximum degradation effect, J Med Chem. 2020 Sep 10; 63 (17): 9977-9989.), this type of degrader can achieve rapid degradation of PRMT5 in two days, and exhibits significant anti-proliferative activity in triple-negative breast cancer cells, which can effectively induce cell cycle arrest and apoptosis. Mechanism studies have shown that this type of degrader can regulate the expression of the "difficult to target" transcription factor KLF5, which is also very important for breast cancer treatment, by degrading PRMT5 protein. In addition, in organoids derived from TNBC patients, anti-proliferative activity can also be effectively achieved. Certain anti-tumor activity was achieved in mice, and good tolerability was shown. In summary, the present invention provides a class of PROTACs molecular entities that can effectively degrade PRMT5 and is expected to prepare a drug targeting PRMT5-related malignant tumors.

[0007] The present invention proposes a class of PROTACs compounds for targeted degradation of PRMT5, which is a compound represented by formula (I) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, metabolites, pharmaceutically acceptable salts or prodrugs:

[0008]

[0009] Wherein, X represents a ligand that binds to the PRMT5 protein, Z represents a ligand of the E3 ubiquitin ligase, and Y represents a linker group connecting X and Z.

[0010] According to an embodiment of the present invention, the above compound may further include at least one of the following additional technical features:

[0011] In the formula (I) of the present invention, according to an embodiment of the present invention, representative X includes structures as described in formula (II) and formula (III):

[0012]

[0013]

[0014] In the formula (I) of the present invention, Z is selected from the structures described in the following formulas (IV) to (VI):

[0015]

[0016] in:

[0017] A is selected from any one of CH2, C(O), SO, SO2;

[0018] R is selected from any one of F or H;

[0019] M is selected from Any one of;

[0020] P is selected from any one of the following groups: H, F or CF3;

[0021] Q is selected from any one of the following groups: -NH- or -CH2-.

[0022] In the structural formulas (IV) to (VI) of the present invention, according to embodiments of the present invention, representative Z includes:

[0023]

[0024] In the formula (I) of the present invention, Y is selected from the following structures:

[0025]

[0026] in:

[0027] m is an integer between 0 and 12; preferably, 0, 2, 3, 4, 5, 6, 7, 8, 9, 10;

[0028] n is an integer between 1 and 10; preferably, 1, 2, 3, 4, 5;

[0029] q is an integer between 0 and 8; preferably, 0 or 2;

[0030] t is an integer between 0 and 8; preferably, it is 5.

[0031] The present invention provides PROTACs compounds that target PRMT5 degradation as shown in the above formulas (I)-(VI), including but not limited to the following compounds:

[0032] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0033] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0034] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0035] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-3,6,9,12-tetraoxopentadecane-15-yl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0036] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-3,6,9,12,15-pentaoxaoctadecane-18-yl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0037] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0038] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)pentanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0039] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)hexanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0040] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0041] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0042] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(9-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)nonanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0043] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0044] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0045] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)ethoxy)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0046] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0047] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0048] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(9-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)nonanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0049] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0050] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(11-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)undecanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0051] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(12-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)dodecanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0052] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((2-(2,6-dioxopiperidin-3-yl)-6-fluoro-1,3-dioxoisoindolin-5-yl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0053] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0054] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)octyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0055] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)octyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0056] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)octyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0057] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)pentyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0058] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0059] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0060] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0061] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)pentanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0062] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(6-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)hexanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0063] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0064] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0065] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oct-7-ynyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0066] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(11-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)undec-10-ynyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0067] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(2-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)acetyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0068] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(4-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)butyl)piperazin-1-yl)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0069] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(1-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperidin-4-carbonyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0070] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(1-(3-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)propyl)piperidin-4-carbonyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0071] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)benzoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0072] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0073] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-(4-(3-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)propyl)piperazin-1-yl)benzoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0074] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(5-(7-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,7-diazaspiro[3.5]nonan-2-yl)pentanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0075] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-(9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-3,9-diazaspiro[5.5]undec-3-yl)butanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0076] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0077] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(11-(4-(2,6-dioxopiperidin-3-yl)phenoxy)undecanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0078] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((4-(2,6-dioxopiperidin-3-yl)phenyl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0079] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-(4-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperazin-1-yl)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0080] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(2-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)hexyl)amino)-2-oxoethyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0081] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetylamino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0082] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetamido)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0083] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(6-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetamido)hexanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0084] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)acetylamino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0085] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)acetamido)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0086] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(2-(3-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)acetyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide;

[0087] N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((4-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)decanamido)cyclohexyl)amino)pyrimidine-4-carboxamide.

[0088] The present invention also provides a pharmaceutical composition, comprising the PROTACs compound targeting the degradation of PRMT5.

[0089] As a further description of the above scheme: the pharmaceutical composition further comprises a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle or a combination thereof.

[0090] The present invention also provides use of the PROTACs compound or the pharmaceutical composition in the preparation of a drug for degrading or inhibiting PRMT5.

[0091] The present invention also provides use of the PROTACs compound or the pharmaceutical composition in the preparation of a drug for preventing or treating tumors.

[0092] Preferably, the tumor is at least one of breast cancer, cervical cancer, non-small cell lung cancer, B cell lymphoma, mantle cell lymphoma and melanoma.

[0093] The preparation methods of several representative compounds provided by the present invention are as follows:

[0094] Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-18):

[0095] Dissolve 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione and 1-2 molar multiples of tert-butyl 10-aminodecanoate in DMSO, add 1.5-2.5 molar multiples of DIEA, protect with nitrogen, and stir overnight at 80-95°C. Detect the reaction by TLC. After the reaction is completed, extract with ethyl acetate and water, collect the organic phase, concentrate, separate and purify by column chromatography (DCM:MeOH=80-120:1 elution) to obtain the intermediate;

[0096] The intermediate was dissolved in DCM, TFA was added, and the mixture was stirred at room temperature overnight for ester decomposition reaction. After the reaction was completed, the mixture was distilled under reduced pressure to obtain the intermediate after de-tert-butylation.

[0097] The intermediate after de-tert-butoxylation and 0.5-1.5 molar multiples of intermediate P1 are dissolved in DMSO, and 1-2 molar multiples of HATU and 2-5 molar multiples of DIEA are added in sequence. The reaction is stirred at room temperature, extracted with EA and water, and the organic phase is collected. After concentration, the target product YZ-18 is obtained by column chromatography separation and purification (DCM:MeOH=30-80:1).

[0098] Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-41):

[0099] Dissolve 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione and 0.4-0.8 molar multiple of tert-butyl 4-(4-(piperazin-1-ylmethyl)piperidin-1-yl)benzoate in DMSO, add 0.8-1.5 molar multiple of DIEA, and react at 80-95° C. under nitrogen protection overnight. The mixture was extracted with EA and water, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, concentrated, separated and purified by column chromatography (DCM:MeOH=30-80:1) to obtain the target product 4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoic acid tert-butyl ester; the target product was dissolved in DCM, TFA was added, the mixture was stirred at room temperature overnight to remove the tert-butoxy group, and the mixture was distilled under reduced pressure to obtain the intermediate 4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoic acid;

[0100] 4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazine-1-yl)methyl)piperidin-1-yl)benzoic acid and 0.5-1.5 molar times of intermediate P1 are dissolved in DMSO, and 1.5-3 molar times of HATU and 3-5 molar times of DIEA are added in sequence and stirred at room temperature. After the reaction is completed, extraction is performed, the organic phase is collected, concentrated, and separated and purified by column chromatography (DCM:MeOH=30-80:1) to obtain the target product YZ-41.

[0101] Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(11-(4-(2,6-dioxopiperidin-3-yl)phenoxy)undecanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-46):

[0102] Dissolve 2,6-bis(benzyloxy)-3-bromopyridine, 1-2 molar times of 4-hydroxyphenylboronic acid pinacol ester and 2-5 molar times of potassium carbonate in 1,4-dioxane and water and stir at room temperature, add a catalyst of bistriphenylphosphine palladium dichloride, stir overnight at 80-110° C. under nitrogen protection, filter, distill under reduced pressure, separate and purify by column chromatography (PE:EA=3-10:1) to obtain an intermediate 4-(2,6-bis(benzyloxy)pyridin-3-yl)phenol;

[0103] The intermediate 4-(2,6-bis(benzyloxy)pyridin-3-yl)phenol and 1-2 molar times of tert-butyl 11-bromodecaneate are dissolved in DMF, 3-5 molar times of cesium carbonate are added, and the mixture is stirred at room temperature; EA and water are extracted, the organic phase is collected, and after concentration, the organic phase is separated and purified by column chromatography (PE:EA=5-12:1) to obtain the intermediate tert-butyl 11-(4-(2,6-bis(benzyloxy)pyridin-3-yl)phenoxy)undecanoate.

[0104] The intermediate 11-(4-(2,6-bis(benzyloxy)pyridin-3-yl)phenoxy)undecanoic acid tert-butyl ester was dissolved in methanol, and an appropriate amount of Pd / C was added under nitrogen protection, and the nitrogen was replaced with hydrogen again, and the mixture was stirred at room temperature overnight. After the reaction was completed, the mixture was filtered and distilled under reduced pressure to obtain a crude product 11-(4-(2,6-dioxopiperidin-3-yl)phenoxy)undecanoic acid tert-butyl ester; the crude product was dissolved in DCM, TFA was added, and the mixture was stirred at room temperature overnight. After the reaction was completed, the mixture was distilled under reduced pressure to obtain a crude product 11-(4-(2,6-dioxopiperidin-3-yl)phenoxy)undecanoic acid.

[0105] Dissolve 11-(4-(2,6-dioxopiperidin-3-yl)phenoxy)undecanoic acid and 0.5-2 molar times of intermediate P1 in DMSO, add 1-2 molar times of HTAU and 2-5 molar times of DIEA in sequence, stir at room temperature, and monitor the reaction by TLC. Extract with EA and water, collect the organic phase, concentrate, separate and purify by column chromatography (DCM:MeOH=30-80:1) to obtain the target product YZ-46.

[0106] Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((4-(2,6-dioxopiperidin-3-yl)phenyl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-47):

[0107] Dissolve 2,6-bis(benzyloxy)-3-bromopyridine, 0.5-1.5 molar multiples of 4-aminophenylboronic acid pinacol ester and 2-4 molar multiples of potassium carbonate in 1,4-dioxane and water and stir at room temperature, add the catalyst bistriphenylphosphine palladium dichloride under nitrogen protection, and stir again at 95-110°C overnight under nitrogen protection. Filter by suction, distill under reduced pressure, separate and purify by column chromatography (PE:EA=1-3:1) to obtain the intermediate 4-(2,6-bis(benzyloxy)pyridin-3-yl)aniline;

[0108] The intermediate 4-(2,6-bis(benzyloxy)pyridin-3-yl)aniline and 1-2 molar times of tert-butyl 10-bromodecanoate were dissolved in DMF, 3-5 molar times of cesium carbonate were added, and the mixture was stirred at 80-95° C. overnight. The mixture was extracted with EA and water, and the organic phase was collected, concentrated, and purified by column chromatography (PE:EA=3-10:1) to obtain the intermediate tert-butyl 10-((4-(2,6-bis(benzyloxy)pyridin-3-yl)phenyl)amino)decanoate.

[0109] The intermediate tert-butyl 10-((4-(2,6-bis(benzyloxy)pyridin-3-yl)phenyl)amino)decanoate was dissolved in ethanol, an appropriate amount of Pd / C was added, nitrogen was protected, hydrogen was replaced, and the mixture was stirred at room temperature overnight. After the reaction was completed, suction was filtered and vacuum distilled to obtain the crude product tert-butyl 10-((4-(2,6-dioxopiperidin-3-yl)phenyl)amino)decanoate, which was then dissolved in DCM, TFA was added, and the mixture was stirred at room temperature overnight. After the reaction was completed, vacuum distillation was performed to obtain the crude product 10-((4-(2,6-dioxopiperidin-3-yl)phenyl)amino)decanoate

[0110] Dissolve 10-((4-(2,6-dioxopiperidin-3-yl)phenyl)amino)decanoic acid and 0.8-1.5 molar times of intermediate P1 in DMSO, add 1-2 molar times of HATU and 2-5 molar times of DIEA in sequence and stir at room temperature. Extract with EA and water, collect the organic phase, concentrate and separate and purify by column chromatography (DCM:MeOH=30-80:1) to obtain the target product YZ-47.

[0111] The term "pharmaceutically acceptable salt" as used in the present invention refers to a salt of a compound of the present invention, prepared from a compound of the present invention having a specific substituent and a relatively non-toxic acid or base. When the compound of the present invention contains a relatively acidic functional group, a base addition salt can be obtained by contacting a neutral form of such compound with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, amine, organic amine or magnesium salts or similar salts. When the compound of the present invention contains a relatively basic functional group, an acid addition salt can be obtained by contacting a neutral form of such compound with a sufficient amount of acid in a pure solution or a suitable inert solvent. The implementation of pharmaceutically acceptable acid addition salts includes inorganic acid salts, such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, bisulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid and methanesulfonic acid, etc.; also includes salts of amino acids (such as arginine, etc.), and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain acidic and basic functional groups, so that they can be converted into any base or acid addition salts.

[0112] In addition to the salt form, the compounds provided by the present invention also exist in the form of prodrugs. Prodrugs of the compounds described in the present invention are easily chemically changed under physiological conditions to be converted into the compounds of the present invention. Any compound that can be converted in vivo to provide a biologically active substance (i.e., a compound that conforms to the compounds shown in formula (I) to (VI)) is a prodrug within the scope and spirit of the present invention. For example, a compound containing a carboxyl group can form a physiologically hydrolyzable ester, which can be hydrolyzed in vivo to obtain a compound that conforms to the compounds shown in formula (I) to (VI) itself and act as a prodrug.

[0113] Certain compounds of the present invention may have asymmetric carbon atoms (optical centers) or double bonds. Racemates, diastereomers, geometric isomers and individual isomers are included within the scope of the present invention, including cis and trans isomers, (-)- and (+)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, all of which are included within the scope of the present invention as well as their mixtures.

[0114] The compounds of the invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that make up the compound. For example, the compounds may be labeled with isotopes, such as deuterium ( 2 H), tritium ( 3 H) or C-14( 14C) All isotopic variations of the compounds of the present invention, whether radioactive or not, are encompassed within the scope of the present invention.

[0115] The present invention provides a pharmaceutical composition, which contains a therapeutic dose of the aforementioned compound. "Pharmaceutical composition" means a mixture containing one or more compounds of the present invention or their physiologically / pharmaceutically acceptable salts or prodrugs and other chemical components, or other components such as physiological saline / pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to promote administration to an organism, facilitate the absorption of the active ingredient, and thus exert biological activity.

[0116] The pharmaceutical composition is formulated into an injectable fluid, an aerosol, a cream, a gel, a pill, a capsule, a syrup, a transdermal patch or an excipient. According to a specific embodiment of the present invention, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier, an excipient, a diluent, an adjuvant, a vehicle or a combination thereof.

[0117] The term "excipient" generally refers to carriers, diluents and / or vehicles required to formulate an effective pharmaceutical composition.

[0118] The present invention relates to a method for degrading PRMT5 protein by using bifunctional small molecules, comprising administering any one of the above compounds to a patient or subject, optionally in combination with another bioactive agent for the treatment of a disease, wherein the disease type is mainly cancer.

[0119] The present invention proposes the use of the above-mentioned PROTACs compounds and related analogs or pharmaceutically acceptable salts, metabolites or prodrugs for targeting the degradation of PRMT5 or the pharmaceutical composition in the preparation of PRMT5 regulators and anti-tumor drugs, wherein the drug is used to degrade PRMT5 protein through bifunctional small molecules.

[0120] The PROTACs compounds and related analogs or pharmaceutically acceptable salts, metabolites or prodrugs targeting PRMT5 degradation of the present invention, or the pharmaceutical composition is used to inhibit the proliferation, growth, migration, infiltration, cloning and metastasis of cancer cells, promote apoptosis of cancer cells, promote autophagy of tumor cells, prevent or overcome BTK inhibitor resistance, and / or prolong the survival of tumor patients. Wherein, the tumor includes at least one of breast cancer, cervical cancer, non-small cell lung cancer, B cell lymphoma, mantle cell lymphoma and melanoma. BRIEF DESCRIPTION OF THE DRAWINGS

[0121] Figure 1 The degradation activity of some compounds of the present invention in triple-negative breast cancer cells HCC1806 and HCC1937;

[0122] Figure 2The effect of compound YZ-18 on the proliferation of HCC1806, HCC1937, MDA-MB-231, MDA-MB-468, MCF10A and 184A1 cells;

[0123] Figure 3 The ability of compound YZ-18 to induce cell cycle in breast cancer cells;

[0124] Figure 4 is the ability of compound YZ-18 to induce apoptosis in triple-negative breast cancer cells;

[0125] Figure 5 The effect of compound YZ-18 on the survival rate of cancer cells;

[0126] Figure 6 The inhibitory activity of compound YZ-18 on the proliferation of triple-negative breast cancer organoid cells;

[0127] Figure 7 The inhibitory ability of compound YZ-18 on the growth of HCC1806 triple-negative breast cancer xenograft tumors in nude mice;

[0128] Figure 8 The inhibitory effect of compound YZ-18 on the weight of HCC1806 triple-negative breast cancer xenograft tumor tissue in nude mice;

[0129] Fig. 9 The figure shows the effect of compound YZ-18 alone on the body weight of nude mice. DETAILED DESCRIPTION

[0130] The present invention is further described in detail below in conjunction with specific embodiments and drawings, and the protection content of the present invention is not limited to the following embodiments.

[0131] The present invention proposes a bifunctional small molecule and its derivatives or pharmaceutically acceptable salts, and a pharmaceutical composition containing such compounds. The present invention relates to a protein degradation targeting chimera (PROTACs) bifunctional small molecule, which can selectively induce PRMT5 protein degradation. The present invention successfully prepared a PROTACs compound that targets and degrades PRMT5 by connecting a PRMT5 small molecule ligand to a ligand of a CRBN E3 ubiquitin ligase using linkers of different types and lengths.

[0132] The purity results of the compounds obtained in the present invention are all from Agilent 1200 series LC system high performance liquid chromatograph, the ultraviolet detection wavelength is 254nm; the nuclear magnetic resonance spectrometer is Bruker 500 type, and the solvent used is DMSO-d6; the purification of reaction intermediates and final products uses chromatography columns (silica gel 200-300 mesh), and the silica gel used is purchased from Qingdao Ocean Chemical Plant. Unless otherwise specified, all reactions are tracked by TLC. The term "react overnight" or "stir overnight" generally refers to the reaction for 8-18 hours; the term "room temperature" generally refers to 10-40°C.

[0133] Example 1: Preparation of the compounds of the present invention

[0134] Preparation of the key intermediate (S)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-(piperidin-4-ylamino)pyrimidine-4-carboxamide (P1):

[0135]

[0136] Dissolve 6-chloropyrimidine-4-carboxylic acid (2.0 g, 12.36 mmol) in 39 mL of DCM (dichloromethane), stir at 40 °C, slowly add 4.6 mL of (COCl)2 (oxalyl chloride), then add 3 drops of DMF (N,N-dimethylformamide), stir for 3 h, and detect the reaction by thin layer chromatography. After the reaction is completed, distill under reduced pressure, and the obtained crude 6-chloropyrimidine-4-carbonyl chloride can be directly used in the next step reaction.

[0137] The intermediate 6-chloropyrimidine-4-carbonyl chloride (12.36 mmol) and (S)-1-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)propan-2-ol (3.0 g, 12.36 mmol) were dissolved in DCM (15 mL), and TEA (triethylamine) (4.1 g, 37.08 mmol) was slowly added dropwise and stirred at room temperature. After the reaction was completed, the mixture was evaporated under reduced pressure and purified by silica gel column chromatography (DCM: MeOH = 80:1) to obtain the intermediate (S)-6-chloro-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)pyrimidine-4-carboxamide (2.6 g, 7.49 mmol, yield 61%).

[0138] The intermediate (S)-6-chloro-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)pyrimidine-4-carboxamide (2.6 g, 7.49 mmol) and tert-butyl 4-amino-1-piperidinecarboxylate (3.0 g, 14.97 mmol) were dissolved in 10 mL of i-PrOH (isopropanol), triethylamine (1.5 g, 14.97 mmol) was added, and the mixture was stirred at 80°C overnight. A large amount of isopropanol was removed by distillation under reduced pressure, and the intermediate (S)-tert-butyl 4-((6-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)carbamoyl)pyrimidin-4-yl)amino)piperidine-1-carboxylate (3.6 g, 7.03 mmol, yield 99%) was obtained by silica gel column chromatography (DCM:MeOH=80:1).

[0139] The intermediate (S)-tert-butyl 4-((6-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)carbamoyl)pyrimidin-4-yl)amino)piperidine-1-carboxylate (3.6 g, 7.03 mmol) was dissolved in DCM (15 mL), trifluoroacetic acid (TFA) (3 mL) was added, and stirred at room temperature overnight. After the reaction was completed, the mixture was evaporated under reduced pressure, and the intermediate (S)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-(piperidin-4-ylamino)pyrimidine-4-carboxamide (P1) obtained can be directly used in subsequent reactions.

[0140] Preparation of the key intermediate (S)-6-((4-aminocyclohexyl)amino)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)pyrimidine-4-carboxamide (P2)

[0141]

[0142] The intermediate (S)-6-chloro-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)pyrimidine-4-carboxamide (430 mg, 1.2 mmol) and N-Boc-1,4-cyclohexanediamine (532 mg, 2.5 mmol) were dissolved in n-butanol (n-BuOH, 5 mL), TEA (251 mg, 2.5 mmol) was added, and the mixture was stirred at 80°C overnight. The mixture was evaporated under reduced pressure and purified by silica gel column chromatography (DCM:MeOH=50:1) to obtain the intermediate (S)-tert-butyl (4-((6-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)carbamoyl)pyrimidin-4-yl)amino)cyclohexyl)carbamate (610 mg, 1.2 mmol, yield 97%).

[0143] Tert-butyl (S)-(4-((6-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)carbamoyl)pyrimidin-4-yl)amino)cyclohexyl)carbamate was dissolved in DCM (10 mL), and a hydrochloric acid dioxane solution (3 mL) was added, and the mixture was stirred at room temperature overnight. After the reaction was completed, the mixture was distilled under reduced pressure, and the resulting crude product was used directly as an intermediate (S)-6-((4-aminocyclohexyl)amino)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)pyrimidine-4-carboxamide without further purification and was used in the next step.

[0144]

[0145]

[0146] Preparation of the key intermediate 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetic acid

[0147]

[0148] 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (1 g, 3.6 mmol) and tert-butyl bromoacetate (854 mg, 4.4 mmol) were dissolved in DMF (5 mL), potassium carbonate (756 mg, 5.5 mmol) was added, and the mixture was stirred at room temperature overnight. The mixture was extracted with ethyl acetate (EA) and water, and the organic phase was collected, washed with saturated NaCl solution, combined, dried over anhydrous sodium sulfate, distilled under reduced pressure, and separated and purified by column chromatography (DCM: MeOH = 100: 1) to obtain tert-butyl 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)oxy)acetate (890 mg, yield 64%).

[0149] Tert-butyl 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetate (890 mg, 2.3 mmol) was dissolved in DCM (10 mL), TFA (2 mL) was added, and the mixture was stirred at room temperature overnight. After the reaction was completed, the mixture was evaporated under reduced pressure, and the crude product was used as the intermediate 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetic acid without further purification and was directly used in the next step.

[0150] Preparation of the key intermediate (2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)glycine

[0151]

[0152] 4-Amino-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (1g, 3.7mmol) and tert-butyl bromoacetate (856mg, 4.4mmol) were dissolved in DMF (6mL), potassium carbonate (760mg, 5.5mmol) was added, and stirred at 50°C. The reaction was detected by thin layer chromatography (TLC), extracted with ethyl acetate and water, the organic phase was collected, washed with saturated NaCl solution, combined with organic phases, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM: MeOH = 100: 1) gave the intermediate (2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)glycine tert-butyl ester (1.2g, yield: 84%).

[0153] The intermediate (2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)glycine tert-butyl ester (1.2 g, 3.1 mmol) was dissolved in DCM (10 mL), TFA (2 mL) was added, and the mixture was stirred at room temperature. After the reaction was completed, the mixture was distilled under reduced pressure, and the obtained crude product was used as the intermediate (2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)glycine without further purification and was directly used in the next step.

[0154] Example 1-1. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-1):

[0155]

[0156] 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (200 mg, 0.72 mmol) and tert-butyl 3-(2-aminoethoxy)propionate (205 mg, 1.09 mmol) were dissolved in dimethyl sulfoxide (DMSO) (3 mL), and N,N-diisopropylethylamine (DIEA) (187 mg, 1.45 mmol) was added. The mixture was protected with nitrogen for 3 times and stirred at 90 ° C overnight. The reaction was detected by TLC. After the reaction was completed, the mixture was extracted with ethyl acetate and water, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM:MeOH=100:1) gave the intermediate tert-butyl 3-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)propanoate (92 mg, yield 28%).

[0157] The intermediate tert-butyl 3-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)propanoate (45 mg, 0.10 mmol) was dissolved in DCM (3 mL), TFA (1 mL) was added, and the mixture was stirred at room temperature overnight. The reaction was monitored by TLC. After the reaction was completed, the mixture was evaporated under reduced pressure to obtain the intermediate 3-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)propanoic acid.

[0158] 3-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)propionic acid (0.10 mmol) and intermediate P1 (0.10 mmol) were dissolved in DMSO (2 mL), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) (50 mg, 0.13 mmol) and DIEA (40 mg, 0.31 mmol) were added in sequence, and stirred at room temperature. EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM:MeOH=50:1) gave the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (13 mg, yield 17%) (YZ-1). 1HNMR(500MHz,DMSO-d6)δ11.09(d,J=4.1Hz,1H),8.77-8.73(m,1H),8.28(s,1H),7.73(d,J=7.4Hz,1H),7.61-7.55(m,1H) ,7.14(d,J=8.7Hz,1H),7.09(d,J=6.1Hz,3H),7.06-7.00(m,3H),6.61-6.56(m,1H),5.07-5.02(m,1H),4.97(d,J=4.7Hz, 1H),4.22(d,J=12.9Hz,1H),4.07(s,1H),3.91-3.81(m,2H),3.70-3.66(m,2H),3.63-3.58(m,4H),3.49-3.40(m,3H),3.1 9-3.11(m,1H),2.91-2.65(m,7H),2.62-2.52(m,4H),2.05-1.98(m,1H),1.88(s,2H),1.40-1.20(m,4H); HRMS(ESI):calcd for C 44 H 60 N7O6[M+H] + :782.4600,found782.4614. Example 1-2. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-2):

[0159] Similar to Example 1-1, but replacing tert-butyl 3-(2-aminoethoxy)propionate with tert-butyl 3-(2-(2-aminoethoxy)ethoxy)propionate, the target product obtained by silica gel column chromatography is N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-2). 1H NMR (500MHz, DMSO-d6) δ11.09(s,1H),8.79-8.72(m,1H),8.29(s,1H),7.75(d,J=7.5Hz,1H),7.61-7.56(m,1H),7.14(d,J=8.7 Hz,1H),7.10(d,J=6.1Hz,3H),7.07-7.00(m,3H),6.63-6.59(m,1H),5.09-5.03(m,1H),4.97(d,J=4.7Hz,1H),4.22(d,J=13.1 Hz,1H),4.07(s,1H),3.92-3.87(m,1H),3.83(d,J=13.7Hz,1H),3.64-3.60(m,6H),3.58-3.55(m,2H),3.54-3.51(m,2H),3.48 -3.45(m,2H),3.17-3.10(m,1H),2.93-2.65(m,8H),2.56(d,J=7.3Hz,2H),2.06-2.00(m,1H),1.88(s,2H),1.38-1.21(m,6H).

[0160] Example 1-3. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-3):

[0161] Similar to Example 1-1, except that tert-butyl 3-(2-aminoethoxy)propionate was replaced with tert-butyl 3-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)propionate, the target product obtained by silica gel column chromatography was N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-3): 1H NMR (500MHz, DMSO-d6) δ11.09(s,1H),8.77-8.72(m,1H),8.28(s,1H),7.74(d,J=7.4Hz,1H),7.61-7.55(m,1H),7.14(d,J=8.7Hz,1H) ,7.09(d,J=6.0Hz,3H),7.06-7.00(m,3H),6.64-6.57(m,1H),5.09-5.01(m,1H),4.96(d,J=4.7Hz,1H),4.21(d,J=12.9Hz,1H),4.07(s ,1H),3.88(d,J=6.0Hz,1H),3.83(d,J=13.9Hz,1H),3.63-3.59(m,6H),3.57-3.54(m,2H),3.54-3.52(m,2H),3.51-3.46(m,6H),3.18- 3.11(m,1H),2.93-2.65(m,8H),2.56(d,J=6.3Hz,2H),2.05-1.99(m,1H),1.90(d,J=17.2Hz,2H),1.40-1.20(m,6H); HRMS(ESI):calcd for C 44 H 55 N9NaO 10 [M+Na] + :892.3964, found 892.3979.

[0162] Example 1-4. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-3,6,9,12-tetraoxopentadecane-15-yl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-4):

[0163] Similar to Example 1-1, but replacing tert-butyl 3-(2-aminoethoxy)propionate with tert-butyl 15-amino-4,7,10,13-tetraoxapentadecanoate, the target product obtained by silica gel column chromatography is N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-3,6,9,12-tetraoxapentadecan-15-yl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-4): 1HNMR(500MHz,DMSO-d6)δ11.09(s,1H),8.77-8.73(m,1H),8.31(s,1H),7.75(d,J=7.5Hz,1H),7.61-7.54(m,1H),7.1 4(d,J=8.6Hz,1H),7.11(d,J=6.5Hz,3H),7.07-7.01(m,3H),6.62-6.57(m,1H),5.09-5.00(m,2H),4.22(d,J=12.9Hz ,1H),4.08(s,1H),3.91(s,1H),3.84(d,J=13.6Hz,1H),3.63-3.59(m,4H),3.57-3.51(m,5H),3.51-3.45(m,10H),3. 20-3.11(m,1H),2.93-2.76(m,6H),2.64-2.52(m,6H),2.06-1.99(m,1H),1.90(d,J=17.1Hz,2H),1.44-1.17(m,5H).

[0164] Example 1-5. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-3,6,9,12,15-pentaoxaoctadecane-18-yl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-5):

[0165] Similar to Example 1-1, but replacing tert-butyl 3-(2-aminoethoxy)propanoate with tert-butyl 1-amino-3,6,9,12,15-pentaoxaoctadecane-18-olate, the target product obtained by silica gel column chromatography is N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-3,6,9,12,15-pentaoxaoctadecane-18-yl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-5). 1H NMR(500MHz,DMSO-d6)δ11.09(s,1H),8.76-8.73(m,1H),8.29(s,1H),7.75(d,J=7.5Hz,1H),7.61-7.55(m, 1H),7.14(d,J=8.6Hz,1H),7.09(d,J=7.1Hz,3H),7.07-7.00(m,3H),6.62-6.58(m,1H),5.08-5.02(m,1H),4 .96(s,1H),4.22(d,J=13.1Hz,1H),4.08(s,1H),3.91-3.81(m,2H),3.63-3.59(m,5H),3.56-3.52(m,4H),3. 50-3.47(m,12H),3.18-3.12(m,1H),2.92-2.53(m,12H),2.06-1.98(m,2H),1.88(s,2H),1.39-1.19(m,6H).

[0166] Example 1-6. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butyryl)piperidin-4-yl)amino)pyrimidine-4-carboxamide:

[0167] Similar to Example 1-1, but tert-butyl 3-(2-aminoethoxy)propionate was replaced with tert-butyl 4-aminobutyrate. The target product obtained by silica gel column chromatography was N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-6). 1H NMR(500MHz,DMSO-d6)δ11.01(s,1H),8.80-8.73(m,1H),8.29(s,1H),7.76(d, J=7.5Hz,1H),7.32-7.26(m,1H),7.12-7.08(m,3H),7.06(d,J=1.2Hz,1H),7.03 -6.99(m,1H),6.93(d,J=7.4Hz,1H),6.79(d,J=8.0Hz,1H),5.71-5.66(m,1H),5 .14-5.08(m,1H),4.98(d,J=4.6Hz,1H),4.23(d,J=17.2Hz,2H),4.12(d,J=17.1 Hz,2H),3.89(d,J=5.6Hz,1H),3.83(d,J=13.8Hz,1H),3.61(d,J=9.2Hz,2H),3 .45-3.39(m,1H),3.18-3.13(m,3H),2.84-2.80(m,3H),2.77-2.72(m,1H),2.71 -2.66(m,1H),2.65-2.62(m,1H),2.47-2.43(m,2H),2.32-2.26(m,1H),2.06-2. 00(m,1H),1.89(s,2H),1.83-1.79(m,2H),1.34(d,J=10.2Hz,2H),1.23(s,2H).

[0168] Example 1-7. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)pentanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-7):

[0169] Similar to Example 1-1, but tert-butyl 3-(2-aminoethoxy)propionate was replaced with tert-butyl 5-aminopentanoate. The target product obtained by silica gel column chromatography was N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)pentanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-7). 1H NMR (500MHz, DMSO-d6) δ11.11(s,1H),8.77(s,1H),8.30(s,1H),7.77(d,J=7.4Hz,1H),7.61-7.55(m,1H ),7.15-7.08(m,4H),7.08-7.01(m,3H),6.60-6.56(m,1H),5.09-4.94(m,2H),4.23(d,J=13.0Hz,1H),4. 09(s,1H),3.93-3.79(m,2H),3.62(d,J=6.7Hz,1H),3.44-3.38(m,2H),3.19-3.11(m,1H),2.89-2.77(m, 4H), 2.38 (d, J = 7.2Hz, 2H), 2.05-1.99 (m, 1H), 1.94-1.83 (m, 2H), 1.64-1.53 ​​(m, 5H), 1.39-1.14 (m, 10H).

[0170] Example 1-8. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)hexanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-8):

[0171] Similar to Example 1-1, but tert-butyl 3-(2-aminoethoxy)propionate was replaced with tert-butyl 6-aminohexanoate. The target product obtained by silica gel column chromatography was N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)hexanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-8). 1H NMR(500MHz,DMSO-d6)δ11.11(s,1H),8.77(s,1H),8.29(s,1H),7.76(s,1H),7.62-7.55(m,1H),7.13-7.0 9(m,4H),7.06(s,1H),7.04-7.01(m,2H),6.58-6.54(m,1H),5.09-5.02(m,1H),4.99(s,1H),4.25(s,1H),4 .09(s,1H),3.93-3.81(m,2H),3.67-3.55(m,2H),3.20-3.13(m,1H),2.87-2.54(m,8H),2.36-2.31(m,2H) ,2.06-1.97(m,1H),1.89(s,2H),1.63-1.51(m,3H),1.40-1.32(m,1H),1.24(s,10H).HRMS(ESI):calcdfor C 41 H 49 N9NaO7[M+Na] + :802.3647,found802.3653.

[0172] Example 1-9. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-9):

[0173] Similar to Example 1-1, but tert-butyl 3-(2-aminoethoxy)propionate was replaced with tert-butyl 7-aminoheptanoate. The target product obtained by silica gel column chromatography was N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-9). 1H NMR (500MHz, DMSO-d6) δ11.09(s,1H),8.78-8.72(m,1H),8.29(s,1H),7.75(d,J=7.4Hz,1H),7.61-7.57(m,1H),7.09(d ,J=8.4Hz,4H),7.05(s,1H),7.02(d,J=7.1Hz,2H),6.57-6.50(m,1H),5.09-5.01(m,1H),4.97(s,1H),4.23(d,J=12.9Hz ,1H),4.09(s,1H),3.95-3.78(m,3H),3.62(s,2H),3.45-3.38(m,1H),3.19-3.10(m,1H),2.89-2.72(m,6H),2.33-2.28 (m,2H),2.07-1.98(m,1H),1.88(s,3H),1.61-1.45(m,5H),1.39-1.29(m,6H),1.22(d,J=17.9Hz,4H).HRMS(ESI):calcd for C 42 H 52 N9O7[M+H] + :794.3984, found 794.3973.

[0174] Example 1-10. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-10):

[0175] Similar to Example 1-1, but replacing tert-butyl 3-(2-aminoethoxy)propionate with tert-butyl 8-aminooctanoate, the target product obtained by silica gel column chromatography is N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-10). 1H NMR (500MHz, DMSO-d6) δ11.09(s,1H),8.76(d,J=6.5Hz,1H),8.30(s,1H),7.75(d,J=7.5Hz,1H),7.61-7.56(m,1H),7.13-7. 08(m,4H),7.07-7.00(m,3H),6.55-6.51(m,1H),5.08-5.01(m,1H),4.97(s,1H),4.23(d,J=13.1Hz,1H),4.09(s,1H),3.93- 3.78(m,2H),3.62(s,2H),3.42(d,J=13.6Hz,1H),3.19-3.11(m,1H),2.92-2.66(m,4H),2.58(d,J=19.2Hz,1H),2.33-2.27( m,2H),2.06-1.99(m,1H),1.91(d,J=1.7Hz,4H),1.59-1.56(m,2H),1.52-1.46(m,2H),1.38-1.20(m,14H).HRMS(ESI):calcd for C 43 H 54 N9O7[M+H] + :808.4141, found 808.4164.

[0176] Example 1-11. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(9-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)nonanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-11):

[0177] Similar to Example 1-1, but replacing tert-butyl 3-(2-aminoethoxy)propionate with tert-butyl 9-aminononanoate, the target product obtained by silica gel column chromatography is N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(9-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)nonanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-11). 1H NMR(500MHz,DMSO-d6)δ11.05(s,1H),8.78-8.72(m,1H),8.29(s,1H),7.75(d,J=7.5Hz,1H),7.55(d,J=8.4Hz,1H),7 .14-6.99(m,7H),6.93(d,J=2.1Hz,1H),6.86-6.81(m,1H),5.05-4.94(m,2H),4.23(d,J=13.0Hz,1H),3.89(s,1H),3 .82(d,J=13.7Hz,1H),3.62(s,2H),3.46-3.39(m,1H),3.15(d,J=5.8Hz,2H),2.88-2.73(m,6H),2.32-2.27(m,2H),2 .01-1.96(m,1H),1.87(d,J=18.1Hz,2H),1.59-1.54(m,2H),1.47(s,2H),1.34(d,J=7.8Hz,2H),1.30-1.20(m,14H).

[0178] Example 1-12. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-12):

[0179] Similar to Example 1-1, but replacing tert-butyl 3-(2-aminoethoxy)propionate with tert-butyl 10-aminodecanoate, the target product obtained by silica gel column chromatography is N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-12). 1H NMR (500MHz, DMSO-d6) δ11.08(s,1H),8.78-8.72(m,1H),8.29(s,1H),7.75(d,J=7.5Hz,1H),7.62-7.54(m,1H),7.14-7.07(m, 4H),7.05(s,1H),7.01(d,J=7.0Hz,2H),6.55-6.49(m,1H),5.09-4.93(m,2H),4.24(s,1H),4.09(s,1H),3.89(s,1H),3.82(d, J=13.6Hz,1H),3.62(s,2H),3.42(d,J=13.4Hz,2H),3.29(d,J=6.4Hz,2H),3.19-3.10(m,1H),2.93-2.72(m,6H),2.60(s,1H), 2.56(s,1H),2.32-2.25(m,2H),2.07-1.99(m,1H),1.60-1.54(m,2H),1.50-1.44(m,2H),1.36-1.23(m,16H).HRMS(ESI):calcd for C 45 H 58 N9O7[M+H] + :836.4454, found 836.4479.

[0180] Example 1-13. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-13):

[0181]

[0182] 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (200 mg, 0.72 mmol) and tert-butyl 3-(2-aminoethoxy)propionate (205 mg, 1.09 mmol) were dissolved in DMSO (3 mL), DIEA (187 mg, 1.45 mmol) was added, nitrogen was protected for 3 times, and stirred at 90 ° C overnight. TLC was used to detect the reaction. After the reaction was completed, ethyl acetate and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phase was combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM: MeOH = 100: 1) gave the intermediate tert-butyl 3-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)amino)ethoxy)propionate (92 mg, yield 28%).

[0183] The intermediate tert-butyl 3-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)ethoxy)propanoate (45 mg, 0.10 mmol) was dissolved in DCM (3 mL), TFA (1 mL) was added, and the mixture was stirred at room temperature overnight. The reaction was monitored by TLC. After the reaction was completed, the mixture was evaporated under reduced pressure to obtain the intermediate 3-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)ethoxy)propanoic acid.

[0184] 3-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)ethoxy)propionic acid (0.10 mmol) and intermediate P1 (0.10 mmol) were dissolved in DMSO (2 mL), HATU (50 mg, 0.13 mmol) and DIEA (40 mg, 0.31 mmol) were added in sequence, and stirred at room temperature. EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM:MeOH=50:1) gave the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (12 mg, yield 15%) (YZ-13). 1H NMR (500MHz, DMSO-d6) δ11.05(s,1H),8.77-8.73(m,1H),8.29(s,1H),7.74(d,J=7.5Hz,1H),7.56(d,J=8.4Hz,1H),7.15-7. 11(m,1H),7.09(d,J=6.3Hz,3H),7.06-7.00(m,3H),6.92-6.87(m,1H),5.06-5.00(m,1H),4.97(s,1H),4.23(d,J=13.1Hz,1H ),4.12-4.03(m,1H),3.89(d,J=6.5Hz,1H),3.83(d,J=13.4Hz,1H),3.69-3.65(m,2H),3.63-3.58(m,4H),3.20-3.14(m,1H) ,2.92-2.79(m,5H),2.72(d,J=27.5Hz,2H),2.63-2.54(m,5H),2.02-1.96(m,1H),1.90(d,J=13.3Hz,2H),1.42-1.20(m,6H).

[0185] Example 1-14. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)ethoxy)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-14):

[0186] Similar to Example 1-13, but tert-butyl 3-(2-aminoethoxy)propionate was replaced with tert-butyl 3-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)propionate. The target product obtained by silica gel column chromatography was N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)ethoxy)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-14). 1H NMR(500MHz,DMSO-d6)δ11.09(s,1H),8.78-8.72(m,1H),8.29(s,1H),7.74(d,J=7.5Hz,1H),7.60-7.55(m,1H),7.14 (d,J=8.6Hz,1H),7.11-7.08(m,3H),7.07-7.00(m,3H),6.63-6.57(m,1H),5.09-5.02(m,1H),4.97(s,1H),4.21(d,J= 11.1Hz,1H),4.07(s,1H),3.92-3.86(m,1H),3.83(d,J=13.8Hz,1H),3.64-3.58(m,6H),3.57-3.52(m,4H),3.52-3.3 8(m,9H),3.18-3.10(m,1H),2.86-2.72(m,5H),2.58-2.53(m,3H),2.06-1.98(m,1H),1.88(s,2H),1.40-1.20(m,5H).

[0187] Example 1-15. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-15):

[0188] Similar to Example 1-13, but tert-butyl 3-(2-aminoethoxy)propionate was replaced with tert-butyl 7-aminoheptanoate. The target product obtained by silica gel column chromatography was N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-15). 1H NMR (500MHz, DMSO-d6) δ11.05(d,J=2.7Hz,1H),8.75(d,J=6.4Hz,1H),8.29(d,J=2.7Hz,1H),7.75(d,J=7.5Hz,1H), 7.58-7.53(m,1H),7.13-6.99(m,7H),6.94(s,1H),6.84(d,J=8.2Hz,1H),5.06-4.99(m,1H),4.97(s,1H),4.23(s,1H ),4.09(s,1H),3.89(s,1H),3.82(d,J=13.3Hz,1H),3.61(d,J=8.8Hz,2H),3.42(d,J=13.5Hz,1H),3.17-3.13(m,4H ),2.85-2.73(m,4H),2.34-2.29(m,2H),2.02-1.84(m,5H),1.60-1.46(m,5H),1.39-1.32(m,4H),1.27-1.22(m,4H).

[0189] Example 1-16. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-16):

[0190] Similar to Example 1-13, but tert-butyl 3-(2-aminoethoxy)propionate was replaced with tert-butyl 8-aminooctanoate. The target product obtained by silica gel column chromatography was N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-16). 1H NMR(500MHz,DMSO-d6)δ11.05(s,1H),8.77-8.73(m,1H),8.29(s,1H),7.75(d,J=7.4Hz, 1H),7.55(d,J=8.3Hz,1H),7.09(d,J=6.0Hz,4H),7.05(d,J=1.2Hz,1H),7.01(d,J=6.5H z,1H),6.94(d,J=2.2Hz,1H),6.86-6.82(m,1H),5.06-5.00(m,1H),4.97(s,1H),4.23(d ,J=12.6Hz,1H),3.89(d,J=5.3Hz,1H),3.82(d,J=13.5Hz,1H),3.67-3.56(m,2H),3.45-3 .39(m,1H),3.29(s,1H),3.15(d,J=5.4Hz,2H),2.89-2.65(m,7H),2.57(d,J=18.9Hz,2H ),2.33-2.28(m,2H),2.01-1.96(m,1H),1.88(d,J=24.9Hz,2H),1.60-1.46(m,5H),1.39 -1.27 (m, 8H), 1.23 (s, 2H). Example 1-17. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(9-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)nonanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-17):

[0191] Similar to Example 1-13, but tert-butyl 3-(2-aminoethoxy)propionate was replaced with tert-butyl 9-aminononanoate. The target product obtained by silica gel column chromatography was N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(9-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)nonanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-17). 1H NMR (500MHz, DMSO-d6) δ11.05(s,1H),8.77-8.72(m,1H),8.29(s,1H),7.75(d,J=7.5Hz,1H),7.55(d,J=8.3Hz,1H),7.09(d,J=5 .4Hz,4H),7.05(s,1H),7.01(d,J=7.2Hz,1H),6.94(s,1H),6.86-6.81(m,1H),5.05-5.00(m,1H),4.97(s,1H),4.23(d,J=12.7H z,1H),3.93-3.87(m,1H),3.82(d,J=13.5Hz,1H),3.62(s,2H),3.16-3.11(m,3H),2.89-2.72(m,6H),2.32-2.27(m,2H),2.02-1 .96(m,1H),1.89(d,J=12.3Hz,3H),1.60-1.52(m,3H),1.52-1.44(m,3H),1.36(d,J=8.8Hz,4H),1.32-1.26(m,8H),1.23(s,2H).

[0192] Example 1-18. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-18):

[0193] Similar to Example 1-13, but tert-butyl 3-(2-aminoethoxy)propionate was replaced with tert-butyl 10-aminodecanoate. The target product obtained by silica gel column chromatography was N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-18). 1H NMR(500MHz,DMSO-d6)δ11.05(s,1H),8.77-8.73(m,1H),8.29(d,J=1.2Hz,1H), 7.75(d,J=7.4Hz,1H),7.55(d,J=8.4Hz,1H),7.11-7.07(m,4H),7.05(d,J=1.2Hz ,1H),7.01(d,J=6.4Hz,1H),6.93(d,J=2.1Hz,1H),6.85-6.82(m,1H),5.02(dd,J =12.7,5.4Hz,1H),4.96(d,J=4.7Hz,1H),4.23(d,J=13.1Hz,1H),4.12-4.06(m,2 H),3.91-3.86(m,1H),3.82(d,J=13.5Hz,1H),3.65-3.57(m,2H),3.46-3.39(m,1 H),3.17(d,J=5.2Hz,2H),3.14(d,J=5.9Hz,2H),2.84-2.81(m,2H),2.79-2.72(m ,2H),2.71-2.66(m,1H),2.29(t,J=7.4Hz,2H),2.03-1.95(m,1H),1.95-1.83(m, 2H),1.60-1.53(m,2H),1.51-1.45(m,2H),1.39-1.21(m,16H); HRMS(ESI):calcd for C 45 H 58 N9O7[M+H] + :836.4454,found 836.4456.

[0194] Example 1-19. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(11-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)undecanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-19):

[0195] Similar to Example 1-13, but tert-butyl 3-(2-aminoethoxy)propionate was replaced with tert-butyl 11-aminoundecanoate. The target product obtained by silica gel column chromatography was N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(11-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)undecanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-19). 1H NMR (500MHz, DMSO-d6) δ11.05 (s, 1H), 8.77-8.72 (m, 1H), 8.29 (d, J = 1.2Hz, 1H),7.75(d,J=7.4Hz,1H),7.55(d,J=8.4Hz,1H),7.11-7.07(m,4H),7.05(d ,J=1.2Hz,1H),7.03-6.99(m,1H),6.93(d,J=2.2Hz,1H),6.85-6.82(m,1H) ,5.05-4.99(m,1H),4.96(d,J=4.8Hz,1H),4.23(d,J=13.0Hz,1H),4.12-4.0 4(m,1H),3.91-3.86(m,1H),3.82(d,J=13.7Hz,1H),3.45-3.39(m,1H),3.1 8-3.11(m,4H),2.84-2.81(m,2H),2.76-2.72(m,1H),2.71-2.66(m,1H),2.3 1-2.27(m,2H),2.01-1.96(m,1H),1.87(d,J=17.1Hz,3H),1.59-1.54(m,2H) ,1.47(s,3H),1.38-1.29(m,6H),1.25(d,J=15.3Hz,14H); HRMS(ESI):calcd for C 46 H 60 N9O7[M+H] + :850.4610, found 850.4613. Example 1-20. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(12-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)dodecanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-20):

[0196] Similar to Example 1-13, but tert-butyl 3-(2-aminoethoxy)propionate was replaced with tert-butyl 12-aminododecanoate. The target product obtained by silica gel column chromatography was N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(12-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)dodecanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-20). 1H NMR (500MHz, DMSO-d6) δ11.05(s,1H),8.77-8.72(m,1H),8.29(s,1H),7.75(d,J=7.5Hz,1H),7.55(d,J=8.4Hz,1H),7.14-6.99(m,6H) ,6.93(d,J=2.1Hz,1H),6.86-6.81(m,1H),5.06-4.94(m,2H),4.23(d,J=12.7Hz,1H),4.09(s,1H),3.89(s,1H),3.82(d,J=13.5Hz,1H) ,3.62(s,2H),3.41(d,J=6.0Hz,1H),3.29(s,1H),3.14(d,J=6.4Hz,2H),2.84(d,J=6.3Hz,2H),2.77(d,J=13.2Hz,2H),2.69(s,1H),2 .59(d,J=3.9Hz,1H),2.55(d,J=4.9Hz,1H),2.32-2.26(m,2H),2.01-1.96(m,1H),1.60-1.52(m,2H),1.47(s,2H),1.39-1.21(m,20H).

[0197] Example 1-21. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((2-(2,6-dioxopiperidin-3-yl)-6-fluoro-1,3-dioxoisoindolin-5-yl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-21):

[0198]

[0199] 2-(2,6-dioxopiperidin-3-yl)-5,6-difluoroisoindoline-1,3-dione (300 mg, 1.0 mmol) and 10-aminodecanoic acid (230 mg, 1.2 mmol) were dissolved in DMSO, DIEA (263 mg, 2.0 mmol) was added, nitrogen was protected, and stirred at 90 ° C for 6 h. TLC was used to detect the reaction. After the reaction was completed, EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phase was combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM: MeOH = 100: 1) gave the intermediate 10-((2-(2,6-dioxopiperidin-3-yl)-6-fluoro-1,3-dioxoisoindoline-5-yl)amino)decanoic acid (90 mg, yield 20%).

[0200] The intermediate 10-((2-(2,6-dioxopiperidin-3-yl)-6-fluoro-1,3-dioxoisoindolin-5-yl)amino)decanoic acid (90 mg, 0.20 mmol) and P1 (80 mg, 0.20 mmol) were dissolved in DMSO (2 mL), and HATU (96 mg, 0.25 mmol) and DIEA (75 mg, 0.59 mmol) were added in sequence and stirred at room temperature. After the reaction was completed, EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM:MeOH=100:1) gave the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((2-(2,6-dioxopiperidin-3-yl)-6-fluoro-1,3-dioxoisoindolin-5-yl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-21) (23 mg, yield 13%). 1 H NMR(500MHz,DMSO-d6)δ11.07(s,1H),8.77-8.72(m,1H),8.29(d,J=1.1Hz ,1H),7.75(d,J=7.6Hz,1H),7.55(d,J=10.4Hz,1H),7.12-7.03(m,5H),7.0 3-6.99(m,1H),6.92-6.87(m,1H),5.08-5.02(m,1H),4.97(d,J=4.7Hz,1H ),4.23(d,J=12.9Hz,1H),4.09(d,J=5.2Hz,1H),3.88(d,J=5.8Hz,1H),3.8 2(d,J=13.3Hz,1H),3.66-3.56(m,2H),3.46-3.38(m,1H),3.28-3.22(m,2 H),3.19-3.10(m,3H),2.84-2.81(m,2H),2.79-2.65(m,3H),2.61-2.55(m, 1H),2.32-2.26(m,2H),2.04-1.97(m,1H),1.89(s,2H),1.61-1.53(m,2H) ,1.47(s,3H),1.36-1.29(m,5H),1.25(d,J=18.0Hz,9H).HRMS(ESI):calcd for C 45 H 57 FN9O7[M+H] + :854.4359, found 854.4364.

[0201] Example 1-22. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-22):

[0202]

[0203] 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (200 mg, 0.73 mmol) and 8-bromooctanoic acid tert-butyl ester (244 mg, 0.88 mmol) were dissolved in DMF (3 mL), DIEA (190 mg, 1.5 mmol) was added, nitrogen was protected for 3-4 times, and stirred at 90 ° C overnight. After the reaction was completed, EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phase was combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM: MeOH = 100: 1) gave the intermediate 8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)oxy)octanoic acid tert-butyl ester (120 mg, yield 35%).

[0204] The intermediate tert-butyl 8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)octanoate was dissolved in DCM (5 mL), TFA (1 mL) was added, and the mixture was stirred at room temperature overnight. The reaction was monitored by TLC. After the reaction was completed, the mixture was evaporated under reduced pressure, and the obtained crude product 8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)octanoic acid was directly used in the next step.

[0205] 8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)octanoic acid (54 mg, 0.12 mmol) and intermediate P1 (60 mg, 0.12 mmol) were dissolved in DMSO (2 mL), HATU (60 mg, 0.16 mmol) and DIEA (50 mg, 0.36 mmol) were added in sequence, and stirred at room temperature. After the reaction was completed, EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM:MeOH=50:1) gave the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-22) (12 mg, yield 12%). 1 H NMR (500MHz, DMSO-d6) δ11.09(s,1H),8.79-8.71(m,1H),8.29(d,J=1.1Hz,1H),7.83-7.78(m,1H),7.75(d,J=7.4Hz,1H),7.51(d,J=8.6Hz, 1H),7.43(d,J=7.2Hz,1H),7.14-6.99(m,6H),5.11-5.05(m,1H),4.96(d,J=4.8Hz,1H),4.23-4.18(m,2H),4.12-4.05(m,1H),3.92-3.86(m ,1H),3.82(d,J=13.4Hz,1H),3.61(d,J=8.8Hz,2H),3.45-3.40(m,1H),3.18-3.11(m,1H),2.86-2.80(m,3H),2.77-2.65(m,3H),2.62-2.55 (m,2H),2.34-2.27(m,2H),2.07-2.00(m,1H),1.89(s,2H),1.79-1.72(m,2H),1.54-1.42(m,5H),1.39-1.28(m,6H),1.24(d,J=7.2Hz,2H).

[0206] Example 1-23. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)octyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-23):

[0207]

[0208] 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (200 mg, 0.72 mmol) and 8-amino-1-octanol (137 mg, 0.94 mmol) were dissolved in DMSO (2 mL), DIEA (280 mg, 2.2 mmol) was added, nitrogen was protected, and stirred at 90 ° C overnight. TLC was used to detect the reaction. After the reaction was completed, ethyl acetate and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phase was combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM: MeOH = 100: 1) gave the intermediate 2-(2,6-dioxopiperidin-3-yl)-4-((8-hydroxyoctyl)amino)isoindoline-1,3-dione (169 mg, yield: 58%).

[0209] Under ice bath conditions, the intermediate 2-(2,6-dioxopiperidin-3-yl)-4-((8-hydroxyoctyl)amino)isoindoline-1,3-dione (169 mg, 0.42 mmol) and p-toluenesulfonyl chloride (240 mg, 1.3 mmol) were dissolved in DCM (10 mL), TEA (140 mg, 1.3 mmol) was added, and stirred overnight. TLC was used to detect the reaction. After the reaction was completed, DCM and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM: MeOH = 100: 1) gave the intermediate 8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)amino)octyl 4-methylbenzenesulfonate (190 mg, yield: 82%).

[0210] 8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)octyl 4-methylbenzenesulfonate (140 mg, 0.35 mmol) and intermediate P1 (110 mg, 0.27 mmol) were dissolved in DMF (3 mL), DIEA (103 mg, 0.80 mmol) was added, and stirred at 90 ° C overnight. After the reaction was completed, ethyl acetate and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM:MeOH=50:1) gave N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)octyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-23) (32 mg, yield: 12%). 1H NMR(500MHz,DMSO-d6)δ11.10(s,1H),8.75(s,1H),8.28(s,1H),7.74(s,1H),7. 61-7.55(m,1H),7.12-7.00(m,7H),6.55-6.51(m,1H),5.08-5.01(m,1H),4.98( s,1H),3.88(s,2H),3.61(d,J=8.8Hz,2H),2.92-2.81(m,3H),2.72(d,J=31.5Hz ,1H),2.08-1.97(m,1H),1.56(d,J=7.3Hz,2H),1.46(s,7H),1.38-1.19(m,22H).

[0211] Example 1-24. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)octyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-24):

[0212] Similar to Example 1-23, but replacing 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione with 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione, the target product obtained by silica gel column chromatography is N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)oxy)octyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-24). 1H NMR (500MHz, DMSO-d6) δ11.11(s,1H),8.76(s,1H),8.31(s,1H),7.84-7.79(m,2H),7.52(d,J=8.5 Hz,1H),7.45(d,J=7.3Hz,1H),7.09(d,J=22.1Hz,4H),7.03(s,1H),5.11-5.06(m,1H),4.98(s,1H) ,4.23-4.19(m,2H),3.90(s,1H),3.63(s,2H),3.46-3.39(m,2H),2.93-2.87(m,2H),2.87-2.80(m, 3H),2.07-1.94(m,5H),1.80-1.73(m,3H),1.46(d,J=8.5Hz,4H),1.40-1.28(m,10H),1.24(s,6H).

[0213] Example 1-25. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)octyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-25):

[0214] Similar to Example 1-23, but replacing 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione with 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione, the target product obtained by silica gel column chromatography is N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)amino)octyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-25). 1H NMR (500MHz, DMSO-d6) δ11.05(s,1H),8.75-8.70(m,1H),8.28(s,1H),7.70(d,J=7.5Hz,1H),7.55(d,J=8.4Hz,1H),7.11-7.08(m,4H),7. 05(s,1H),7.02-6.99(m,1H),6.94(d,J=2.2Hz,1H),6.86-6.82(m,1H),5.06-4.99(m,1H),4.96(d,J=4.7Hz,1H),3.90-3.86(m,1H),3.82( s,1H),3.61(d,J=8.3Hz,2H),3.45-3.39(m,1H),3.17-3.13(m,2H),2.92-2.85(m,1H),2.86-2.80(m,4H),2.77-2.64(m,3H),2.60-2.53( m,2H),2.30-2.22(m,2H),2.02-1.94(m,3H),1.85(s,2H),1.59-1.54(m,2H),1.45-1.39(m,4H),1.36(d,J=8.0Hz,2H),1.33-1.24(m,8H).

[0215] Example 1-26. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)pentyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-26):

[0216]

[0217] 3-(4-amino-1-oxoisoindolin-2-yl)piperidine-2,6-dione (200 mg, 0.77 mmol) and 1,5-dibromopentane (355 mg, 1.5 mmol) were dissolved in N-methylpyrrolidone (NMP) (3 mL), DIEA (300 mg, 2.3 mmol) was added, nitrogen was protected for 3-4 times, and stirred at 90 ° C overnight. TLC was used to detect the reaction. After the reaction was completed, ethyl acetate and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phase was combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM: MeOH = 100: 1) gave the intermediate 3-(4-((5-bromopentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (250 mg, yield: 80%).

[0218] The intermediate 3-(4-((5-bromopentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (50 mg, 0.12 mmol) and the intermediate P1 (50 mg, 0.12 mmol) were dissolved in DMF (2 mL), TEA (99 mg, 0.98 mmol) was added, nitrogen was protected for 3-4 times, and stirred at 90 ° C overnight. TLC was used to monitor the reaction. After the reaction was completed, ethyl acetate and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM:MeOH=50:1) gave N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)pentyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-26) (11 mg, yield: 12%). 1 H NMR (500MHz, DMSO-d6) δ11.10(s,1H),8.76(s,1H),8.30(s,1H),7.85-7.78(m,2H),7.53(d,J=8.6Hz,1H),7. 45(d,J=7.2Hz,1H),7.09(d,J=20.4Hz,5H),7.02(s,1H),5.10-5.05(m,1H),4.98(s,1H),4.25-4.20(m,2H),4 .12-4.06(m,1H),3.90(s,1H),3.62(s,2H),3.46-3.39(m,2H),3.17(d,J=5.0Hz,1H),2.93-2.80(m,5H),2.74 (s,4H),2.58(d,J=18.7Hz,3H),2.06-1.98(m,3H),1.86-1.76(m,4H),1.69(s,2H),1.49(s,4H),1.23(s,2H).

[0219] Example 1-27. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-27):

[0220]

[0221] 3-(4-amino-1-oxoisoindolin-2-yl)piperidine-2,6-dione (200 mg, 0.77 mmol) and 8-bromooctanoic acid tert-butyl ester (280 mg, 1.0 mmol) were dissolved in NMP (2 mL), DIEA was added, nitrogen protection was applied 3-4 times, and the mixture was stirred at 90 ° C overnight. The reaction was detected by TLC. After the reaction was completed, ethyl acetate and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phase was combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography was separated and purified (DCM: MeOH = 100: 1) to obtain the intermediate 8-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)octanoic acid tert-butyl ester (200 mg, yield: 57%).

[0222] The intermediate tert-butyl 8-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)octanoate (65 mg, 0.14 mmol) was dissolved in DCM (3 mL), TFA (1 mL) was added, and the mixture was stirred at room temperature overnight. After the reaction was completed, the mixture was evaporated under reduced pressure, and the crude product 8-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)octanoic acid was directly used in the next step.

[0223] 8-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)octanoic acid (0.14 mmol) and intermediate P1 (58 mg, 0.14 mmol) were dissolved in DMSO (2 mL), HATU (70 mg, 0.18 mmol) and DIEA (55 mg, 0.43 mmol) were added in sequence, and stirred at room temperature. After the reaction was completed, ethyl acetate and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM:MeOH=50:1) gave N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-27) (17 mg, yield: 15%). 1H NMR (500MHz, DMSO-d6) δ11.00(s,1H),8.77-8.73(m,1H),8.29(s,1H),7.75(d,J=7.6Hz,1H),7.31-7.24(m,1H),7.16-6.84( m,7H),6.74(d,J=8.1Hz,1H),5.57-5.52(m,1H),5.14-5.07(m,1H),4.97(s,1H),4.30-4.05(m,3H),3.95-3.76(m,3H),3.68- 3.56(m,2H),3.45-3.39(m,1H),3.15-3.08(m,4H),2.97-2.86(m,1H),2.82(d,J=6.3Hz,2H),2.77(d,J=12.9Hz,1H),2.71-2 .58(m,1H),2.36-2.23(m,4H),2.10-1.99(m,1H),1.91(d,J=19.1Hz,3H),1.64-1.44(m,6H),1.35-1.29(m,4H),1.23(s,3H).

[0224] Example 1-28. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-28):

[0225] Similar to Example 1-27, but replacing tert-butyl 8-bromooctanoate with tert-butyl 3-(2-(2-bromoethoxy)ethoxy)propanoate, the target product obtained by silica gel column chromatography is N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-28). 1H NMR(500MHz,DMSO-d6)δ11.00(s,1H),10.00(s,1H),9.44(s,1H),8.77(s,1H),7.80(s,1H),7.31-7.05(m,6H),6.9 4(d,J=7.4Hz,1H),6.80(d,J=8.1Hz,1H),5.60-5.55(m,1H),5.11(dd,J=13.3,5.0Hz,1H),4.23(d,J=17.1Hz,2H), 4.16-4.06(m,2H),3.83(d,J=13.3Hz,1H),3.66-3.57(m,4H),3.56-3.50(m,3H),3.40(s,2H),3.13-3.04(m,6H),2 .96-2.87(m,2H),2.84-2.77(m,1H),2.56(d,J=6.0Hz,6H),2.03(d,J=13.4Hz,1H),1.89(s,2H),1.40-1.22(m,6H).

[0226] Example 1-29. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-29):

[0227] Similar to Example 1-27, but replacing tert-butyl 8-bromooctanoate with tert-butyl 3-(2-(2-(2-bromoethoxy)ethoxy)ethoxy)propanoate, the target product obtained by silica gel column chromatography is N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-29). 1HNMR(500MHz,DMSO-d6)δ11.00(s,1H),8.75(s,1H),8.29(s,1H),7.75(d,J=7.4Hz,1H),7.30-7.25(m,1H),7.14-6.99(m,6H),6.9 4(d,J=7.3Hz,1H),6.80(d,J=8.1Hz,1H),5.59-5.55(m,1H),5.14-5.08(m,1H),4.96(s,1H),4.23(d,J=17.1Hz,2H),4.12(d,J=17 .0Hz,2H),3.93-3.79(m,3H),3.45-3.38(m,2H),3.17(d,J=5.2Hz,1H),3.13(d,J=12.5Hz,1H),2.96-2.87(m,2H),2.81(d,J=14.4 Hz,3H),2.78-2.65(m,3H),2.62(s,1H),2.60-2.54(m,3H),2.06-1.99(m,2H),1.88(s,3H),1.43-1.18(m,14H); HRMS(ESI):calcd forC 44 H 57 N9NaO9[M+Na] + :878.4171, found 878.4173.

[0228] Example 1-30. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)pentanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-30):

[0229] Similar to Example 1-27, but replacing 8-bromooctanoic acid tert-butyl ester with 5-bromovalerate, the target product obtained by silica gel column chromatography is N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)pentanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-30). 1H NMR(500MHz,DMSO-d6)δ11.00(s,1H),8.78-8.72(m,1H),8.29(s,1H),7.75(d,J=7.5Hz,1H),7.31-7.26(m,1H),7.13-6.99(m,6H) ,6.93(d,J=7.4Hz,1H),6.79(d,J=8.1Hz,1H),5.69-5.64(m,1H),5.14-5.08(m,1H),4.97(s,1H),4.23(d,J=17.1Hz,2H),4.12(d,J =17.1Hz,2H),3.94-3.79(m,3H),3.66-3.56(m,2H),3.45-3.39(m,1H),3.19-3.11(m,3H),2.86-2.78(m,4H),2.71(d,J=27.6Hz,3H ),2.60(d,J=3.8Hz,1H),2.32-2.26(m,1H),2.07-1.99(m,2H),1.89(s,2H),1.84-1.78(m,2H),1.34(d,J=7.5Hz,2H),1.23(s,3H).

[0230] Example 1-31. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(6-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)hexanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-31):

[0231] Similar to Example 1-27, but replacing 8-bromooctanoic acid tert-butyl ester with 6-bromohexanoic acid tert-butyl ester, the target product obtained by silica gel column chromatography is N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(6-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)hexanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-31). 1H NMR(500MHz,DMSO-d6)δ11.03(s,1H),10.07(s,2H),8.77(d,J=6.4Hz,1H),8.38(s,1H),7.87(s,1H),7.32-7.23(m,1H),7.23-7 .12(m,3H),7.08(s,2H),6.92(d,J=7.4Hz,1H),6.74(d,J=8.0Hz,1H),5.63-5.58(m,1H),5.15-5.08(m,1H),4.23(d,J=17.3Hz,3 H),4.12(d,J=17.1Hz,3H),3.83(d,J=13.4Hz,1H),3.20-3.08(m,5H),2.97-2.88(m,3H),2.83-2.75(m,2H),2.61(d,J=17.8Hz, 1H),2.38-2.23(m,4H),2.13-2.07(m,1H),2.03(d,J=12.7Hz,1H),1.89(d,J=18.3Hz,2H),1.63-1.50(m,5H),1.42-1.30(m,4H).

[0232] Example 1-32. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyryl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-32):

[0233] Similar to Example 1-27, but replacing tert-butyl 8-bromooctanoate with tert-butyl 4-bromobutyrate, the target product obtained by silica gel column chromatography is N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-32). 1H NMR (500MHz, DMSO-d6) δ11.02(s,1H),8.80-8.74(m,1H),8.30(s,1H),7.77(d,J=7.5Hz,1H),7.32-7.26(m,1H),7.10(d,J=6.9Hz,3H),7.06(s ,1H),7.02(d,J=7.1Hz,1H),6.93(d,J=7.4Hz,1H),6.76(d,J=8.1Hz,1 H),5.64-5.58(m,1H),5.16-5.08(m,1H),4.99(s,1H),4.23(d,J=17.1H z,2H),4.17-4.08(m,2H),3.90(s,1H),3.85(d,J=13.6Hz,1H),3.62(s ,1H),3.43(d,J=13.5Hz,1H),3.31(s,2H),3.20-3.12(m,4H),2.98-2.8 7(m,2H),2.83(s,2H),2.61(d,J=17.8Hz,2H),2.37(s,2H),2.35-2.24( m,2H),2.07-2.00(m,1H),1.89(s,1H),1.64-1.58(m,4H),1.24(s,2H).

[0234] Example 1-33. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-33):

[0235] Similar to Example 1-27, but replacing tert-butyl 8-bromooctanoate with tert-butyl 10-bromodecanoate, the target product obtained by silica gel column chromatography is N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-33). 1H NMR (500MHz, DMSO-d6) δ11.00(s,1H),8.78-8.71(m,1H),8.29(s,1H),7.75(d,J=7.4Hz,1H),7.31-7.24(m,1H),7.13-6.98(m,6H),6. 92(d,J=7.4Hz,1H),6.73(d,J=8.0Hz,1H),5.57-5.52(m,1H),5.15-5.07(m,1H),4.96(d,J=4.7Hz,1H),4.22(d,J=17.2Hz,2H),4.12(d ,J=17.1Hz,2H),3.92-3.78(m,3H),3.61(d,J=8.8Hz,2H),3.13-3.08(m,2H),2.86-2.80(m,3H),2.80-2.65(m,4H),2.32-2.27(m,3H) ,2.07-2.00(m,1H),1.89(d,J=18.4Hz,2H),1.60-1.55(m,2H),1.50-1.45(m,2H),1.40-1.32(m,4H),1.32-1.25(m,10H),1.23(s,2H).

[0236] Example 1-34. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oct-7-ynyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-34):

[0237]

[0238] The compound 3-(4-bromo-1-oxyylidene-2,3-dihydro-1H-isoindol-2-yl)piperidin-2,6-dione (500 mg, 1.5 mmol), 7-octynoic acid (260 mg, 1.9 mmol) and cuprous iodide (CuI) (60 mg, 0.31 mmol) were dissolved in anhydrous DMF (2 mL), nitrogen protection was applied 3-4 times, triethylamine (470 mg, 4.6 mmol) was added, nitrogen protection was applied again, catalyst bis(triphenylphosphine)palladium dichloride (109 mg, 0.15 mmol) was added, nitrogen protection was applied again three times, and then heated and stirred at 90°C overnight. The mixture was distilled under reduced pressure, filtered with EA, extracted with EA and water, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, and dried over anhydrous sodium sulfate. The intermediate 8-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oct-7-ynoic acid was obtained by column chromatography separation and purification (DCM:MeOH=80:1).

[0239] 8-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oct-7-ynoic acid (64 mg, 0.17 mmol) and intermediate P1 (85 mg, 0.17 mmol) were dissolved in DMSO (2 mL), HATU (83 mg, 0.22 mmol) and DIEA (65 mg, 0.50 mmol) were added in sequence, and stirred at room temperature. EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM:MeOH=50:1) gave the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oct-7-ynyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-34) (24 mg, yield: 20%). 1 H NMR (500MHz, DMSO-d6) δ10.99(s,1H),8.78-8.72(m,1H),8.30(s,1H),7.75(d,J=7.4Hz,1H),7.70(d,J=7.5Hz,1H),7.63(d,J=7.6Hz, 1H),7.55-7.48(m,1H),7.10(s,3H),7.06(s,1H),7.02(s,1H),5.17-5.11(m,1H),4.97(s,1H),4.46(d,J=17.7Hz,1H),4.32(d,J=17. 7Hz,1H),4.23(s,1H),4.08(s,1H),3.89(s,1H),3.83(d,J=13.6Hz,1H),3.62(s,2H),3.44-3.39(m,1H),3.18-3.12(m,1H),2.89-2.6 6(m,7H),2.36-2.32(m,2H),2.05-1.97(m,2H),1.89(s,3H),1.61-1.53(m,4H),1.48-1.43(m,2H),1.33(s,2H),1.24(d,J=7.0Hz,4H).

[0240] Example 1-35. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(11-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)undec-10-ynyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-35):

[0241]

[0242] The compound 3-(5-bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione (300 mg, 0.93 mmol), 10-undecynoic acid (340 mg, 1.9 mmol) and CuI (35 mg, 0.19 mmol) were dissolved in DMF (5 mL), nitrogen protection was applied 3-4 times, triethylamine (375 mg, 3.7 mmol) was added, nitrogen protection was applied again, catalyst bis(triphenylphosphine)palladium dichloride (260 mg, 0.37 mmol) was added, nitrogen protection was applied again three times, and then heated and stirred at 90°C overnight. EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM:MeOH=80:1) gave the intermediate 1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)undec-10-ynoic acid (180 mg, yield: 45%).

[0243] The intermediate 1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)undec-10-ynoic acid (60 mg, 0.14 mmol) and intermediate P1 (58 mg, 0.14 mmol) were dissolved in DMSO (3 mL), and HATU (70 mg, 0.18 mmol) and DIEA (54 mg, 0.42 mmol) were added in sequence, and stirred at room temperature. EA and water were extracted, and the organic phase was collected, washed with saturated NaCl solution, and the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM:MeOH=50:1) gave the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(11-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)undec-10-ynyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (23 mg, yield 20%) (YZ-35). 1H NMR(500MHz,DMSO-d6)δ10.99(s,1H),8.77-8.73(m,1H),8.30(s,1H),7.75(d,J=7.4Hz,1H),7.70-7.57(m,3H),7.52-7.44(m,1H),7.16-6 .99(m,6H),5.14-5.07(m,1H),4.47-4.40(m,2H),4.31(d,J=17.5Hz,1H),4.27-4.19(m,1H),4.08(s,1H),3.90(d,J=8.2Hz,1H),3.82(d,J =13.5Hz,1H),3.70-3.58(m,2H),3.50(d,J=6.6Hz,1H),3.45-3.37(m,1H),3.19-3.10(m,1H),2.87-2.73(m,4H),2.63-2.52(m,1H),2.33- 2.24(m,3H),2.04-1.96(m,1H),1.94-1.82(m,2H),1.61-1.52(m,2H),1.51-1.37(m,4H),1.32-1.28(m,8H),1.23(s,4H).HRMS(ESI):calcd for C 46 H 57 N8O6[M+H] + :817.4396,found817.4399.

[0244] Example 1-36. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(2-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)acetyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-36):

[0245]

[0246] 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (300 mg, 1.1 mmol) and tert-butyl 4-(piperazin-1-ylmethyl)piperidine-1-carboxylate (370 mg, 1.3 mmol) were dissolved in DMSO (3 mL), DIEA (280 mg, 2.2 mmol) was added, nitrogen was protected for 3-4 times, and the reaction was heated at 90 ° C for 6 h. EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phase was combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM: MeOH = 80: 1) gave the intermediate tert-butyl 4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)piperazin-1-yl)methyl)piperidine-1-carboxylate (450 mg, yield 76%).

[0247] The intermediate tert-butyl 4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidine-1-carboxylate (450 mg, 0.83 mmol) was dissolved in DCM (10 mL), TFA (2 mL) was added, and the mixture was stirred at room temperature overnight. The mixture was evaporated under reduced pressure to obtain a crude product 4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidine-1-carboxylic acid, which was directly used in the next step.

[0248] 4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidine-1-carboxylic acid (190 mg, 0.43 mmol) was dissolved in DMF (3 mL), tert-butyl bromoacetate (170 mg, 0.87 mmol) and DIEA (168 mg, 1.3 mmol) were added, and stirred at room temperature. After the reaction was completed, EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM: MeOH = 100: 1) gave the intermediate tert-butyl 2-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)acetate (55 mg, yield 23%).

[0249] Tert-butyl 2-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)acetate (55 mg, 0.099 mmol) was dissolved in DCM (3 mL), TFA (1 mL) was added, and the mixture was stirred at room temperature overnight. The mixture was evaporated under reduced pressure to obtain a crude product, 2-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)acetic acid, which was directly used in the next step.

[0250] 2-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazine-1-yl)methyl)piperidin-1-yl)acetic acid (0.099 mmol) and intermediate P1 (40 mg, 0.099 mmol) were dissolved in DMSO (3 mL), HATU (49 mg, 0.13 mmol) and DIEA (38 mg, 0.30 mmol) were added in sequence, and stirred at room temperature. EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM:MeOH=50:1) gave the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(2-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)acetyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-36) (12 mg, yield 14%). 1H NMR (500MHz, DMSO-d6) δ11.08(s,1H),8.78-8.72(m,1H),8.30(s,1H),7.76(d,J=7.4Hz,1H),7.68(d,J=8.5Hz,1H),7.34(d,J=2.3Hz,1H),7.27-7 .23(m,1H),7.10(d,J=6.3Hz,3H),7.06(s,1H),7.02(d,J=6.3Hz,1H),5. 12-5.03(m,1H),4.97(d,J=4.7Hz,1H),4.25(d,J=13.0Hz,1H),4.12-4.0 7(m,1H),3.92-3.87(m,1H),3.84(d,J=13.4Hz,1H),3.66-3.56(m,2H),3 .45-3.41(m,4H),3.17(d,J=5.3Hz,2H),2.93-2.87(m,1H),2.86-2.78(m ,3H),2.78-2.66(m,3H),2.59(d,J=19.0Hz,2H),2.35-2.28(m,4H),2.05 -1.99(m,1H),1.95-1.84(m,2H),1.54-1.42(m,5H),1.34-1.22(m,12H).

[0251] Example 1-37. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(4-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)butyl)piperazin-1-yl)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-37):

[0252]

[0253] 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (300 mg, 1.1 mmol) and tert-butyl 4-(3-aminopropyl)piperazine-1-carboxylate (320 mg, 1.3 mmol) were dissolved in DMSO (2 mL), DIEA (280 mg, 2.2 mmol) was added, nitrogen was protected for 3-4 times, and stirred at 90°C overnight. EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM: MeOH = 80: 1) gave tert-butyl 4-(3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)amino)propyl)piperazine-1-carboxylate (230 mg, yield 42%).

[0254] Dissolve tert-butyl 4-(3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)propyl)piperazine-1-carboxylate (230 mg, 0.46 mmol) in DCM (5 mL), add dioxane hydrochloride solution (1.5 mL), and stir at room temperature overnight. Extract with DCM and saturated sodium bicarbonate, collect the organic phase, and distill under reduced pressure to obtain the crude product 2-(2,6-dioxopiperidin-3-yl)-5-((3-(piperazin-1-yl)propyl)amino)isoindolin-1,3-dione (120 mg, yield 65%).

[0255] Dissolve 2-(2,6-dioxopiperidin-3-yl)-5-((3-(piperazin-1-yl)propyl)amino)isoindoline-1,3-dione (120 mg, 0.30 mmol) in DCM (5 mL), add tert-butyl 4-oxobutyrate (57 mg, 0.36 mmol), add sodium triacetoxyborohydride (190 mg, 0.90 mmol), add acetic acid (27 mg, 0.45 mmol) dropwise, and stir at room temperature overnight. Distill under reduced pressure. Separate and purify by column chromatography (DCM: MeOH = 50: 1) to obtain tert-butyl 4-(4-(3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)amino)propyl)piperazin-1-yl)butyrate (100 mg, yield 62%).

[0256] Tert-butyl 4-(4-(3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)propyl)piperazin-1-yl)butanoate (100 mg, 0.19 mmol) was dissolved in DCM (10 mL), TFA (2 mL) was added, and the mixture was stirred at room temperature overnight. The mixture was evaporated under reduced pressure, and the crude product 4-(4-(3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)propyl)piperazin-1-yl)butanoic acid was directly used for the next step without purification.

[0257] 4-(4-(3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)propyl)piperazin-1-yl)butanoic acid (80 mg, 0.19 mmol) and intermediate P1 (92 mg, 0.19 mmol) were dissolved in DMSO (3 mL), and HATU (91 mg, 0.24 mmol) and DIEA (72 mg, 0.56 mmol) were added in sequence and stirred at room temperature. The reaction was monitored by TLC. The mixture was extracted with EA and water, and the organic phase was collected and washed with saturated NaCl solution. The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=50:1) to give the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(4-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolyl-5-yl)amino)butyl)piperazin-1-yl)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-37) (52 mg, yield 32%). 1 H NMR(500MHz,DMSO-d6)δ11.05(s,1H),8.78-8.72(m,1H),8.29(d,J=1.1Hz,1H),7.7 5(d,J=7.4Hz,1H),7.55(d,J=8.3Hz,1H),7.17-7.12(m,1H),7.11-7.08(m,3H),7.0 6(d,J=1.2Hz,1H),7.03-6.99(m,1H),6.95(d,J=2.1Hz,1H),6.87-6.82(m,1H),5.0 6-5.00(m,1H),4.97(d,J=4.8Hz,1H),4.24(d,J=12.9Hz,1H),4.09(s,1H),3.89(d, J=5.8Hz,1H),3.83(d,J=13.5Hz,1H),3.66-3.56(m,2H),3.45-3.38(m,1H),3.22-3 .18(m,2H),2.92-2.86(m,1H),2.85-2.80(m,3H),2.78-2.66(m,3H),2.61-2.57(m, 1H),2.56(d,J=5.0Hz,1H),2.39-2.29(m,9H),2.27(d,J=8.8Hz,3H),2.01-1.96(m, 1H),1.89(d,J=14.2Hz,2H),1.75-1.61(m,5H),1.40-1.20(m,6H); HRMS(ESI):calcd for C 46 H 60 N 11O7[M+H] + :878.4672, found 878.4665.

[0258] Example 1-38. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(1-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperidin-4-carbonyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-38):

[0259]

[0260] 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (300 mg, 1.1 mmol) and 4-piperidinol (160 mg, 1.4 mmol) were dissolved in DMSO (2 mL), DIEA (420 mg, 3.3 mmol) was added, nitrogen was protected for 3-4 times, and stirred at 90°C overnight. EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM: MeOH = 100: 1) gave 2-(2,6-dioxopiperidin-3-yl)-5-(4-(hydroxymethyl)piperidin-1-yl)isoindoline-1,3-dione (350 mg, yield 85%).

[0261] Under ice bath conditions, 2-(2,6-dioxopiperidin-3-yl)-5-(4-(hydroxymethyl)piperidin-1-yl)isoindoline-1,3-dione (350 mg, 0.94 mmol) and p-toluenesulfonyl chloride (540 mg, 2.8 mmol) were dissolved in DCM (10 mL), TEA (310 mg, 2.8 mmol) was added, and stirred overnight. TLC was used to detect the reaction. DCM and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (DCM: MeOH = 100: 1) gave (1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)piperidin-4-yl)4-methylbenzenesulfonate (300 mg, yield 61%).

[0262] Dissolve (1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)4-methylbenzenesulfonic acid methyl ester (150 mg, 0.29 mmol) and tert-butyl 4-piperidincarboxylate (70 mg, 0.37 mmol) in DMF (3 mL), add DIEA (110 mg, 0.86 mmol), and stir at 70°C overnight. Extract with EA and water, collect the organic phase, wash with saturated NaCl solution, combine the organic phases, dry over anhydrous sodium sulfate, and distill under reduced pressure. Separate and purify by column chromatography (DCM:MeOH=50:1) to obtain tert-butyl 1-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperidin-4-carboxylate (90 mg, yield 58%).

[0263] 1-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperidine-4-carboxylic acid tert-butyl ester (90 mg, 0.17 mmol) was dissolved in DCM (5 mL), TFA (1 mL) was added, and the mixture was stirred at room temperature overnight. After the reaction was completed, the mixture was evaporated under reduced pressure, and the obtained crude product 1-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperidine-4-carboxylic acid was directly used in the next step.

[0264] 1-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperidine-4-carboxylic acid (0.17 mmol) and intermediate P1 (80 mg, 0.19 mmol) were dissolved in DMSO (2 mL), and HATU (96 mg, 0.25 mmol) and DIEA (75 mg, 0.58 mmol) were added in sequence and stirred at room temperature. The reaction was monitored by TLC. The mixture was extracted with EA and water, and the organic phase was collected and washed with saturated NaCl solution. The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=50:1) to give the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(1-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperidin-4-carbonyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-38) (22 mg, yield 15%). 1H NMR (500MHz, DMSO-d6) δ11.07(s,1H),8.78-8.73(m,1H),8.30(s,1H),7.76(d,J=7.4Hz,1H),7.66(d,J=8.5Hz,1H),7.29-7.2 3(m,1H),7.23-7.16(m,1H),7.14-7.00(m,6H),5.09-5.04(m,1H),4.97(d,J=4.7Hz,1H),4.27(s,1H),4.11(s,1H),3.67-3.5 6(m,2H),3.46-3.39(m,1H),3.24-3.16(m,1H),3.14-3.07(m,1H),2.92-2.73(m,8H),2.71-2.65(m,1H),2.62-2.54(m,3H),2 .28-2.21(m,1H),2.09(d,J=12.4Hz,1H),2.04-1.98(m,2H),1.96-1.76(m,6H),1.68(s,3H),1.63-1.45(m,5H),1.24(s,6H).

[0265] Example 1-39. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(1-(3-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)propyl)piperidin-4-carbonyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-39):

[0266] Similar to Example 1-38, but replacing 4-piperidinemethanol with 4-(3-hydroxypropyl)-piperidine, the target product obtained by silica gel column chromatography is N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(1-(3-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)propyl)piperidin-4-carbonyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-39). 1H NMR(500MHz,DMSO-d6)δ11.07(s,1H),8.78-8.72(m,1H),8.30(s,1H),7.76(d, J=7.4Hz,1H),7.65(d,J=8.5Hz,1H),7.30(d,J=2.3Hz,1H),7.25-7.21(m,1H),7 .15-7.07(m,3H),7.06(d,J=1.1Hz,1H),7.04-6.99(m,1H),5.10-5.03(m,1H),4 .97(d,J=4.7Hz,1H),4.25(s,1H),4.04(d,J=13.1Hz,2H),3.89(d,J=9.1Hz,2H) ,3.67-3.56(m,3H),3.45-3.39(m,1H),3.23-3.15(m,1H),2.98-2.91(m,3H),2 .90-2.86(m,2H),2.85-2.82(m,2H),2.78-2.67(m,3H),2.58-2.53(m,2H),2.25 (s,2H),2.04-1.99(m,1H),1.92(d,J=8.7Hz,4H),1.78-1.72(m,2H),1.58(s,5H ),1.46(s,1H),1.23(d,J=9.1Hz,8H),1.15(d,J=10.6Hz,2H); HRMS(ESI):calcd for C 49 H 62 N 10 NaO7[M+Na] + :925.4695,found 925.4691.

[0267] Example 1-40. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)benzoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-40):

[0268]

[0269] Methyl (1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)4-methylbenzenesulfonate (290 mg, 0.56 mmol) and tert-butyl 4-(piperazin-1-yl)benzoate (220 mg, 0.84 mmol) were dissolved in DMF (3 mL), TEA (122 mg, 1.1 mmol) was added, and the mixture was stirred at 90 °C overnight. The mixture was extracted with EA and water, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=50:1) to give tert-butyl 4-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)benzoate (100 mg, yield 29%).

[0270] Tert-butyl 4-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)benzoate (100 mg, 0.16 mmol) was dissolved in DCM (5 mL), TFA (1.5 mL) was added, and the mixture was stirred at room temperature overnight. The mixture was evaporated under reduced pressure, and the intermediate 4-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)benzoic acid was directly used in the next step without further purification.

[0271] The intermediate 4-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)benzoic acid (0.16 mmol) and the intermediate P1 (80 mg, 0.16 mmol) were dissolved in DMSO (2 mL), and HATU (80 mg, 0.21 mmol) and DIEA (63 mg, 0.49 mmol) were added in sequence and stirred at room temperature. The reaction was monitored by TLC. The mixture was extracted with EA and water, and the organic phase was collected and washed with saturated NaCl solution. The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=50:1) to give the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)benzoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-40) (12 mg, yield: 8.4%). 1H NMR(500MHz,DMSO-d6)δ11.06(s,1H),8.78-8.72(m,1H),8.29(d,J=1.1Hz,1H),7.78 (d,J=7.3Hz,1H),7.64(d,J=8.5Hz,1H),7.27(d,J=8.4Hz,2H),7.13-7.04(m,4H),7.0 4-6.98(m,1H),6.95(d,J=8.5Hz,2H),6.91(d,J=2.2Hz,1H),6.83-6.77(m,1H),5.08 -5.01(m,1H),4.96(d,J=4.8Hz,1H),4.13(s,3H),3.92-3.84(m,2H),3.66-3.58(m,3H ),3.54-3.47(m,1H),3.46-3.34(m,3H),3.25-3.19(m,4H),3.16-3.01(m,3H),2.93- 2.87(m,1H),2.86-2.80(m,3H),2.77-2.72(m,1H),2.71-2.65(m,1H),2.61-2.55(m,1 H),2.45-2.39(m,2H),2.38-2.31(m,1H),2.22-2.15(m,1H),2.04-1.97(m,1H),1.91( s,2H),1.73-1.59(m,4H),1.44-1.31(m,3H),1.29-1.19(m,3H).HRMS(ESI):calcdfor C 52 H 62 N 11 O7[M+H] + :952.4828, found 952.4839.

[0272] Example 1-41. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-41):

[0273]

[0274] 2-(2,6-Dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (77 mg, 0.28 mmol) and tert-butyl 4-(4-(piperazin-1-ylmethyl)piperidin-1-yl)benzoate (49 mg, 0.14 mmol) were dissolved in DMSO (2 mL), DIEA (36 mg, 0.28 mmol) was added, and the mixture was stirred at 90 °C overnight under nitrogen protection. The mixture was extracted with EA and water, and the organic phase was collected and washed with saturated NaCl solution. The organic phases were combined and dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=50:1) to give the target product 4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoic acid tert-butyl ester (34 mg, yield: 20%).

[0275] Tert-butyl 4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoate (34 mg, 0.05 mmol) was dissolved in DCM (5 mL), TFA (1 mL) was added, and the mixture was stirred at room temperature overnight. The mixture was evaporated under reduced pressure to obtain the intermediate 4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoic acid, which was directly used for the next step without further purification.

[0276] 4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoic acid (0.05 mmol) and intermediate P1 (30 mg, 0.05 mmol) were dissolved in DMSO (2 mL), and HATU (30 mg, 0.08 mmol) and DIEA (25 mg, 0.19 mmol) were added in sequence and stirred at room temperature. The reaction was monitored by TLC. The mixture was extracted with EA and water, and the organic phase was collected and washed with saturated NaCl solution. The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=50:1) to give the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolyl-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-41) (13 mg, yield: 21%). 1HNMR(500MHz,DMSO-d6)δ11.08(s,1H),8.78-8.72(m,1H),8.29(s,1H),7. 79(d,J=7.2Hz,1H),7.68(d,J=8.5Hz,1H),7.34(d,J=2.2Hz,1H),7.27-7.2 4(m,3H),7.11-7.08(m,3H),7.06(d,J=1.2Hz,1H),7.01(d,J=7.1Hz,1H), 6.94(d,J=8.6Hz,2H),5.11-5.04(m,1H),4.97(s,1H),4.08(d,J=51.4Hz,3 H),3.89(s,1H),3.80(d,J=12.3Hz,2H),3.61(d,J=9.0Hz,2H),3.44(s,6H ),3.11(s,2H),2.93-2.86(m,1H),2.82(d,J=5.3Hz,2H),2.78-2.72(m,3H) ,2.69(s,1H),2.58(d,J=18.3Hz,2H),2.22(d,J=6.9Hz,2H),2.04-1.99(m ,1H),1.84-1.73(m,4H),1.42-1.33(m,3H),1.23(s,8H).HRMS(ESI):calcd for C 52 H 62 N 11 O7[M+H] + :952.4828,found952.4826.

[0277] Example 1-42. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-(4-(3-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)propyl)piperazin-1-yl)benzoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-42):

[0278]

[0279] 3-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)propyl 4-methylbenzenesulfonate (190 mg, 0.34 mmol) and tert-butyl 4-(1-piperazinyl)benzoate (110 mg, 0.41 mmol) were dissolved in DMF (3 mL), TEA (89 mg, 0.69 mmol) was added and stirred at 90 °C overnight. The mixture was extracted with EA and water, and the organic phase was collected and washed with saturated NaCl solution. The organic phases were combined and dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=100:1) to give tert-butyl 4-(4-(3-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)propyl)piperazin-1-yl)benzoate (190 mg, yield 45%).

[0280] Dissolve tert-butyl 4-(4-(3-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)propyl)piperazin-1-yl)benzoate (100 mg, 0.16 mmol) in DCM (5 mL), add TFA (1 mL), and stir at room temperature. Monitor the reaction by TLC. After the reaction is completed, evaporate under reduced pressure, and the resulting crude product 4-(4-(3-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)propyl)piperazin-1-yl)benzoic acid is directly used in the next step.

[0281] 4-(4-(3-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)propyl)piperazin-1-yl)benzoic acid (0.16 mmol) and intermediate P1 (65 mg, 0.16 mmol) were dissolved in DMSO (2 mL), and HATU (77 mg, 0.20 mmol) and DIEA (60 mg, 0.47 mmol) were added in sequence and stirred at room temperature. The reaction was monitored by TLC. The mixture was extracted with EA and water, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=50:1) to give the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-(4-(3-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)propyl)piperazin-1-yl)benzoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-42). 1HNMR(500MHz,DMSO-d6)δ11.07(s,1H),8.78-8.72(m,1H),8.29(d,J=1.2Hz,1H),7.78 (d,J=7.3Hz,1H),7.64(d,J=8.6Hz,1H),7.30(d,J=2.3Hz,1H),7.27(d,J=8.7Hz,2H),7 .24-7.21(m,1H),7.11-7.08(m,3H),7.06(d,J=1.2Hz,1H),7.01(d,J=6.2Hz,1H),6.9 4(d,J=8.7Hz,2H),5.09-5.02(m,1H),4.97(d,J=4.6Hz,1H),4.18-4.00(m,6H),3.89(d ,J=5.9Hz,1H),3.66-3.56(m,2H),3.46-3.38(m,1H),3.29(d,J=7.1Hz,1H),3.23-3.1 8(m,4H),3.17(d,J=5.2Hz,4H),2.97-2.91(m,2H),2.85-2.81(m,2H),2.78-2.65(m,2H ),2.58(d,J=18.5Hz,1H),2.30(s,2H),2.04-1.97(m,1H),1.91(s,2H),1.76(d,J=12.4 Hz,2H),1.50(d,J=7.8Hz,4H),1.42-1.34(m,2H),1.28-1.10(m,8H); HRMS(ESI):calcd for C 54 H 66 N 11 O7[M+H] + :980.5141,found 980.5139. Example 1-43. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(5-(7-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,7-diazaspiro[3.5]nonane-2-yl)pentanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-43):

[0282]

[0283] 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (250 mg, 0.91 mmol) and tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate (250 mg, 1.1 mmol) were dissolved in DMSO (3 mL), DIEA (240 mg, 1.8 mmol) was added, nitrogen was protected, and the mixture was stirred at 90°C overnight. EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, evaporated under reduced pressure, and purified by column chromatography (DCM:MeOH=100:1) to obtain tert-butyl 7-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-2,7-diazaspiro[3.5]nonane-2-carboxylate (325 mg, yield: 74%).

[0284] 7-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,7-diazaspiro[3.5]nonane-2-carboxylic acid tert-butyl ester (325 mg, 0.66 mmol) was dissolved in DCM (10 mL), and dioxane hydrochloride solution (2 mL) was added. After the reaction was completed, the reaction was evaporated under reduced pressure to obtain the crude product 2-(2,6-dioxopiperidin-3-yl)-5-(2,7-diazaspiro[3.5]nonan-7-yl)isoindole-1,3-dione without further purification and directly used in the next step.

[0285] 2-(2,6-dioxopiperidin-3-yl)-5-(2,7-diazaspiro[3.5]nonan-7-yl)isoindole-1,3-dione (0.66 mmol) was dissolved in DMF (3 mL), tert-butyl 5-bromopentanoate (314 mg, 1.3 mmol) and TEA (145 mg, 1.3 mmol) were added, and the mixture was stirred at 90°C overnight. The mixture was extracted with EA and water, and the organic phase was collected, washed with saturated NaCl solution, combined, dried over anhydrous sodium sulfate, evaporated under reduced pressure, and separated and purified by column chromatography (DCM:MeOH=80:1) to obtain the intermediate tert-butyl 5-(7-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,7-diazaspiro[3.5]nonan-2-yl)pentanoate (195 mg, yield: 61%).

[0286] Tert-butyl 5-(7-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,7-diazaspiro[3.5]nonan-2-yl)pentanoate (195 mg, 0.36 mmol) was dissolved in DCM (5 mL), and a dioxane hydrochloride solution (2 mL) was added. After the reaction was completed, the mixture was evaporated under reduced pressure to obtain a crude product, 5-(7-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,7-diazaspiro[3.5]nonan-2-yl)pentanoic acid.

[0287] 5-(7-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,7-diazaspiro[3.5]non-2-yl)pentanoic acid (85 mg, 0.19 mmol) and intermediate P1 (80 mg, 0.19 mmol) were dissolved in DMSO (3 mL), and HATU (96 mg, 0.25 mmol) and DIEA (75 mg, 0.58 mmol) were added in sequence and stirred at room temperature. The reaction was monitored by TLC. The mixture was extracted with EA and water, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=50:1) to give the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(5-(7-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolyl-5-yl)-2,7-diazaspiro[3.5]nonan-2-yl)pentanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-43) (32 mg, yield: 20%). 1H NMR(500MHz,DMSO-d6)δ11.07(s,1H),10.94(s,1H),8.80-8.72(m,1H),8.35(s,1H),7.84(d,J=20.7Hz,1H),7.66 (d,J=8.5Hz,1H),7.40-7.35(m,1H),7.30-7.25(m,1H),7.18-7.03(m,5H),5.11-5.01(m,1H),4.24(d,J=11.2Hz,1 H),4.16-4.05(m,3H),3.92(s,2H),3.87-3.75(m,4H),3.52-3.39(m,5H),2.93-2.78(m,5H),2.62-2.55(m,2H),2. 37(d,J=6.3Hz,2H),2.05-1.79(m,8H),1.58-1.45(m,5H),1.42-1.32(m,2H),1.30-1.22(m,5H).HRMS(ESI):calcd for C 47 H 59 N 10 O7[M+H] + :875.4563, found 875.4551.

[0288] Example 1-44. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-(9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-3,9-diazaspiro[5.5]undecane-3-yl)butanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-44):

[0289]

[0290] 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (250 mg, 0.91 mmol) and tert-butyl 3,9-diazaspiro[5.5]undecane-3-carboxylate (276 mg, 1.1 mmol) were dissolved in DMSO (3 mL), DIEA (240 mg, 1.8 mmol) was added, nitrogen was protected, and the mixture was stirred at 90°C overnight. EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, evaporated under reduced pressure, and purified by column chromatography (DCM:MeOH=80:1) to obtain tert-butyl 9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-3,9-diazaspiro[5.5]undecane-3-carboxylate (515 mg, yield: 99%).

[0291] 9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-3,9-diazaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (515 mg, 1.0 mmol) was dissolved in DCM (10 mL), and dioxane hydrochloride solution (1.5 mL) was added. After the reaction was completed, the reaction was evaporated under reduced pressure to obtain the crude product 2-(2,6-dioxopiperidin-3-yl)-5-(-3,9-diazaspiro[5.5]undecane-3-yl)isoindole-1,3-dione without further purification and directly used in the next step.

[0292] 2-(2,6-dioxopiperidin-3-yl)-5-(-3,9-diazaspiro[5.5]undecane-3-yl)isoindole-1,3-dione (0.55 mmol) was dissolved in DMF (2 mL), tert-butyl 4-bromobutyrate (245 mg, 1.1 mmol) and TEA (120 mg, 1.1 mmol) were added, and the mixture was stirred at 90°C overnight. The mixture was extracted with EA and water, and the organic phase was collected, washed with saturated NaCl solution, combined, dried over anhydrous sodium sulfate, evaporated under reduced pressure, and separated and purified by column chromatography (DCM:MeOH=50:1) to obtain the intermediate tert-butyl 4-(9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-3,9-diazaspiro[5.5]undecane-3-yl)butyrate (225 mg, yield: 66%).

[0293] Tert-butyl 4-(9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-3,9-diazaspiro[5.5]undec-3-yl)butanoate (225 mg, 0.41 mmol) was dissolved in DCM (5 mL), and a dioxane hydrochloride solution (1.5 mL) was added. After the reaction was completed, the mixture was evaporated under reduced pressure to obtain a crude product 4-(9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-3,9-diazaspiro[5.5]undec-3-yl)butanoic acid.

[0294] 4-(9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-3,9-diazaspiro[5.5]undecane-3-yl)butanoic acid (80 mg, 0.16 mmol) and intermediate P1 (66 mg, 0.16 mmol) were dissolved in DMSO (2 mL), and HATU (80 mg, 0.21 mmol) and DIEA (62 mg, 0.48 mmol) were added in sequence and stirred at room temperature. The reaction was monitored by TLC. The mixture was extracted with EA and water, and the organic phase was collected and washed with saturated NaCl solution. The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=50:1) to give the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-(9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolyl-5-yl)-3,9-diazaspiro[5.5]undecane-3-yl)butanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-44) (35 mg, yield: 24%). 1 H NMR (500MHz, DMSO-d6) δ11.07(s,1H),8.80-8.71(m,1H),8.29(s,1H),7.76(d,J=7.2Hz,1H),7.64(d,J=8.4Hz,1H),7.30(s,1H),7. 21(d,J=8.8Hz,1H),7.08(d,J=17.5Hz,4H),7.01(d,J=7.2Hz,1H),5.10-5.02(m,1H),4.97(s,1H),4.24(d,J=12.5Hz,1H),4.09(s,1 H),3.93-3.79(m,3H),3.62(t,J=11.9Hz,3H),3.51-3.40(m,6H),3.17(d,J=12.0Hz,1H),2.87-2.79(m,4H),2.76-2.66(m,3H),2.35 (d,J=15.1Hz,4H),2.00(d,J=14.0Hz,1H),1.96-1.85(m,2H),1.72-1.63(m,2H),1.51(s,9H),1.41-1.31(m,2H),1.29-1.20(m,4H).

[0295] Example 1-45. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-45):

[0296]

[0297] 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (500 mg, 1.8 mmol) and 1-Boc piperazine (405 mg, 2.2 mmol) were dissolved in DMSO (5 mL), DIEA (470 mg, 3.6 mmol) was added, and the mixture was stirred at 90°C overnight under nitrogen protection. The organic phase was extracted with EA and water, and the organic phase was collected, washed with saturated NaCl solution, combined, dried over anhydrous sodium sulfate, distilled under reduced pressure, and separated and purified by column chromatography (DCM: MeOH = 150: 1) to obtain tert-butyl 4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)piperazine-1-carboxylate (850 mg, yield 95%).

[0298] Dissolve tert-butyl 4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazine-1-carboxylate (850 mg, 1.9 mmol) in DCM (10 mL), add TFA (2 mL), and stir at room temperature. Detect the reaction by TLC. After the reaction is completed, distill under reduced pressure to obtain the crude product 2-(2,6-dioxopiperidin-3-yl)-5-(piperazine-1-yl)isoindolin-1,3-dione, which is directly used for the next step without further purification.

[0299] 2-(2,6-dioxopiperidin-3-yl)-5-(piperazine-1-yl)isoindoline-1,3-dione (100 mg, 0.29 mmol) and tert-butyl 7-bromoheptanoate (93 mg, 0.35 mmol) were dissolved in DMF (3 mL), DIEA (75 mg, 0.58 mmol) was added, and the mixture was stirred at room temperature overnight. EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, distilled under reduced pressure, and purified by column chromatography (DCM: MeOH = 50: 1) to obtain tert-butyl 7-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)piperazine-1-yl)heptanoate (120 mg, yield 79%).

[0300] Dissolve tert-butyl 7-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)heptanoate (120 mg, 0.23 mmol) in DCM (10 mL), add TFA (1 mL), and stir at room temperature. Monitor the reaction by TLC. After the reaction is completed, distill under reduced pressure to obtain the crude product 7-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)heptanoic acid, which is directly used in the next step without further purification.

[0301] 7-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)heptanoic acid (75 mg, 0.16 mmol) and intermediate P1 (65 mg, 0.16 mmol) were dissolved in DMSO (3 mL), and HATU (78 mg, 0.21 mmol) and DIEA (61 mg, 0.48 mmol) were added in sequence and stirred at room temperature. The reaction was monitored by TLC. The mixture was extracted with EA and water, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=50:1) to give the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolyl-5-yl)piperazin-1-yl)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-45) (12 mg, yield 8.7%). 1H NMR(500MHz,DMSO-d6)δ11.07(s,1H),8.81-8.73(m,1H),8.30(s,1H),7.80-7.74(m,1H),7.70-7.61(m,1H),7.32(d,J=8.6Hz,1H), 7.27-7.22(m,1H),7.17(d,J=9.1Hz,1H),7.13-7.05(m,4H),7.01(d,J=5.1Hz,1H),5.09-5.03(m,1H),4.97(d,J=4.6Hz,1H),4.21( s,1H),4.11(d,J=38.5Hz,3H),3.89(s,1H),3.65-3.57(m,2H),3.44-3.37(m,5H),3.26-3.11(m,3H),2.90-2.80(m,4H),2.78-2.67 (m,3H),2.20(d,J=41.0Hz,3H),2.04-1.88(m,4H),1.80(s,2H),1.62(s,3H),1.46(s,3H),1.24(d,J=7.5Hz,6H).HRMS(ESI):calcd for C 46 H 59 N 10 O7[M+H] + :863.4563, found863.4563. Example 1-46. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(11-(4-(2,6-dioxopiperidin-3-yl)phenoxy)undecanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-46):

[0302]

[0303] Dissolve 2,6-bis(benzyloxy)-3-bromopyridine (1g, 2.7mmol), 4-hydroxyphenylboronic acid pinacol ester (770mg, 3.5mmol) and potassium carbonate (1.1g, 8.1mmol) in 1,4-dioxane (10mL) and water (3mL) and stir at room temperature. Protect with nitrogen for 3-4 times. Add catalyst bistriphenylphosphine palladium dichloride, protect with nitrogen for 3-4 times again, and stir at 100℃ overnight. Filter with suction and evaporate under reduced pressure. Separate and purify by column chromatography (PE:EA=5:1) to obtain the intermediate 4-(2,6-bis(benzyloxy)pyridin-3-yl)phenol (3g, yield 95%).

[0304] The intermediate 4-(2,6-bis(benzyloxy)pyridin-3-yl)phenol (150 mg, 0.39 mmol) and 11-bromondecanoic acid tert-butyl ester (150 mg, 0.47 mmol) were dissolved in DMF (2 mL), cesium carbonate (380 mg, 1.2 mmol) was added, and stirred at room temperature. EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (PE: EA = 8: 1) gave the intermediate 11-(4-(2,6-bis(benzyloxy)pyridin-3-yl)phenoxy)undecanoic acid tert-butyl ester (220 mg, yield 90%).

[0305] The intermediate 11-(4-(2,6-bis(benzyloxy)pyridin-3-yl)phenoxy)undecanoic acid tert-butyl ester (220 mg, 0.35 mmol) was dissolved in methanol (20 mL), nitrogen protection was performed 3-4 times, an appropriate amount of Pd / C was added, nitrogen protection was performed again 3-4 times, hydrogen was replaced, and the mixture was stirred at room temperature overnight. After the reaction was completed, suction filtration was performed, and vacuum distillation was performed. The crude product 11-(4-(2,6-dioxopiperidin-3-yl)phenoxy)undecanoic acid tert-butyl ester (170 mg, yield 99%) was directly used for the next step.

[0306] Dissolve tert-butyl 11-(4-(2,6-dioxopiperidin-3-yl)phenoxy)undecanoate (170 mg, 0.38 mmol) in DCM (5 mL), add TFA (1 mL), and stir at room temperature overnight. After the reaction is completed, evaporate under reduced pressure, and the resulting crude product 11-(4-(2,6-dioxopiperidin-3-yl)phenoxy)undecanoic acid is directly used in the next step.

[0307] 11-(4-(2,6-dioxopiperidin-3-yl)phenoxy)undecanoic acid (80 mg, 0.21 mmol) and intermediate P1 (86 mg, 0.21 mmol) were dissolved in DMSO (2 mL), HTAU (100 mg, 0.27 mmol) and DIEA (80 mg, 0.62 mmol) were added in sequence and stirred at room temperature. The reaction was monitored by TLC. The mixture was extracted with EA and water, and the organic phase was collected and washed with saturated NaCl solution. The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=50:1) to give the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(11-(4-(2,6-dioxopiperidin-3-yl)phenoxy)undecanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-46) (37 mg, yield 23%). 1H NMR(500MHz,DMSO-d6)δ10.78(s,1H),8.77-8.72(m,1H),8.29(d,J=1.2Hz,1 H),7.75(d,J=7.4Hz,1H),7.12-7.08(m,5H),7.06(d,J=1.2Hz,1H),7.02-7.0 0(m,1H),6.88-6.85(m,2H),4.97(d,J=4.6Hz,1H),4.23(d,J=12.8Hz,1H),4 .08(s,1H),3.95-3.91(m,2H),3.91-3.87(m,1H),3.82(d,J=13.5Hz,1H),3.7 9-3.74(m,1H),3.66-3.56(m,2H),3.45-3.39(m,1H),3.20-3.10(m,1H),2.8 4-2.81(m,2H),2.79-2.72(m,2H),2.71-2.66(m,1H),2.65-2.60(m,1H),2.32 -2.27(m,2H),2.19-2.10(m,1H),2.04-1.96(m,1H),1.88(d,J=24.6Hz,2H), 1.72-1.65(m,2H),1.50-1.45(m,2H),1.41-1.37(m,2H),1.35-1.22(m,16H).

[0308] Example 1-47. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((4-(2,6-dioxopiperidin-3-yl)phenyl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-47):

[0309]

[0310] Dissolve 2,6-bis(benzyloxy)-3-bromopyridine (4.4g, 11mmol), 4-aminophenylboronic acid pinacol ester (2g, 9.1mmol) and potassium carbonate (3.8g, 27mmol) in 1,4-dioxane (20mL) and water (6mL) and stir at room temperature. Protect with nitrogen for 3-4 times. Add catalyst bistriphenylphosphine palladium dichloride, protect with nitrogen for 3-4 times again, and stir at 100℃ overnight. Filter with suction and evaporate under reduced pressure. Separate and purify by column chromatography (PE:EA=2:1) to obtain the intermediate 4-(2,6-bis(benzyloxy)pyridin-3-yl)aniline (2.5g, yield 72%).

[0311] The intermediate 4-(2,6-bis(benzyloxy)pyridin-3-yl)aniline (150 mg, 0.39 mmol) and tert-butyl 10-bromodecanoate (180 mg, 0.59 mmol) were dissolved in DMF (2 mL), cesium carbonate (380 mg, 1.2 mmol) was added, and stirred at 90°C overnight. EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Column chromatography separation and purification (PE:EA=6:1) gave the intermediate tert-butyl 10-((4-(2,6-bis(benzyloxy)pyridin-3-yl)phenyl)amino)decanoate (76 mg, yield 32%).

[0312] The intermediate tert-butyl 10-((4-(2,6-bis(benzyloxy)pyridin-3-yl)phenyl)amino)decanoate (76 mg, 0.12 mmol) was dissolved in ethanol (20 mL), nitrogen protection was performed for 3-4 times, an appropriate amount of Pd / C was added, nitrogen protection was performed again for 3-4 times, hydrogen was replaced, and the mixture was stirred at room temperature overnight. After the reaction was completed, suction filtration was performed, and vacuum distillation was performed. The crude product tert-butyl 10-((4-(2,6-dioxopiperidin-3-yl)phenyl)amino)decanoate (43 mg, yield 80%) was directly used for the next step.

[0313] Dissolve tert-butyl 10-((4-(2,6-dioxopiperidin-3-yl)phenyl)amino)decanoate (43 mg, 0.10 mmol) in DCM (5 mL), add TFA (1 mL), and stir at room temperature overnight. After the reaction is completed, evaporate under reduced pressure, and the resulting crude product 10-((4-(2,6-dioxopiperidin-3-yl)phenyl)amino)decanoic acid is directly used in the next step.

[0314] 10-((4-(2,6-dioxopiperidin-3-yl)phenyl)amino)decanoic acid (0.10 mmol) and intermediate P1 (41 mg, 0.10 mmol) were dissolved in DMSO (2 mL), HATU (49 mg, 0.13 mmol) and DIEA (39 mg, 0.30 mmol) were added in sequence and stirred at room temperature. The reaction was monitored by TLC. The mixture was extracted with EA and water, and the organic phase was collected and washed with saturated NaCl solution. The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=50:1) to give the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((4-(2,6-dioxopiperidin-3-yl)phenyl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-47) (28 mg, yield 13%). 1H NMR (500MHz, DMSO-d6) δ10.72(s,1H),8.80-8.71(m,1H),8.30(s,1H),7.75(d,J=7.5Hz,1H),7.15-7.00(m,5H),6.88(d,J=8.3Hz,2H),6.4 9(d,J=8.3Hz,2H),5.51-5.40(m,1H),4.98(s,1H),4.24(d,J=12.8Hz,1H),4.09(s,1H),3.90(s,1H),3.82(d,J=13.5Hz,1H),3.71-3.55(m, 3H),3.46-3.38(m,1H),3.20-3.11(m,1H),3.00-2.91(m,2H),2.83(d,J=6.1Hz,2H),2.77(d,J=12.1Hz,1H),2.71(s,1H),2.65-2.55(m,2H) ,2.34-2.25(m,2H),2.12-2.03(m,1H),2.03-1.96(m,1H),1.91(d,J=22.0Hz,2H),1.55-1.45(m,4H),1.40-1.18(m,16H).HRMS(ESI):calcd for C 43 H 59 N8O5[M+H] + :767.4603, found 767.4600.

[0315] Example 1-48. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-(4-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperazin-1-yl)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-48):

[0316]

[0317] 4-(4-BOC-piperazin-1-yl)-3-fluoroaniline (1g, 3.4mmol) and 3-bromopiperidine-2,6-dione (1.95g, 10mmol) were dissolved in DMF (5mL), sodium bicarbonate (2.8g, 34mmol) was added, and the mixture was stirred at 80°C overnight. The mixture was extracted with EA and water, and the organic phase was collected, washed with saturated NaCl solution, combined, dried over anhydrous sodium sulfate, evaporated under reduced pressure, and purified by column chromatography (DCM:MeOH=100:1) to obtain tert-butyl 4-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperazine-1-carboxylate (300mg, yield 22%).

[0318] Dissolve tert-butyl 4-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperazine-1-carboxylate (300 mg, 0.74 mmol) in DCM (10 mL), add dioxane hydrochloride solution (2 mL), and stir at room temperature overnight. After the reaction is completed, evaporate under reduced pressure to obtain the crude product 3-((3-fluoro-4-(piperazin-1-yl)phenyl)amino)piperidine-2,6-dione without purification and directly use it in the next step.

[0319] 3-((3-fluoro-4-(piperazin-1-yl)phenyl)amino)piperidine-2,6-dione (300 mg, 0.98 mmol) was dissolved in DMF (5 mL), tert-butyl 7-bromoheptanoate (514 mg, 1.9 mmol) and DIEA (376 mg, 2.9 mmol) were added in sequence, and the mixture was stirred at room temperature overnight. The mixture was extracted with EA and water, and the organic phase was collected, washed with saturated NaCl solution, combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The intermediate was separated and purified by column chromatography (DCM:MeOH=50:1) to obtain tert-butyl 7-(4-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperazin-1-yl)heptanoate (89 mg, yield: 18%).

[0320] Dissolve tert-butyl 7-(4-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperazin-1-yl)heptanoate (89 mg, 0.18 mmol) in DCM (5 mL), add TFA (1 mL), and stir at room temperature overnight. After the reaction is completed, distill under reduced pressure to obtain the crude product 7-(4-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperazin-1-yl)heptanoic acid without purification and directly use it in the next step.

[0321] 7-(4-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperazin-1-yl)heptanoic acid (0.18 mmol) and intermediate P1 (73 mg, 0.18 mmol) were dissolved in DMSO (3 mL), and HATU (88 mg, 0.23 mmol) and DIEA (70 mg, 0.54 mmol) were added in sequence and stirred at room temperature. The reaction was monitored by TLC. The mixture was extracted with EA and water, and the organic phase was collected and washed with saturated NaCl solution. The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=50:1) to give the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-(4-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperazin-1-yl)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-48) (22 mg, yield: 15%). 1 H NMR (500MHz, DMSO-d6) δ10.77(s,1H),8.78-8.72(m,1H),8.29(s,1H),7.76(d,J=7.5Hz,1H),7.10(d,J=6.2Hz,3H),7.06(s,1H),7 .01(d,J=7.9Hz,1H),6.85-6.78(m,1H),6.53-6.47(m,1H),6.44-6.39(m,1H),5.80(d,J=7.5Hz,1H),4.98(s,1H),4.29-4.20(m,2H ),4.09(s,1H),3.93-3.87(m,1H),3.83(d,J=13.7Hz,1H),3.68-3.54(m,3H),3.46-3.37(m,2H),3.19-3.11(m,1H),2.90-2.68(m,1 2H),2.34-2.28(m,3H),2.11-2.05(m,1H),1.96-1.81(m,4H),1.53-1.41(m,5H),1.32-1.27(m,5H),1.23(s,3H).HRMS(ESI):calcd for C 44 H 59 FN 10 NaO5[M+H] + :849.4546, found 849.4569.

[0322] Example 1-49. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(2-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)hexyl)amino)-2-oxoethyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-49):

[0323]

[0324] The intermediate (S)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-(piperidin-4-ylamino)pyrimidine-4-carboxamide (100 mg, 0.24 mmol) was dissolved in tetrahydrofuran (15 mL), cesium carbonate (154 mg, 0.47 mmol) was added, and then tert-butyl bromoacetate (92 mg, 0.47 mmol) was added, and stirred at 70°C overnight. The reaction was monitored by TLC, and the mixture was evaporated under reduced pressure. The intermediate (S)-tert-butyl 2-(4-((6-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)carbamoyl)pyrimidin-4-yl)amino)piperidin-1-yl)acetate (83 mg, yield 66%) was obtained by silica gel column chromatography (DCM:MeOH=80:1).

[0325] The intermediate (S)-2-(4-((6-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)carbamoyl)pyrimidin-4-yl)amino)piperidin-1-yl)acetic acid tert-butyl ester (83 mg, 0.16 mmol) was dissolved in DCM (5 mL), TFA (1 mL) was added, and the mixture was stirred at room temperature overnight. After the reaction was completed, the mixture was distilled under reduced pressure, and the resulting crude product was used as the intermediate (S)-2-(4-((6-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)carbamoyl)pyrimidin-4-yl)amino)piperidin-1-yl)acetic acid without further purification.

[0326] The intermediate (S)-2-(4-((6-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)carbamoyl)pyrimidin-4-yl)amino)piperidin-1-yl)acetic acid (0.16 mmol) and 4-((6-aminohexyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (60 mg, 0.16 mmol) were dissolved in DMSO (2 mL), and HATU (80 mg, 0.21 mmol) and DIEA (62 mg, 0.48 mmol) were added in sequence and stirred at room temperature. The reaction was monitored by TLC. The mixture was extracted with EA and water, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=50:1) to give the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(2-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolyl-4-yl)amino)hexyl)amino)-2-oxoethyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-49) (12 mg, yield: 9%). 1 H NMR(500MHz,DMSO-d6)δ11.10(s,1H),8.75(d,J=6.2Hz,1H),8.28(s,1H),7.75-7.68(m,2H),7.58-7.53(m,1H ),7.11-7.06(m,4H),7.03-6.99(m,2H),6.56-6.51(m,1H),5.07-5.02(m,1H),4.99(s,1H),3.88(s,1H),3.82( s,1H),3.61(d,J=9.2Hz,2H),3.11-3.05(m,3H),2.88(d,J=6.7Hz,4H),2.83(d,J=6.1Hz,2H),2.75(s,2H),2.1 7(s,2H),1.86(s,2H),1.58-1.47(m,6H),1.45-1.38(m,4H),1.36-1.27(m,6H),1.23(s,4H).HRMS(ESI):calcd for C 43 H 55 N 10 O7[M+H] + :823.4250, found 823.4267.

[0327] Example 1-50. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetylamino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-50):

[0328]

[0329] The intermediate P1 (40 mg, 0.098 mmol) and 8-((tert-butyloxycarbonyl)amino)octanoic acid (25 mg, 0.098 mmol) were dissolved in DMF (2 mL), and HATU (49 mg, 0.13 mmol) and DIEA (38 mg, 0.30 mmol) were added in sequence, and the mixture was stirred at room temperature. The mixture was extracted with EA and water, and the organic phase was collected, washed with saturated NaCl solution, combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM: MeOH = 100: 1) to obtain (S)-(8-(4-((6-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)carbamoyl)pyrimidin-4-yl)amino)piperidin-1-yl)-8-oxooctyl)carbamic acid tert-butyl ester (38 mg, yield 59%).

[0330] (S)-tert-butyl (8-(4-((6-((3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)carbamoyl)pyrimidin-4-yl)amino)piperidin-1-yl)-8-oxooctyl)carbamate (38 mg, 0.058 mmol) was dissolved in DCM (3 mL), TFA (1 mL) was added, and the mixture was stirred at room temperature overnight. After the reaction was completed, the mixture was evaporated under reduced pressure, and the crude product was used as the intermediate (S)-6-((1-(8-aminooctanoyl)piperidin-4-yl)amino)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)pyrimidine-4-carboxamide, which was directly used for the next step without further purification.

[0331] The intermediate (S)-6-((1-(8-aminooctanoyl)piperidin-4-yl)amino)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)pyrimidine-4-carboxamide (0.058 mmol) and the intermediate 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetic acid (20 mg, 0.058 mmol) were dissolved in DMF (2 mL), and HATU (30 mg, 0.075 mmol) and DIEA (22 mg, 0.17 mmol) were added in sequence, and the mixture was stirred at room temperature. The mixture was extracted with EA and water, and the organic phase was collected and washed with saturated NaCl solution. The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=50:1) to give N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolyl-4-yl)oxy)acetylamino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-50) (8 mg, yield 16%). 1 H NMR (500MHz, DMSO-d6) δ11.12(s,1H),8.79-8.73(m,1H),8.29(s,1H),7.96-7.90(m,1H),7.84-7.78(m,1H),7.76(d,J=7.5Hz,1H),7.49( d,J=7.3Hz,1H),7.39(d,J=8.6Hz,1H),7.09(d,J=6.2Hz,3H),7.05(s,1H),7.01(d,J=8.1Hz,1H),5.16-5.09(m,1H),4.98(s,1H),4.23(d ,J=14.4Hz,1H),4.08(s,1H),3.89(s,1H),3.82(d,J=13.4Hz,1H),3.61(d,J=9.1Hz,2H),3.45-3.39(m,1H),3.18-3.11(m,4H),2.86-2.8 0(m,3H),2.76(d,J=13.7Hz,2H),2.69(s,1H),2.31-2.27(m,2H),2.07-2.01(m,1H),1.90(s,2H),1.49-1.42(m,6H),1.37-1.21(m,12H).

[0332] Example 1-51. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetamido)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-51):

[0333] Similar to Example 1-50, except that 8-((tert-butoxycarbonyl)amino)octanoic acid was replaced with 7-((tert-butoxycarbonyl)amino)heptanoic acid, the target product obtained by silica gel column chromatography was N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetamido)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-51). 1 H NMR(500MHz,DMSO-d6)δ11.12(s,1H),8.78-8.73(m,1H),8.29(s,1H),7.95 -7.91(m,1H),7.83-7.79(m,1H),7.76(d,J=7.5Hz,1H),7.49(d,J=7.3Hz,1 H),7.40(d,J=8.5Hz,1H),7.09(d,J=6.3Hz,3H),7.05(s,1H),7.01(d,J=7. 1Hz,1H),5.15-5.09(m,1H),4.98(d,J=4.8Hz,1H),4.77(s,2H),4.23(d,J=1 4.0Hz,1H),4.08(s,1H),3.92-3.86(m,1H),3.82(d,J=13.6Hz,1H),3.61(d ,J=9.2Hz,2H),3.46-3.38(m,2H),3.17-3.12(m,3H),2.82(d,J=6.0Hz,2H), 2.79-2.73(m,2H),2.68(d,J=11.4Hz,1H),2.63-2.56(m,2H),2.32-2.27(m ,2H),2.07-2.01(m,1H),1.89(s,2H),1.50-1.41(m,5H),1.32-1.22(m,8H).

[0334] Example 1-52. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(6-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetamido)hexanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-52):

[0335] Similar to Example 1-50, except that 8-((tert-butoxycarbonyl)amino)octanoic acid is replaced with 6-((tert-butoxycarbonyl)amino)hexanoic acid, the target product obtained by silica gel column chromatography is N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(6-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetamido)hexanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide. 1 H NMR(500MHz,DMSO-d6)δ11.11(s,1H),8.78-8.72(m,1H),8.29(s,1H),7.98-7.93(m,1H),7.84-7.79(m,1H),7.75(d,J=7.5Hz,1H),7 .50(d,J=7.3Hz,1H),7.39(d,J=8.6Hz,1H),7.09(d,J=6.9Hz,3H),7.05(s,1H),7.01(d,J=7.1Hz,1H),5.16-5.08(m,1H),4.99(s,1H) ,4.77(s,2H),4.30-4.19(m,2H),4.09(s,1H),3.89(s,1H),3.81(d,J=12.4Hz,1H),3.63(d,J=7.8Hz,2H),3.17-3.12(m,3H),2.83(d, J=5.8Hz,2H),2.77(d,J=12.7Hz,2H),2.32-2.25(m,3H),2.05-2.00(m,1H),1.90(s,2H),1.50-1.43(m,4H),1.25(d,J=16.0Hz,10H).

[0336] Example 1-53. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)acetylamino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-53):

[0337]

[0338] (S)-6-((1-(8-aminooctanoyl)piperidin-4-yl)amino)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)pyrimidine-4-carboxamide (23 mg, 0.058 mmol) and intermediate (2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)glycine (20 mg, 0.058 mmol) were dissolved in DMF (2 mL), and HATU (30 mg, 0.076 mmol) and DIEA (22 mg, 0.17 mmol) were added in sequence, and the mixture was stirred at room temperature. The reaction mixture was extracted with EA and water, and the organic phase was collected and washed with saturated NaCl solution. The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=50:1) to give the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolyl-4-yl)amino)acetylamino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-53) (6 mg, yield: 12%). 1 H NMR (500MHz, DMSO-d6) δ8.78-8.72(m,1H),8.29(s,1H),7.91-7.85(m,1H),7.75(d,J=7.5Hz,1H),7.50-7.45(m,1H),7.12-7.07(m,3 H),7.05(d,J=1.2Hz,1H),7.04-6.99(m,3H),6.53(s,2H),5.19-5.12(m,1H),4.97(s,1H),4.21(d,J=4.1Hz,2H),4.09(s,1H),3.89(s ,1H),3.82(d,J=13.4Hz,1H),3.62(s,2H),3.18-3.11(m,1H),3.08-2.96(m,4H),2.82(d,J=5.6Hz,2H),2.79-2.74(m,2H),2.71-2.5 8(m,1H),2.31-2.25(m,2H),2.11-2.04(m,1H),1.88(d,J=29.1Hz,2H),1.47(d,J=9.0Hz,2H),1.40-1.33(m,4H),1.30-1.18(m,12H).

[0339] Example 1-54. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)acetamido)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-54):

[0340]

[0341] (S)-6-((1-(7-aminoheptanoyl)piperidin-4-yl)amino)-N-(3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)pyrimidine-4-carboxamide (35 mg, 0.067 mmol) and intermediate (2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)glycine (22 mg, 0.067 mmol) were dissolved in DMF (2 mL), and HATU (33 mg, 0.087 mmol) and DIEA (26 mg, 0.20 mmol) were added in sequence, and the mixture was stirred at room temperature. The mixture was extracted with EA and water, and the organic phase was collected and washed with saturated NaCl solution. The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=50:1) to give the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)acetamido)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-54) (8 mg, yield: 14%). 1H NMR(500MHz,DMSO-d6)δ11.12(s,1H),8.78-8.73(m,1H),8.29(s,1H),7.96-7.91(m,1H),7.84-7.78(m,1H),7.76(d,J=7.5Hz,1H),7.4 9(d,J=7.3Hz,1H),7.40(d,J=8.5Hz,1H),7.09(d,J=6.3Hz,4H),7.05(s,1H),7.01(d,J=7.1Hz,1H),5.15-5.09(m,1H),4.98(d,J=4.8Hz ,1H),4.77(s,2H),4.23(d,J=14.0Hz,1H),4.08(s,1H),3.92-3.78(m,3H),3.61(d,J=9.2Hz,2H),3.18-3.11(m,4H),2.82(d,J=6.0Hz,2 H),2.78-2.73(m,2H),2.68(d,J=11.4Hz,1H),2.31-2.27(m,2H),2.06-2.01(m,1H),1.89(s,3H),1.51-1.39(m,6H),1.31-1.22(m,8H).

[0342] Example 1-55. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(2-(3-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)acetyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-55):

[0343]

[0344] 2-(2,6-dioxopiperidin-3-yl)-5-(piperazine-1-yl)isoindoline-1,3-dione (300 mg, 0.88 mmol) and 1-Boc-piperidine-3-carboxaldehyde (374 mg, 1.8 mmol) were dissolved in DMF (5 mL), sodium triacetoxyborohydride (370 mg, 1.8 mmol) was added, and the mixture was stirred at room temperature overnight. EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phases were combined, dried over anhydrous sodium sulfate, evaporated under reduced pressure, and purified by column chromatography (DCM: MeOH = 100: 1) to obtain tert-butyl 3-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)piperazine-1-yl)methyl)piperidine-1-carboxylate (500 mg, yield 99%).

[0345] tert-Butyl 3-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidine-1-carboxylate (500 mg, 0.93 mmol) was dissolved in DCM (15 mL), and a dioxane hydrochloride solution (3 mL) was added. After the reaction was completed, the reaction was evaporated under reduced pressure. The crude product obtained as the intermediate 2-(2,6-dioxopiperidin-3-yl)-5-(4-(piperidin-3-ylmethyl)piperazin-1-yl)isoindolin-1,3-dione was directly used for the next step reaction without purification.

[0346] 2-(2,6-dioxopiperidin-3-yl)-5-(4-(piperidin-3-ylmethyl)piperazin-1-yl)isoindoline-1,3-dione (240 mg, 0.56 mmol) and tert-butyl bromoacetate (220 mg, 1.1 mmol) were dissolved in DMF (5 mL), DIEA (215 mg, 1.7 mmol) was added, and the mixture was stirred at room temperature overnight. EA and water were extracted, the organic phase was collected, washed with saturated NaCl solution, the organic phase was combined, dried over anhydrous sodium sulfate, evaporated under reduced pressure, and purified by column chromatography (DCM: MeOH = 50: 1) to obtain tert-butyl 2-(3-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)acetate (162 mg, yield: 53%).

[0347] Tert-butyl 2-(3-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)acetate (162 mg, 0.29 mmol) was dissolved in DCM (5 mL), TFA (2 mL) was added, and the mixture was stirred at room temperature overnight. After the reaction was completed, the mixture was evaporated under reduced pressure, and the obtained crude product was used as the intermediate 2-(3-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)acetic acid, which was directly used for the next step without further purification.

[0348] 2-(3-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)acetic acid (66 mg, 0.15 mmol) and intermediate P1 (60 mg, 0.15 mmol) were dissolved in DMSO (3 mL), and HATU (72 mg, 0.19 mmol) and DIEA (56 mg, 0.44 mmol) were added in sequence, and the mixture was stirred at room temperature. The mixture was extracted with EA and water, and the organic phase was collected and washed with saturated NaCl solution. The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=50:1) to give the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(2-(3-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolyl-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)acetyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide (YZ-55) (18 mg, yield: 13%). 1 H NMR(500MHz,DMSO-d6)δ11.07(s,1H),8.80-8.73(m,1H),8.29(s,1H),7.80-7.74(m,1H),7.69-7.61(m,1H),7.35-7.30(m,1H), 7.27-7.21(m,1H),7.12-7.06(m,4H),7.01(d,J=4.5Hz,1H),5.09-5.04(m,1H),4.97(d,J=4.6Hz,1H),4.27-3.96(m,4H),3.89( s,1H),3.61(d,J=8.6Hz,2H),3.47-3.37(m,6H),3.26-3.10(m,3H),2.85-2.80(m,3H),2.79-2.66(m,4H),2.44-2.39(m,2H),2. 29-2.12(m,3H),2.06-1.88(m,5H),1.80(s,2H),1.65(d,J=26.0Hz,2H),1.46(s,2H),1.25(d,J=11.9Hz,6H).HRMS(ESI):calcd for C 47 H 60 N 11 O7[M+H] + :890.4672,found890.4671.

[0349] Example 1-56. Preparation of N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((4-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)decylamido)cyclohexyl)amino)pyrimidine-4-carboxamide (YZ-56):

[0350]

[0351] Intermediate P2 (72 mg, 0.17 mmol) and 10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)decanoic acid (75 mg, 0.17 mmol) were dissolved in DMSO (3 mL), and HATU (84 mg, 0.22 mmol) and DIEA (66 mg, 0.51 mmol) were added in sequence, and the mixture was stirred at room temperature. The mixture was extracted with EA and water, and the organic phase was collected and washed with saturated NaCl solution. The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The mixture was separated and purified by column chromatography (DCM:MeOH=50:1) to give the target product N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((4-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)decylamido)cyclohexyl)amino)pyrimidine-4-carboxamide (YZ-56) (23 mg, yield: 16%). 1 H NMR (500MHz, DMSO-d6) δ11.05(s,1H),8.76-8.70(m,1H),8.29-8.25(m,1H),7.65(s,2H),7.55(d,J=8.3Hz,1H),7.18(d,J=12.0Hz,1H ),7.09(d,J=6.2Hz,4H),7.01(d,J=6.3Hz,1H),6.93(d,J=2.2Hz,1H),6.85-6.80(m,1H),5.05-4.99(m,1H),4.97(d,J=4.8Hz,1H),3. 93-3.85(m,2H),3.68-3.59(m,3H),3.47-3.40(m,2H),3.16-3.10(m,2H),2.85-2.81(m,2H),2.78-2.71(m,1H),2.71-2.66(m,1H),2. 09-2.05(m,2H),2.02-1.96(m,2H),1.69-1.60(m,4H),1.56(d,J=7.7Hz,6H),1.48(d,J=10.2Hz,2H),1.35(s,2H),1.30-1.21(m,12H).

[0352] Example 2: Evaluation of the effect of the compounds of the present invention on inducing PRMT5 degradation in breast cancer cells

[0353] Example 2-1. Cell culture

[0354] The breast cancer cells HCC1806 and HCC1937 and human normal tissue cells HEK-293T (derived from normal human embryonic kidney) used in the present invention were purchased from the American Type Culture Collection (ATCC).

[0355] The cells were cultured in a medium containing 10% fetal bovine serum (ExCell Bio) and 100 units / ml penicillin and 100 μg / ml streptomycin (VivaCell) in a 37°C constant temperature cell culture incubator (humidity 95%, CO2 concentration 5%). RPMI-1640 (Gibco) and DMEM high glucose medium (Gibco) were used for cell culture.

[0356] Example 2-2. Evaluation of the activity of the compounds of the present invention in inducing PRMT5 degradation

[0357] A series of PROTACs (YZ-1 to YZ-56) targeting degradation of PRMT5 described in Example 1 were dissolved in dimethyl sulfoxide (DMSO) to a final concentration of 4 μmol / L. An equal amount of DMSO was added to the control group and incubated with breast cancer cells HCC1806 and HCC1937 for 48 h. The cells were collected by centrifugation and denatured and lysed to collect total cell protein. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was then used to analyze the degradation of PRMT5 by the compounds.

[0358] Image J software was used to analyze the gray values ​​of PRMT5 and GAPDH, β-actin or Vinculin in different lanes. The relative gray ratio of PRMT5 / GAPDH, β-actin or Vinculin corresponding to each compound and the relative gray ratio of PRMT5 / GAPDH, β-actin or Vinculin in the control group were calculated in Excel. The protein degradation rate (%) of PRMT5 was calculated according to the following formula.

[0359]

[0360] The degradation ability of the compounds YZ-1 to YZ-56 of the present invention on PRMT5 in breast cancer cells is expressed by the PRMT5 protein degradation rate (%) of the compounds under a concentration of 4 μmol / L. Among them, "A", "B", "C" and "D" respectively indicate that the degradation rate of PRMT5 in tumor cells by the compounds of the present invention under the conditions of the concentration is less than 25%, between 25% and 50%, between 50% and 75% and greater than 75%, and "NT" indicates that the ability of the compound to induce PRMT5 degradation under the specified concentration conditions has not been evaluated. The evaluation results of the degradation efficiency of the compounds of the present invention on PRMT5 in breast cancer cells HCC1806 and HCC1937 are shown in Table 1. The degradation effect of some compounds of the present invention on inducing PRMT5 in breast cancer cells HCC1806 and HCC1937 under specified concentration conditions is shown in Table 1. Figure 1 shown.

[0361] Table 1 The degradation activity of PRMT5 induced by compounds YZ-1 to YZ-56 of the present invention in breast cancer cells

[0362]

[0363]

[0364] The results showed that some of the compounds YZ-1 to YZ-56 of the present invention exhibited certain degradation activity in breast cancer cells, among which YZ-18 had the best degradation activity. At a concentration of 4 μmol / L, it could effectively induce the degradation of more than 50% of the PRMT5 protein level in breast cancer cells HCC1806 and HCC1937 and could significantly downregulate the protein expression of KLF5.

[0365] Example 3: Evaluation of the antiproliferative activity of the compounds of the present invention in breast cancer cells

[0366] Example 3-1. Determination of the proliferation inhibitory activity of compounds on breast cancer cells by SRB method

[0367] HCC1806 and HCC1937 cells were cultured at 5×10 3 pcs / hole and 2.5×10 3Cells were inoculated at a density of 100 μl / well in a 96-well plate. After conventional culture for 16 hours, different concentrations of the compounds of the present invention were added in sequence, with a concentration of 4 μmol / L. The control group was added with an equal amount of DMSO. After continued culture for 48 hours, pre-cooled trichloroacetic acid (50%, w / V) was added, 100 μl / well, and incubated overnight at 4°C to fix the cells. After fixation, the cells were washed 5 times with distilled water and air-dried. 100 μl of SRB dye solution (4%, w / V) was added to each well and incubated at room temperature for 30 minutes for staining. The dye solution was aspirated and washed 5-6 times with 1% acetic acid to remove unbound dye. After air drying, 100 μl of Tris base solution with a concentration of 10 mmol / L was added to each well, and the bound SRB dye was dissolved by shaking. The 96-well plate was placed in an ELISA instrument, and the OD value was measured at a wavelength of 530 nm. The cell survival rate (%) was calculated according to the following formula.

[0368]

[0369] The effects of the compounds of the present invention on the proliferation of breast cancer cells were statistically analyzed, and the evaluation results are shown in Table 2.

[0370] The inhibitory activity of the compounds YZ-1 to YZ-56 of the present invention on the viability of breast cancer cells is expressed by the inhibition rate (%) of the compounds under a concentration of 4 micromol / L. Among them, "+", "++", "+++" and "++++" respectively indicate that the inhibition rate of the compound is less than 25%, between 25% and 50%, between 50% and 75% and greater than 75%, and "NT" indicates that the growth inhibitory activity of the compound under the specified concentration conditions has not been evaluated. The inhibitory activity of the compounds of the present invention on breast cancer cells HCC1806 and HCC1937 is shown in Table 2.

[0371] Table 2 Results of tumor inhibition activity of compounds YZ-1 to YZ-56 of the present invention in breast cancer cells

[0372]

[0373]

[0374] The results showed that the compounds of the present invention exhibited certain proliferation inhibition activity against breast cancer cells HCC1806 and HCC1937. Among them, YZ-18, YZ-41, YZ-46, and YZ-47 all exhibited proliferation inhibition activity greater than 50% in both breast cancer cells. These results indicate that the compounds of the present invention have a good anti-proliferation effect on breast cancer cells.

[0375] Example 3-2. Proliferation inhibitory activity of representative compounds in the examples in breast cancer cells and normal breast epithelial cells.

[0376] In order to further test the proliferation inhibitory activity of the compounds of the present invention in breast cancer cells and their toxicity to normal breast epithelial cells, the representative compounds in the examples of the present invention continued to use the SRB method to determine the proliferation inhibitory activity of the compounds on breast cancer cells and normal breast epithelial cells, and statistically analyzed the effects of the representative compounds of the present invention on the proliferation of breast cancer cells under different concentration conditions. Prism v.8.0 (GraphPad) software was used for the IC 50 The test results of YZ-18 are shown in Table 3 and Figure 2 .

[0377] Table 3 IC values ​​of representative compound YZ-18 in breast cancer cells and normal breast epithelial cells 50

[0378]

[0379] The results showed that the representative compound YZ-18 exhibited weak cytotoxicity to normal breast epithelial cells, and breast cancer cells HCC1806 and HCC1937 were more sensitive to compound YZ-18.

[0380] Example 3-3. Evaluation of the ability of the representative compound YZ-18 in the examples to induce cell cycle and apoptosis in breast cancer cells.

[0381] Cells were treated with different concentrations of YZ-18 for 48 hours, then digested with trypsin and fixed with 75% ethanol overnight at 4°C. Cells were then treated with 100 μg / mL RNase solution, stained with propidium iodide (PI) for 30 min, and finally analyzed for DNA content using a BDLSRFortessa flow cytometer. Cells treated with different concentrations of compounds, including floating cells and adherent cells, were collected and then centrifuged. Cells were washed with PBS and stained with FITC / Annexin V and propidium iodide using the Annexin detection kit (1133534, BD Pharmingen). The cell apoptosis rate was detected using an Accuri C6 flow cytometer (BD Biosciences). Figure 3 , Figure 4 The representative compound YZ-18 in this example has the ability to induce cell cycle and apoptosis in breast cancer cells.

[0382] The results showed that the representative compound YZ-18 in the examples of the present invention could induce a concentration-dependent increase in the number of cells in the G1 phase and a concentration-dependent decrease in the number of cells in the S phase, and significantly increased the percentage of Annexin-V positive cells, indicating that the representative compound YZ-18 could effectively induce cell cycle arrest and the occurrence of cell apoptosis.

[0383] Example 3-4. Evaluation of the antiproliferative activity of the representative compound YZ-18 in human breast cancer organoids

[0384] Tumor tissues were obtained from patients undergoing breast cancer surgery at Yunnan Cancer Hospital, and all participants and / or their legal guardians gave informed consent. After surgical collection, fresh breast cancer tissues were immediately placed in DMEM / F12 medium containing 50 μg / mL antibiotics and transported to the laboratory. The tissues were rinsed with sterile 1× PBS, minced with surgical scissors, and digested in a medium containing collagenase I, III, and IV (0.5-1 mg / mL), hyaluronidase (0.05-0.1 mg / mL), DMEM / F12, 10 mM HEPES, 2% BSA, 0.48 μg / mL hydrocortisone, 5 μM Y-27632, and 50 μg / mL primocin (37°C, 1-2 h). The cells were passed through a 100 μM filter, centrifuged at 500 g for 3 minutes, resuspended in 1× PBS, and treated with red blood cell lysis buffer for 3-5 minutes. After rinsing once more with 1× PBS, the cells were resuspended in basement membrane matrix and seeded into 24-well plates. The cells were allowed to solidify for 30 min before adding organoid culture medium. The growth of organoids was monitored and the culture medium was changed regularly. Once the organoids reached sufficient size and number, they were digested, counted, and placed in 96-well plates and treated with representative compounds. The viability of organoid cells after 2 days of treatment with different concentrations of representative compounds was assessed using an ATPase activity assay kit (Promega). This study has been approved by the Clinical Research Ethics Committee of Yunnan Cancer Hospital (KYCS2023-078).

[0385] Figure 5 , Figure 6 The inhibitory activity of compound YZ-18 on breast cancer organoid cell proliferation. Microscopic observation showed that the DMSO control group retained a coherent and orderly structure, while the YZ-18-treated group split into smaller aggregates and showed signs of necrosis. In addition, ATPase activity assays showed that the viability of organoids was significantly reduced after YZ-18 treatment.

[0386] Example 4: Evaluation of the antiproliferative activity of the compounds of the present invention in other tumor cells

[0387] The cervical cancer cells Hela, human non-small cell lung cancer cells A549, human B lymphoma cells SUDHL-4, human mantle cell lymphoma cells Z-138, and human melanoma cells A375 used in the present invention were purchased from the American Type Culture Collection (ATCC).

[0388] The cells were cultured in a medium containing 10% fetal bovine serum (ExCell Bio) and 100 units / ml penicillin and 100 μg / ml streptomycin (VivaCell) in a 37°C constant temperature cell culture incubator (humidity 95%, CO2 concentration 5%). RPMI-1640 (Gibco) and DMEM high glucose medium (Gibco) were used for cell culture.

[0389] Hela, A549, SUDHL-4, Z-138 and A375 cells were respectively cultured at 5×10 3 pcs / hole and 2.5×10 3 Cells were inoculated at a density of 100 μl / well in a 96-well plate. After conventional culture for 16 hours, different concentrations of the compounds of the present invention were added in sequence, with a concentration of 4 μmol / L. The control group was added with an equal amount of DMSO. After continued culture for 48 hours, pre-cooled trichloroacetic acid (50%, w / V) was added, 100 μl / well, and incubated overnight at 4°C to fix the cells. After fixation, the cells were washed 5 times with distilled water and air-dried. 100 μl of SRB dye solution (4%, w / V) was added to each well and incubated at room temperature for 30 minutes for staining. The dye solution was aspirated and washed 5-6 times with 1% acetic acid to remove unbound dye. After air drying, 100 μl of Tris base solution with a concentration of 10 mmol / L was added to each well, and the bound SRB dye was dissolved by shaking. The 96-well plate was placed in an ELISA instrument, and the OD value was measured at a wavelength of 530 nm. The cell survival rate (%) was calculated according to the following formula.

[0390]

[0391] The effects of the compounds of the present invention on the proliferation of Hela, A549, SUDHL-4, Z-138 and A375 cells were statistically analyzed, and the evaluation results are shown in Table 4.

[0392] The inhibitory activity of the compounds of the present invention on the viability of various tumor cells is expressed by the inhibition rate (%) of the compounds at a concentration of 4 μmol / L, wherein "+", "++", "+++" and "++++" respectively indicate that the inhibition rate of the compound is less than 25%, between 25% and 50%, between 50% and 75% and greater than 75%.

[0393] Table 4 Results of the cell inhibition activity of the compounds of the present invention in various tumor cells

[0394]

[0395] The results showed that some of the compounds of the present invention exhibited certain proliferation inhibition activity on Hela, A549, SUDHL-4, Z-138 and A375 cells. These results showed that the compounds of the present invention had good antiproliferative effects on other tumor cells including cervical cancer cells, human non-small cell lung cancer cells, human B lymphoma cells, human mantle cell lymphoma cells, and human melanoma cells.

[0396] Example 5: Evaluation of the inhibitory activity of the representative compound YZ-18 of the present invention on the growth of breast cancer cell HCC1806 xenograft tumors in nude mice

[0397] Nude mice (about 6 weeks old) were purchased from SJA Experimental Animal Co., Ltd. (Changsha, China) and housed in the specific pathogen-free (SPF) animal facility of the Kunming Institute of Zoology, Chinese Academy of Sciences. HCC1806 cells (1×10 6 cells / mouse) were injected bilaterally into the fourth mammary fat pad of nude mice. 3 At 14:00, the nude mice were randomly divided into a control group and a treatment group. The treatment group was intraperitoneally injected with YZ-18 (50 mg / kg) once every other day for a total of 4 injections. The tumor size was measured with a vernier caliper, and the weight of the nude mice was recorded with a weighing scale every 2 days. After 15 days, tumor samples were collected for analysis. This animal experiment was approved by the Animal Ethics Committee of Kunming Medical University (kmmu20240729).

[0398]

[0399] The use and welfare of experimental animals were carried out in accordance with the regulations of the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). The health status and mortality of the animals were monitored daily, and routine examinations included observing the effects of the test substances and drugs on the daily behavior of the animals, such as behavioral activities, weight changes, and physical signs.

[0400] The results showed that compound YZ-18 alone had a certain inhibitory effect on the growth of HCC1806 breast cancer xenograft tumors in nude mice. Figure 7-Figure 8 The inhibitory effect of YZ-18 on the growth of HCC1806 breast cancer xenograft tumors in nude mice, such as Figure 7 and Figure 8 As shown, compound YZ-18 inhibited the growth of HCC1806 breast cancer xenograft tumors in nude mice to a certain extent, and had no significant effect on the body weight of nude mice ( Fig. 9 ).

[0401] In summary, the compounds of the present invention not only show significant activity in inducing PRMT5 protein degradation, but also show obvious anti-proliferation effect, and show a certain degree of tumor growth inhibition effect in vivo, and are a class of compounds with potential for treating PRMT5-related malignant tumors.

Claims

1. A PROTACs compound targeting degradation of PRMT5, characterized in that: The compound comprises a compound as represented by formula (I), a related analog of the compound as represented by formula (I), or a pharmaceutically acceptable salt, metabolite or prodrug thereof: The analogs include stereoisomers, geometric isomers, tautomers, nitrogen oxides, solvates and pharmaceutically acceptable salts; Wherein, X represents a ligand that binds to the PRMT5 protein, Z represents a ligand of the E3 ubiquitin ligase, and Y represents a linker group that connects X and Z; The X is a structure as described in formula (II) or formula (III): or The structure of Z is shown in the following formula (IV) to formula (VI): or or in: A is selected from any one of CH2, C(O), SO, SO2; R is selected from any one of F or H; M is selected from Any one of; P is selected from any one of H, F or CF3; Q is selected from any one of -NH- or -CH2-; The Y is an optional structure as shown below: in: m is an integer between 0 and 12; n is an integer between 1 and 10; q is an integer between 0 and 8; t is an integer between 0 and 8.

2. The PROTACs compound targeting PRMT5 degradation according to claim 1, characterized in that The ligand Z of the E3 ubiquitin ligase is selected from one of the following compounds:

3. The PROTACs compound targeting degradation of PRMT5 according to claim 1, characterized in that include: N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-3,6,9,12-tetraoxopentadecane-15-yl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-3,6,9,12,15-pentaoxaoctadecane-18-yl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)pentanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)hexanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(9-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)nonanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)ethoxy)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(9-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)nonanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(11-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)undecanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(12-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)dodecanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((2-(2,6-dioxopiperidin-3-yl)-6-fluoro-1,3-dioxoisoindolin-5-yl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)octyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)octyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)octyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)pentyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethoxy)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)pentanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(6-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)hexanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oct-7-ynyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(11-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)undec-10-ynyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(2-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)acetyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(3-(4-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)butyl)piperazin-1-yl)propanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(1-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperidin-4-carbonyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(1-(3-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)propyl)piperidin-4-carbonyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)benzoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-(4-(3-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)propyl)piperazin-1-yl)benzoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(5-(7-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,7-diazaspiro[3.5]nonan-2-yl)pentanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(4-(9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-3,9-diazaspiro[5.5]undec-3-yl)butanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(11-(4-(2,6-dioxopiperidin-3-yl)phenoxy)undecanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(10-((4-(2,6-dioxopiperidin-3-yl)phenyl)amino)decanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-(4-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperazin-1-yl)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(2-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)hexyl)amino)-2-oxoethyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetylamino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetamido)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(6-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetamido)hexanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(8-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)acetylamino)octanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(7-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)acetamido)heptanoyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((1-(2-(3-((4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)acetyl)piperidin-4-yl)amino)pyrimidine-4-carboxamide; N-((S)-3-(3,4-dihydroisoquinolin-2(1H)-yl)-2-hydroxypropyl)-6-((4-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)decanamido)cyclohexyl)amino)pyrimidine-4-carboxamide.

4. A pharmaceutical composition, characterized in that A PROTACs compound targeting PRMT5 degradation according to any one of claims 1 to 3.

5. The pharmaceutical composition according to claim 4, characterized in that The pharmaceutical composition further comprises a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle or a combination thereof.

6. Use of the PROTACs compound according to any one of claims 1-3 or the pharmaceutical composition according to any one of claims 4-5 in the preparation of a drug for degrading or inhibiting PRMT5.

7. Use of the PROTACs compound according to any one of claims 1 to 3 or the pharmaceutical composition according to any one of claims 4 to 5 in the preparation of a drug for preventing or treating tumors.

8. The use according to claim 7, characterized in that: The tumor is at least one of breast cancer, cervical cancer, non-small cell lung cancer, B cell lymphoma, mantle cell lymphoma and melanoma.

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