Human tumor therapeutics targeting eRF3a protein degradation

Abstract

Provided are a targeted protein degradation compound, TPB-L-E3B, ​​its synthesis method, and uses. Such compound can treat human tumor diseases through the mechanism of eRF3a targeted degradation, and in vitro experiments have shown great potential in treating such diseases, particularly prostate cancer, ovarian cancer, liver cancer, cervical cancer, leukemia, and breast cancer.

Description

[Technical Field]

[0001] The present invention relates to the field of medicinal chemistry, and in particular to targeted protein degradation compounds, methods for their synthesis, and uses in drug preparation. [Background technology]

[0002] PROTACs (Protein Degradation Targeting Chimeras) are bifunctional molecules that utilize the cell's own ubiquitin proteasome system to tag and promote the degradation of target proteins. They generally consist of three parts: an E3 ligase (i.e., E3 ubiquitin ligase) binding part, a linker part, and a target protein binding part. PROTACs have been under development for over 20 years, and many have now entered the clinical stage, demonstrating certain clinical advantages.

[0003] Molecular gels (MGs) are small molecules that induce the interaction of E3 ligases with target proteins, leading to their degradation by the cellular ubiquitin proteasome system. Representative drugs include thalidomide, pomalidomide, and lenalidomide.

[0004] Both PROTACs and MGs rely on the ubiquitin proteasome system to degrade target proteins, but their mechanisms differ. The ability of PROTACs to degrade target proteins often depends on their ability to bind to E3 ligases and target proteins, respectively, whereas the ability of MGs to degrade target proteins often depends on the ability of E3 ligases to bind to target proteins after the MG molecule is added. PROTACs generally use linear or rigid ring structures with a large number of atoms as linkers, whereas MGs contain linkers with fewer atoms or no linkers at all. While PROTACs currently have a wide range of target proteins that can be degraded, which is related to their unique mechanisms, only a limited number of degrading proteins are known for molecular gels, typically including Ikaros, Aiolos, eRF3a (GSPT1), and CK1α.

[0005] GSPT1 (G1-to-S phase transition 1 gene) is located on the long arm of human chromosome 16, region 13. It spans a total of 7,141 base pairs and consists of 15 exons. The protein-coding region, spanning regions 237–2150, encodes a total of 637 amino acids. GSPT1 (eukaryotic peptide chain release factor 3, eRF3a) is an important member of the peptide chain release factor family and is involved in various important biological processes, including protein translation termination, apoptosis, cell cycle regulation, cytoskeleton assembly, and tumorigenesis. Numerous studies have demonstrated that eRF3a is closely related to the development and progression of various human tumors. Currently, drugs targeting eRF3a have entered clinical trials, but no approved drugs have yet been approved. Increasing research has demonstrated that targeted degradation of eRF3a has beneficial therapeutic effects on many human tumors.

[0006] Steroid 5α-reductase (SRD5A) is an important enzyme system in the human body. It has three isozymes: SRD5A1, SRD5A2, and SRD5A3. All of these are membrane proteases of NADPH-dependent reduced coenzyme II (NADPH) located in the endoplasmic reticulum. They are widely distributed in various tissues of the human body with varying levels of expression. The main function of SRD5A1 and SRD5A2 is to reduce certain steroid substrates, such as testosterone to dihydrotestosterone. SRD5A3 not only reduces certain steroid substrates, but also plays an important role in the N-glycosylation modification of certain proteins.

[0007] Various literature reports that many human diseases are associated with abnormally high expression or activity of steroid 5α-reductase, especially human tumor diseases such as breast cancer, testicular cancer, ovarian cancer, thyroid cancer, lung cancer, esophageal cancer, liver cancer, and prostate cancer. Many researchers believe that SRD5A1, SRD5A2, and SRD5A3 are all potential biomarkers for some tumors and are associated with tumor progression and prognosis. Therefore, directly or indirectly targeting or downregulating the biological function of steroid 5α-reductase may be a potential mechanism for treating tumors.

[0008] Finasteride and dutasteride are known steroid 5α-reductase inhibitors and have been clinically used as specific SRD5A2 inhibitors to treat human diseases associated with abnormal expression of SRD5A2, such as benign prostatic hyperplasia and androgen-dependent alopecia. Currently, there are no drugs that effectively inhibit the biological functions of SRD5A1 and SRD5A3 to clinically treat human diseases associated with abnormal expression of SRD5A1 or SRD5A3.

[0009] Prostate cancer is one of the most common tumors in the male urinary system. The androgen receptor (AR) plays a key role in the development and progression of prostate cancer. After binding and activation by androgen, it can regulate the expression of downstream target genes, thereby promoting the progression and metastasis of prostate cancer. Androgen deprivation therapy (ADT) is the primary treatment for late-stage prostate cancer, but most patients develop drug resistance after 18–24 months of treatment and progress to castration-resistant prostate cancer (CRPC). In CRPC, androgen levels are suppressed, but the AR signaling pathway plays a key role in CRPC. Current research suggests that the drug resistance mechanisms of abiraterone and enzalutamide are primarily due to AR gene mutations, including AR amplification, AR mutations, and AR shearing variants (AR-Vs). AR-Vs are AR cutoff variants. Currently, more than 20 AR-Vs have been reported, of which AR-V7 is the most frequently detected and most widely studied shearing variant in CRPC. Recent studies have shown that AR-V7 plays an important role in the progression of CRPC and the development of drug resistance, and can be used as a molecular marker to guide drug selection for CRPC patients.

[0010] The present invention provides compounds that degrade eRF3a as MG, which have shown great potential in the treatment of various human tumors in in vitro experiments, such as prostate cancer, ovarian cancer, liver cancer, breast cancer, pancreatic cancer, glioma, cervical cancer, leukemia, gastric cancer, and colon cancer, particularly most human prostate cancers. Such compounds inhibit the activity of SRD5A1 and SRD5A3 in some prostate cancers, or downregulate the expression of SRD5A1 and SRD5A3 proteins, while simultaneously degrading eRF3a, thereby affecting the expression of proteins such as AR and AR-V7 that are necessary for the survival and proliferation of some tumor cells, thereby achieving the goal of treating human prostate cancer. Summary of the Invention

[0011] The present invention provides pharmaceutical targeted protein degrading compounds, compositions comprising said compounds, methods for their preparation and uses. An object of the present invention is to provide a compound having the structure TPB-L-E3B, ​​where TPB is the target protein binding moiety, L is the linker moiety, and E3B is the E3 ligase binding moiety.

[0012] Specifically, in the compound TPB-L-E3B, ​​TPB is represented by the following structure (I). JPEG0007796456000001.jpg34170L has the following structures (IIA) and (IIB): It is one selected from JPEG0007796456000002.jpg19170. X is CH2, O, NH, or CF2, n is an integer of 2-3, Y is NH or O, and Z is NH or O. E3B has the following structure: It is one selected from JPEG0007796456000003.jpg21170. JPEG0007796456000004.jpg8170 indicates that C is in the R configuration, S configuration, or RS mixed configuration. R1 indicates that H at any one of the other three positions on the benzene ring is substituted with F. Preferably, L has the following structure (IIA) or (IIB): JPEG0007796456000005.jpg17170. X is CH2, O, NH, or CF2, n is an integer of 2 to 3, Y is NH or O, and Z is NH. E3B has the following structures (IIIB) and (IIIC): It is one of the following: JPEG0007796456000006.jpg22170. JPEG0007796456000007.jpg8170 indicates that C is in the R arrangement, the S arrangement, or the RS mixed arrangement. More preferably, L has the following structure (IIA) or (IIB): JPEG0007796456000008.jpg19170. X is CH2, n is the number 3, Y is NH, and Z is NH. E3B has the structure (IIIB): Represented by JPEG0007796456000009.jpg21170. JPEG0007796456000010.jpg8170 indicates that C is in the R arrangement, the S arrangement, or the RS mixed arrangement.

[0013] Additionally, preferred compounds of the present invention include, but are not limited to, the following structures: Another object of the present invention is to provide a method for synthesizing the compound TPB-L-E3B. The compound according to the present invention can be synthesized by several methods described below. 1. When Z is NH, (1)E3B is JPEG0007796456000014.jpg21170, and the compound TPB-L-E3B has the following structure CA: Represented as JPEG0007796456000015.jpg41170. Here, R2 is JPEG0007796456000016.jpg17170, where X is CH2, O, NH, or CF2, n is an integer of 2-3, Y is NH or O, R3 is hydrogen or R1, and R1 indicates that H at any one of the other three positions on the benzene ring is replaced with F. In this case, compound TPB-L-E3B can be synthesized by the following Method 1.

[0014] Method 1 JPEG0007796456000017.jpg33170JPEG0007796456000018.jpg43170JPEG0007796456000019.jpg25170, where X is CH2, O, NH, or CF2, n is an integer of 2-3, Y is NH or O, R3 is hydrogen or R1, R1 indicates that H at any one of the other three positions on the benzene ring is substituted with F, R4 is H or a Boc group, and X1 is a halogen.

[0015] (2) E3B is JPEG0007796456000020.jpg22170, and the compound TPB-L-E3B has the following structure CA1: It can be represented as JPEG0007796456000021.jpg42170. Here, R2 is JPEG0007796456000022.jpg18170, where X is CH2, O, NH, or CF2, n is an integer of 2-3, and Y is NH or O. In this case, the compound TPB-L-E3B can be synthesized by Method 2 below.

[0016] Method 2 JPEG0007796456000023.jpg42170JPEG0007796456000024.jpg40170JPEG0007796456000025.jpg23170, where X is CH2, O, NH, or CF2, n is an integer of 2-3, and Y is NH or O.

[0017] 2. When Z is O, (1)E3B is JPEG0007796456000026.jpg21170, and the compound TPB-L-E3B has the following structure CA2: It can be represented as JPEG0007796456000027.jpg36170. Y is NH or O, n is an integer of 2-3, R3 is hydrogen or R1, and R1 indicates that H at any one of the other three positions on the benzene ring is replaced with F. In this case, compound TPB-L-E3B can be synthesized by the following Method 3.

[0018] Method 3: JPEG0007796456000028.jpg29170JPEG0007796456000029.jpg39170Y is NH or O, n is an integer of 2-3, R3 is hydrogen or R1, and R1 indicates that H at any one of the other three positions on the benzene ring is replaced with F.

[0019] (2) E3B is JPEG0007796456000030.jpg21170, and compound TPB-L-E3B has the structure CA3: It can be represented as JPEG0007796456000031.jpg37170. Y is NH or O, and n is an integer of 2-3. In this case, the compound TPB-L-E3B can be synthesized by Method 4 below.

[0020] Method 4: JPEG0007796456000032.jpg32170JPEG0007796456000033.jpg33170Y is NH or O, and n is an integer of 2-3.

[0021] In another aspect, the present invention provides the use of the compound TPB-L-E3B in the preparation of a tumor therapeutic drug. Drugs containing TPB-L-E3B can treat human tumor diseases through the eRF3a target protein degradation mechanism. The tumors include, but are not limited to, prostate cancer, ovarian cancer, liver cancer, esophageal squamous cell carcinoma, breast cancer, pancreatic cancer, cervical cancer, lung cancer, leukemia, gastric cancer, or colon cancer. Preferably, the tumor is prostate cancer, ovarian cancer, liver cancer, breast cancer, pancreatic cancer, cervical cancer, or lung cancer. More preferably, the tumor is prostate cancer. [Brief explanation of the drawings]

[0022] [Figure 1] FIG. 10 is a graph showing the weight data of mice administered C29 intraperitoneally (IP). [Figure 2] FIG. 1 shows tumor volume data for mice administered C29 intraperitoneally (IP). [Figure 3] FIG. 1 shows the weight data of C59 intraperitoneally injected (IP) and oral gavage (PO) administered mice. [Figure 4] FIG. 1 shows tumor volume data for C59 intraperitoneally injected (IP) and oral gavage (PO) administered mice. [Figure 5] This is a graph showing the weight data of C63 mice administered orally (PO). [Figure 6] This is a graph showing tumor volume data for mice given C63 by oral gavage (PO). [Figure 7] FIG. 10 is a WB image of downregulation of related proteins in 22Rv1 cells after 24 h by different concentrations of C59. [Figure 8] Figure 10 shows WB images of down-regulation of related proteins in 22Rv1 cells at different times by C59 at a concentration of 3 nM. [Figure 9] FIG. 10 is a WB image of downregulation of related proteins in PC-3 cells after 24 h by different concentrations of C59. [Figure 10] Figure 10 is a WB image of down-regulation of related proteins in HepG2 cells after 24 h by different concentrations of C59. DETAILED DESCRIPTION OF THE INVENTION

[0023] The present invention will be further described below with reference to specific examples, which are provided for illustrative purposes only and are not intended to limit the scope of the present invention.

[0024] Starting materials according to the present invention can be obtained commercially, synthesized by methods known in the art, or synthesized by the methods described herein.

[0025] The English abbreviations used herein are as follows: Boc: tert-butoxycarbonyl pyBoP: benzotriazol-1-yl-oxytripyrrolidinylphosphonium hexafluorophosphate DIPEA: N,N-diisopropylethylamine DMF: N,N-dimethylformamide STAB: sodium triacetoxyborohydride DME: 1,2-dichloroethane XPhoS: 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl Cbz: benzyloxycarbonyl V / V: volume ratio SPF: Specific pathogen free IVC System: Independently Ventilated Cage System CCK8: Cell viability detection reagent L-Glutamine PBS buffer: phosphate buffer RIPA Lysate: Radioimmunoprecipitation Assay Buffer PVDF membrane: Polyvinylidene fluoride membrane TBST solution: a buffered salt solution of trihydroxymethylaminomethane and hydrochloric acid containing 2% Tween

[0026] Abbreviations used herein not included above represent their meanings generally accepted in the art.

[0027] Example 1: Synthesis of N-(2-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)ethyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C1) JPEG0007796456000034.jpg41170 Step 1: Synthesis of N-(2-(Boc-amino)ethyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (IV-1) JPEG0007796456000035.jpg35170JPEG0007796456000036.jpg34170At room temperature, 1.00 g of 3-oxo-4-aza-5α-androst-1-ene-17β-carboxylic acid (I-1), 0.74 g of N-Boc ethylenediamine hydrochloride, 2.00 g of pyBoP, and 20 ml of DMF were sequentially added to a 100 ml three-neck flask and stirred for 5 minutes. 1.22 g of DIPEA was added dropwise to the three-neck flask and the reaction was allowed to proceed overnight. 50 ml of water was added to quench the reaction, yielding a reaction solution. The reaction solution was extracted three times with 50 ml of ethyl acetate. The combined organic phases were washed successively with 50 ml of 1 mol / L aqueous hydrochloric acid, 50 ml of saturated aqueous sodium carbonate, and 50 ml of saturated aqueous sodium chloride. The organic phases were then dried over anhydrous sodium sulfate and concentrated to dryness to obtain 2.56 g of a black oil. The oil was purified by column chromatography (eluent: dichloromethane:methanol=20:1) to obtain 0.70 g of an off-white solid (i.e., compound IV-1).

[0028] Step 2: Synthesis of N-(2-aminoethyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (V-1) 0.70 g of compound IV-1 and 10 ml of dichloromethane were added to a 50 ml three-neck flask at room temperature, followed by the dropwise addition of 10 ml of trifluoroacetic acid. After reacting for 10 minutes, the reaction mixture was concentrated to dryness to obtain an oil. The oil (i.e., the crude product of compound V-1) was used in the next reaction without further purification.

[0029] Step 3: Synthesis of N-(2-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)ethyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C1) JPEG0007796456000038.jpg43170JPEG0007796456000039.jpg35170The oil obtained in Step 2 (i.e., the crude product of Compound V-1), 0.15 g of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione (III-1), 0.20 g of DIPEA, and 5 ml of DMF were sequentially added to a reaction flask at room temperature. The reaction flask was placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 20 ml of water was added to the reaction flask, and the mixture was extracted three times with dichloromethane (20 ml each time). The organic phases were combined, washed with 20 ml of water, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain an oil. This oil was purified by column chromatography (eluent: dichloromethane:methanol=10:1) to give 40 mg of compound C1.

[0030] Mass spectrum data of compound C1: MS (ESI): (M+H) + ,616.3 The nuclear magnetic resonance data of compound C1 is as follows: HNMR(400MHz,CDCl3)δ:8.45-8.47(m,1H),7.48-7.50(m,1H),7.00-7.03(m,1H ),6.78-6.81(m,1H),6.62-6.65(m,1H),5.73-5.76(m,2H),5.57-5.61(m,1H), 4.89-4.92(m,1H),3.10-3.22(m,5H),2.65-2.80(m,3H),2.01-2.15(m,2H),1. 5-1.73(m,7H),1.15-1.47(m,6H),0.99-1.05(m,2H),0.93(s,3H),0.70(s,3H)

[0031] Example 2: Synthesis of N-(2-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)ethyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C2) JPEG0007796456000040.jpg34170JPEG0007796456000041.jpg37170In Step 3 of Example 1, 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione was replaced with 5-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione (i.e., compound III-2), and the synthesis was carried out using the same synthetic method as in Example 1.

[0032] Mass spectrum data of compound C2: MS (ESI): (M+H) + ,616.3 The nuclear magnetic resonance data of compound C2 is as follows: HNMR(400MHz,CDCl3)δ:8.85(s,1H),7.45-7.48(m,1H),6.91(s,1H),6.69-6.71(m,2H),5.73-5.77(m,2H),5.41-5.45(m,1H),4.92-4.97(m,1 H),3.08-3.27(m,5H),2.65-2.75(m,3H),2.01-2.15(m,2H),1.58-1.93 (m,7H),1.15-1.47(m,6H),0.95-1.01(m,2H),0.94(s,3H),0.71(s,3H)

[0033] Example 3: Synthesis of N-(3-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)propyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C3) JPEG0007796456000042.jpg40170 The compound was synthesized by the same synthesis method as in Example 1, except that N-Boc-1,3-propylenediamine hydrochloride was used in Step 1 of Example 1 instead of N-Boc-ethylenediamine hydrochloride.

[0034] Mass spectrum data of compound C3: MS (ESI): (M+H) + ,630.3 The nuclear magnetic resonance data of compound C3 is as follows: HNMR(400MHz,CDCl3)δ:8.47-8.49(m,1H),7.46-7.48(m,1H),7.01-7.03(m,1H ),6.78-6.83(m,1H),6.62-6.65(m,1H),5.72-5.76(m,2H),5.57-5.60(m,1H), 4.89-4.92(m,1H),3.10-3.22(m,5H),2.65-2.80(m,3H),2.01-2.15(m,2H),1. 73-1.98(m,9H),1.15-1.47(m,6H),0.99-1.05(m,2H),0.92(s,3H),0.68(s,3H)

[0035] Example 4: Synthesis of N-(3-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)propyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C4) JPEG0007796456000043.jpg34170 The synthesis was carried out in the same manner as in Example 1, except that N-Boc ethylenediamine hydrochloride in Step 1 of Example 1 was replaced with N-Boc-1,3-propylenediamine hydrochloride, and 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione in Step 3 was replaced with 5-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione.

[0036] Mass spectrum data of compound C4: MS (ESI): (M+H) + ,630.3 The nuclear magnetic resonance data of compound C4 is as follows: HNMR(400MHz,CDCl3)δ:8.65-8.68(m,1H),7.42-7.46(m,1H),6.88(s,1H),6. 65-6.68(m,2H),5.73-5.77(m,2H),5.39-5.42(m,1H),4.85-4.90(m,1H),3.2 5-3.27(m,2H),3.10-3.15(m,3H),2.63-2.75(m,3H),1.91-2.10(m,5H),1.89 -1.73(m,6H),1.15-1.47(m,6H),0.95-1.01(m,2H),0.91(s,3H),0.68(s,3H)

[0037] Example 5: Synthesis of N-(4-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)butyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C5) JPEG0007796456000044.jpg43170 The synthesis was carried out in the same manner as in Example 1, except that N-Boc-1,4-butanediamine hydrochloride was used in Step 1 of Example 1 instead of N-Boc-ethylenediamine hydrochloride.

[0038] Mass spectrum data of compound C5: MS (ESI): (M+H) + ,644.3 The nuclear magnetic resonance data of compound C5 is as follows: HNMR(400MHz,CDCl3)δ:8.77-8.79(m,1H),7.42-7.45(m,1H),7.02-7.06(m,1H),6. 77-6.83(m,1H),6.62-6.68(m,1H),5.72-5.76(m,2H),5.58-5.60(m,1H),4.89-4.9 2(m,1H),3.11-3.28(m,5H),2.64-2.80(m,3H),2.01-2.15(m,3H),1.89-1.97(m,2H ),1.51-1.73(m,7H),1.15-1.47(m,7H),0.93-1.08(m,2H),0.90(s,3H),0.65(s,3H)

[0039] Example 6: Synthesis of N-(4-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)butyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C6) JPEG0007796456000045.jpg31170 The synthesis was carried out in the same manner as in Example 1, except that in Step 1 of Example 1, N-Boc ethylenediamine hydrochloride was replaced with N-Boc-1,4-butanediamine hydrochloride, and in Step 3, 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione was replaced with 5-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione.

[0040] Mass spectrum data of compound C6: MS (ESI): (M+H) + ,644.3 The nuclear magnetic resonance data of compound C6 is as follows: HNMR(400MHz,CDCl3)δ:8.71-8.74(m,1H),7.42-7.48(m,1H),6.85(s,1H),6. 62-6.68(m,2H),5.73-5.76(m,2H),5.38-5.42(m,1H),4.83-4.85(m,1H),3.2 4-3.27(m,2H),3.10-3.15(m,3H),2.63-2.79(m,3H),2.05-2.17(m,3H),1.89 -1.97(m,2H),1.17-1.75(m,14H),0.95-1.01(m,2H),0.91(s,3H),0.68(s,3H)

[0041] Example 7: Synthesis of N-(5-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)pentyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C7) JPEG0007796456000046.jpg33170 The synthesis was carried out in the same manner as in Example 1, except that N-Boc-1,5-pentylenediamine hydrochloride was used in Step 1 of Example 1 instead of N-Boc-ethylenediamine hydrochloride.

[0042] Mass spectrum data of compound C7: MS(ESI):(M+H) + ,658.4 The nuclear magnetic resonance data of compound C7 is as follows: HNMR(400MHz,CDCl3)δ:8.60-8.62(m,1H),7.44-7.48(m,1H),7.03-7.06(m,1H),6.77-6 .82(m,1H),6.62-6.68(m,1H),5.71-5.76(m,2H),5.54-5.56(m,1H),4.85-4.88(m,1H), 3.29-3.31(m,2H),3.11-3.16(m,3H),2.64-2.80(m,3H),2.01-2.15(m,5H),1.89-1.97( m,2H),1.51-1.73(m,5H),1.15-1.47(m,9H),0.93-1.08(m,2H),0.90(s,3H),0.65(s,3H)

[0043] Example 8: Synthesis of N-(5-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)pentyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C8) JPEG0007796456000047.jpg34170 The synthesis was carried out in the same manner as in Example 1, except that N-Boc ethylenediamine hydrochloride in Step 1 of Example 1 was replaced with N-Boc-1,5-pentylenediamine hydrochloride, and 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione in Step 3 was replaced with 5-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione.

[0044] Mass spectrum data of compound C8: MS(ESI):(M+H) + ,658.4 The nuclear magnetic resonance data of compound C8 is as follows: HNMR(400MHz,CDCl3)δ:8.65-8.67(m,1H),7.46-7.49(m,1H),6.84(s,1H),6.62-6 .68(m,2H),5.71-5.76(m,2H),5.38-5.43(m,1H),4.81-4.85(m,1H),3.24-3.27(m ,2H),3.10-3.15(m,3H),2.63-2.79(m,3H),2.02-2.17(m,5H),1.89-1.97(m,2H), 1.51-1.75(m,5H),1.17-1.47(m,9H),0.93-1.08(m,2H),0.90(s,3H),0.67(s,3H)

[0045] Example 9: Synthesis of N-(6-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)hexyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C9) JPEG0007796456000048.jpg42170 The compound was synthesized in the same manner as in Example 1, except that N-Boc-1,6-hexamethylenediamine hydrochloride was used in Step 1 of Example 1 instead of N-Boc-ethylenediamine hydrochloride.

[0046] Mass spectrum data of compound C9: MS(ESI):(M+H) + ,672.4 The nuclear magnetic resonance data of compound C9 is as follows: HNMR(400MHz,CDCl3)δ:8.56-8.57(m,1H),7.42-7.48(m,1H),7.02-7.05(m,1H),6.75-6 .82(m,1H),6.62-6.68(m,1H),5.70-5.76(m,2H),5.54-5.56(m,1H),4.88-4.91(m,1H),3 .28-3.33(m,2H),3.11-3.17(m,3H),2.64-2.81(m,3H),2.05-2.17(m,5H),1.89-1.97(m ,2H),1.51-1.73(m,5H),1.15-1.47(m,10H),0.93-1.08(m,3H),0.89(s,3H),0.65(s,3H)

[0047] Example 10: Synthesis of N-(6-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)hexyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C10) JPEG0007796456000049.jpg29170 Step 1: Synthesis of N-(6-(Boc-amino)hexyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (IV-10) JPEG0007796456000050.jpg30170JPEG0007796456000051.jpg33170At room temperature, 2.00 g of compound I-1, 1.92 g of N-Boc-1,6-hexamethylenediamine hydrochloride, 4.00 g of pyBoP, and 30 mL of DMF were sequentially added to a reaction flask and stirred for 5 minutes. 2.44 g of DIPEA was added dropwise to the three-neck flask and allowed to react overnight. The next day, 70 mL of water was added to obtain a reaction solution. The reaction solution was extracted three times (70 mL each time) with ethyl acetate, and the organic phases were combined. The organic phase was washed sequentially with 70 mL of 1 mol / L aqueous hydrochloric acid, 70 mL of saturated aqueous sodium carbonate, and 70 mL of saturated aqueous sodium chloride. The mixture was then dried over anhydrous sodium sulfate and concentrated to dryness to obtain 5.29 g of a black oil. The oil was purified by column chromatography (eluent: dichloromethane:methanol=20:1) to give 1.48 g of compound IV-10.

[0048] Step 2: Synthesis of N-(6-aminohexyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (V-10) 1.40 g of compound IV-10 and 15 ml of dichloromethane were added to a reaction flask at room temperature, followed by dropwise addition of 15 ml of trifluoroacetic acid. After reacting for 10 minutes, the reaction mixture was concentrated to dryness to obtain an oil. The oil (i.e., the crude product of compound V-10) was used in the next reaction without further purification.

[0049] Step 3: Synthesis of N-(6-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)hexyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C10) JPEG0007796456000053.jpg29170JPEG0007796456000054.jpg20170The oil obtained in Step 2 (i.e., the crude product of Compound V-10), 0.66 g of Compound 5-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione (III-2), 0.86 g of DIPEA, and 20 ml of DMF were sequentially added to a reaction flask at room temperature. The reaction flask was placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 30 ml of water was added to the reaction flask, and the mixture was extracted three times with dichloromethane (30 ml each time). The organic phases were combined, washed with 30 ml of water, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain an oil. The oil was purified by column chromatography (eluent: dichloromethane:methanol=10:1) to give 0.46 g of compound C10.

[0050] Mass spectrum data of compound C10: MS (ESI): (M+H) + ,672.4 The nuclear magnetic resonance data of compound C10 is as follows: HNMR(400MHz,CDCl3)δ:8.66-8.67(m,1H),7.48-7.50(m,1H),6.84(s,1H),6.62-6 .68(m,2H),5.70-5.76(m,2H),5.38-5.41(m,1H),4.82-4.86(m,1H),3.24-3.28(m ,2H),3.11-3.18(m,3H),2.63-2.79(m,3H),2.01-2.17(m,5H),1.89-1.97(m,2H), 1.51-1.70(m,5H),1.20-1.47(m,10H),0.93-1.08(m,3H),0.88(s,3H),0.64(s,3H)

[0051] Example 11: Synthesis of N-(7-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)heptyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C11) JPEG0007796456000055.jpg32170 The synthesis was carried out using the same synthetic method as in Example 10, except that N-Boc-1,6-hexamethylenediamine hydrochloride in step 1 of Example 10 was replaced with N-Boc-1,7-heptanediamine hydrochloride, and 5-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione in step 3 was replaced with 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione.

[0052] Mass spectrum data of compound C11: MS (ESI): (M+H) + ,686.4 The nuclear magnetic resonance data of compound C11 is as follows: HNMR(400MHz,CDCl3)δ:8.55-8.57(m,1H),7.44-7.48(m,1H),7.06-7.08(m,1H),6.75-6 .82(m,1H),6.62-6.68(m,1H),5.69-5.73(m,2H),5.53-5.56(m,1H),4.85-4.91(m,1H),3 .25-3.33(m,2H),3.11-3.17(m,3H),2.64-2.81(m,3H),2.05-2.17(m,4H),1.89-1.97(m ,2H),1.51-1.83(m,8H),1.17-1.47(m,10H),0.99-1.08(m,3H),0.88(s,3H),0.66(s,3H)

[0053] Example 12: Synthesis of N-(7-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)heptyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C12) JPEG0007796456000056.jpg50170 The synthesis was carried out in the same manner as in Example 10, except that N-Boc-1,6-hexamethylenediamine hydrochloride in Step 1 of Example 10 was replaced with N-Boc-1,7-heptanediamine hydrochloride.

[0054] Mass spectrum data of compound C12: MS (ESI): (M+H) + ,686.4 The nuclear magnetic resonance data of compound C12 is as follows: HNMR(400MHz,CDCl3)δ:8.09-8.38(m,1H),7.52-7.55(m,1H),6.84(s,1H),6.63-6 .75(m,2H),5.65-5.74(m,2H),5.27-5.30(m,1H),4.71-4.86(m,1H),3.20-3.28(m ,2H),3.10-3.16(m,3H),2.61-2.82(m,3H),2.02-2.16(m,5H),1.90-1.96(m,2H), 1.54-1.72(m,7H),1.17-1.43(m,10H),0.94-1.10(m,3H),0.97(s,3H),0.61(s,3H)

[0055] Example 13: Synthesis of 1-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-4-(4-(1-(3-oxo-4-aza-5α-androst-1-ene-17β-formyl)piperidine)methyl)-piperazine (C13) JPEG0007796456000057.jpg37170 Step 1: Synthesis of 4-(piperidin-4-yl-methyl)-1-Boc piperazine (II-13) a. Synthesis of 4-(4-(1-Cbzpiperidine)methyl)-1-Bocpiperazine (II-13-1) 3.96 g of 1-Cbz-4-piperidineformaldehyde, 3.16 g of N-Boc piperazine, 40 ml of DME, and 0.72 g of glacial acetic acid were sequentially added to a 100 ml three-neck flask at room temperature. 6.90 g of STAB was added with stirring, and the mixture was heated to 40 °C and reacted for 3 hours. The mixture was concentrated to dryness to obtain an oily product. The oily product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 10:1) to obtain 2.71 g of compound II-13-1.

[0056] b. Synthesis of 4-(piperidin-4-yl-methyl)-1-Boc piperazine (II-13) 2.71 g of compound II-13-1, 40 ml of methanol, 270 mg of 5% palladium carbon, and 0.77 g of glacial acetic acid were sequentially added to a 100 ml hydrogenation reactor at room temperature, and the mixture was hydrogenated at 60°C for 4 hours. After completion of the reaction, the mixture was concentrated to dryness to obtain a black oil. The oil was purified by column chromatography (eluent: dichloromethane:methanol = 10:1) to obtain 1.39 g of a gray oil (i.e., crude compound II-13).

[0057] Step 2: Synthesis of 4-(4-(1-(3-oxo-4-aza-5α-androst-1-ene-17β-formyl)piperidine)methyl)-1-Boc piperazine (IV-13) JPEG0007796456000060.jpg31170JPEG0007796456000061.jpg33170At room temperature, 1.11 g of compound I-1, 1.39 g of the crude product of compound II-13, 6.55 g of pyBoP, and 50 ml of DMF were sequentially added to a 100 ml three-neck flask. The mixture was stirred for 5 minutes, and 2.44 g of DIPEA was added dropwise. The mixture was allowed to react overnight. The next day, 50 ml of water was added, and the mixture was extracted three times with ethyl acetate (50 ml each time). The organic phases were combined and washed once with 50 ml of 1 mol / L aqueous hydrochloric acid, 50 ml of saturated aqueous sodium carbonate, and 50 ml of saturated aqueous sodium chloride. The organic phase was then dried over anhydrous sodium sulfate and concentrated to dryness to obtain 2.56 g of a black oil. The oil was purified by column chromatography (eluent: dichloromethane:methanol=20:1) to give 1.52 g of compound IV-13.

[0058] Step 3: Synthesis of 4-(4-(1-(3-oxo-4-aza-5α-androst-1-ene-17β-formyl)piperidine)methyl)-piperazine (V-13) 1.52 g of compound IV-13 and 15 ml of dichloromethane were added to a 50 ml three-neck flask at room temperature, and then 15 ml of trifluoroacetic acid was added dropwise and reacted for 10 minutes. The mixture was concentrated to dryness to obtain an oily product. The oily product (i.e., the crude product of compound V-13) was used in the next reaction without further purification.

[0059] Step 4: Synthesis of 1-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-4-(4-(1-(3-oxo-4-aza-5α-androst-1-ene-17β-formyl)piperidine)methyl)-piperazine (C13) The oil obtained in Step 3 (i.e., the crude product of Compound V-13), 0.17 g of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 0.16 g of DIPEA, and 15 mL of DMF were sequentially added to a reaction flask at room temperature. The reaction flask was then placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 20 mL of water was added, followed by extraction with dichloromethane three times (20 mL each time). The organic phases were combined, washed once with 20 mL of water, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 10:1) to obtain 40 mg of Compound C13.

[0060] Mass spectrum data of compound C13: MS (ESI): (M+H) + ,739.4 The nuclear magnetic resonance data of compound C13 is as follows: HNMR(400MHz,CDCl3)δ:8.57-8.59(m,1H),7.55-7.57(m,1H),7.09-7.13(m,1H),6.87-6 .89(m,1H),6.65-6.67(m,1H),5.73-5.76(m,2H),4.89-4.92(m,1H),3.53-3.59(m,1H),3 .33-3.52(m,3H),3.11-3.28(m,5H),2.51-2.80(m,5H),2.28-2.38(m,4H),2.07-2.21(m ,2H),1.50-1.93(m,10H),1.15-1.47(m,8H),0.99-1.05(m,2H),0.93(s,3H),0.70(s,3H)

[0061] Example 14: Synthesis of 1-(5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-4-(4-(1-(3-oxo-4-aza-5α-androst-1-ene-17β-formyl)piperidine)methyl)-piperazine (C14) JPEG0007796456000064.jpg32170 The synthesis was carried out in the same manner as in Example 13, except that 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione in Step 4 of Example 13 was replaced with 5-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione. Mass spectrum data of compound C14: MS (ESI): (M+H) + ,739.4 The nuclear magnetic resonance data of compound C14 is as follows: HNMR(400MHz,CDCl3)δ:8.77(s,1H),7.49-7.50(m,1H),6.96(s,1H),6.71 -6.72(m,1H),5.88(s,1H),5.73-5.75(m,1H),5.41-5.45(m,1H),4.92-4.9 7(m,1H),3.15-3.49(m,9H),2.59-2.75(m,5H),2.07-2.35(m,6H),1.58-1. 99(m,10H),1.11-1.44(m,8H),0.95-1.01(m,2H),0.94(s,3H),0.71(s,3H)

[0062] Example 15: Synthesis of N-(4-((2-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)ethyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C15) JPEG0007796456000065.jpg38170 The compound was synthesized in the same manner as in Example 17, except that 1,3-dibromopropane in Step 1 of Example 17 was replaced with 1,2-dibromoethane. Mass spectrum data of compound C15: MS (ESI): (M+H) + ,708.3 The nuclear magnetic resonance data of compound C15 is as follows: HNMR(400MHz,CDCl3)δ:8.65(s,1H),7.53-7.57(m,1H),7.36-7.40(m,2H),7.17-7.23(m,3H),7.02-7.03(m,1 H),6.79-6.82(d,1H,J=12Hz),6.69-6.73(m,1H),6.61-6.64(m,1H),5.75(s,1H),5.65-5.68(d,1H,J=12Hz),5 .05-5.09(m,1H),4.02-4.06(q,2H,J=8Hz),3.44-3.48(m,2H),3.18-3.22(m,1H),2.80-2.92(m,1H),2.50-2.5 7(m,1H),2.41-2.44(m,1H),2.01-2.09(m,2H),1.58-1.93(m,6H),1.01-1.48(m,8H),0.97(s,3H),0.80(s,3H)

[0063] Example 16: Synthesis of N-(4-((2-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)ethyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C16) JPEG0007796456000066.jpg45170 The synthesis was carried out in the same manner as in Example 17, except that 1,3-dibromopropane in step 1 of Example 17 was replaced with 1,2-dibromoethane, and 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione in step 4 was replaced with 5-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione.

[0064] Mass spectrum data of compound C16: MS(ESI):(M+H) + ,708.3 The nuclear magnetic resonance data of compound C16 is as follows: HNMR(400MHz,CDCl3)δ:8.85(s,1H),7.45-7.48(m,1H),7.36-7.40(m,1H),7.17-7.23(m ,2H),7.02-7.03(m,2H)6.91(s,1H),6.69-6.71(m,2H),5.73-5.77(m,1H),5.55(s,1H), 4.92-4.97(m,1H),3.99-4.03(m,2H)3.19-3.27(m,3H),2.65-2.75(m,3H),2.01-2.15(m ,2H),1.57-1.93(m,7H),1.15-1.47(m,6H),0.95-1.01(m,2H),0.97(s,3H),0.72(s,3H)

[0065] Example 17: Synthesis of N-(4-((3-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)propyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C17) JPEG0007796456000067.jpg45170Step 1: Synthesis of 4-((3-(N,N-diBocamino)propyl)oxy)aniline (II-17) a. Synthesis of (3-bromopropyl)-N,N-diBoc amine (II-17-1) 4.65 g of 1,3-dibromopropane, 1.00 g of N,N-diBoc amine, 3.18 g of potassium carbonate, and 50 mL of DMF were added sequentially to a 250 mL three-neck flask at room temperature. The resulting reaction solution was heated to 60 °C and reacted for 3 hours. The reaction solution was extracted three times with dichloromethane (40 mL each time), washed once with 40 mL of water, and the combined organic phases were dried over anhydrous sodium sulfate and then concentrated to dryness to obtain a crude product. The crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 15:1) to obtain 0.82 g of a colorless, transparent oil (i.e., Compound II-17-1).

[0066] b. Synthesis of 4-((3-(N,N-diBocamino)propyl)oxy)nitrobenzene (II-17-2) 0.50 g of compound II-17-1, 0.20 g of 4-nitrophenol, 0.40 g of potassium carbonate, and 30 ml of DMF were added sequentially to a 100 ml three-neck flask at room temperature. The resulting reaction solution was heated to 60 °C and reacted for 3 hours. The reaction solution was extracted three times with dichloromethane (40 ml each time), washed once with 40 ml of water, and the combined organic phases were dried over anhydrous sodium sulfate and then concentrated to dryness to obtain a crude product. The crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 5:1) to obtain 0.30 g of a colorless, transparent oil (i.e., compound II-17-2).

[0067] c. Synthesis of 4-((3-(N,N-diBocamino)propyl)oxy)aniline (II-17) JPEG0007796456000070.jpg21170At room temperature, 0.30 g of compound II-17-2, 15 ml of methanol, and 30 mg of 5% palladium on carbon were added sequentially to a 100 ml hydrogenation reactor, and then the hydrogenation reaction was carried out at 25°C for 1 hour. After the reaction was completed, the reaction solution was concentrated to dryness to obtain 0.28 g of a gray-black oil (i.e., the crude product of compound II-17).

[0068] Step 2: Synthesis of N-(4-((3-(N,N-diBocamino)propyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (IV-17) JPEG0007796456000071.jpg37170JPEG0007796456000072.jpg33170At room temperature, 0.26 g of compound I-1, 0.28 g of the crude product of compound II-17, 0.79 g of pyBoP, and 20 ml of DMF were sequentially added to a 50 ml three-neck flask and stirred for 5 minutes. 0.49 g of DIPEA was added dropwise to the reaction solution and allowed to react overnight. The next day, 50 ml of water was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (50 ml each time). The organic phases were combined, washed once with 50 ml of 1 mol / L aqueous hydrochloric acid, 50 ml of saturated aqueous sodium carbonate, and 50 ml of saturated aqueous sodium chloride, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain 2.56 g of a black oil. The oil was purified by column chromatography (eluent: dichloromethane:methanol=20:1) to give 0.21 g of compound IV-17.

[0069] Step 3: Synthesis of N-(4-((3-aminopropyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (V-17) 0.21 g of compound IV-17 and 10 ml of dichloromethane were added sequentially to a 50 ml three-neck flask at room temperature, followed by the dropwise addition of 10 ml of trifluoroacetic acid, and the mixture was allowed to react for 10 minutes. The mixture was concentrated to dryness to obtain an oily product. The oily product (i.e., the crude product of compound V-17) was used in the next reaction without further purification.

[0070] Step 4: Synthesis of N-(4-((3-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)propyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C17) The oil obtained in Step 3 (i.e., the crude product of Compound V-17), 0.09 g of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 0.14 g of DIPEA, and 3 ml of DMF were added to a reaction flask at room temperature. The reaction flask was then placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 15 ml of water was added to the reaction flask, followed by extraction with dichloromethane three times (15 ml each time). The combined organic phase was washed once with 15 ml of water, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 10:1) to obtain 35 mg of Compound C17.

[0071] Mass spectrum data of compound C17: MS (ESI): (M+H) + ,722.3 The nuclear magnetic resonance data of compound C17 is as follows: HNMR(400MHz,CDCl3)δ:8.49(s,1H),7.52-7.56(m,1H),7.35-7.39(m,2H),7.16-7.22(m,3H),7.02-7.03(m,1 H),6.79-6.82(d,1H,J=12Hz),6.69-6.73(m,1H),6.61-6.64(m,1H),5.75(s,1H),5.65-5.68(d,1H,J=12Hz),5 .05-5.09(m,1H),4.02-4.06(q,2H,J=8Hz),3.44-3.48(m,2H),3.18-3.22(m,1H),2.80-2.92(m,1H),2.50-2.5 7(m,1H),2.41-2.44(m,1H),2.01-2.09(m,4H),1.58-1.79(m,6H),0.97-1.48(m,8H),0.93(s,3H),0.81(s,3H)

[0072] Example 18: Synthesis of N-(4-((3-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)propyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C18) JPEG0007796456000075.jpg33170 The synthesis was carried out in the same manner as in Example 17, except that 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione was replaced with 5-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione in Step 4 of Example 17.

[0073] Mass spectrum data of compound C18: MS (ESI): (M+H) + ,722.3 The nuclear magnetic resonance data of compound C18 is as follows: HNMR(400MHz,CDCl3)δ:8.55(s,1H),7.44-7.45(m,1H),7.37-7.40(m,1H),7.17-7.22(m,2H ),7.01-7.03(m,2H)6.95(s,1H),6.70-6.75(m,2H),5.75-5.77(m,1H),5.57(s,1H),4.92-4. 97(m,1H),4.01-4.04(m,2H)3.25-3.33(m,3H),2.65-2.75(m,3H),2.01-2.15(m,2H),1.78-1 .97(m,4H)1.57-1.73(m,5H),1.15-1.47(m,6H),0.95-1.01(m,2H),0.97(s,3H),0.72(s,3H)

[0074] Example 19: Synthesis of N-(4-((5-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)pentyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C19) JPEG0007796456000076.jpg43170 The synthesis was carried out in the same manner as in Example 17, except that 1,3-dibromopropane in Step 1 of Example 17 was replaced with 1,5-dibromopentane.

[0075] Mass spectrum data of compound C19: MS (ESI): (M+H) + ,750.4 The nuclear magnetic resonance data of compound C19 is as follows: HNMR(400MHz,CDCl3)δ:8.65(s,1H),7.53-7.56(m,1H),7.36-7.43(m,2H),7.15-7.22(m,3H),7.02-7.03(m,1H),6. 79-6.82(d,1H,J=12Hz),6.69-6.73(m,1H),6.65-6.67(m,1H),5.75(s,1H),5.65-5.68(d,1H,J=12Hz),5.05-5.09( m,1H),4.02-4.06(q,2H,J=8Hz),3.44-3.48(m,2H),3.18-3.22(m,1H),2.80-2.92(m,1H),2.50-2.57(m,1H),2.41- 2.44(m,1H),2.01-2.09(m,4H),1.58-1.91(m,10H),0.97-1.11(m,2H),1.15-1.48(m,6H),0.91(s,3H),0.77(s,3H)

[0076] Example 20: Synthesis of N-(4-((5-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)pentyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C20) JPEG0007796456000077.jpg32170 The synthesis was carried out in the same manner as in Example 17, except that 1,3-dibromopropane in step 1 of Example 17 was replaced with 1,5-dibromopentane, and 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione in step 4 was replaced with 5-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione.

[0077] Mass spectrum data of compound C20: MS (ESI): (M+H) + ,750.4 The nuclear magnetic resonance data of compound C20 is as follows: HNMR(400MHz,CDCl3)δ:8.57(s,1H),7.43-7.45(m,1H),7.37-7.41(m,1H),7.17-7.22(m ,2H),7.02-7.05(m,2H),6.95(s,1H),6.71-6.75(m,2H),5.75-5.77(m,1H),5.58(s,1H), 4.92-4.97(m,1H),4.00-4.03(m,2H)3.18-3.31(m,3H),2.61-2.75(m,3H),2.05-2.15(m ,2H),1.61-1.99(m,12H),1.15-1.55(m,7H),0.92-1.01(m,2H),0.96(s,3H),0.72(s,3H)

[0078] Example 21: Synthesis of N-(4-((6-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)hexyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C21) JPEG0007796456000078.jpg37170 The synthesis was carried out in the same manner as in Example 17, except that 1,3-dibromopropane in Step 1 of Example 17 was replaced with 1,6-dibromohexane.

[0079] Mass spectrum data of compound C21: MS (ESI): (M+H) + ,764.4 The nuclear magnetic resonance data of compound C21 is as follows: HNMR(400MHz,CDCl3)δ:8.65(s,1H),7.53-7.56(m,1H),7.36-7.43(m,2H),7.15-7.22(m,3H),7.02-7.03(m,1H),6.79-6 .82(d,1H,J=12Hz),6.69-6.73(m,1H),6.65-6.67(m,1H),5.75(s,1H),5.65-5.68(d,1H,J=12Hz),5.05-5.09(m,1H),4. 02-4.06(q,2H,J=8Hz),3.44-3.48(m,2H),3.18-3.22(m,1H),2.80-2.92(m,1H),2.50-2.57(m,1H),2.41-2.44(m,1H),2 .01-2.09(m,4H),1.85-1.99(m,2H),1.58-1.79(m,10H),0.97-1.11(m,2H),1.15-1.48(m,6H),0.91(s,3H),0.76(s,3H)

[0080] Example 22: Synthesis of N-(4-((6-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)hexyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C22) JPEG0007796456000079.jpg43170 The synthesis was carried out in the same manner as in Example 17, except that 1,3-dibromopropane in step 1 of Example 17 was replaced with 1,6-dibromohexane, and 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione in step 4 was replaced with 5-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione.

[0081] Mass spectrum data of compound C22: MS(ESI):(M+H) + ,764.4 The nuclear magnetic resonance data of compound C22 is as follows: HNMR(400MHz,CDCl3)δ:8.65(s,1H),7.41-7.45(m,1H),7.37-7.39(m,1H),7.18-7.22(m ,2H),7.02-7.05(m,2H),7.01(s,1H),6.71-6.75(m,2H),5.74-5.77(m,1H),5.45(s,1H), 4.88-4.91(m,1H),3.99-4.03(m,2H)3.21-3.35(m,3H),2.61-2.90(m,3H),2.01-2.08(m, 2H),1.71-1.99(m,12H),1.11-1.65(m,11H),0.98-1.08(m,2H),0.93(s,3H),0.70(s,3H)

[0082] Example 23: Synthesis of N-(4-((7-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)heptyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C23) JPEG0007796456000080.jpg43170 The synthesis was carried out in the same manner as in Example 17, except that 1,3-dibromopropane in Step 1 of Example 17 was replaced with 1,7-dibromoheptane.

[0083] Mass spectrum data of compound C23: MS (ESI): (M+H) + ,778.4 The nuclear magnetic resonance data of compound C23 is as follows: HNMR(400MHz,CDCl3)δ:8.55(s,1H),7.53-7.57(m,1H),7.36-7.43(m,2H),7.16-7.22(m,3H),7.01-7.03(m,1H) ),6.80-6.82(d,1H,J=12Hz),6.69-6.74(m,1H),6.65-6.67(m,1H),5.77(s,1H),5.70-5.72(d,1H,J=12Hz),5. 05-5.09(m,1H),4.04-4.06(q,2H,J=8Hz),3.45-3.48(m,2H),2.98-3.15(m,3H),2.44-2.55(m,2H),2.01-2.09 (m,5H),1.85-1.99(m,3H),1.58-1.79(m,10H),0.97-1.11(m,2H),1.15-1.48(m,6H),0.89(s,3H),0.75(s,3H)

[0084] Example 24: Synthesis of N-(4-((7-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)heptyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C24) JPEG0007796456000081.jpg33170 The synthesis was carried out in the same manner as in Example 17, except that 1,3-dibromopropane in step 1 of Example 17 was replaced with 1,7-dibromoheptane, and 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione in step 4 was replaced with 5-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione.

[0085] Mass spectrum data of compound C24: MS (ESI): (M+H) + ,778.4 The nuclear magnetic resonance data of compound C24 is as follows: HNMR(400MHz,CDCl3)δ:8.68(s,1H),7.39-7.42(m,1H),7.37-7.39(m,1H),7.18-7. 25(m,2H),7.01-7.05(m,2H),6.99(s,1H),6.71-6.74(m,2H),5.74-5.77(m,1H),5.6 6(s,1H),4.89-4.95(m,1H),3.99-4.03(m,2H)3.23-3.41(m,3H),2.61-2.77(m,3H), 1.71-2.11(m,16H),1.11-1.65(m,11H),0.98-1.08(m,2H),0.89(s,3H),0.69(s,3H)

[0086] Example 25: Synthesis of N-(4-((8-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)octyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C25) JPEG0007796456000082.jpg38170 The synthesis was carried out in the same manner as in Example 17, except that 1,3-dibromopropane in Step 1 of Example 17 was replaced with 1,8-dibromooctane.

[0087] Mass spectrum data of compound C25: MS (ESI): (M+H) + ,792.4 The nuclear magnetic resonance data of compound C25 is as follows: HNMR(400MHz,CDCl3)δ:8.55(s,1H),7.53-7.57(m,1H),7.36-7.43(m,2H),7.16-7.22(m,3H),7.01-7.03( m,1H),6.80-6.82(d,1H,J=12Hz),6.69-6.74(m,1H),6.65-6.67(m,1H),5.77(s,1H),5.70-5.72(d,1H,J= 12Hz),5.05-5.09(m,1H),4.04-4.06(q,2H,J=8Hz),3.45-3.48(m,2H),2.98-3.15(m,3H),2.44-2.55(m,2 H),2.01-2.09(m,5H),1.58-1.97(m,15H),0.97-1.11(m,2H),1.15-1.48(m,6H),0.90(s,3H),0.71(s,3H)

[0088] Example 26: Synthesis of N-(4-((8-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)octyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C26) JPEG0007796456000083.jpg44170 The synthesis was carried out in the same manner as in Example 17, except that 1,3-dibromopropane in step 1 of Example 17 was replaced with 1,8-dibromooctane, and 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione in step 4 was replaced with 5-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione.

[0089] Mass spectrum data of compound C26: MS (ESI): (M+H) + ,792.4 The nuclear magnetic resonance data of compound C26 is as follows: HNMR(400MHz,CDCl3)δ:8.69(s,1H),7.39-7.42(m,1H),7.35-7.39(m,1H),7.1 9-7.25(m,2H),7.01-7.04(m,2H),6.95(s,1H),6.71-6.74(m,2H),5.74-5.77(m ,1H),5.72(s,1H),4.85-4.93(m,1H),4.00-4.03(m,2H)3.21-3.38(m,3H),2.60 -2.78(m,3H),1.55-2.11(m,20H),1.11-1.47(m,11H),0.86(s,3H),0.68(s,3H)

[0090] Example 27: Synthesis of N-(3-((2-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)ethyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C27) JPEG0007796456000084.jpg45170Step 1: Synthesis of 3-((2-(N,N-diBocamino)ethyl)oxy)aniline (II-27) a. Synthesis of (2-bromoethyl)-N,N-diBoc amine (II-27-1) 4.32 g of 1,2-dibromoethane, 1.00 g of N,N-diBoc amine, 3.18 g of potassium carbonate, and 50 mL of DMF were added sequentially to a 250 mL three-neck flask at room temperature. The resulting reaction solution was heated to 60 °C and reacted for 3 hours. The reaction solution was extracted three times with dichloromethane (40 mL each time), washed once with 40 mL of water, and the combined organic phases were dried over anhydrous sodium sulfate and then concentrated to dryness to obtain a crude product. The crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 15:1) to obtain 0.76 g of a colorless, transparent oil (i.e., Compound II-27-1).

[0091] b. Synthesis of 3-((2-(N,N-diBocamino)ethyl)oxy)nitrobenzene (II-27-2) 0.70 g of compound II-27-1, 0.29 g of 3-nitrophenol, 0.58 g of potassium carbonate, and 30 ml of DMF were sequentially added to a 100 ml three-neck flask at room temperature. The resulting reaction solution was heated to 60 °C and reacted for 3 hours. The reaction solution was extracted three times with dichloromethane (40 ml each time), washed once with 40 ml of water, and the combined organic phases were dried over anhydrous sodium sulfate and then concentrated to dryness to obtain a crude product. The crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 5:1) to obtain 0.46 g of a colorless, transparent oil (i.e., compound II-27-2).

[0092] c. Synthesis of 3-((2-(N,N-diBocamino)ethyl)oxy)aniline (II-27) JPEG0007796456000087.jpg15170 At room temperature, 0.46 g of compound II-27-2, 15 ml of methanol, and 46 mg of 5% palladium on carbon were added sequentially to a 100 ml hydrogenation reactor, and then the hydrogenation reaction was carried out at 25°C for 1 hour. After the reaction was completed, the reaction solution was concentrated to dryness to obtain 0.44 g of a gray-black oil (i.e., the crude product of compound II-27).

[0093] Step 2: Synthesis of N-(3-((2-(N,N-diBocamino)ethyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (IV-27) JPEG0007796456000088.jpg38170JPEG0007796456000089.jpg34170At room temperature, 0.38 g of compound I-1, 0.44 g of the crude product of compound II-27, 1.25 g of pyBoP, and 25 ml of DMF were sequentially added to a reaction flask and stirred for 5 minutes. 0.70 g of DIPEA was added dropwise to the reaction solution and allowed to react overnight. The next day, 50 ml of water was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (50 ml each time). The organic phases were combined and washed once with 50 ml of 1 mol / L aqueous hydrochloric acid, 50 ml of saturated aqueous sodium carbonate, and 50 ml of saturated aqueous sodium chloride. The organic phase was then dried over anhydrous sodium sulfate and concentrated to dryness to obtain 2.49 g of a black oil. The oil was purified by column chromatography (eluent: dichloromethane:methanol=20:1) to give 0.40 g of compound IV-27.

[0094] Step 3: Synthesis of N-(3-((2-aminoethyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (V-27) 0.40 g of compound IV-27 and 15 ml of dichloromethane were added to a reaction flask at room temperature, followed by the dropwise addition of 15 ml of trifluoroacetic acid, and the mixture was allowed to react for 10 minutes. The mixture was concentrated to dryness to obtain an oily product. The oily product (i.e., the crude product of compound V-27) was used in the next reaction without further purification.

[0095] Step 4: Synthesis of N-(3-((2-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)ethyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C27) The oil obtained in Step 3 (i.e., the crude product of Compound V-27), 0.23 g of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 0.36 g of DIPEA, and 6 ml of DMF were sequentially added to a reaction flask at room temperature. The reaction flask was placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 20 ml of water was added to the reaction flask, followed by extraction with dichloromethane three times (20 ml each time). The organic phases were combined, washed once with 20 ml of water, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 10:1) to obtain 0.13 g of Compound C27.

[0096] Mass spectrum data of compound C27: MS (ESI): (M+H) + ,708.3 The nuclear magnetic resonance data of compound C27 is as follows: HNMR(400MHz,CDCl3)δ:8.95-8.96(m,1H),7.51-7.52(m,1H),7.35-7.37(m,1H),7.09-7.12(m,1H),6.95-7.02 (m,2H),6.82-6.86(m,2H),6.70-6.74(m,1H),6.59-6.61(m,1H),6.38-6.42(m,1H),5.75(s,1H),5.71-5.73(m ,1H),4.75-4.83(m,1H),4.01-4.03(m,2H),3.41-3.44(m,2H),3.25-3.28(m,1H),2.60-2.79(m,3H),2.18-2.2 5(m,2H),1.65-2.10(m,6H),1.48-1.53(m,2H),1.16-1.37(m,5H),0.92-0.95(m,2H),0.91(s,3H),0.78(s,3H)

[0097] Example 28: Synthesis of N-(3-((2-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)ethyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C28) JPEG0007796456000092.jpg31170 The synthesis was carried out in the same manner as in Example 27, except that 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione in Step 4 of Example 27 was replaced with 5-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione.

[0098] Mass spectrum data of compound C28: MS (ESI): (M+H) + ,708.3 The nuclear magnetic resonance data of compound C28 is as follows: HNMR(400MHz,CDCl3)δ:8.88-8.90(m,1H),7.45-7.47(m,1H),7.28-7.29(m,1H),7.09-7.12(m,1H),7.00-7.02(m ,2H),6.91(s,1H),6.79-6.83(m,2H),6.70-6.72(m,1H),6.55-6.57(m,1H),6.38-6.42(m,1H),5.75(s,1H),5.71- 5.73(m,1H),4.73-4.83(m,1H),4.01-4.03(m,2H),3.41-3.44(m,2H),3.25-3.28(m,1H),2.60-2.79(m,3H),2.10- 2.25(m,2H),1.65-2.01(m,6H),1.48-1.55(m,2H),1.11-1.37(m,5H),0.92-0.95(m,2H),0.91(s,3H),0.76(s,3H)

[0099] Example 29: Synthesis of N-(3-((3-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)propyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C29) JPEG0007796456000093.jpg36170 Step 1: Synthesis of 3-((3-(N,N-diBocamino)propyl)oxy)aniline (II-29) a. Synthesis of 3-((3-(N,N-diBocamino)propyl)oxy)nitrobenzene (II-29-1) 0.80 g of compound II-17-1, 0.31 g of 3-nitrophenol, 0.64 g of potassium carbonate, and 30 ml of DMF were sequentially added to a 100 ml three-neck flask at room temperature. The resulting reaction solution was heated to 60 °C and reacted for 3 hours. The reaction solution was extracted three times with dichloromethane (40 ml each time), washed once with 40 ml of water, and the combined organic phases were dried over anhydrous sodium sulfate and then concentrated to dryness to obtain a crude product. The crude product was purified by column chromatography (eluent: petroleum ether: ethyl acetate = 5:1) to obtain 0.52 g of a colorless, transparent oil (i.e., compound II-29-1).

[0100] b. Synthesis of 3-((3-(N,N-diBocamino)propyl)oxy)aniline (II-29) JPEG0007796456000095.jpg21170At room temperature, 0.52 g of compound II-29-1, 15 ml of methanol, and 52 mg of 5% palladium on carbon were sequentially added to a 100 ml hydrogenation reactor, and then the hydrogenation reaction was carried out at 25°C for 1 hour. After the reaction was completed, the reaction solution was concentrated to dryness to obtain 0.46 g of a gray-black oil (i.e., the crude product of compound II-29).

[0101] Step 2: Synthesis of N-(3-((3-(N,N-diBocamino)propyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (IV-29) JPEG0007796456000096.jpg38170JPEG0007796456000097.jpg321700.41 g of compound I-1, 0.46 g of the crude product of compound II-29, 1.30 g of pyBoP, and 25 ml of DMF were sequentially added to a reaction flask at room temperature and stirred for 5 minutes. 0.72 g of DIPEA was added dropwise to the reaction solution and allowed to react overnight. The next day, 50 ml of water was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (50 ml each time). The organic phases were combined and washed once with 50 ml of 1 mol / L aqueous hydrochloric acid, 50 ml of saturated aqueous sodium carbonate, and 50 ml of saturated aqueous sodium chloride. The organic phase was then dried over anhydrous sodium sulfate and concentrated to dryness to obtain 2.56 g of a black oil. The oil was purified by column chromatography (eluent: dichloromethane:methanol=20:1) to give 0.52 g of compound IV-29.

[0102] Step 3: Synthesis of N-(3-((3-aminopropyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (V-29) 0.52 g of compound IV-29 and 15 ml of dichloromethane were added sequentially to a reaction flask at room temperature, followed by dropwise addition of 15 ml of trifluoroacetic acid, and the mixture was allowed to react for 10 minutes. The mixture was concentrated to dryness to obtain an oily product. The oily product (i.e., the crude product of compound V-29) was used in the next reaction without further purification.

[0103] Step 4: Synthesis of N-(3-((3-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)propyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C29) The oil obtained in Step 3 (i.e., the crude product of Compound V-29), 0.30 g of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 0.47 g of DIPEA, and 10 ml of DMF were sequentially added to a reaction flask at room temperature. The reaction flask was placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 20 ml of water was added to the reaction flask, followed by extraction with dichloromethane three times (20 ml each time). The combined organic phase was washed once with 20 ml of water, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 10:1) to obtain 0.18 g of Compound C29.

[0104] Mass spectrum data of compound C29: MS(ESI):(M+H) + ,722.3 The nuclear magnetic resonance data of compound C29 is as follows: HNMR(400MHz,CDCl3)δ:8.82-8.94(m,1H),7.38-7.41(m,1H),7.32-7.34(m,1H),7.08-7.12(m,1H),6.98-7.02(m,2 H),6.81-6.88(m,2H),6.70-6.75(m,1H),6.59-6.61(m,1H),6.38-6.45(m,1H),5.85(s,1H),5.72-5.75(m,1H),4.7 4-4.83(m,1H),4.01-4.02(m,2H),3.41-3.44(m,2H),3.22-3.26(m,1H),2.61-2.79(m,3H),2.18-2.22(m,2H),1.80 -2.10(m,5H),1.61-1.75(m,3H),1.48-1.53(m,2H),1.16-1.37(m,5H),0.92-0.95(m,2H),0.90(s,3H),0.78(s,3H)

[0105] Example 30: Synthesis of N-(3-((3-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)propyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C30) JPEG0007796456000100.jpg39170 In step 4 of Example 29, 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione was replaced with 5-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, and the synthesis was carried out using the same synthetic method as in Example 29.

[0106] Mass spectrum data of compound C30: MS (ESI): (M+H) + ,722.3 The nuclear magnetic resonance data of compound C30 is as follows: HNMR(400MHz,CDCl3)δ:8.85(s,1H),7.45-7.47(m,1H),7.28-7.30(m,1H),7.10-7.12(m,1H),6.98-7.02(m,2H) ,6.89(s,1H),6.79-6.83(m,2H),6.70-6.72(m,1H),6.54-6.57(m,1H),6.38-6.42(m,1H),5.76(s,1H),5.70-5.7 2(m,1H),4.75-4.81(m,1H),3.99-4.01(m,2H),3.39-3.43(m,2H),3.25-3.28(m,1H),2.65-2.81(m,3H),2.10-2 .25(m,2H),1.65-2.01(m,8H),1.48-1.55(m,2H),1.11-1.37(m,5H),0.90-0.95(m,2H),0.90(s,3H),0.76(s,3H)

[0107] Example 31: Synthesis of N-(3-((4-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)butyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C31) JPEG0007796456000101.jpg52170 Composite using the same synthesis method as in Example 29.

[0108] Mass spectrum data of compound C31: MS (ESI): (M+H) + ,736.4 The nuclear magnetic resonance data of compound C31 is as follows: HNMR(400MHz,CDCl3)δ:8.86-8.88(m,1H),7.40-7.42(m,1H),7.34-7.36(m,1H),7.08-7.12(m,1H),6.98-7.02(m, 2H),6.81-6.87(m,2H),6.70-6.73(m,1H),6.59-6.61(m,1H),6.38-6.40(m,1H),5.83(s,1H),5.72-5.75(m,1H),4. 78-4.83(m,1H),4.00-4.02(m,2H),3.41-3.45(m,2H),3.22-3.26(m,1H),2.61-2.79(m,3H),2.18-2.22(m,2H),2.0 2-2.10(m,2H),1.80-1.95(m,3H)1.48-1.75(m,7H),1.18-1.45(m,5H),0.92-0.99(m,2H),0.89(s,3H),0.76(s,3H)

[0109] Example 32: Synthesis of N-(3-((4-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)butyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C32) JPEG0007796456000102.jpg33170 Composite using the same synthesis method as in Example 29.

[0110] Mass spectrum data of compound C32: MS (ESI): (M+H) + ,736.4 The nuclear magnetic resonance data of compound C32 is as follows: HNMR(400MHz,CDCl3)δ:8.80(s,1H),7.47-7.48(m,1H),7.28-7.30(m,1H),7.10-7.12(m,1H),6.98-7.02(m ,2H),6.87(s,1H),6.80-6.83(m,2H),6.70-6.73(m,1H),6.54-6.57(m,1H),6.38-6.42(m,1H),5.77(s,1H), 5.70-5.72(m,1H),4.75-4.81(m,1H),3.99-4.01(m,2H),3.39-3.43(m,2H),3.25-3.28(m,1H),2.65-2.81(m ,3H),2.05-2.20(m,2H),1.55-1.99(m,12H),1.11-1.37(m,5H),0.95-1.01(m,2H),0.89(s,3H),0.75(s,3H)

[0111] Example 33: Synthesis of N-(3-((5-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)pentyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C33) JPEG0007796456000103.jpg34170 Composite using the same synthesis method as in Example 29.

[0112] Mass spectrum data of compound C33: MS (ESI): (M+H) + ,750.4 The nuclear magnetic resonance data of compound C33 is as follows: HNMR(400MHz,CDCl3)δ:8.90-8.91(m,1H),7.43-7.45(m,1H),7.34-7.37(m,1H),7.09-7.12(m,1H),6.98-7.02 (m,2H),6.83-6.87(m,2H),6.71-6.74(m,1H),6.59-6.62(m,1H),6.38-6.41(m,1H),5.83(s,1H),5.72-5.75(m ,1H),4.79-4.83(m,1H),4.00-4.03(m,2H),3.41-3.45(m,2H),3.22-3.26(m,1H),2.61-2.79(m,3H),2.18-2.2 2(m,2H),2.02-2.10(m,2H),1.57-1.95(m,11H),1.15-1.55(m,6H),0.92-1.00(m,2H),0.87(s,3H),0.75(s,3H)

[0113] Example 34: Synthesis of N-(3-((5-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)pentyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C34) JPEG0007796456000104.jpg41170 Composite using the same synthesis method as in Example 29.

[0114] Mass spectrum data of compound C34: MS (ESI): (M+H) + ,750.4 The nuclear magnetic resonance data of compound C34 is as follows: HNMR(400MHz,CDCl3)δ:8.75(s,1H),7.47-7.48(m,1H),7.25-7.28(m,1H),7.10-7.12(m,1H),6.98-7. 02(m,2H),6.88(s,1H),6.80-6.83(m,2H),6.71-6.73(m,1H),6.54-6.57(m,1H),6.39-6.42(m,1H),5.9 3(s,1H),5.70-5.72(m,1H),4.81-4.83(m,1H),4.00-4.03(m,2H),3.38-3.41(m,2H),3.25-3.28(m,1H) ,2.62-2.78(m,3H),1.71-2.11(m,14H),1.11-1.58(m,9H),0.95-1.03(m,2H),0.88(s,3H),0.74(s,3H)

[0115] Example 35: Synthesis of N-(3-((6-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)hexyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C35) JPEG0007796456000105.jpg47170 Composite using the same synthesis method as in Example 29.

[0116] Mass spectrum data of compound C35: MS (ESI): (M+H) + ,764.4 The nuclear magnetic resonance data of compound C35 is as follows: HNMR(400MHz,CDCl3)δ:8.85-8.86(m,1H),7.45-7.47(m,1H),7.35-7.37(m,1H),7.09-7.12(m,1H),6.98-7.02 (m,2H),6.84-6.87(m,2H),6.71-6.75(m,1H),6.59-6.62(m,1H),6.39-6.41(m,1H),5.57(s,1H),5.72-5.75(m ,1H),4.79-4.82(m,1H),4.03-4.05(m,2H),3.41-3.45(m,2H),3.23-3.26(m,1H),2.65-2.79(m,3H),2.18-2.2 1(m,2H),2.05-2.10(m,2H),1.71-1.95(m,11H),1.15-1.57(m,8H),0.92-1.01(m,2H),0.89(s,3H),0.75(s,3H)

[0117] Example 36: Synthesis of N-(3-((6-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)hexyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C36) JPEG0007796456000106.jpg38170 Composite image was created using the same synthesis method as in Example 29.

[0118] Mass spectrum data of compound C36: MS (ESI): (M+H) + ,764.4 The nuclear magnetic resonance data of compound C36 is as follows: HNMR(400MHz,CDCl3)δ:8.55(s,1H),7.48-7.50(m,1H),7.25-7.28(m,1H),7.12-7.15(m,1H),6.99-7.0 5(m,2H),6.84(s,1H),6.79-6.83(m,2H),6.71-6.73(m,1H),6.54-6.56(m,1H),6.38-6.40(m,1H),5.72 -5.74(m,1H),5.70(s,1H),4.91-4.95(m,1H),4.00-4.02(m,2H),3.35-3.38(m,2H),3.27-3.30(m,1H), 2.62-2.78(m,3H),2.00-2.11(m,2H),1.55-1.97(m,15H),1.01-1.45(m,10H),0.87(s,3H),0.74(s,3H)

[0119] Example 37: Synthesis of N-(3-((7-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)heptyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C37) JPEG0007796456000107.jpg36170 Composite using the same synthesis method as in Example 29.

[0120] Mass spectrum data of compound C37: MS (ESI): (M+H) + ,778.4 The nuclear magnetic resonance data of compound C37 is as follows: HNMR(400MHz,CDCl3)δ:8.85-8.87(m,1H),7.45-7.47(m,1H),7.34-7.37(m,1H),7.09-7.13(m,1H),6.98-7.02 (m,2H),6.84-6.87(m,2H),6.71-6.75(m,1H),6.58-6.63(m,1H),6.39-6.42(m,1H),5.57(s,1H),5.72-5.76(m, 1H),4.78-4.82(m,1H),4.03-4.06(m,2H),3.41-3.46(m,2H),3.22-3.26(m,1H),2.65-2.79(m,3H),2.18-2.22 (m,2H),1.97-2.10(m,5H),1.55-1.91(m,10H),1.15-1.52(m,10H),0.95-1.01(m,2H),0.87(s,3H),0.71(s,3H)

[0121] Example 38: Synthesis of N-(3-((7-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)heptyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C38) JPEG0007796456000108.jpg43170 Composite using the same synthesis method as in Example 29.

[0122] Mass spectrum data of compound C38: MS (ESI): (M+H) + ,778.4 The nuclear magnetic resonance data of compound C38 is as follows: HNMR(400MHz,CDCl3)δ:8.59(s,1H),7.49-7.51(m,1H),7.26-7.28(m,1H),7.13-7.15(m,1H),7.03 -7.08(m,2H),6.85(s,1H),6.79-6.84(m,2H),6.71-6.73(m,1H),6.54-6.57(m,1H),6.37-6.40(m, 1H),5.79(s,1H),5.72-5.74(m,1H),4.91-4.95(m,1H),4.00-4.02(m,2H),3.22-3.38(m,3H),2.66 -2.79(m,3H),1.59-2.15(m,17H),1.15-1.49(m,12H),1.02-1.07(m,2H),0.86(s,3H),0.72(s,3H)

[0123] Example 39: Synthesis of N-(3-((8-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)octyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C39) JPEG0007796456000109.jpg45170 Composite using the same synthesis method as in Example 29.

[0124] Mass spectrum data of compound C39: MS (ESI): (M+H) + ,792.4 The nuclear magnetic resonance data of compound C39 is as follows: HNMR(400MHz,CDCl3)δ:8.72-8.77(m,1H),7.43-7.45(m,1H),7.34-7.36(m,1H),7.09-7.13(m,1H),6.98- 7.02(m,2H),6.85-6.87(m,2H),6.71-6.74(m,1H),6.58-6.62(m,1H),6.39-6.42(m,1H),5.76(s,1H),5.72 -5.76(m,1H),4.74-4.81(m,1H),4.01-4.04(m,2H),3.41-3.45(m,2H),3.21-3.25(m,1H),2.65-2.78(m,3 H),2.18-2.22(m,2H),1.97-2.10(m,5H),1.45-1.89(m,13H),1.01-1.37(m,11H),0.85(s,3H),0.69(s,3H)

[0125] Example 40: Synthesis of N-(3-((8-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)octyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C40) JPEG0007796456000110.jpg34170 Composite using the same synthesis method as in Example 29.

[0126] Mass spectrum data of compound C40: MS (ESI): (M+H) + ,792.4 The nuclear magnetic resonance data of compound C40 is as follows: HNMR(400MHz,CDCl3)δ:8.71(s,1H),7.50-7.52(m,1H),7.28-7.31(m,1H),7.05-7.13(m,3H),6.81 (s,1H),6.77-6.82(m,2H),6.71-6.73(m,1H),6.54-6.57(m,1H),6.38-6.41(m,1H),5.93(s,1H),5 .72-5.74(m,1H),4.95-4.99(m,1H),4.00-4.03(m,2H),3.24-3.38(m,3H),2.65-2.82(m,3H),1.88 -2.15(m,9H),1.55-1.71(m,12H),1.15-1.45(m,12H),1.03-1.09(m,2H),0.81(s,3H),0.69(s,3H)

[0127] Example 41: Synthesis of N-(3-((3-((4-(2-(3-(2,6-dioxopiperidine))-1-oxo-2,3-dihydro-1H-isoindole))amino)propyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C41) Step 1: Synthesis of N-(3-((3-((4-(2-(3-(2,6-dioxopiperidine))-3-hydroxy-1-oxo-2,3-dihydro-1H-isoindole))amino)propyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (VIII-41) 1.00 g of compound C29, 0.45 g of zinc powder, and 10 ml of glacial acetic acid were added to a reaction flask at room temperature, and the mixture was heated to 70°C and reacted for 2 hours. After hot filtration, the filtrate was concentrated to dryness to obtain a pale yellow solid, i.e., compound VIII-41.

[0128] Step 2: Synthesis of N-(3-((3-((4-(2-(3-(2,6-dioxopiperidine))-1-oxo-2,3-dihydro-1H-isoindole))amino)propyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C41) Compound VIII-41 obtained in step 1, 5 ml of triethylsilane, and 10 ml of trifluoroacetic acid were added to a reaction flask, and the mixture was heated to 60°C and reacted for 3 hours. The reaction solution was purified by column chromatography (eluent: dichloromethane:methanol = 10:1) to obtain 20 mg of compound C41.

[0129] Mass spectrum data of compound C41: MS (ESI): (M+H) + ,708.4 The nuclear magnetic resonance data of compound C41 is as follows: HNMR(400MHz,CDCl3)δ:8.91-8.94(m,1H),7.28-7.34(m,2H),7.05-7.07(m,1H),6.98-7.05(m,2H),6.81-6.85 (m,2H),6.72-6.75(m,1H),6.59-6.61(m,1H),6.38-6.40(m,1H),5.88(s,1H),5.72-5.75(m,1H),4.83-4.86(m ,1H),4.23-4.38(m,2H),3.95-3.97(m,2H),3.39-3.41(m,2H),3.20-3.22(m,1H),2.55-2.75(m,3H),2.10-2.1 7(m,2H),1.88-2.05(m,5H),1.67-1.75(m,3H),1.14-1.53(m,7H),0.92-0.95(m,2H),0.90(s,3H),0.78(s,3H)

[0130] Example 42: Synthesis of N-(3-((4-methyl-1-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-piperidinyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C42) JPEG0007796456000113.jpg47170 Composite using the same synthesis method as in Example 29.

[0131] Mass spectrum data of compound C42: MS(ESI):(M+H) + ,762.4 The nuclear magnetic resonance data of compound C42 is as follows: HNMR(400MHz,CDCl3)δ:8.84-8.86(m,1H),7.45-7.48(m,1H),7.25-7.27(m,1H),7.10-7.13(m,1H),6.95- 7.03(m,2H),6.82-6.85(m,2H),6.70-6.73(m,1H),6.59-6.61(m,1H),6.38-6.41(m,1H),5.79(s,1H),5.7 1-5.74(m,1H),4.79-4.83(m,1H),4.01-4.03(m,2H),3.25-3.47(m,5H),2.60-2.79(m,3H),2.15-2.25(m, 2H),1.65-2.10(m,9H),1.48-1.53(m,2H),1.16-1.37(m,7H),0.95-0.99(m,2H),0.91(s,3H),0.75(s,3H)

[0132] Example 43: Synthesis of N-(3-((4-methyl-1-(5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-piperidinyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C43) JPEG0007796456000114.jpg33170 Composite using the same synthesis method as in Example 29.

[0133] Mass spectrum data of compound C43: MS(ESI):(M+H) + ,762.4 The nuclear magnetic resonance data of compound C43 is as follows: HNMR(400MHz,CDCl3)δ:8.81-8.85(m,1H),7.33-7.35(m,1H),7.21-7.23(m,1H),7.09-7.12(m,1H),7.00 -7.02(m,2H),6.93(s,1H),6.80-6.84(m,2H),6.71-6.73(m,1H),6.55-6.57(m,1H),6.38-6.42(m,1H),5 .71-5.73(m,1H),5.65(s,1H),4.77-4.80(m,1H),4.05-4.07(m,2H),3.19-3.43(m,5H),2.60-2.79(m,3H) ),2.07-2.22(m,2H),1.48-2.01(m,11H),1.10-1.37(m,7H),0.96-1.00(m,2H),0.90(s,3H),0.75(s,3H)

[0134] Example 44: Synthesis of 1-(4-(1-(3-(3-oxo-4-aza-5α-androst-1-ene-17β-formamido)phenyl)piperidine))methyl-4-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))piperazine (C44) JPEG0007796456000115.jpg38170 Step 1: Synthesis of N-(3-hydroxyphenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (VI-44) JPEG0007796456000116.jpg33170JPEG0007796456000117.jpg32170 2.00 g of compound I-1, 0.72 g of 3-hydroxyaniline, 6.55 g of pyBoP, and 50 mL of DMF were sequentially added to a reaction flask at room temperature. The mixture was stirred for 5 minutes, and 2.44 g of DIPEA was added dropwise. The mixture was allowed to react overnight. The next day, 50 mL of water was added to the reaction flask, and the mixture was extracted three times with ethyl acetate (50 mL each time). The mixture was then washed once with 50 mL of 1 mol / L aqueous hydrochloric acid, 50 mL of saturated aqueous sodium carbonate, and 50 mL of saturated aqueous sodium chloride. The mixture was then dried over anhydrous sodium sulfate and concentrated to dryness to yield 2.56 g of a black oil. The oil was purified by column chromatography (eluent: dichloromethane:methanol = 20:1) to yield 1.51 g of compound VI-44.

[0135] Step 2: Synthesis of 3-(3-oxo-4-aza-5α-androst-1-ene-17β-formamido)-nonafluorobutane-1-sulfonic acid phenyl ester (VII-44) 1.00 g of compound VI-44, 1.00 g of nonafluoro-1-butanesulfonyl fluoride, 2.15 g of cesium carbonate, and 50 mL of DMF were sequentially added to a reaction flask at room temperature under nitrogen protection, and the mixture was allowed to react at 70 °C for 5 hours. The reaction mixture was extracted three times with ethyl acetate (50 mL each time), washed once with 50 mL of water and once with 50 mL of saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain a black oil. The oil was purified by column chromatography (eluent: dichloromethane:methanol = 20:1) to obtain 1.24 g of compound VII-44.

[0136] Step 3: Synthesis of 1-(4-(1-(3-(3-oxo-4-aza-5α-androst-1-ene-17β-formamido)phenyl)piperidine))methyl-4-Boc piperazine (IV-44) 1.00 g of compound VII-44, 1.00 g of compound II-13, 1.00 g of cesium carbonate, 0.4 g of XPhoS, 0.1 g of palladium acetate, and 20 mL of DMF were sequentially added to a reaction flask at room temperature under nitrogen protection. The reaction flask was placed in a microwave reactor and heated at 60 Hz for 8 minutes. The mixture was then extracted three times with ethyl acetate (50 mL each time), washed once with 50 mL of water and once with 50 mL of saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain a black oil. The oil was purified by column chromatography (eluent: dichloromethane:methanol = 20:1) to obtain 40 mg of compound IV-44.

[0137] Step 4: Synthesis of 1-(4-(1-(3-(3-oxo-4-aza-5α-androst-1-ene-17β-formamido)phenyl)piperidine))methyl-piperazine (V-44) 40 mg of compound IV-44 and 5 ml of dichloromethane were added to a reaction flask at room temperature, followed by dropwise addition of 5 ml of trifluoroacetic acid. The mixture was reacted for 10 minutes and then concentrated to dryness to obtain an oily product. The oily product (i.e., the crude product of compound V-44) was used in the next reaction without further purification.

[0138] Step 5: Synthesis of 1-(4-(1-(3-(3-oxo-4-aza-5α-androst-1-ene-17β-formamido)phenyl)piperidine))methyl-4-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))piperazine (C44) The oil obtained in Step 4 (i.e., the crude product of Compound V-44), 20 mg of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 20 mg of DIPEA, and 5 mL of DMF were sequentially added to a reaction flask at room temperature. The reaction flask was placed in a microwave reactor and reacted at 50 Hz for 10 minutes. The reaction was quenched by adding 20 mL of water to the reaction flask, extracted three times with dichloromethane (20 mL each time), washed once with 20 mL of water, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 10:1) to obtain 2 mg of Compound C44.

[0139] Mass spectrum data of compound C44: MS (ESI): (M+H) + ,830.5 The nuclear magnetic resonance data of compound C44 is as follows: HNMR(400MHz,CDCl3)δ:8.85(s,1H),7.39-7.41(m,1H),7.10-7.12(m,2H),6.94-7.00(m,2 H),6.76-6.79(m,1H),6.68-6.70(m,1H),6.21-6.24(m,1H),5.87(s,1H),5.70-5.73(d,1H ),4.89-4.92(m,1H),3.41-3.59(m,4H),3.10-3.37(m,5H),2.62-2.80(m,5H),2.07-2.38( m,6H),1.50-1.93(m,10H),1.15-1.49(m,8H),0.98-1.06(m,2H),0.93(s,3H),0.70(s,3H)

[0140] Example 45: Synthesis of 1-(4-(1-(3-(3-oxo-4-aza-5α-androst-1-ene-17β-formamido)phenyl)piperidine)methyl)-4-(5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))piperazine (C45) JPEG0007796456000122.jpg39170 In step 5 of Example 44, 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione was replaced with 5-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, and the synthesis was carried out using the same synthetic method as in Example 44.

[0141] Mass spectrum data of compound C45: MS (ESI): (M+H) + ,708.3 The nuclear magnetic resonance data of compound C45 is as follows: HNMR(400MHz,CDCl3)δ:8.75-8.79(m,1H),7.35-7.37(m,1H),7.08-7.09(m,1H),7.09-7.12(m,1H),7.00 -7.02(m,2H),6.91(s,1H),6.79-6.83(m,2H),6.70-6.72(m,1H),6.38-6.42(m,1H),5.75(s,1H),5.71-5 .73(m,1H),4.83-4.85(m,1H),3.45-3.64(m,2H),3.15-3.49(m,7H),2.59-2.75(m,5H),2.07-2.35(m,4H) ),2.07-2.15(m,2H),1.55-2.01(m,10H),1.07-1.45(m,8H),0.95-1.01(m,2H),0.95(s,3H),0.71(s,3H)

[0142] Example 46: Synthesis of N-(3-((3-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))oxy)propyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C46) JPEG0007796456000123.jpg36170 Step 1: Synthesis of N-(3-hydroxyphenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (X-46) JPEG0007796456000124.jpg30170JPEG0007796456000125.jpg30170At room temperature under nitrogen protection, 2.00 g of compound VI-44, 2.00 g of 1,3-dibromopropane, 3.20 g of cesium carbonate, and 50 mL of DMF were sequentially added to a reaction flask and reacted at 70 °C for 5 hours. The reaction solution was extracted three times with ethyl acetate (100 mL each), washed once with 50 mL of water and once with 50 mL of saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain a black oil. The oil was purified by silica gel column chromatography (eluent: dichloromethane:methanol = 20:1) to obtain 1 g of compound X-46.

[0143] Step 2: Synthesis of N-(3-((3-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))oxy)propyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C46) 1 g of compound X-46, 0.57 g of 4-hydroxy-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 0.92 g of cesium carbonate, 0.3 g of potassium iodide, and 50 mL of DMF were added to a reaction flask at room temperature under nitrogen protection. The mixture was reacted at 70 °C for 5 hours, extracted three times with ethyl acetate (100 mL each time), washed once with 50 mL of water and once with 50 mL of saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain a black oil. The oil was purified by silica gel column chromatography (eluent: dichloromethane:methanol = 20:1) to obtain 0.2 g of compound C46.

[0144] Mass spectrum data of compound C46: MS(ESI):(M+H) + ,723.3 The nuclear magnetic resonance data of compound C46 is as follows: HNMR(400MHz,CDCl3)δ:7.48-7.60(m,2H),7.35-7.42(m,1H),6.91-7.17(m,3H),6.66-6.69( m,1H),6.52-6.58(m,1H),6.25-6.28(m,1H),5.59-5.61(m,1H),4.83-4.86(m,1H),4.05-4.2 6(m,2H),3.85-3.88(m,2H),3.93-3.96(m,1H),2.65-2.87(m,3H),2.17-2.04(m,3H),1.89-1 .99(m,3H),1.55-1.63(m,5H),1.18-1.40(m,5H),0.95-1.05(m,3H),0.91(s,3H),0.69(s,3H)

[0145] Example 47: Synthesis of N-(5-(2-((2-((5-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)ethyl)oxy)pyridine))-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C47) JPEG0007796456000127.jpg49170 Composite using the same synthesis method as in Example 17.

[0146] Mass spectrum data of compound C47: MS(ESI):(M+H) + ,709.3 The nuclear magnetic resonance data of compound C47 is as follows: HNMR(400MHz,CDCl3)δ:8.95-8.96(m,1H),7.99(s,1H),7.45-7.51(m,2H),7.17-7.19(m,1H) ,7.02-7.05(m,2H)6.95(s,1H),6.69-6.71(m,2H),5.75-5.77(m,1H),5.55(s,1H),4.94-4.9 7(m,1H),3.99-4.02(m,2H)3.25-3.31(m,2H),3.19-3.20(m,1H),2.69-2.77(m,3H),2.01-2. 11(m,2H),1.57-1.93(m,7H),1.15-1.47(m,6H),0.95-1.03(m,2H),0.99(s,3H),0.75(s,3H)

[0147] Example 48: Synthesis of N-(3-(4-(((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)methyl)piperidine)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C48) JPEG0007796456000128.jpg50170 Composite using the same synthesis method as in Example 56.

[0148] Mass spectrum data of compound C48: MS(ESI):(M+H) + ,761.4 The nuclear magnetic resonance data of compound C48 is as follows: HNMR(400MHz,CDCl3)δ:8.71-8.73(m,1H),7.21-7.25(m,1H),7.10-7.13(m,2H),6.95-7.00(m,2 H),6.77-6.80(m,1H),6.68-6.70(m,1H),6.57-6.59(m,1H),6.21-6.24(m,2H),5.88(s,1H),5.7 1-5.74(m,1H),4.83-4.86(m,1H),3.07-3.38(m,7H),2.60-2.78(m,3H),2.15-2.20(m,2H),1.96 -2.06(m,2H),1.60-1.88(m,10H),1.48-1.55(m,2H),1.08-1.40(m,8H),0.88(s,3H),0.70(s,3H)

[0149] Example 49: Synthesis of N-(3-((3-((4-(2-(3-(1-pivaloyloxymethyl-2,6-dioxopiperidine))-isoindole-1,3-dione))amino)propyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C49) JPEG0007796456000129.jpg53170JPEG0007796456000130.jpg38170At room temperature under nitrogen protection, 0.50 g of compound C29, 0.12 g of chloromethyl 2,2-dimethylpropionate, 0.11 g of potassium iodide, 0.19 g of potassium carbonate, and 15 mL of DMF were sequentially added to a reaction flask and reacted at 70 °C for 5 hours. After the reaction was completed, the mixture was extracted three times with ethyl acetate (50 mL each time). The aqueous phase was washed once with 50 mL of water and once with 50 mL of saturated aqueous sodium chloride solution. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain an oil. The oil was purified by column chromatography (eluent: dichloromethane:methanol = 15:1) to obtain 82 mg of compound C49.

[0150] Mass spectrum data of compound C49: MS(ESI):(M+H) + ,835.4 The nuclear magnetic resonance data of compound C49 is as follows: HNMR(400MHz,CDCl3)δ:8.88-8.90(m,1H),7.43-7.45(m,1H),7.33-7.34(m,1H),7.09-7.12(m,1H),6.98-7.01( m,2H),6.84-6.89(m,2H),6.71-6.74(m,1H),6.59-6.61(m,1H),6.38-6.45(m,1H),5.89(s,1H),5.72-5.75(m,1 H),4.95-4.99(m,1H),4.85(s,2H),4.01-4.02(m,2H),3.41-3.44(m,2H),3.22-3.26(m,1H),2.61-2.79(m,3H), 2.11-2.21(m,3H),1.66-2.05(m,7H),1.17-1.55(m,7H),1.11(s,9H)0.95-1.02(m,2H),0.91(s,3H),0.77(s,3H)

[0151] Example 50: Synthesis of N-(3-((3-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)propyl)amino)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C50) JPEG0007796456000131.jpg36170Step 1: Synthesis of 3-((3-(N-Bocamino)propyl)amino)aniline (II-50) a. Synthesis of 3-((3-(N-Bocamino)propyl)amino)nitrobenzene (II-50-1) 2.00 g of 3-nitroaniline, 3.01 g of N-Boc-3-aminopropionaldehyde, 40 ml of DME, and 0.65 g of glacial acetic acid were sequentially added to a 100 ml three-neck flask at room temperature. 6.14 g of STAB was added with stirring, and the reaction mixture was heated to 40 °C and reacted for 3 hours. The mixture was concentrated to dryness to obtain a yellow solid. This solid was purified by column chromatography (eluent: petroleum ether: methyl tert-butyl ether = 3:1) to obtain 1.81 g of compound II-50-1.

[0152] b. Synthesis of 3-((3-(N-Bocamino)propyl)amino)aniline (II-50) JPEG0007796456000133.jpg17170At room temperature, 1.81 g of compound II-50-1, 10 ml of methanol, and 100 mg of 5% palladium on carbon were added sequentially to a 100 ml hydrogenation reactor, and the hydrogenation reaction was carried out at 25°C for 1 hour. After the reaction was completed, the mixture was concentrated to dryness to obtain 1.54 g of a gray oil (i.e., the crude product of compound II-50).

[0153] Step 2: Synthesis of N-(3-((3-(N-Bocamino)propyl)amino)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (IV-50) JPEG0007796456000134.jpg32170JPEG0007796456000135.jpg34170At room temperature under nitrogen protection, 2.19 g of compound I-1, 1.54 g of crude compound II-50, 5.41 g of pyBoP, and 50 mL of DMF were sequentially added to a 100 mL three-neck flask. After stirring for 5 minutes, 2.68 g of DIPEA was added dropwise and allowed to react overnight. 70 mL of water was added to the reaction mixture, followed by three extractions with ethyl acetate (70 mL each time). The combined organic phases were washed once with 70 mL of 1 mol / L aqueous hydrochloric acid, 70 mL of saturated aqueous sodium carbonate, and 70 mL of saturated aqueous sodium chloride, dried over anhydrous sodium sulfate, and concentrated to dryness to yield 4.73 g of a black oil (i.e., crude compound IV-50).

[0154] Step 3: Synthesis of N-(3-((3-aminopropyl)amino)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (V-50) 4.73 g of the crude product of Compound IV-50 and 20 ml of dichloromethane were added to a 50 ml three-neck flask at room temperature, and then 20 ml of trifluoroacetic acid was added dropwise. The mixture was concentrated to dryness to obtain an oil. The oil (i.e., the crude product of Compound V-50) was used in the next reaction without further purification.

[0155] Step 4: Synthesis of N-(3-((3-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)propyl)amino)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C50) The oil obtained in Step 3 (i.e., the crude product of Compound V-50), 2.15 g of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 3.36 g of DIPEA, and 20 ml of DMF were sequentially added to a reaction flask at room temperature. The reaction flask was then placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 30 ml of water was added to the reaction flask, and the mixture was extracted three times with dichloromethane (30 ml each time). The combined organic phase was washed once with 30 ml of water, dried over anhydrous sodium sulfate, and then concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 10:1) to obtain 1.87 g of Compound C50.

[0156] Mass spectrum data of compound C50: MS (ESI): (M+H) + ,721.4 The nuclear magnetic resonance data of compound C50 is as follows: HNMR(400MHz,CDCl3)δ:8.85-8.87(m,1H),7.37-7.40(m,1H),7.09-7.11(m,2H),6.94-7.00(m,2H),6.76 -6.79(d,1H,J=12Hz),6.68-6.70(d,1H,J=12Hz),6.56-6.58(m,1H),6.21-6.24(m,2H),5.81(s,1H),5.71 -5.74(m,1H),4.81-4.85(m,1H),3.05-3.15(m,5H),2.60-2.78(m,3H),2.15-2.20(m,2H),1.96-2.06(m, 2H),1.83-1.88(m,2H),1.60-1.73(m,5H),1.48-1.55(m,2H),1.08-1.40(m,6H),0.83(s,3H),0.68(s,3H)

[0157] Example 51: Synthesis of N-(3-((3-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)propyl)oxy)-4-chloro-phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C51) JPEG0007796456000138.jpg35170 Composite image was created using the same synthesis method as in Example 29.

[0158] Mass spectrum data of compound C51: MS (ESI): (M+H) + ,756.3 The nuclear magnetic resonance data of compound C51 is as follows: HNMR(400MHz,CDCl3)δ:8.69(s,1H),7.41-7.43(m,1H),7.32-7.34(m,1H),6.98-7.05(m,2H),6.82-6.86(m, 2H),6.70-6.73(m,1H),6.59-6.61(m,1H),6.38-6.41(m,1H),5.83(s,1H),5.73-5.75(m,1H),4.84-4.87(m,1 H),4.00-4.02(m,2H),3.42-3.45(m,2H),3.23-3.26(m,1H),2.63-2.76(m,3H),2.12-2.22(m,2H),1.80-2.10 (m,5H),1.61-1.75(m,3H),1.49-1.53(m,2H),1.13-1.37(m,5H),0.92-0.99(m,2H),0.91(s,3H),0.78(s,3H)

[0159] Example 52: Synthesis of N-(3-((3-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)propyl)oxy)-4-methyl-phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C52) JPEG0007796456000139.jpg36170 Composite using the same synthesis method as in Example 29.

[0160] Mass spectrum data of compound C52: MS (ESI): (M+H) + ,736.4 The nuclear magnetic resonance data of compound C52 is as follows: HNMR(400MHz,CDCl3)δ:8.82-8.94(m,1H),7.40-7.43(m,1H),7.32-7.35(m,1H),7.02-7.10(m,2H),6.82-6. 88(m,2H),6.70-6.74(m,1H),6.59-6.61(m,1H),6.38-6.42(m,1H),5.89(s,1H),5.75-5.77(m,1H),4.79-4.8 3(m,1H),4.03-4.05(m,2H),3.41-3.44(m,2H),3.21-3.28(m,1H),2.59-2.75(m,3H),2.38(s,3H),2.16-2.22 (m,2H),1.80-2.10(m,5H),1.61-1.75(m,3H),1.16-1.55(m,7H),0.92-0.95(m,2H),0.89(s,3H),0.74(s,3H)

[0161] Example 53: Synthesis of N-(2-(4-((3-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)propyl)oxy)-pyridine))-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C53) JPEG0007796456000140.jpg36170 Composite using the same synthesis method as in Example 29.

[0162] Mass spectrum data of compound C53: MS (ESI): (M+H) + ,723.3 The nuclear magnetic resonance data of compound C53 is as follows: HNMR(400MHz,CDCl3)δ:8.82-8.94(m,1H),8.05-8.07(m,1H),7.75-7.78(m,1H),7.49-7.51(m,1H),7.18-7.12(m,1 H),6.98-7.02(m,1H),6.81-6.83(m,1H),6.77-6.78(m,1H),6.59-6.61(m,1H),6.41-6.43(m,1H),5.98(s,1H),5.8 5-5.88(m,1H),4.94-4.98(m,1H),4.12-4.14(m,2H),3.47-3.49(m,2H),3.25-3.27(m,1H),2.61-2.77(m,3H),2.15 -2.25(m,2H),1.80-2.10(m,5H),1.48-1.77(m,5H),1.16-1.37(m,5H),0.95-1.01(m,2H),0.93(s,3H),0.79(s,3H)

[0163] Example 54: Synthesis of N-(3-((4-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)cyclohexyl)amino)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C54) JPEG0007796456000141.jpg54170 Composite using the same synthesis method as in Example 56.

[0164] Mass spectrum data of compound C54: MS (ESI): (M+H) + ,761.4 The nuclear magnetic resonance data of compound C54 is as follows: HNMR(400MHz,CDCl3)δ:8.54-8.64(m,1H),7.46-7.49(m,1H),7.28-7.35(m,1H),6.97-7.18(m ,3H),6.83-6.89(m,1H),6.41-6.55(m,1H),6.21-6.35(m,2H),6.15-6.18(m,1H)5.83(m,1H),5 .61(s,1H),4.88-4.91(m,1H),3.65-3.75(m,1H),3.31-3.47(m,2H),2.62-2.87(m,3H),2.01- 2.31(m,5H),1.75-1.95(m,8H),1.15-1.66(m,9H),1.02-1.09(m,2H),0.96(s,3H),0.77(s,3H)

[0165] Example 55: Synthesis of N-(3-((3-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)propyl)oxy)-5-chloro-phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C55) JPEG0007796456000142.jpg37170 Composite using the same synthesis method as in Example 29.

[0166] Mass spectrum data of compound C55: MS (ESI): (M+H) + ,756.3 The nuclear magnetic resonance data of compound C55 is as follows: HNMR(400MHz,CDCl3)δ:8.67(s,1H),7.41-7.43(m,1H),7.20-7.23(m,1H),7.02-7.09(m,2H),6.81 -6.86(m,2H),6.71-6.74(m,1H),6.55-6.58(m,1H),6.39-6.43(m,1H),5.89(s,1H),5.78-5.81(m,1 H),4.87-4.50(m,1H),4.05-4.07(m,2H),3.43-3.46(m,2H),3.23-3.26(m,1H),2.65-2.79(m,3H),1 .95-2.15(m,7H),1.71-1.81(m,3H),1.13-1.59(m,7H),0.95-1.05(m,2H),0.95(s,3H),0.79(s,3H)

[0167] Example 56: Synthesis of N-(3-((2-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)ethyl)amino)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C56) JPEG0007796456000143.jpg48170Step 1: Synthesis of 3-((2-(N-Bocamino)ethyl)amino)aniline (II-56) a. Synthesis of 3-((2-(N-Bocamino)ethyl)amino)nitrobenzene (II-56-1) 2.00 g of 3-nitroaniline, 2.77 g of N-Boc-2-aminoacetaldehyde, 40 ml of DME, and 0.65 g of glacial acetic acid were sequentially added to a 100 ml three-neck flask at room temperature. 6.14 g of STAB was added with stirring, and the reaction mixture was heated to 40 °C and reacted for 3 hours. The mixture was concentrated to dryness to obtain a yellow solid. This solid was purified by column chromatography (eluent: petroleum ether: methyl tert-butyl ether = 3:1) to obtain 1.70 g of compound II-56-1.

[0168] b. Synthesis of 3-((2-(N-Bocamino)ethyl)amino)aniline (II-56) JPEG0007796456000145.jpg17170At room temperature, 1.70 g of compound II-56-1, 10 ml of methanol, and 100 mg of 5% palladium on carbon were added sequentially to a 100 ml hydrogenation reactor, and the hydrogenation reaction was carried out at 25°C for 1 hour. After the reaction was completed, the mixture was concentrated to dryness to obtain 1.38 g of a gray oil (i.e., the crude product of compound II-56).

[0169] Step 2: Synthesis of N-(3-((2-(N-Bocamino)ethyl)amino)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (IV-56) JPEG0007796456000146.jpg31170JPEG0007796456000147.jpg33170At room temperature under nitrogen protection, 2.09 g of compound I-1, 1.38 g of crude compound II-56, 5.14 g of pyBoP, and 50 mL of DMF were sequentially added to a 100 mL three-neck flask. After stirring for 5 minutes, 2.38 g of DIPEA was added dropwise and allowed to react overnight. 70 mL of water was added to the reaction mixture, followed by three 70 mL extractions with ethyl acetate. The combined organic phases were washed once with 70 mL of 1 mol / L aqueous hydrochloric acid, 70 mL of saturated aqueous sodium carbonate, and 70 mL of saturated aqueous sodium chloride. The organic phase was then dried over anhydrous sodium sulfate and concentrated to dryness to yield 4.35 g of a black oil (i.e., crude compound IV-56).

[0170] Step 3: Synthesis of N-(3-((2-aminoethyl)amino)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (V-56) 4.35 g of the crude product of Compound IV-56 and 20 mL of dichloromethane were added to a 50 mL three-neck flask at room temperature, and then 20 mL of trifluoroacetic acid was added dropwise. The mixture was concentrated to dryness to obtain an oily product. The oily product (i.e., the crude product of Compound V-56) was directly subjected to the next reaction without purification.

[0171] Step 4: Synthesis of N-(3-((2-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)ethyl)amino)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C56) The oil obtained in Step 3 (i.e., the crude product of Compound V-56), 1.98 g of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 3.08 g of DIPEA, and 20 mL of DMF were sequentially added to a reaction flask at room temperature. The reaction flask was then placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 30 mL of water was added to the reaction flask, and the mixture was extracted three times with dichloromethane (30 mL each time). The combined organic phase was washed once with 30 mL of water, dried over anhydrous sodium sulfate, and then concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 10:1) to obtain 1.52 g of Compound C56.

[0172] Mass spectrum data of compound C56: MS (ESI): (M+H) + ,707.3 The nuclear magnetic resonance data of compound C56 is as follows: HNMR(400MHz,CDCl3)δ:8.80-8.82(m,1H),7.38-7.41(m,1H),7.10-7.13(m,2H),6.98-7.01(m,2H ),6.76-6.79(d,1H,J=12Hz),6.55-6.58(m,1H),6.56-6.59(m,1H),6.21-6.24(m,2H),5.88(s,1H) ,5.70-5.73(m,1H),4.82-4.87(m,1H),3.05-3.15(m,5H),2.60-2.78(m,3H),2.10-2.18(m,2H),1. 88-2.06(m,2H),1.60-1.83(m,5H),1.45-1.55(m,2H),1.10-1.40(m,6H),0.84(s,3H),0.69(s,3H)

[0173] Example 57: Synthesis of N-(3-((3-((4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))amino)cyclopentyl)oxy)phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C57) JPEG0007796456000150.jpg35170 Composite image was created using the same synthesis method as in Example 29.

[0174] Mass spectrum data of compound C57: MS(ESI):(M+H) + ,748.4 The nuclear magnetic resonance data of compound C57 is as follows: HNMR(400MHz,CDCl3)δ:8.87-8.89(m,1H),7.39-7.41(m,1H),7.31-7.34(m,1H),7.09-7.13(m,1H),6.97-7.01(m,2H),6 .80-6.87(m,2H),6.71-6.75(m,1H),6.59-6.61(m,1H),6.39-6.45(m,1H),5.95(s,1H),5.71-5.75(m,1H),4.79-4.82(m ,1H),4.15-4.20(m,1H),3.48-3.53(m,1H),3.23-3.27(m,1H),2.65-2.85(m,3H),2.28-2.36(m,2H),2.14-2.24(m,4H), 1.80-2.10(m,5H),1.61-1.75(m,3H),1.45-1.53(m,2H),1.10-1.37(m,5H),0.92-0.99(m,2H),0.89(s,3H),0.77(s,3H)

[0175] Example 58: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C58) JPEG0007796456000151.jpg36170 Composite using the same synthesis method as in Example 59.

[0176] Mass spectrum data of compound C58: MS (ESI): (M+H) + ,761.4 The nuclear magnetic resonance data of compound C58 is as follows: HNMR(400MHz,CDCl3)δ:8.37-8.48(m,1H),7.42-7.48(m,1H),6.96-7.21(m,5H),6.73- 6.81(m,2H),6.43-6.57(m,1H),6.16-6.35(m,2H),5.80-5.83(m,1H),5.55(s,1H),4.8 8-4.91(m,1H),3.73-3.80(m,1H),3.42-3.55(m,1H),3.28-3.35(m,1H),2.67-2.91(m, 3H),2.21-2.33(m,2H),2.00-2.12(m,2H),1.01-1.93(m,21H),0.96(s,3H),0.75(s,3H)

[0177] Example 59: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C59) JPEG0007796456000152.jpg36170 Step 1: Synthesis of 3-(N-(N'-Boc-1R,3R-cyclohexanediamine))aniline (Compound II-59) a. Synthesis of 3-(N-(N'-Boc-1R,3R-cyclohexanediamine))nitrobenzene (Compound II-59-1) 5.00 g of (1R)-N-Boc-3-oxocyclohexylamine, 3.24 g of 3-nitroaniline, 30 ml of DME, and 1.05 g of glacial acetic acid were sequentially added to a 250 ml three-neck flask at room temperature. 9.96 g of STAB was added with stirring, and the mixture was heated to 40 °C and reacted for 3 hours. The mixture was concentrated to dryness to obtain a yellow solid. The yellow solid was purified by column chromatography (eluent: petroleum ether: methyl tert-butyl ether = 5:1) to obtain 3.11 g of compound II-59-1 and 0.82 g of compound II-59-2.

[0178] b. Synthesis of 3-(N-(N'-Boc-1R,3R-cyclohexanediamine))aniline (Compound II-59) JPEG0007796456000154.jpg20170At room temperature, 1.00 g of compound II-59-1, 10 ml of methanol, and 100 mg of 5% palladium on carbon were sequentially added to a 100 ml hydrogenation reactor, and the hydrogenation reaction was carried out at 25°C for 1 hour. After the reaction was completed, the reaction solution was concentrated to dryness to obtain 0.90 g of a gray-black oil (i.e., the crude product of compound II-59).

[0179] Step 2: Synthesis of N-(3-(N-(N'-Boc-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (Compound IV-59) JPEG0007796456000155.jpg35170JPEG0007796456000156.jpg34170At room temperature under nitrogen protection, 1.14 g of compound I-1, 0.90 g of crude compound II-59, 3.41 g of pyBoP, and 30 mL of DMF were sequentially added to a 100 mL three-neck flask. After stirring for 5 minutes, 1.27 g of DIPEA was added dropwise to the reaction solution and allowed to react overnight. 50 mL of water was added to the reaction solution, followed by three 50 mL extractions with ethyl acetate. The combined organic phases were washed once with 50 mL of 1 mol / L aqueous hydrochloric acid, 50 mL of saturated aqueous sodium carbonate, and 50 mL of saturated aqueous sodium chloride, dried over anhydrous sodium sulfate, and concentrated to dryness to yield 2.56 g of a black oil (i.e., crude compound IV-59).

[0180] Step 3: Synthesis of N-(3-(N-(1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (Compound V-59) 2.56 g of the crude product of compound IV-59 and 10 ml of dichloromethane were added to a 100 ml three-neck flask at room temperature. 20 ml of trifluoroacetic acid was then added dropwise. The mixture was allowed to react for 10 minutes and then concentrated to dryness to obtain an oil. 100 ml of water was added, the pH was adjusted to 11 with 2 mol / L aqueous sodium hydroxide, and 100 ml of isobutanol was added for extraction. The organic phase was concentrated to dryness, and the residue was separated by column chromatography (eluent: dichloromethane:methanol = 8:1) to obtain 1.21 g of an off-white solid (i.e., compound V-59).

[0181] Step 4: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C59) 1.21 g of compound V-59, 0.98 g of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 1.53 g of DIPEA, and 10 ml of DMF were added to a reaction flask at room temperature. The reaction flask was then placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 20 ml of water was added to the reaction flask, and the mixture was extracted three times with dichloromethane (20 ml each time). The combined organic phase was washed once with 20 ml of water, dried over anhydrous sodium sulfate, and then concentrated to dryness to obtain a crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 10:1) to obtain 0.70 g of compound C59.

[0182] Mass spectrum data of compound C59: MS (ESI): (M+H) + ,761.4 The nuclear magnetic resonance data of compound C59 is as follows: HNMR(400MHz,CDCl3)δ:8.48-8.50(m,1H),7.42-7.48(m,1H),7.15-7.21(m,1H),7.08-7.12(m,3 H),6.73-6.81(m,2H),6.51-6.57(m,1H),6.28-6.35(m,2H),5.80(d,1H,J=12Hz),5.60(s,1H),4. 88-4.91(m,1H),3.73-3.80(m,2H),3.28-3.33(m,1H),2.67-2.90(m,3H),2.21-2.33(m,2H),2.00 -2.12(m,2H),1.90-1.71(m,8H),1.58-1.15(m,11H),1.01-1.11(m,2H),0.96(s,3H),0.75(s,3H)

[0183] Example 60: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-1S,3S-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C60) JPEG0007796456000159.jpg40170 Composite using the same synthesis method as in Example 59.

[0184] Mass spectrum data of compound C60: MS (ESI): (M+H) + ,761.4 The nuclear magnetic resonance data of compound C60 is as follows: HNMR(400MHz,CDCl3)δ:8.36-8.41(m,1H),7.40-7.42(m,1H),7.18-7.26(m,1H),7.05-7.09(m,3H),6 .94-6.97(m,1H),6.76-6.83(m,2H),6.53-6.57(m,1H),6.28-6.34(m,2H),5.80-5.83(m,1H),5.49(s ,1H),4.89-4.92(m,1H),3.74-3.82(m,2H),3.31-3.35(m,1H),2.72-2.90(m,3H),2.23-2.27(m,2H), 2.01-2.13(m,2H),1.71-1.90(m,8H),1.20-1.58(m,11H),1.01-1.08(m,2H),0.96(s,3H),0.75(m,3H)

[0185] Example 61: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-1S,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C61) JPEG0007796456000160.jpg39170Step 1: Synthesis of 3-(N-(N'-Boc-1S,3R-cyclohexanediamine))aniline (Compound II-61) JPEG0007796456000161.jpg40170At room temperature, 0.72 g of compound II-59-2, 10 ml of methanol, and 72 mg of 5% palladium on carbon were added sequentially to a 100 ml hydrogenation reactor, and the hydrogenation reaction was carried out at 25°C for 1 hour. After the reaction was completed, the reaction solution was concentrated to dryness to obtain 0.68 g of a gray-black oil (i.e., the crude product of compound II-61).

[0186] Step 2: Synthesis of N-(3-(N-(N'-Boc-1S,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (Compound IV-61) JPEG0007796456000162.jpg35170JPEG0007796456000163.jpg32170At room temperature under nitrogen protection, 0.85 g of compound I-1, 0.68 g of the crude product of compound II-61, 2.51 g of pyBoP, and 30 mL of DMF were sequentially added to a 100 mL three-neck flask. After stirring for 5 minutes, 0.94 g of DIPEA was added dropwise to the reaction solution and allowed to react overnight. 50 mL of the reaction solution was added, followed by three 50 mL extractions with ethyl acetate. The combined organic phases were washed once with 50 mL of 1 mol / L aqueous hydrochloric acid, 50 mL of saturated aqueous sodium carbonate, and 50 mL of saturated aqueous sodium chloride, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain 2.34 g of a black oil (i.e., crude product of compound IV-61).

[0187] Step 3: Synthesis of N-(3-(N-(1S,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (Compound V-61) 2.34 g of the crude product of compound IV-61 and 10 ml of dichloromethane were added to a 100 ml three-neck flask at room temperature. 20 ml of trifluoroacetic acid was then added dropwise. The mixture was allowed to react for 10 minutes and then concentrated to dryness to obtain an oil. 100 ml of water was added, the pH was adjusted to 11 with 2 mol / L aqueous sodium hydroxide, and 100 ml of isobutanol was added for extraction. The organic phase was concentrated to dryness, and the residue was separated by column chromatography (eluent: dichloromethane:methanol = 8:1) to obtain 1.06 g of an off-white solid (i.e., compound V-61).

[0188] Step 4: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-1S,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C61) 1.06 g of compound V-61, 0.87 g of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 1.36 g of DIPEA, and 10 ml of DMF were added to a reaction flask at room temperature. The reaction flask was then placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 20 ml of water was added to the reaction flask, and the mixture was extracted three times with dichloromethane (20 ml each time). The organic phases were combined, washed once with 20 ml of water, dried over anhydrous sodium sulfate, and then concentrated to dryness to obtain a crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 10:1) to obtain 0.56 g of compound C61.

[0189] Mass spectrum data of compound C61: MS (ESI): (M+H) + ,761.4 The nuclear magnetic resonance data of compound C61 is as follows: HNMR(400MHz,CDCl3)δ:8.62-8.64(m,1H),7.44-7.48(m,1H),7.35-7.38(m,1H),7.25-7.28(m,1H),7 .18-7.24(m,2H),6.83-6.87(m,2H),6.45-6.47(m,1H),6.26-6.35(m,2H),5.80(d,1H,J=12Hz),5.70( s,1H),4.85-4.90(m,1H),3.65-3.68(m,1H),3.80-3.83(m,1H),3.30-3.35(m,1H),2.67-2.90(m,3H), 2.05-2.33(m,4H),1.95-1.70(m,13H),1.58-1.15(m,6H),1.03-1.11(m,2H),0.95(s,3H),0.77(s,3H)

[0190] Example 62: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-1-oxo-2,3-dihydro-1H-isoindole))-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C62) JPEG0007796456000166.jpg42170 Step 1: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-1-oxo-3-hydroxy-2,3-dihydro-1H-isoindole))-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (VIII-62) 1.00 g of compound C59, 0.44 g of zinc powder, and 10 ml of glacial acetic acid were added to a reaction flask at room temperature, and the mixture was heated to 70°C and reacted for 2 hours. The mixture was then filtered while hot, and the filtrate was concentrated to dryness to obtain compound VIII-62 as a pale yellow solid.

[0191] Step 2: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-1-oxo-2,3-dihydro-1H-isoindole))-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C62) The pale yellow solid obtained in Step 1 (i.e., Compound VIII-62), 5 ml of triethylsilane, and 10 ml of trifluoroacetic acid were added to a reaction flask, and the mixture was heated to 60°C and reacted for 3 hours. The reaction solution was purified by column chromatography (eluent: dichloromethane:methanol = 10:1) to obtain 15 mg of Compound C62.

[0192] Mass spectrum data of compound C62: MS (ESI): (M+H) + ,747.4 The nuclear magnetic resonance data of compound C62 is as follows: HNMR(400MHz,CDCl3)δ:9.00(s,1H),7.28-7.33(m,1H),7.11-7.21(m,1H),7.00-7.12(m,3H),6.81-6.84(m,1H),6 .73-6.87(m,1H),6.44-6.50(m,1H),6.20-6.31(m,2H),5.85(d,1H,J=12Hz),5.67(s,1H),5.02-5.07(m,1H),4.23 -4.38(m,2H),3.75-3.85(m,2H),3.28-3.33(m,1H),2.65-2.69(m,1H),2.45-2.48(m,1H),2.38-2.41(m,1H)2.10- 2.28(m,2H),2.01-2.11(m,2H),1.90-1.43(m,14H),1.35-1.15(m,5H),1.08-1.11(m,2H),0.92(s,3H),0.75(s,3H)

[0193] Example 63: Preparation of N-(3-(N-(N'-(4-(2-(3S-(2,6-dioxopiperidine))-isoindole-1,3-dione))-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C63) JPEG0007796456000169.jpg37170 Compound C59 was separated by preparative chromatography and the second eluting component was collected to give compound C63. The separation conditions are as follows: Column: Daicel CHIRALPAK IA (5.0cm ID×25cm L, 10μm) Mobile phase: methanol / dichloromethane = 60 / 40 (V / V) Flow rate: 60 ml / min Wavelength: 254nm Column temperature: 38℃ Mass spectrum data for compound C63: MS(ESI): (M+H)+, 761.4 The nuclear magnetic resonance data of compound C63 is as follows: HNMR(400MHz,CDCl3)δ:8.56(s,1H),7.39-7.43(m,1H),7.29(s,1H),7.04-7.08(m,3H),6.77-6.8 3(m,2H),6.57-6.59(m,1H),6.37-6.39(m,1H),6.26-6.28(m,1H),5.80-5.82(m,1H),5.74(s,1H), 4.89-4.93(m,1H),3.73-3.83(m,2H),3.30-3.34(m,1H),2.72-2.90(m,3H),2.23-2.25(m,2H),2.1 0-2.14(m,2H),1.72-1.93(m,8H),1.25-1.60(m,11H),1.04-1.07(m,2H),0.95(s,3H),0.74(s,3H)

[0194] Example 64: Preparation of N-(3-(N-(N'-(4-(2-(3R-(2,6-dioxopiperidine))-isoindole-1,3-dione))-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C64) Compound C59 was separated by preparative chromatography, and the first eluted component was collected to obtain compound C64. The preparation conditions were the same as in Example 63.

[0195] Mass spectrum data of compound C64: MS (ESI): (M+H) + ,761.4 The nuclear magnetic resonance data of compound C64 is as follows: HNMR(400MHz,CDCl3)δ:8.43(s,1H),7.40-7.44(m,1H),7.21(s,1H),7.05-7.10(m,2H),7.00(s,1H), 6.78-6.83(m,2H),6.57-6.59(m,1H),6.36-6.38(m,1H),6.26-6.28(m,1H),5.80-5.83(m,1H),5.67(s ,1H),4.88-4.94(m,1H),3.71-3.82(m,2H),3.30-3.34(m,1H),2.71-2.90(m,3H),2.25-2.26(m,2H), 2.10-2.12(m,2H),1.71-1.91(m,8H),1.27-1.60(m,11H),1.02-1.06(m,2H),0.96(s,3H),0.74(s,3H)

[0196] Example 65: Synthesis of N-(3-(N-(N'-(4-(2-(3S-(2,6-dioxopiperidine))-1-oxo-2,3-dihydro-1H-isoindole))-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C65) JPEG0007796456000171.jpg37170 Step 1: Synthesis of N-(3-(N-(N'-(4-(2-(3S-(2,6-dioxopiperidine))-1-oxo-3-hydroxy-2,3-dihydro-1H-isoindole))-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (VIII-65) 1.00 g of compound C63, 0.44 g of zinc powder, and 10 ml of glacial acetic acid were added to a reaction flask at room temperature, and the mixture was heated to 70°C and reacted for 2 hours. The mixture was then filtered while hot, and the filtrate was concentrated to dryness to obtain a crude product of compound VIII-65.

[0197] Step 2: Synthesis of N-(3-(N-(N'-(4-(2-(3S-(2,6-dioxopiperidine))-1-oxo-2,3-dihydro-1H-isoindole))-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C65) 5 ml of triethylsilane and 10 ml of trifluoroacetic acid were added to the crude product of compound VIII-65 obtained in step 1, and the mixture was heated to 60°C and reacted for 3 hours. The reaction solution was purified by column chromatography (eluent: dichloromethane:methanol = 10:1) to obtain 14 mg of compound C65.

[0198] Mass spectrum data of compound C65: MS (ESI): (M+H) + ,747.4 The nuclear magnetic resonance data of compound C65 is as follows: HNMR(400MHz,CDCl3)δ:9.01(s,1H),7.29-7.33(m,1H),7.11-7.21(m,1H),7.00-7.12(m,3H),6.82-6.85(m,1H), 6.82-6.85(m,1H),6.45-6.50(m,1H),6.22-6.25(m,2H),5.85-5.86(m,1H),5.78(s,1H),5.02-5.07(m,1H),4.24- 4.37(m,2H),3.76-3.80(m,2H),3.28-3.33(m,1H),2.65-2.69(m,1H),2.45-2.48(m,1H),2.38-2.41(m,1H)2.10-2 .28(m,2H),2.01-2.12(m,2H),1.91-1.43(m,14H),1.35-1.15(m,5H),1.08-1.11(m,2H),0.93(s,3H),0.76(s,3H)

[0199] Example 66: Synthesis of N-(3-(N-(N'-(4-(2-(3R-(2,6-dioxopiperidine))-1-oxo-2,3-dihydro-1H-isoindole))-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C66) JPEG0007796456000174.jpg38170 Step 1: Synthesis of N-(3-(N-(N'-(4-(2-(3R-(2,6-dioxopiperidine))-1-oxo-3-hydroxy-2,3-dihydro-1H-isoindole))-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (VIII-66) 1.00 g of compound C64, 0.44 g of zinc powder, and 10 ml of glacial acetic acid were added to a reaction flask at room temperature, and the mixture was heated to 70°C and reacted for 2 hours. The mixture was then filtered while hot, and the filtrate was concentrated to dryness to obtain a crude product of compound VIII-66.

[0200] Step 2: Synthesis of N-(3-(N-(N'-(4-(2-(3R-(2,6-dioxopiperidine))-1-oxo-2,3-dihydro-1H-isoindole))-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C66) 5 ml of triethylsilane and 10 ml of trifluoroacetic acid were added to the crude product of compound VIII-66 obtained in step 1, and the mixture was heated to 60°C and reacted for 3 hours. The reaction solution was purified by column chromatography (eluent: dichloromethane:methanol = 10:1) to obtain 14.5 mg of compound C66.

[0201] Mass spectrum data of compound C66: MS(ESI):(M+H) + ,747.4 The nuclear magnetic resonance data of compound C66 is as follows: HNMR(400MHz,CDCl3)δ:9.02(s,1H),7.28-7.33(m,1H),7.01-7.20(m,4H),6.81-6.84(m,1H),6.81-6.84(m,1 H),6.44-6.50(m,1H),6.29-6.31(m,2H),5.89-5.90(m,1H),5.67(s,1H),5.02-5.07(m,1H),4.23-4.38(m,2H) ),3.79-3.84(m,2H),3.29-3.34(m,1H),2.66-2.70(m,1H),2.46-2.49(m,1H),2.39-2.42(m,1H),2.12-2.26( m,2H),2.01-2.11(m,2H),1.89-1.45(m,14H),1.35-1.11(m,5H),1.04-1.09(m,2H),0.92(s,3H),0.75(s,3H)

[0202] Example 67: Synthesis of N-(3-(N-(N'-(4-(2-(3S-(1-pivaloyloxymethyl-2,6-dioxopiperidine))-isoindole-1,3-dione))-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C67) JPEG0007796456000177.jpg55170 Composite using the same synthesis method as in Example 49.

[0203] Mass spectrum data of compound C67: MS(ESI):(M+H) + ,875.5 The nuclear magnetic resonance data of compound C67 is as follows: HNMR(400MHz,CDCl3)δ:8.61-8.63(m,1H),7.45-7.48(m,1H),7.19-7.23(m,1H),7.08-7.14(m,3H), 6.75-6.82(m,2H),6.52-6.58(m,1H),6.27-6.34(m,2H),5.79-5.82(m,1H),5.70(s,1H),4.87-4.91 (m,1H),4.81(s,2H),3.73-3.81(m,2H),3.28-3.33(m,1H),2.65-2.89(m,3H),2.20-2.32(m,2H),2. 01-2.12(m,2H),1.90-1.73(m,8H),1.58-1.15(m,11H),1.01-1.11(m,5H),0.95(s,3H),0.74(s,3H)

[0204] Example 68: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-1,3-dideutero-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C68) JPEG0007796456000178.jpg35170Step 1: Synthesis of 3-(N-(N'-Boc-1,3-dideutero-1R,3R-cyclohexanediamine))aniline (II-68) a. Synthesis of 3-(N-(1,3-dideutero-1R,3R-cyclohexanediamine))nitrobenzene (II-68-1) 100 mg of m-fluoronitrobenzene and 820 mg of 1,3-dideutero-1R,3R-cyclohexanediamine were placed in a three-neck flask and heated to 100°C under nitrogen protection. The mixture was stirred for 12 hours. The mixture was cooled to room temperature and concentrated to dryness to obtain a crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 6:1) to obtain 21 mg of a pale yellow solid (i.e., compound II-68-1).

[0205] MS(ESI):(M+H) +,238.1 b. Synthesis of 3-(N-(N'-Boc-1,3-dideutero-1R,3R-cyclohexanediamine))nitrobenzene (Compound II-68-2) 21 mg of compound II-68-1, 9.8 mg of triethylamine, 21 mg of Boc anhydride, and 2 mL of dichloromethane were sequentially added to a reaction flask and allowed to react for 30 minutes with stirring at room temperature. The mixture was washed once with 0.5 mL of 1 mol / L aqueous hydrochloric acid and once with 0.5 mL of water. The aqueous phase was extracted twice with dichloromethane (0.5 mL each time). The combined organic phase was dried over anhydrous sodium sulfate and concentrated to dryness to yield 29 mg of crude compound II-68-2.

[0206] MS(ESI):(M+H) + ,338.2 c. Synthesis of 3-(N-(N'-Boc-1,3-dideutero-1R,3R-cyclohexanediamine))aniline (II-68) 29 mg of crude compound II-68-2, 2 ml of methanol, and 2.9 mg of 5% palladium on carbon were added to a 100 ml hydrogenation reactor, and the mixture was maintained at 25°C and 50 psi. Hydrogen gas was introduced until the reaction was complete. The reaction mixture was concentrated to dryness to obtain 24 mg of crude compound II-68.

[0207] MS(ESI):(M+H) + ,308.2 Step 2: Synthesis of N-(3-(N-(N'-Boc-1,3-dideutero-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (IV-68) Under nitrogen protection, 11 mg of compound I-1, 15 mg of crude compound II-68, 32.8 mg of pyBoP, 12.2 mg of DIPEA, and 0.5 mL of DMF were added to a reaction flask and stirred overnight until the reaction was complete. 2.5 mL of water was added to the reaction flask, followed by extraction with ethyl acetate three times (2.5 mL each time). The combined organic phase was washed once with 1 mL of 1 mol / L aqueous hydrochloric acid and once with 1 mL of saturated aqueous sodium carbonate, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 20:1) to obtain 19 mg of compound IV-68.

[0208] Step 3: Synthesis of N-(3-(N-(1,3-dideutero-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (V-68) JPEG0007796456000184.jpg34170 19 mg of compound IV-68 and 1 ml of dichloromethane were added to a reaction flask at room temperature, and then 0.1 ml of trifluoroacetic acid was added dropwise. The mixture was reacted for 10 minutes and concentrated to dryness to obtain 16 mg of crude product of compound V-68.

[0209] Step 4: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-1,3-dideutero-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C68) 16 mg of the crude product of compound V-68, 13 mg of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 30 mg of DIPEA, and 1 ml of DMF were sequentially added to a reaction flask, and the reaction flask was placed in a microwave reactor and reacted at 50 Hz for 10 minutes. Extraction was performed three times (2.5 ml each time) using water and dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 15:1) to obtain 10 mg of compound C68.

[0210] Mass spectrum data of compound C68: MS(ESI):(M+H) + ,763.4 The nuclear magnetic resonance data of compound C68 is as follows: HNMR(400MHz,CDCl3)δ:8.48-8.50(m,1H),7.42-7.48(m,1H),7.15-7.21(m,1H),7.08-7.12 (m,3H),6.73-6.81(m,2H),6.43-6.57(m,1H),6.16-6.35(m,2H),5.80(d,1H,J=12Hz),5.60( s,1H),4.88-4.91(m,1H),3.40-3.57(m,1H),2.67-2.90(m,3H),2.21-2.33(m,2H),2.00-2.1 2(m,2H),1.90-1.71(m,8H),1.58-1.15(m,11H),1.01-1.11(m,2H),0.96(s,3H),0.75(s,3H)

[0211] Example 69: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-2-methyl-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C69) Step 1: Synthesis of 3-(N-(N'-Boc-2-methyl-1R,3R-cyclohexanediamine))aniline (II-69) a. Synthesis of 3-(N-(2-methyl-1R,3R-cyclohexanediamine))nitrobenzene (Compound II-69-1) 100 mg of m-fluoronitrobenzene and 900 mg of 2-methyl-1R,3R-cyclohexanediamine were placed in a three-neck flask and heated to 100 °C under nitrogen protection. The mixture was stirred for 12 hours, cooled to room temperature, and concentrated to dryness to obtain a crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 6:1) to obtain 26 mg of a pale yellow solid (i.e., Compound II-69-1).

[0212] MS(ESI):(M+H) + ,250.1 b. Synthesis of 3-(N-(N'-Boc-2-methyl-1R,3R-cyclohexanediamine))nitrobenzene (II-69-2) JPEG0007796456000187.jpg19170 26 mg of compound II-69-1, 14.2 mg of triethylamine, 30 mg of Boc anhydride, and 2 ml of dichloromethane were added sequentially to a reaction flask, and the mixture was allowed to react for 30 minutes with stirring at room temperature. The mixture was then washed once with 0.5 ml of 1 mol / L aqueous hydrochloric acid solution and once with 0.5 ml of water. The aqueous phase was extracted twice with dichloromethane (2 ml each time). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain 36 mg of crude compound II-69-2.

[0213] MS(ESI):(M+H) + ,350.2 c. Synthesis of 3-(N-(N'-Boc-2-methyl-1R,3R-cyclohexanediamine))aniline (II-69) JPEG0007796456000188.jpg1817036 mg of crude compound II-69-2, 2 ml of methanol, and 3.6 mg of 5% palladium on carbon were placed in a 100 ml hydrogenation reactor, maintained at 25°C and 50 psi, and hydrogen was introduced until the reaction was complete. The reaction solution was concentrated to dryness to obtain 30 mg of crude compound II-69.

[0214] Step 2: Synthesis of N-(3-(N-(N'-Boc-2-methyl-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (Compound IV-69) Under nitrogen protection, 11 mg of compound I-1, 16 mg of crude compound II-69, 32.8 mg of pyBoP, 12.2 mg of DIPEA, and 0.5 mL of DMF were sequentially added to a reaction flask and stirred overnight to complete the reaction. 2.5 mL of water was added to the reaction flask, followed by three extractions with ethyl acetate (2.5 mL each time). The combined organic phase was washed once with 1 mL of 1 mol / L aqueous hydrochloric acid and once with 1 mL of saturated aqueous sodium carbonate, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 20:1) to obtain 19.5 mg of compound IV-69.

[0215] MS(ESI):(M+H) + ,619.4 Step 3: Synthesis of N-(3-(N-(2-methyl-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (V-69) JPEG0007796456000191.jpg34170 At room temperature, 19.5 mg of compound IV-69 and 1 ml of dichloromethane were added to a reaction flask, and then 0.1 ml of trifluoroacetic acid was added dropwise. The mixture was reacted for 10 minutes and concentrated to dryness to obtain 16 mg of crude product of compound V-69.

[0216] Step 4: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-2-methyl-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (compound C69) 16 mg of the crude product of compound V-69, 13 mg of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 30 mg of DIPEA, and 1 ml of DMF were sequentially added to a reaction flask, and the reaction flask was placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 2.5 ml of water was added, and the mixture was extracted with dichloromethane three times (2.5 ml each time). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 15:1) to obtain 9.5 mg of compound C69.

[0217] Mass spectrum data of compound C69: MS(ESI):(M+H) + ,775.4 The nuclear magnetic resonance data of compound C69 is as follows: HNMR(400MHz,CDCl3)δ:8.53-8.55(m,1H),7.40-7.47(m,1H),7.17-7.20(m,1H),7.12-7.18 (m,3H),6.72-6.81(m,2H),6.51-6.57(m,1H),6.29-6.35(m,2H),5.80(d,1H,J=12Hz),5.71( s,1H),4.81-4.85(m,1H),3.85-3.75(m,2H),3.40-3.59(m,1H),2.77-2.90(m,3H),2.11-2.3 3(m,4H),1.71-2.00(m,7H),1.23-1.58(m,14H),1.03-1.19(m,2H),1.05(s,3H),0.88(s,3H)

[0218] Example 70: Synthesis of a mixture of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-6S-methyl-1S,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide and N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-4R-methyl-1R,3S-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C70) JPEG0007796456000193.jpg37170JPEG0007796456000194.jpg39170Step 1: Synthesis of 3-(N-(N'-Boc-6R-methyl-1S,3R-cyclohexanediamine))aniline and 3-(N-(N'-Boc-4R-methyl-1R,3S-cyclohexanediamine))aniline mixture (II-70) a. Synthesis of 3-(N-(6R-methyl-1S,3R-cyclohexanediamine))nitrobenzene and 3-(N-(4R-methyl-1R,3S-cyclohexanediamine))nitrobenzene mixture (II-70-1) 100 mg of m-fluoronitrobenzene and 900 mg of 6R-methyl-1S,3R-cyclohexanediamine were placed in a three-neck flask and heated to 100 °C under nitrogen protection and stirred for 12 hours. The mixture was cooled to room temperature and concentrated to dryness to obtain a crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 6:1) to obtain 24 mg of a pale yellow solid (i.e., Compound II-70-1).

[0219] MS(ESI):(M+H) + ,250.1 b. Synthesis of 3-(N-(N'-Boc-6R-methyl-1S,3R-cyclohexanediamine))nitrobenzene and 3-(N-(N'-Boc-4R-methyl-1R,3S-cyclohexanediamine))nitrobenzene mixture (II-70-2) 24 mg of compound II-70-1, 13.2 mg of triethylamine, 28 mg of Boc anhydride, and 2 mL of dichloromethane were added to a reaction flask and stirred at room temperature for 30 minutes. The reaction mixture was washed once with 0.5 mL of 1 mol / L aqueous hydrochloric acid and once with 0.5 mL of water. The aqueous phase was extracted twice with dichloromethane (2.5 mL each time). The combined organic phases were dried over anhydrous sodium sulfate and concentrated to dryness to yield 36 mg of crude compound II-70-2.

[0220] MS(ESI):(M+H) + ,350.2 c. Synthesis of 3-(N-(N'-Boc-6R-methyl-1S,3R-cyclohexanediamine))aniline and 3-(N-(N'-Boc-4R-methyl-1R,3S-cyclohexanediamine))aniline mixture (II-70) JPEG0007796456000197.jpg2317036 mg of crude compound II-70-2, 2 ml of methanol, and 3.6 mg of 5% palladium on carbon were placed in a 00 ml hydrogenation reactor, maintained at 25°C and 50 psi, and hydrogen gas was introduced until the reaction was complete. The reaction solution was then concentrated to dryness to obtain 33 mg of crude compound II-70.

[0221] Step 2: Synthesis of N-(3-(N-(N'-Boc-6R-methyl-1S,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide and N-(3-(N-(N'-Boc-4R-methyl-1R,3S-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide mixture (IV-70) JPEG0007796456000198.jpg36170JPEG0007796456000199.jpg33170Under nitrogen protection, 11 mg of compound I-1, 10 mg of the crude compound II-70, 32.8 mg of pyBoP, 12.2 mg of DIPEA, and 0.5 mL of DMF were sequentially added to a reaction flask and stirred overnight to complete the reaction. 2.5 mL of water was added to the reaction flask, and the mixture was extracted three times with ethyl acetate (2.5 mL each time). The combined organic phase was washed once with 1 mL of 1 mol / L aqueous hydrochloric acid and once with 1 mL of saturated aqueous sodium carbonate, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 20:1) to obtain 18.6 mg of compound IV-70.

[0222] MS(ESI):(M+H) + ,619.4 Step 3: Synthesis of N-(3-(N-(6R-methyl-1S,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide and N-(3-(N-(4R-methyl-1R,3S-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide mixture (V-70) JPEG0007796456000200.jpg35170 At room temperature, 18.6 mg of compound IV-70 and 1 ml of dichloromethane were added to a reaction flask, and then 0.1 ml of trifluoroacetic acid was added dropwise. The mixture was reacted for 10 minutes and concentrated to dryness to obtain 14.5 mg of crude product of compound V-70.

[0223] MS(ESI):(M+H) + ,519.4 Step 4: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-6R-methyl-1S,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide and N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-4R-methyl-1R,3S-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide mixture (C70) JPEG0007796456000201.jpg36170JPEG0007796456000202.jpg37170 14.5 mg of the crude product of compound V-70, 11.3 mg of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 26.3 mg of DIPEA, and 1 mL of DMF were sequentially added to a reaction flask, and the reaction flask was placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 2.5 mL of water was added, and the mixture was extracted with dichloromethane three times (2.5 mL each time). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by silica gel column chromatography (eluent: dichloromethane:methanol = 15:1) to obtain 9.6 mg of compound C70.

[0224] Mass spectrum data of compound C70: MS (ESI): (M+H) + ,775.4

[0225] Example 71: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-(1R,4S)-bicyclo[2,2,1]heptane-2R,6R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide and N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-(1S,4R)-bicyclo[2,2,1]heptane-2R,6R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide mixture (C71) JPEG0007796456000203.jpg32170JPEG0007796456000204.jpg34170Step 1: Synthesis of 3-(N-(N'-Boc-(1R,4S)-bicyclo[2,2,1]heptane-2R,6R-diamine))aniline and 3-(N-(N'-Boc-(1S,4R)-bicyclo[2,2,1]heptane-2R,6R-diamine))aniline Mixture (II-71) a. Synthesis of 3-(N-((1R,4S)-bicyclo[2,2,1]heptane-2R,6R-diamine))nitrobenzene and 3-(N-((1S,4R)-bicyclo[2,2,1]heptane-2R,6R-diamine))nitrobenzene mixture (II-71-1) 100 mg of m-fluoronitrobenzene and 893 mg of (1R,4S)-bicyclo[2,2,1]heptane-2R,6R-diamine were placed in a three-neck flask and heated to 100°C under nitrogen protection. The mixture was stirred for 12 hours. The mixture was cooled to room temperature and concentrated to dryness to obtain a crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 6:1) to obtain 21 mg of compound II-71-1.

[0226] MS(ESI):(M+H) + ,248.1 b. Synthesis of 3-(N-(N'-Boc-(1R,4S)-bicyclo[2,2,1]heptane-2R,6R-diamine))nitrobenzene and 3-(N-(N'-Boc-(1S,4R)-bicyclo[2,2,1]heptane-2R,6R-diamine))nitrobenzene mixture (II-71-2) 21 mg of compound II-71-1, 9.4 mg of triethylamine, 20.4 mg of Boc anhydride, and 2 mL of dichloromethane were added to a reaction flask and stirred at room temperature for 30 minutes. The reaction mixture was washed once with 0.5 mL of 1 mol / L aqueous hydrochloric acid and once with 0.5 mL of water. The aqueous phase was extracted twice with dichloromethane (2.5 mL each time). The combined organic phases were dried over anhydrous sodium sulfate and concentrated to dryness to yield 29.5 mg of crude compound II-71-2.

[0227] MS(ESI):(M+H) + ,348.2 c. Synthesis of 3-(N-(N'-Boc-(1R,4S)-bicyclo[2,2,1]heptane-2R,6R-diamine))aniline and 3-(N-(N'-Boc-(1S,4R)-bicyclo[2,2,1]heptane-2R,6R-diamine))aniline mixture (II-71) JPEG0007796456000207.jpg2217029.5 mg of crude compound II-71-2, 2 ml of methanol, and 3 mg of 5% palladium on carbon were placed in a 100 ml hydrogenation reactor, maintained at 25°C and 50 psi, and hydrogen gas was introduced until the reaction was complete. The reaction solution was then concentrated to dryness to obtain 27 mg of crude compound II-71.

[0228] MS(ESI):(M+H) + ,318.2 Step 2: Synthesis of N-(3-(N-(N'-Boc-(1R,4S)-bicyclo[2,2,1]heptane-2R,6R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide and N-(3-(N-(N'-Boc-(1S,4R)-bicyclo[2,2,1]heptane-2R,6R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide mixture (IV-71) JPEG0007796456000208.jpg37170JPEG0007796456000209.jpg33170Under nitrogen protection, 11 mg of compound I-1, 10 mg of crude compound II-71, 32.8 mg of pyBoP, 12.2 mg of DIPEA, and 0.5 mL of DMF were sequentially added to a reaction flask and stirred overnight to complete the reaction. 2.5 mL of water was added to the reaction flask, and the mixture was extracted three times with ethyl acetate (2.5 mL each time). The combined organic phase was washed once with 1 mL of 1 mol / L aqueous hydrochloric acid and once with 1 mL of saturated aqueous sodium carbonate, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 20:1) to obtain 17 mg of compound IV-71.

[0229] MS(ESI):(M+H) + ,617.4 Step 3: Synthesis of N-(3-(N-((1R,4S)-bicyclo[2,2,1]heptane-2R,6R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide and N-(3-(N-((1S,4R)-bicyclo[2,2,1]heptane-2R,6R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide mixture (V-71) JPEG0007796456000210.jpg37170 17 mg of compound IV-71 and 1 ml of dichloromethane were added to a reaction flask at room temperature, and then 0.1 ml of trifluoroacetic acid was added dropwise. The mixture was reacted for 10 minutes and concentrated to dryness to obtain 14 mg of crude product of compound V-71.

[0230] MS(ESI):(M+H) + ,517.3

[0231] Step 4: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-(1R,4S)-bicyclo[2,2,1]heptane-2R,6R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide and N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-(1S,4R)-bicyclo[2,2,1]heptane-2R,6R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide mixture (C71) JPEG0007796456000211.jpg33170JPEG0007796456000212.jpg34170 14 mg of the crude product of compound V-71, 13 mg of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 30 mg of DIPEA, and 1 ml of DMF were sequentially added to a reaction flask, and the reaction flask was placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 2.5 ml of water was added, and the mixture was extracted with dichloromethane three times (2.5 ml each time). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 15:1) to obtain 9.2 mg of compound C71.

[0232] Mass spectrum data of compound C71: MS (ESI): (M+H) + ,773.4

[0233] Example 72: Synthesis of a mixture of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-(1R,4S)-bicyclo[2,2,2]octane-2R,6R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide and N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-(1S,4R)-bicyclo[2,2,2]octane-2R,6R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C72) JPEG0007796456000213.jpg32170JPEG0007796456000214.jpg35170Step 1: Synthesis of 3-(N-(N'-Boc-(1R,4S)-bicyclo[2,2,2]octane-2R,6R-diamine))aniline and 3-(N-(N'-Boc-(1S,4R)-bicyclo[2,2,2]octane-2R,6R-diamine))aniline Mixture (II-72) a. Synthesis of 3-(N-((1R,4S)-bicyclo[2,2,2]octane-2R,6R-diamine))nitrobenzene and 3-(N-((1S,4R)-bicyclo[2,2,2]octane-2R,6R-diamine))nitrobenzene mixture (II-72-1) 100 mg of m-fluoronitrobenzene and 985 mg of (1R,4S)-bicyclo[2,2,2]octane-2R,6R-diamine were placed in a three-neck flask and heated to 100°C under nitrogen protection. The mixture was stirred for 12 hours. The mixture was cooled to room temperature and concentrated to dryness to obtain a crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 6:1) to obtain 21 mg of compound II-72-1.

[0234] MS(ESI):(M+H) + ,262.1

[0235] b. Synthesis of 3-(N-(N'-Boc-(1R,4S)-bicyclo[2,2,2]octane-2R,6R-diamine))nitrobenzene and 3-(N-(N'-Boc-(1S,4R)-bicyclo[2,2,2]octane-2R,6R-diamine))nitrobenzene mixture (II-72-2) 21 mg of compound II-72-1, 8.9 mg of triethylamine, 19.3 mg of Boc anhydride, and 2 mL of dichloromethane were added to a reaction flask and allowed to react for 30 minutes with stirring at room temperature. The reaction mixture was washed once with 0.5 mL of 1 mol / L aqueous hydrochloric acid and once with 0.5 mL of water. The aqueous phase was extracted twice with dichloromethane (2.5 mL each time). The combined organic phases were dried over anhydrous sodium sulfate and concentrated to dryness to yield 29 mg of crude compound II-72-2.

[0236] MS(ESI):(M+H) + ,362.2

[0237] c. Synthesis of 3-(N-(N'-Boc-(1R,4S)-bicyclo[2,2,2]octane-2R,6R-diamine))aniline and 3-(N-(N'-Boc-(1S,4R)-bicyclo[2,2,2]octane-2R,6R-diamine))aniline mixture (II-72) JPEG0007796456000217.jpg2217029 mg of crude compound II-72-2, 2 ml of methanol, and 3 mg of 5% palladium on carbon were placed in a 100 ml hydrogenation reactor, maintained at 25°C and 50 psi, and hydrogen gas was introduced until the reaction was complete. The reaction solution was then concentrated to dryness to obtain 26 mg of crude compound II-72.

[0238] MS(ESI):(M+H) + ,332.2

[0239] Step 2: Synthesis of N-(3-(N-(N'-Boc-(1R,4S)-bicyclo[2,2,2]octane-2R,6R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide and N-(3-(N-(N'-Boc-(1S,4R)-bicyclo[2,2,2]octane-2R,6R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide mixture (IV-72) Under nitrogen protection, 11 mg of compound I-1, 10.5 mg of crude compound II-72, 32.8 mg of pyBoP, 12.2 mg of DIPEA, and 0.5 mL of DMF were sequentially added to a reaction flask and stirred overnight to complete the reaction. 2.5 mL of water was added to the reaction flask, followed by three extractions with ethyl acetate (2.5 mL each). The combined organic phases were washed with 1 mL of 1 mol / L aqueous hydrochloric acid and 1 mL of saturated aqueous sodium carbonate, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 20:1) to obtain 17.5 mg of compound IV-72.

[0240] MS(ESI):(M+H) + ,631.4

[0241] Step 3: Synthesis of N-(3-(N-((1R,4S)-bicyclo[2,2,2]octane-2R,6R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide and N-(3-(N-((1S,4R)-bicyclo[2,2,2]octane-2R,6R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide mixture (V-72) JPEG0007796456000220.jpg37170 At room temperature, 17.5 mg of compound IV-72 and 1 ml of dichloromethane were added to a reaction flask, and then 0.1 ml of trifluoroacetic acid was added dropwise. The mixture was reacted for 10 minutes and concentrated to dryness to obtain 14 mg of crude product of compound V-72.

[0242] MS(ESI):(M+H)+,531.4

[0243] Step 4: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-(1R,4S)-bicyclo[2,2,2]octane-2R,6R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide and N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-(1S,4R)-bicyclo[2,2,2]octane-2R,6R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide mixture (C72) 14 mg of the crude product of compound V-72, 13 mg of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 30 mg of DIPEA, and 1 mL of DMF were added to a reaction flask, and the reaction flask was placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 2.5 mL of water was added, and the mixture was extracted with dichloromethane three times (2.5 mL each time). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 15:1) to obtain 7.8 mg of compound C72.

[0244] Mass spectrum data of compound C72: MS (ESI): (M+H) + ,787.4

[0245] Example 73: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-(1R,5S)-bicyclo[3,2,2]nonane-7R,8R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide and N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-(1S,5R)-bicyclo[3,2,2]nonane-7R,8R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide mixture (C73) JPEG0007796456000223.jpg31170JPEG0007796456000224.jpg34170Step 1: Synthesis of 3-(N-(N'-Boc-(1R,5S)-bicyclo[3,2,2]nonane-7R,8R-diamine))aniline and 3-(N-(N'-Boc-(1S,5R)-bicyclo[3,2,2]nonane-7R,8R-diamine))aniline Mixture (II-73) a. Synthesis of 3-(N-((1R,5S)-bicyclo[3,2,2]nonane-7R,8R-diamine))nitrobenzene and 3-(N-((1S,5R)-bicyclo[3,2,2]nonane-7R,8R-diamine))nitrobenzene mixture (II-73-1) 100 mg of m-fluoronitrobenzene and 1.09 g of (1R,5S)-bicyclo[3,2,2]nonane-7R,8R-diamine were placed in a three-neck flask and heated to 100°C under nitrogen protection. The mixture was stirred for 12 hours. The mixture was cooled to room temperature and concentrated to dryness to obtain a crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 6:1) to obtain 19 mg of compound II-73-1.

[0246] MS(ESI):(M+H) + ,276.2

[0247] b. Synthesis of 3-(N-(N'-Boc-(1R,5S)-bicyclo[3,2,2]nonane-7R,8R-diamine))nitrobenzene and 3-(N-(N'-Boc-(1S,5R)-bicyclo[3,2,2]nonane-7R,8R-diamine))nitrobenzene mixture (II-73-2) 19 mg of compound II-73-1, 7.6 mg of triethylamine, 16.6 mg of Boc anhydride, and 2 mL of dichloromethane were added to a reaction flask and allowed to react for 30 minutes with stirring at room temperature. The reaction mixture was washed once with 0.5 mL of 1 mol / L aqueous hydrochloric acid and once with 0.5 mL of water. The aqueous phase was extracted twice with dichloromethane (2.5 mL each time). The combined organic phases were dried over anhydrous sodium sulfate and concentrated to dryness to yield 25 mg of crude compound II-73-2.

[0248] MS(ESI):(M+H) + ,376.2

[0249] c. Synthesis of 3-(N-(N'-Boc-(1R,5S)-bicyclo[3,2,2]nonane-7R,8R-diamine))aniline and 3-(N-(N'-Boc-(1S,5R)-bicyclo[3,2,2]nonane-7R,8R-diamine))aniline mixture (II-73) JPEG0007796456000227.jpg2117025 mg of crude compound II-73-2, 2 ml of methanol, and 2.5 mg of 5% palladium on carbon were placed in a 100 ml hydrogenation reactor, maintained at 25°C and 50 psi, and hydrogen gas was introduced until the reaction was complete. The reaction solution was then concentrated to dryness to obtain 23 mg of crude compound II-73.

[0250] MS(ESI):(M+H) + ,346.2

[0251] Step 2: Synthesis of N-(3-(N-(N'-Boc-(1R,5S)-bicyclo[3,2,2]nonane-7R,8R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide and N-(3-(N-(N'-Boc-(1S,5R)-bicyclo[3,2,2]nonane-7R,8R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide mixture (IV-73) JPEG0007796456000228.jpg37170JPEG0007796456000229.jpg34170Under nitrogen protection, 11 mg of compound I-1, 10.9 mg of crude compound II-73, 32.8 mg of pyBoP, 12.2 mg of DIPEA, and 0.5 mL of DMF were sequentially added to a reaction flask and stirred overnight to complete the reaction. 2.5 mL of water was added to the reaction flask, and the mixture was extracted three times with ethyl acetate (2.5 mL each time). The combined organic phase was washed once with 1 mL of 1 mol / L aqueous hydrochloric acid and once with 1 mL of saturated aqueous sodium carbonate, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 20:1) to obtain 17.2 mg of compound IV-73.

[0252] MS(ESI):(M+H) + ,645.4

[0253] Step 3: Synthesis of N-(3-(N-((1R,5S)-bicyclo[3,2,2]nonane-7R,8R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide and N-(3-(N-((1S,5R)-bicyclo[3,2,2]nonane-7R,8R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide mixture (V-73) JPEG0007796456000230.jpg37170 At room temperature, 17.2 mg of compound IV-73 and 1 ml of dichloromethane were added to a reaction flask, and then 0.1 ml of trifluoroacetic acid was added dropwise. The mixture was reacted for 10 minutes and concentrated to dryness to obtain 14.2 mg of crude product of compound V-73.

[0254] MS(ESI):(M+H)+,545.4

[0255] Step 4: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-(1R,5S)-bicyclo[3,2,2]nonane-7R,8R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide and N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-(1S,5R)-bicyclo[3,2,2]nonane-7R,8R-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide mixture (C73) JPEG0007796456000231.jpg33170JPEG0007796456000232.jpg35170 14.2 mg of the crude product of compound V-73, 13 mg of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 30 mg of DIPEA, and 1 ml of DMF were sequentially added to a reaction flask, and the reaction flask was placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 2.5 ml of water was added, and the mixture was extracted with dichloromethane three times (2.5 ml each time). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 15:1) to obtain 7.6 mg of compound C73.

[0256] Mass spectrum data of compound C73: MS (ESI): (M+H) + ,801.4

[0257] Example 74: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-5,5-difluoro-1S,3S-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C74) JPEG0007796456000233.jpg33170 Step 1: Synthesis of 3-(N-(N'-Boc-5,5-difluoro-1S,3S-cyclohexanediamine))aniline (II-74) a. Synthesis of 3-(N-(5,5-difluoro-1S,3S-cyclohexanediamine))nitrobenzene (II-74-1) 100 mg of m-fluoronitrobenzene and 1.06 g of 5,5-difluoro-1S,3S-cyclohexanediamine were placed in a three-neck flask, heated to 100 °C under nitrogen protection, and reacted for 12 hours with stirring. After cooling to room temperature, the mixture was purified by column chromatography (eluent: dichloromethane:methanol = 5:1) to obtain 20 mg of a pale yellow solid (i.e., Compound II-74-1).

[0258] MS(ESI):(M+H) + ,272.1

[0259] b. Synthesis of 3-(N-(N'-Boc-5,5-difluoro-1S,3S-cyclohexanediamine))nitrobenzene (II-74-2) 20 mg of compound II-74-1, 8.2 mg of triethylamine, 18 mg of Boc anhydride, and 2 mL of dichloromethane were sequentially added to a reaction flask and allowed to react for 30 minutes with stirring at room temperature. The reaction mixture was washed once with 0.5 mL of 1 mol / L aqueous hydrochloric acid and once with 0.5 mL of water. The aqueous phase was extracted twice with dichloromethane (2.5 mL each time). The combined organic phase was dried over anhydrous sodium sulfate and concentrated to dryness to yield 27 mg of crude compound II-74-2.

[0260] MS(ESI):(M+H) + ,372.2

[0261] c. Synthesis of 3-(N-(N'-Boc-5,5-difluoro-1S,3S-cyclohexanediamine))aniline (II-74) JPEG0007796456000236.jpg2117027 mg of crude compound II-74-2, 2 ml of methanol, and 2.7 mg of 5% palladium on carbon were sequentially added to a 100 ml hydrogenation reactor, and the temperature was maintained at 25°C and 50 psi. Hydrogen gas was introduced until the reaction was complete, and the reaction solution was then concentrated to dryness to obtain 24.8 mg of crude compound II-74.

[0262] MS(ESI):(M+H) + ,342.2

[0263] Step 2: Synthesis of N-(3-(N-(N'-Boc-5,5-difluoro-1S,3S-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (IV-74) JPEG0007796456000237.jpg32170JPEG0007796456000238.jpg33170Under nitrogen protection, 11 mg of compound I-1, 10.7 mg of crude compound II-74, 32.8 mg of pyBoP, 12.2 mg of DIPEA, and 0.5 mL of DMF were sequentially added to a reaction flask and stirred overnight to complete the reaction. 2.5 mL of water was added to the reaction flask, and the mixture was extracted three times with ethyl acetate (2.5 mL each time). The combined organic phase was washed once with 1 mL of 1 mol / L aqueous hydrochloric acid and once with 1 mL of saturated aqueous sodium carbonate, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 15:1) to obtain 18.3 mg of compound IV-74.

[0264] MS(ESI):(M+H) + ,641.2

[0265] Step 3: Synthesis of N-(3-(N-(5,5-difluoro-1S,3S-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (V-74) JPEG0007796456000239.jpg32170 At room temperature, 18.3 mg of compound IV-74 and 1 ml of dichloromethane were added to a reaction flask, and then 0.1 ml of trifluoroacetic acid was added dropwise. The mixture was reacted for 10 minutes and concentrated to dryness to obtain 14.8 mg of crude product of compound V-74.

[0266] MS(ESI):(M+H) + ,541.3

[0267] Step 4: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-5,5-difluoro-1S,3S-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C74) 14.8 mg of the crude product of compound V-74, 13 mg of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 30 mg of DIPEA, and 1 ml of DMF were sequentially added to a reaction flask, and the reaction flask was placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 2.5 ml of water was added, and the mixture was extracted with dichloromethane three times (2.5 ml each time). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 15:1) to obtain 7 mg of compound C74.

[0268] Mass spectrum data of compound C74: MS (ESI): (M+H) + ,797.4 The nuclear magnetic resonance data of compound C74 is as follows: HNMR(400MHz,CDCl3)δ:8.78-8.80(m,1H),7.48-7.50(m,1H),7.12-7.28(m,4H),6.75-6.82 (m,2H),6.55-6.59(m,1H),6.25-6.35(m,2H),5.95(d,1H,J=12Hz),5.90(s,1H),5.13-5.18( m,1H),4.02-4.09(m,1H),3.95-3.99(m,1H),3.50-3.59(m,1H),2.80-2.98(m,3H),2.00-2. 43(m,8H),1.71-1.90(m,5H),1.15-1.58(m,8H),1.21-1.31(m,2H),1.10(s,3H),0.92(s,3H)

[0269] Example 75: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-tetrahydropyran-3S,5S-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C75) JPEG0007796456000241.jpg32170 Step 1: Synthesis of 3-(N-(N'-Boc-tetrahydropyran-3S,5S-diamine))aniline (II-75) a. Synthesis of 3-(N-(tetrahydropyran-3S,5S-diamine))nitrobenzene (II-75-1) 100 mg of m-fluoronitrobenzene and 822 mg of tetrahydrofuran-3S,5S-diamine were placed in a three-neck flask, heated to 100°C under nitrogen protection, and reacted for 12 hours with stirring. After cooling to room temperature, the mixture was purified by column chromatography (eluent: dichloromethane:methanol = 5:1) to obtain 12 mg of a pale yellow solid (i.e., Compound II-75-1).

[0270] MS(ESI):(M+H) + ,238.1

[0271] b. Synthesis of 3-(N-(N'-Boc-tetrahydropyran-3S,5S-diamine))nitrobenzene (II-75-2) 12 mg of compound II-75-1, 5.6 mg of triethylamine, 12 mg of Boc anhydride, and 2 mL of dichloromethane were sequentially added to a reaction flask and allowed to react for 30 minutes with stirring at room temperature. The reaction mixture was washed once with 0.5 mL of 1 mol / L aqueous hydrochloric acid and once with 0.5 mL of water. The aqueous phase was extracted twice with dichloromethane (2.5 mL each time). The combined organic phase was dried over anhydrous sodium sulfate and concentrated to dryness to yield 17 mg of crude compound II-75-2.

[0272] MS(ESI):(M+H) + ,338.2

[0273] c. Synthesis of 3-(N-(N'-Boc-tetrahydropyran-3S,5S-diamine))aniline (II-75) JPEG0007796456000244.jpg1717017 mg of crude compound II-75-2, 2 ml of methanol, and 1.7 mg of 5% palladium on carbon were sequentially added to a 100 ml hydrogenation reactor, and the temperature was maintained at 25°C and 50 psi. Hydrogen gas was introduced until the reaction was complete, and the reaction solution was then concentrated to dryness to obtain 15.4 mg of crude compound II-75.

[0274] MS(ESI):(M+H) + ,308.2

[0275] Step 2: Synthesis of N-(3-(N-(N'-Boc-tetrahydropyran-3S,5S-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (IV-75) JPEG0007796456000245.jpg32170JPEG0007796456000246.jpg33170Under nitrogen protection, 11 mg of compound I-1, 9.7 mg of crude compound II-75, 32.8 mg of pyBoP, 12.2 mg of DIPEA, and 0.5 mL of DMF were sequentially added to a reaction flask and stirred overnight to complete the reaction. 2.5 mL of water was added to the reaction flask, and the mixture was extracted three times with ethyl acetate (2.5 mL each time). The combined organic phase was washed once with 1 mL of 1 mol / L aqueous hydrochloric acid and once with 1 mL of saturated aqueous sodium carbonate, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 15:1) to obtain 19 mg of compound IV-75.

[0276] MS(ESI):(M+H) + ,607.4

[0277] Step 3: Synthesis of N-(3-(N-(tetrahydropyran-3S,5S-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (V-75) JPEG0007796456000247.jpg33170 At room temperature, 19 mg of compound IV-75 and 1 ml of dichloromethane were added to a reaction flask, and then 0.1 ml of trifluoroacetic acid was added dropwise. The mixture was reacted for 10 minutes and concentrated to dryness to obtain 15.8 mg of crude product of compound V-75.

[0278] MS(ESI):(M+H) + ,507.3 Step 4: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-tetrahydropyran-3S,5S-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C75) 15.8 mg of the crude product of compound V-75, 13 mg of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 30 mg of DIPEA, and 1 ml of DMF were sequentially added to a reaction flask, and the reaction flask was placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 2.5 ml of water was added, and the mixture was extracted with dichloromethane three times (2.5 ml each time). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 8:1) to obtain 4.8 mg of compound C75.

[0279] Mass spectrum data of compound C75: MS (ESI): (M+H) + ,763.4 The nuclear magnetic resonance data of compound C75 is as follows: HNMR(400MHz,CDCl3)δ:8.71-8.73(m,1H),7.46-7.51(m,1H),7.25-7.29(m,1H),7.12-7.15(m,1H),7.08-7.11 (m,2H),6.77-6.83(m,2H),6.48-6.52(m,1H),6.30-6.38(m,2H),5.95(d,1H,J=12Hz),5.68(s,1H),4.95-5.01( m,1H),4.12-4.15(m,2H),4.04-4.08(m,2H),3.89-4.00(m,2H),3.40-3.57(m,1H),2.75-2.95(m,3H),2.21-2. 48(m,2H),2.00-2.12(m,4H),1.71-1.90(m,4H),1.25-1.58(m,7H),1.10-1.18(m,2H),1.06(s,3H),0.88(s,3H)

[0280] Example 76: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-piperidine-3S,5S-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C76) JPEG0007796456000249.jpg35170 Step 1: Synthesis of 3-(N-(1,N'-bisBoc-piperidine-3S,5S-diamine))aniline (II-76) a. Synthesis of 3-(N-(piperidine-3S,5S-diamine))nitrobenzene (II-76-1) 100 mg of m-fluoronitrobenzene and 815 mg of piperidine-3S,5S-diamine were placed in a three-neck flask, heated to 100°C under nitrogen protection, and reacted for 12 hours with stirring. After cooling to room temperature, the mixture was purified by column chromatography (eluent: dichloromethane:methanol = 5:1) to obtain 9 mg of a pale yellow solid (i.e., Compound II-76-1).

[0281] MS(ESI):(M+H) + ,237.1

[0282] b. Synthesis of 3-(N-(1,N'-bisBoc-piperidine-3S,5S-diamine))nitrobenzene (II-76-2) 9 mg of compound II-76-1, 4.2 mg of triethylamine, 18.2 mg of Boc anhydride, and 2 mL of dichloromethane were sequentially added to a reaction flask and allowed to react for 30 minutes with stirring at room temperature. The reaction mixture was washed once with 0.5 mL of 1 mol / L aqueous hydrochloric acid and once with 0.5 mL of water. The aqueous phase was extracted twice with dichloromethane (2.5 mL each time). The combined organic phase was dried over anhydrous sodium sulfate and concentrated to dryness to yield 16.6 mg of crude compound II-76-2.

[0283] MS(ESI):(M+H) + ,437.2 c. Synthesis of 3-(N-(1,N'-bisBoc-piperidine-3S,5S-diamine))aniline (II-76) JPEG0007796456000252.jpg2417016.6 mg of crude compound II-76-2, 2 ml of methanol, and 1.6 mg of 5% palladium on carbon were sequentially added to a 100 ml hydrogenation reactor, and the temperature was maintained at 25°C and 50 psi. Hydrogen gas was introduced until the reaction was complete, and the reaction solution was then concentrated to dryness to obtain 15.4 mg of crude compound II-76.

[0284] MS(ESI):(M+H) + ,407.3

[0285] Step 2: Synthesis of N-(3-(N-(1,N'-bisBoc-piperidine-3S,5S-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (IV-76) JPEG0007796456000253.jpg32170JPEG0007796456000254.jpg34170Under nitrogen protection, 11 mg of compound I-1, 12.8 mg of crude compound II-76, 32.8 mg of pyBoP, 12.2 mg of DIPEA, and 0.5 mL of DMF were sequentially added to a reaction flask and stirred overnight to complete the reaction. 2.5 mL of water was added to the reaction flask, and the mixture was extracted three times with ethyl acetate (2.5 mL each time). The combined organic phase was washed once with 1 mL of 1 mol / L aqueous hydrochloric acid and once with 1 mL of saturated aqueous sodium carbonate, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 15:1) to obtain 21 mg of compound IV-76.

[0286] MS(ESI):(M+H) + ,706.4

[0287] Step 3: Synthesis of N-(3-(N-(piperidine-3S,5S-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (V-76) JPEG0007796456000255.jpg32170 21 mg of compound IV-76 and 1 ml of dichloromethane were added to a reaction flask at room temperature, and then 0.1 ml of trifluoroacetic acid was added dropwise. The mixture was reacted for 10 minutes and concentrated to dryness to obtain 14.8 mg of crude product of compound V-76.

[0288] MS(ESI):(M+H) + ,506.3

[0289] Step 4: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-piperidine-3S,5S-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C76) 21 mg of the crude product of compound V-76, 13 mg of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 30 mg of DIPEA, and 1 ml of DMF were sequentially added to a reaction flask, and the reaction flask was placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 2.5 ml of water was added, and the mixture was extracted with dichloromethane three times (2.5 ml each time). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 8:1) to obtain 3.3 mg of compound C76.

[0290] Mass spectrum data of compound C76: MS(ESI):(M+H) + ,762.4 The nuclear magnetic resonance data of compound C76 is as follows: HNMR(400MHz,CDCl3)δ:8.57-8.59(m,1H),7.45-7.49(m,1H),7.20-7.25(m,1H),7.12-7.18(m,3H),6.73- 6.81(m,2H),6.57-6.60(m,1H),6.35-6.40(m,2H),5.89(d,1H,J=12Hz),5.71(s,1H),4.91-4.96(m,1H),3. 90-4.02(m,2H),3.82-3.86(m,2H),3.71-3.74(m,2H),3.45-3.62(m,1H),2.67-2.90(m,3H),2.21-2.33(m ,2H),2.05-2.18(m,4H),1.71-1.90(m,4H),1.15-1.58(m,7H),1.01-1.11(m,2H),0.99(s,3H),0.85(s,3H)

[0291] Example 77: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-1-methylpiperidine-3S,5S-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C77) JPEG0007796456000257.jpg32170 Step 1: Synthesis of 3-(N-(N'-Boc-1-methylpiperidine-3S,5S-diamine))aniline (II-77) a. Synthesis of 3-(N-(1-methylpiperidine-3S,5S-diamine))nitrobenzene (II-77-1) 100 mg of m-fluoronitrobenzene and 915 mg of N-methylpiperidine-3S,5S-diamine were placed in a three-neck flask, heated to 100°C under nitrogen protection, and reacted for 12 hours with stirring. After cooling to room temperature, the mixture was purified by column chromatography (eluent: dichloromethane:methanol = 6:1) to obtain 13 mg of a pale yellow solid (i.e., Compound II-77-1).

[0292] MS(ESI):(M+H) + ,251.1

[0293] b. Synthesis of 3-(N-(N'-Boc-1-methylpiperidine-3S,5S-diamine))nitrobenzene (II-77-2) 13 mg of compound II-77-1, 5.8 mg of triethylamine, 11.3 mg of Boc anhydride, and 2 mL of dichloromethane were sequentially added to a reaction flask and allowed to react for 30 minutes with stirring at room temperature. The reaction mixture was washed once with 0.5 mL of 1 mol / L aqueous hydrochloric acid and once with 0.5 mL of water. The aqueous phase was extracted twice with dichloromethane (2.5 mL each time). The combined organic phase was dried over anhydrous sodium sulfate and concentrated to dryness to yield 18 mg of crude compound II-77-2.

[0294] MS(ESI):(M+H) + ,351.2 c. Synthesis of 3-(N-(N'-Boc-1-methylpiperidine-3S,5S-diamine))aniline (II-77) JPEG0007796456000260.jpg2217018 mg of crude compound II-77-2, 2 ml of methanol, and 1.8 mg of 5% palladium on carbon were added sequentially to a 100 ml hydrogenation reactor, and the temperature was maintained at 25°C and 50 psi. Hydrogen gas was introduced until the reaction was complete, and the reaction solution was then concentrated to dryness to obtain 16.4 mg of crude compound II-77.

[0295] MS(ESI):(M+H) + ,321.2

[0296] Step 2: Synthesis of N-(3-(N-(N'-Boc-1-methylpiperidine-3S,5S-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (IV-77) Under nitrogen protection, 11 mg of compound I-1, 10 mg of crude compound II-77, 32.8 mg of pyBoP, 12.2 mg of DIPEA, and 0.5 mL of DMF were sequentially added to a reaction flask and stirred overnight to complete the reaction. 2.5 mL of water was added to the reaction flask, followed by three extractions with ethyl acetate (2.5 mL each). The combined organic phases were washed with 1 mL of 1 mol / L aqueous hydrochloric acid and 1 mL of saturated aqueous sodium carbonate, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 15:1) to obtain 18.7 mg of compound IV-77.

[0297] MS(ESI):(M+H) + ,620.4

[0298] Step 3: Synthesis of N-(3-(N-(1-methylpiperidine-3S,5S-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (V-77) JPEG0007796456000263.jpg33170 At room temperature, 14.8 mg of compound IV-77 and 1 ml of dichloromethane were added to a reaction flask, and then 0.1 ml of trifluoroacetic acid was added dropwise. The mixture was reacted for 10 minutes and concentrated to dryness to obtain 12.4 mg of crude product of compound V-77.

[0299] MS(ESI):(M+H) + ,520.4 Step 4: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-1-methylpiperidine-3S,5S-diamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C77) 16.3 mg of the crude product of compound V-77, 13 mg of 4-fluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 30 mg of DIPEA, and 1 ml of DMF were sequentially added to a reaction flask, and the reaction flask was placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 2.5 ml of water was added, and the mixture was extracted with dichloromethane three times (2.5 ml each time). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol = 10:1) to obtain 5.1 mg of compound C77.

[0300] Mass spectrum data of compound C77: MS(ESI):(M+H) + ,776.4 The nuclear magnetic resonance data of compound C77 is as follows: HNMR(400MHz,CDCl3)δ:8.59-8.61(m,1H),7.47-7.50(m,1H),7.15-7.21(m,1H),7.08-7.12(m,3H),6.73-6.81(m,2H),6.51-6.57(m,1H),6. 35-6.39(m,2H),5.96(d,1H,J=12Hz),5.88(s,1H),5.02-5.10(m,1H), 3.92-3.98(m,2H),3.73-3.84(m,4H),3.58(s,3H),3.40-3.57(m,1H), 2.67-2.90(m,3H),2.28-2.43(m,2H),2.05-2.15(m,4H),1.67-1.95(m,4H),1.15-1.60(m,7H),1.05-1.18(m,2H),0.96(s,3H),0.75(s,3H)

[0301] Example 78: Synthesis of N-(3-(N-(N'-(7-fluoro-4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C78) JPEG0007796456000265.jpg35170JPEG0007796456000266.jpg35170 At room temperature, 1.21 g of compound V-59, 1.04 g of 4,7-difluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 1.53 g of DIPEA, and 10 ml of DMF were added to a reaction flask. The reaction flask was then placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 20 ml of water was added to the reaction flask, and the mixture was extracted three times with dichloromethane (20 ml each time). The organic phases were combined, washed once with 20 ml of water, dried over anhydrous sodium sulfate, and then concentrated to dryness to obtain a crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol=10:1) to give 0.22 g of compound C78.

[0302] Mass spectrum data of compound C78: MS(ESI):(M+H) + ,779.4 The nuclear magnetic resonance data of compound C78 is as follows: HNMR(400MHz,CDCl3)δ: 8.62-8.64(m,1H),7.15-7.18(m,1H),7.20-7.24(m,3H),6.83-6.87(m,2H),6.40-6.47(m, 1H),6.36-6.39(m,2H),5.80(d,1H,J=12Hz),5.70(s,1H),4.85-4.91(m,1H),4.23-4.38(m, 2H)3.75-3.85(m,2H),3.28-3.33(m,1H),2.65-2.81(m,3H),2.15-2.30(m,2H),2.01-2.11 (m,2H),1.90-1.73(m,10H),1.55-1.15(m,9H),1.08-1.11(m,2H),0.92(s,3H),0.75(s,3H)

[0303] Example 79: Synthesis of N-(3-(N-(N'-(6-fluoro-4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C79) JPEG0007796456000267.jpg33170JPEG0007796456000268.jpg34170At room temperature, 1.21 g of compound V-59, 1.04 g of 4,6-difluoro-2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione, 1.53 g of DIPEA, and 10 ml of DMF were added to a reaction flask. The reaction flask was then placed in a microwave reactor and reacted at 50 Hz for 10 minutes. 20 ml of water was added to the reaction flask, and the mixture was extracted three times with dichloromethane (20 ml each time). The combined organic phase was washed once with 20 ml of water, dried over anhydrous sodium sulfate, and then concentrated to dryness to obtain the crude product. The crude product was purified by column chromatography (eluent: dichloromethane:methanol=10:1) to give 0.13 g of compound C79.

[0304] Mass spectrum data of compound C79: MS(ESI):(M+H) + ,779.4 The nuclear magnetic resonance data of compound C79 is as follows: HNMR(400MHz,CDCl3)δ:8.75-8.77(m,1H),7.40-7.47(m,1H),7.17-7.20(m,1H),7.13 -7.18(m,2H),6.72-6.81(m,2H),6.55-6.59(m,1H),6.31-6.35(m,2H),5.80(d,1H,J=1 2Hz),5.71(s,1H),4.88-4.91(m,1H),3.40-3.57(m,1H),2.67-2.90(m,3H),2.21-2.3 3(m,2H),2.08-2.18(m,2H),1.90-1.71(m,8H),1.58-1.15(m,11H),1.01-1.11(m,2H), 0.96(s,3H),0.75(s,3H)

[0305] Example 80: Synthesis of N-(3-(N-(N-methyl-N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C80) JPEG0007796456000269.jpg32170 Step 1 a. Synthesis of 3-(N-(N-methyl-N'-Boc-1R,3R-cyclohexanediamine))nitrobenzene (II-80-1) JPEG0007796456000270.jpg191703-(N-(N'-Boc-1R,3R cyclohexanediamine))nitrobenzene 0.2 g, methylene iodide 127 mg, and potassium carbonate 166 mg were added sequentially to 1.5 mL of N,N-dimethylformamide and stirred overnight at room temperature. After that, 10 mL of water was added and the mixture was extracted three times with ethyl acetate (10 mL each time). The organic phases were combined, washed sequentially with 3 mL of saturated brine and once with 3 mL of water, dried, and the solvent was removed by rotary evaporation to obtain 0.18 g of crude product (i.e., crude product of compound II-80-1), which was directly used in the next reaction.

[0306] b. Synthesis of 3-(N-(N-methyl-N'-Boc-1R,3R-cyclohexanediamine))aniline (II-80) JPEG0007796456000271.jpg181700.18 g of the crude product of compound II-80-1 obtained in the previous step was placed in a 100 ml hydrogenation reactor, followed by the addition of 10 ml of methanol and 18 mg of 5% palladium on carbon. Hydrogen gas was introduced at 25°C and maintained at 50 psi until complete completion. The mixture was then filtered and concentrated to dryness to obtain 0.16 g of a crude product (i.e., the crude product of compound II-80), which was directly used in the next reaction.

[0307] Step 2: Synthesis of N-(3-(N-(N-methyl-N'-Boc-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (IV-80) JPEG0007796456000272.jpg35170JPEG0007796456000273.jpg33170Under nitrogen protection, 36.4 mg of compound I-1, 32 mg of the crude product of compound II-80, 101 mg of pyBoP, 38.7 mg of N,N-diisopropylethylamine, and 1.5 mL of DMF were sequentially added to a reaction flask and stirred overnight. After that, 5 mL of water was added and the mixture was extracted three times with ethyl acetate (10 mL each time). The organic phases were combined and washed once with 3 mL of 1 mol / L dilute hydrochloric acid solution and 3 mL of saturated sodium carbonate solution, dried over anhydrous sodium sulfate, concentrated to dryness, and then purified by column chromatography (eluent: dichloromethane:methanol = 20:1) to obtain 60 mg of the product (i.e., compound IV-80).

[0308] MS(ESI):(M+H)+,620.4

[0309] Step 3: Synthesis of N-(3-(N-(N-methyl-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (V-80) 60 mg of compound IV-80 and 2 mL of dichloromethane were added to a glass bottle at room temperature, and then 0.2 mL of trifluoroacetic acid was slowly added dropwise. The mixture was reacted for 10 minutes and then concentrated to dryness to obtain the target product (i.e., crude product of compound V-80). This was used in the next reaction without separation.

[0310] MS(ESI):(M+H)+,519.4

[0311] Step 4: Synthesis of N-(3-(N-(N-methyl-N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androst-1-ene-17β-formamide (C80) JPEG0007796456000275.jpg32170 The crude product of compound V-80 obtained in step 3, 26 mg of 4-fluoro-2-(2,6-dioxo-3-piperidinyl)-isoindole-1,3-dione, 60 mg of DIPEA, and 1 mL of DMF were placed in a microwave reactor and reacted at 50 Hz for 10 minutes. The mixture was then extracted three times by adding water and dichloromethane, dried over anhydrous sodium sulfate, concentrated, and eluted by column chromatography (eluent: dichloromethane:methanol = 15:1) to obtain 19 mg of compound C80.

[0312] Mass spectrum data of compound C80: MS (ESI): (M+H) + ,775.4 The nuclear magnetic resonance data of compound C80 is as follows: HNMR(400MHz,CDCl3)δ:8.55-8.57(m,1H),7.44-7.48(m,1H),7.16-7.22(m,1H),7.05-7.12(m,3H), 6.73-6.81(m,2H),6.51-6.57(m,1H),6.25-6.35(m,2H),5.80(d,1H,J=12Hz),5.77(s,1H),4.85-4. 92(m,1H),3.70-3.83(m,2H),3.49(s,3H)3.28-3.33(m,1H),2.66-2.90(m,3H),2.21-2.35(m,2H),2 .08-2.20(m,2H),1.91-1.62(m,12H),1.49-1.31(m,7H),1.06-1.12(m,2H),0.99(s,3H),0.73(s,3H)

[0313] Example 81: Synthesis of N-(3-(N-(N'-(4-(2-(3-(2,6-dioxopiperidine))-isoindole-1,3-dione))-1R,3R-cyclohexanediamine))phenyl)-3-oxo-4-aza-5α-androstane-17β-formamide (C81) JPEG0007796456000276.jpg34170JPEG0007796456000277.jpg3517035 mg of compound C59 was added to a 100 mL hydrogenation kettle, 3 mL of methanol and 5 mg of 5% palladium on carbon were added, hydrogen gas was introduced to maintain the pressure at 50 psi, and the reaction was carried out at 50°C for 2 hours. After monitoring to confirm the completion of the reaction, the reaction was filtered, concentrated to dryness, and purified by column chromatography (eluent: dichloromethane:methanol = 10:1) to obtain 33 mg of the product (i.e., compound C81).

[0314] Mass spectrum data of compound C81: MS (ESI): (M+H) + ,763.4 The nuclear magnetic resonance data of compound C81 is as follows: HNMR(400MHz,CDCl3)δ:8.90-8.92(m,1H),7.38-7.41(m,1H),7.05-7.09(m,1H),7.08-7.12(m,3H), 6.73-6.81(m,1H),6.53-6.59(m,1H),6.16-6.35(m,1H),6.06-6.10(m,1H),5.17(s,1H)4.88-4.91( m,1H),3.73-3.80(m,2H),3.28-3.33(m,1H),2.67-2.90(m,3H),2.41(m,2H),2.21-2.33(m,2H),1.9 5-2.10(m,3H),1.91-1.69(m,10H),1.52-1.17(m,10H),0.98-1.11(m,2H),0.91(s,3H),0.70(s,3H)

[0315] Example 82: Growth toxicity test of prostate cancer cell lines Vcap, 22Rv1, and PC-3 (1)Cell culture The Vcap cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: SCSP-5034) was cultured and passaged in a medium (DMEM medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum) in an incubator at 37°C with 5% carbon dioxide. The 22Rv1 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: TCHu100) was cultured and passaged in RPMI-1640 medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum in an incubator at 37°C with 5% carbon dioxide. PC-3 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: SCSP-532) was cultured and passaged in a medium (F12K medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum) in an incubator at 37°C with 5% carbon dioxide.

[0316] (2) Cell inoculation VCap cells were seeded into a 96-well plate at 20,000 cells / well (90 μL) and cultured for 72 hours, after which the old medium was removed and 90 μL of new medium was added. 22Rv1 cells were seeded into a 96-well plate at 5000 cells / well (90 μL) and cultured for 48 hours. PC-3 cells were seeded at 4000 cells / well (90 μL) into a 96-well plate and cultured for 24 hours.

[0317] (3) Preparation of compound stock solutions Compound stock solutions were prepared at 10 mM in DMSO.

[0318] (4) Dilution of stock solution For VCap cells, compound stock solutions were diluted with DMSO to give DMSO solutions 1-10 with the corresponding concentrations: 0 μM, 80 μM, 16 μM, 3.2 μM, 1.5 μM, 0.5 μM, 0.15 μM, 0.05 μM, 0.025 μM, and 0.0005 μM. For 22Rv1 cells and PC-3 cells, the compound stock solutions were diluted with DMSO to give DMSO solutions 11-19 with the corresponding concentrations: 0 μM, 500 μM, 100 μM, 20 μM, 4 μM, 0.8 μM, 0.16 μM, 0.032 μM, and 0.0064 μM.

[0319] (5) Mixing of compounds and medium For Vcap cells, 2 μL of each of the solutions 1-10 was taken and added to 98 μL of Vcap medium to obtain compound-medium mixed solutions 20-29. For 22Rv1 cells, 2 μL of each of the solutions 11-19 was taken and added to 98 μL of 22Rv1 medium to obtain compound-medium mixed solutions 30-38. For PC-3 cells, 2 μL of each of the solutions 11-19 was taken and added to 98 μL of PC-3 medium to obtain mixed solutions 39-47 of the compound and medium.

[0320] (6) Addition of compounds For Vcap cells, 10 μL of each of the solutions 20-29 was added to the inoculated cell solution. The total volume of the solution was 100 μL (90 μL + 10 μL). The concentrations of the corresponding compounds in each well of cells were 0 μM, 0.16 μM, 0.032 μM, 0.006 μM, 0.003 μM, 0.001 μM, 0.0003 μM, 0.0001 μM, 0.00005 μM, and 0.00001 μM, respectively. Each concentration was repeated in triplicate. For 22Rv1 and PC-3 cells, 10 μL of each of solutions 30-38 and 39-47 was added to the inoculated cell solution. The total volume of the solution was 100 μL (90 μL + 10 μL). The concentrations of the corresponding compounds in each well of cells were 0 μM, 1.0 μM, 0.2 μM, 0.04 μM, 0.008 μM, 0.0016 μM, 0.00032 μM, 0.00006 μM, and 0.00001 μM, respectively. Each concentration was replicated in triplicate.

[0321] (7) Cell proliferation toxicity test After the treatment, the cells were cultured for 3 days and then detected with CCK8. 10 μL of CCK8 solution (DOJINDO, catalog number: PF725) was added to each well, placed in a 37°C incubator, and read at 450 nm using a microplate reader after 1-4 hours. Tables 1 to 3 show the growth toxicity data of each compound against Vcap, 22Rv1, and PC-3 cell lines, and IC50 is the half maximal inhibitory concentration. If the IC50 is 0.1 nM or less, it is A; B if the IC50 is 10 nM or less and greater than 0.1 nM; C if IC50 is less than or equal to 1000 nM and greater than 10 nM; If the IC50 is greater than 1000 nM, it is D.

[0322] Table 1: Growth toxicity data for Vcap, 22Rv1, and PC-3 cell lines JPEG0007796456000278.jpg253170JPEG0007796456000279.jpg155170

[0323] Table 2: Growth toxicity data of compounds C29, C50, C59, and C63 in VCaP cell line JPEG0007796456000280.jpg42170

[0324] Table 3: Growth toxicity data of compounds C29, C50, C59, and C63 in the 22Rv1 cell line JPEG0007796456000281.jpg45170

[0325] Example 83: Growth toxicity test of C63 against several tumor cell lines (1)Cell culture PC-3 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: SCSP-532) was cultured and passaged in a medium (F12K medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum) in an incubator at 37°C with 5% carbon dioxide.

[0326] DU 145 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: SCSP-5024) was cultured and passaged in a medium (MEM medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum) in an incubator under conditions of 5% carbon dioxide and 37°C.

[0327] AGS cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: TCHu232) was cultured and passaged in a medium (F12K medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum, 5% carbon dioxide, 37°C) in an incubator.

[0328] HuH-7 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: SCSP-526) was cultured and passaged in a medium (DMEM medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum) in an incubator at 37°C with 5% carbon dioxide.

[0329] The Ca Ski cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: TCHu137) was cultured and passaged in RPMI-1640 medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum in an incubator at 37°C with 5% carbon dioxide.

[0330] C-33 A cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: TCHu176) was cultured and passaged in a medium (MEM medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum) in an incubator at 37°C with 5% carbon dioxide.

[0331] OVCAR-3 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: TCHu228) was cultured and passaged in a medium (RPMI-1640 medium + 1% penicillin-streptomycin double antibody solution + 20% fetal bovine serum + 0.01 mg / ml recombinant human insulin) in an incubator at 37°C with 5% carbon dioxide.

[0332] PANC-1 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: SCSP-535) was cultured and passaged in a medium (DMEM medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum) in an incubator at 37°C with 5% carbon dioxide.

[0333] MCF-7 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: SCSP-531) was cultured and passaged in a medium (MEM medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum + 0.01 mg / ml recombinant human insulin) in an incubator at 37°C with 5% carbon dioxide.

[0334] The MDA-MB-468 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: TCHu136) was cultured and passaged in a medium (L15 medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum) at 37°C in an incubator.

[0335] The MDA-MB-231 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: TCHu227) was cultured and passaged in a 37°C incubator in medium (L15 medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum).

[0336] The NCI-N87 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: TCHu130) was cultured and passaged in RPMI-1640 medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum in an incubator at 37°C with 5% carbon dioxide.

[0337] The BT-549 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: TCHu93) was cultured and passaged in RPMI-1640 medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum + 0.01 mg / ml recombinant human insulin in an incubator at 37°C with 5% carbon dioxide.

[0338] BxPC-3 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: TCHu12) was cultured and passaged in RPMI-1640 medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum in an incubator at 37°C with 5% carbon dioxide.

[0339] The HGC-27 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: TCHu22) was cultured and passaged in RPMI-1640 medium + 1% penicillin-streptomycin double antibody solution + 20% fetal bovine serum in an incubator at 37°C with 5% carbon dioxide.

[0340] DLD-1 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: TCHu134) was cultured and passaged in RPMI-1640 medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum in an incubator at 37°C with 5% carbon dioxide.

[0341] The SNU-1 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: TCHu230) was cultured and passaged in RPMI-1640 medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum in an incubator at 37°C with 5% carbon dioxide.

[0342] The U251 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: TCHu58) was cultured and passaged in a medium (DMEM medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum) in an incubator at 37°C with 5% carbon dioxide.

[0343] The KATOIII cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: TCHu229) was cultured and passaged in an incubator containing IMDM medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum at 37°C with 5% carbon dioxide.

[0344] HepG2 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: TCHu72) was cultured and passaged in a medium (MEM medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum) in an incubator at 37°C with 5% carbon dioxide.

[0345] Hep3B2.1-7 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: TCHu106) was cultured and passaged in a medium (MEM medium + 1% penicillin-streptomycin double antibody solution + 10% fetal bovine serum) in an incubator at 37°C with 5% carbon dioxide.

[0346] The HL-60 cell line (obtained from the Cell Bank of the Chinese Academy of Sciences, catalog number: TCHu23) was cultured and passaged in a medium (IMDM medium + 1% penicillin-streptomycin double antibody solution + 20% fetal bovine serum) in an incubator under conditions of 5% carbon dioxide and 37°C.

[0347] (2) Cell inoculation PC-3 cells were seeded at 4000 cells / well (90 μL) into a 96-well plate and cultured for 24 hours.

[0348] DU 145 cells were seeded into a 96-well plate at 5000 cells / well (90 μL) and cultured for 24 hours.

[0349] AGS cells were seeded into a 96-well plate at 2000 cells / well (90 μL) and cultured for 24 hours.

[0350] HuH-7 cells were seeded at 5000 cells / well (90 μL) into a 96-well plate and cultured for 24 hours.

[0351] Ca Ski cells were seeded into a 96-well plate at 5000 cells / well (90 μL) and cultured for 24 hours.

[0352] C-33 A cells were seeded at 5000 cells / well (90 μL) into a 96-well plate and cultured for 24 hours.

[0353] OVCAR-3 cells were seeded into a 96-well plate at 2000 cells / well (90 μL) and cultured for 24 hours.

[0354] PANC-1 cells were seeded at 5000 cells / well (90 μL) into a 96-well plate and cultured for 24 hours.

[0355] MCF-7 cells were seeded into a 96-well plate at 5000 cells / well (90 μL) and cultured for 24 hours.

[0356] MDA-MB-468 cells were seeded at 5000 cells / well (90 μL) into a 96-well plate and cultured for 24 hours.

[0357] MDA-MB-231 cells were seeded into a 96-well plate at 5000 cells / well (90 μL) and cultured for 24 hours.

[0358] NCI-N87 cells were seeded into a 96-well plate at 8000 cells / well (90 μL) and cultured for 24 hours.

[0359] BT-549 cells were seeded at 5000 cells / well (90 μL) into a 96-well plate and cultured for 24 hours.

[0360] BxPC-3 cells were seeded into a 96-well plate at 3000 cells / well (90 μL) and cultured for 24 hours.

[0361] HGC-27 cells were seeded into a 96-well plate at 3000 cells / well (90 μL) and cultured for 24 hours.

[0362] DLD-1 cells were seeded at 2000 cells / well (90 μL) into a 96-well plate and cultured for 24 hours.

[0363] SNU-1 cells were seeded at 5000 cells / well (90 μL) into a 96-well plate and cultured for 24 hours.

[0364] U251 cells were seeded at 2000 cells / well (90 μL) into a 96-well plate and cultured for 24 hours.

[0365] KATO III cells were seeded into a 96-well plate at 8000 cells / well (90 μL) and cultured for 24 hours.

[0366] Hep G2 cells were seeded into a 96-well plate at 2000 cells / well (90 μL) and cultured for 24 h.

[0367] Hep 3B2.1-7 cells were seeded into a 96-well plate at 2000 cells / well (90 μL) and cultured for 24 hours.

[0368] HL-60 cells were seeded into a 96-well plate at 20,000 cells / well (90 μL) and cultured for 24 hours.

[0369] (3) Preparation of compound stock solutions Compound stock solutions were prepared at 10 mM in DMSO.

[0370] (4) Dilution of stock solution Compound stock solutions were diluted with DMSO to give DMSO solutions 1-9 with the corresponding concentrations: 0 μM, 2500 μM, 500 μM, 100 μM, 20 μM, 4 μM, 0.8 μM, 0.16 μM, and 0.032 μM, respectively.

[0371] (5) Mixing of compounds and medium 2 μL of each of solutions 1-9 was taken and added to 98 μL of the medium to obtain compound-medium mixed solutions 11-19.

[0372] (6) Addition of compounds 10 μL of each of the 11-19 solutions was added to the inoculated cell solution, with the total volume of the solution being 100 μL (90 μL + 10 μL).

[0373] In this case, the corresponding compound concentrations in each well of cells were 0 μM, 5.0 μM, 1.0 μM, 0.2 μM, 0.04 μM, 0.008 μM, 0.0016 μM, 0.00032 μM, and 0.000064 μM, respectively. Each concentration was repeated in triplicate.

[0374] (7) Cell proliferation toxicity test After the treatment, the cells were cultured for 3 days and then detected with CCK8. 10 μL of CCK8 solution (DOJINDO, catalog number: PF725) was added to each well, the cells were placed in a 37°C incubator, and the cells were read at 450 nm using a microplate reader after 1-4 hours.

[0375] JPEG0007796456000282.jpg120170

[0376] Example 83: Experiments of compounds C29, C59, and C63 on tumor transplantation in 22Rv1 mice Breed / Strain: BALB / c nude mouse Gender: Male Weight: 19.9-23.6g Age: 7-8 weeks Animal grade: SPF grade Animal care and environment: The animals were allowed free access to food (cobalt-60 irradiated feed) and water (SPF purified water). Animals were housed in groups of six in cages in an IVC system with a barrier environment. The animal care environment was a temperature of 20-26°C, relative humidity of 40-70%, and a 12-hour light / 12-hour dark regime.

[0377] Cell culture: 22RV1 (human prostate cancer cell line) medium was RPMI1640 (Gibco, product code: C11875500CP), 10% fetal bovine serum (Cellmax, product code: SA211.02), 100 U / ml penicillin and 100 μg / ml streptomycin (Hyclone, product code: SV30010), and 2 mM L-glutamine (Gibco, product code: 25030-081). Cells were digested with 0.25% trypsin and passaged 1:3 every other day. When the cells reached the required cell number and were in logarithmic growth phase, they were harvested and counted, and the cell concentration was adjusted to 5 × 10 7 The cell suspension was adjusted to 2.5 × 10 cells / ml, and the cell suspension and Matrigel (Corning, product number: 354234) were mixed uniformly on ice at a volume ratio of 1:1. The mixture was then stored on ice until use (cell concentration: 2.5 × 10 cells / ml). 7 / ml).

[0378] Tumor cell inoculation: 0.2 ml of cell suspension (5 × 10) was injected subcutaneously into the right back of each mouse. 6 22RV1 cells) were inoculated.

[0379] Selection of tumor-bearing mice: 13 days after inoculation of mice with 22RV1 cells, tumors with a volume of 80 mm 3 -150mm 3 Mice were selected and randomly assigned to groups of 6 mice per group based on tumor volume and body weight.

[0380] Preparation of test samples: DMSO refers to dimethyl sulfoxide, and Captisol refers to sulfobutyl-β-cyclodextrin.

[0381] 15 mg of C29 was weighed and dissolved in 0.375 ml of DMSO solution, and 7.125 ml of 20% Captisol solution was added to obtain 7.5 ml of a 2 mg / ml C29 sample solution.

[0382] 15 mg of C59 was weighed and dissolved in 0.375 ml of DMSO solution, and 7.125 ml of 20% Captisol solution was added to obtain 7.5 ml of a 2 mg / ml C59 sample solution.

[0383] 7.5 mg of C59 was weighed and dissolved in 0.375 ml of DMSO solution, and 7.125 ml of 20% Captisol solution was added to obtain 7.5 ml of a 1 mg / ml C59 sample solution.

[0384] 15 mg of C63 was weighed and dissolved in 0.375 ml of DMSO solution, and 7.125 ml of 20% Captisol solution was added to obtain 7.5 ml of a 2 mg / ml C63 sample solution.

[0385] 7.5 mg of C63 was weighed and dissolved in 0.375 ml of DMSO solution, and 7.125 ml of 20% Captisol solution was added to obtain 7.5 ml of a 1 mg / ml C63 sample solution.

[0386] 3.75 mg of C63 was weighed and dissolved in 0.375 ml of DMSO solution, and 7.125 ml of 20% Captisol solution was added to obtain 7.5 ml of a 0.5 mg / ml C63 sample solution.

[0387] Animal administration: The administration route was intraperitoneal injection (IP) or oral gavage (PO). The administration frequency was once daily for a total of 18-21 days.

[0388] Animal weight measurements: measured twice a week.

[0389] Tumor volume measurement: Tumor volume was measured twice a week, and the long and short diameters of the tumor were measured using a vernier caliper. Tumor volume (TV) = 0.5 × a × b 2 (where a and b represent the long and short diameters of the tumor, respectively).

[0390] The experimental results are shown in the figure below. 1. Data graphs of body weight and tumor volume of mice administered C29 intraperitoneally (IP) are shown in Figures 1 and 2. 2. Data graphs of body weight and tumor volume of C59 intraperitoneally injected (IP) and oral gavage (PO) administered mice are shown in Figures 3 and 4. 3. Data on body weight and tumor volume of C63 mice administered orally (PO) are shown in Figures 5 and 6.

[0391] Example 84: Western blot experiment of compound C59 against 22Rv1, PC-3, and HepG2 (1) Experimental materials: 22Rv1, PC-3, HepG2 cells, Anti-SRD5A1 antibody (ab167606), Anti-SRD5A2 antibody (ab124877), Anti-SRD5A3 antibody (PA5-107029), Anti-Androgen Receptor (AR) antibody (CST, #5153), Anti-GSPT1 antibody (abcam,ab234433,), Anti-GADPH antibody (CST, #5174), Anti-c-Myc antibody (CST, #8583), Anti-α-tublin antibody (CST, #2125), Anti-rabbit IgG, HRP-linked Antibody (CST, #7074), electrophoresis device and membrane transfer device (Beijing Sixty, DYCZ-24DN)

[0392] (2) Experimental method Cell inoculation: 22Rv1 (Chinese Medical University Cell Bank, TCHu100), PC-3 (Chinese Medical University Cell Bank, SCSP-532), and HepG2 cells (Chinese Medical University Cell Bank, TCHu72) were subcultured and then digested with trypsin to give 6 × 10 5 A 1000 / mL cell suspension was prepared, inoculated into a 6-well plate (3 mL / well), and cultured overnight in a 37°C, 5% CO2 cell incubator.

[0393] Compound addition and post-treatment: After adding the test drug at different concentrations, the cells were incubated for 24 hours. After washing twice with ice-cold PBS buffer, 800 μl of RIPA lysis solution was added to each well and lysed on ice for 30 minutes. The lysates were collected in centrifuge tubes and centrifuged at 12,000 rpm for 10 minutes at 4°C. The supernatant was transferred to a new centrifuge tube.

[0394] Protein quantification: Protein was quantified using a BCA protein quantification kit, and the protein concentration was adjusted to a uniform level. The mixture was then added to the loading buffer, mixed, and denatured at 100°C for 5 minutes. The mixture was then stored at -80°C or used directly for Western blot analysis.

[0395] Western blotting experiments (WB): SDS-PAGE gels were prepared at appropriate concentrations based on the molecular weight of the target protein. When the resolving gel concentration was 8%, the stacking gel concentration was 5%. The loading volume was adjusted based on the protein quantification results. The protein samples were loaded at a voltage of 90 V in the stacking gel and 130 V after entering the resolving gel. Electrophoresis was stopped after the bromophenol blue had completely escaped from the PAGE gel. After preparing the transfer sandwich (wet transfer), the transfer apparatus was placed in an ice-water bath and transferred to the membrane at a constant current of 180 mA for 1.2 hours. The PVDF membrane was removed and immersed in TBST solution containing 5% nonfat milk and blocked on a shaker for 1 hour at room temperature. After blocking, the PVDF membrane was washed five times with TBST buffer, incubated overnight at 4°C with the appropriate dilution of primary antibody, and then washed five times (6 min each time) with TBST buffer. A specific dilution of secondary antibody was added, and the membrane was incubated for 2 hours at room temperature, then washed five times (6 minutes each time) with TBST buffer. The membrane was exposed using a gel imager.

[0396] The experimental results are shown in the figure below. 1.C59 down-regulated SRD5A1, SRD5A3, AR, ARV7, and GSPT1 proteins in 22Rv1 cells (Figures 7 and 8). 2. C59 down-regulated SRD5A3 and GSPT1 (eRF3a) proteins in PC-3 and HepG2 (Figures 9 and 10).

Claims

1. A compound having the structure: TPB-L-E3B, wherein TPB is a target protein binding moiety, L is a linker moiety, and E3B is an E3 ligase binding moiety; TPB has the following structure (I): is expressed as L has the following structure (IIA) or (IIB): is one selected from X is CH ２ , O, NH, or CF ２ wherein n is an integer of 2-3, Y is NH or O, and Z is NH or O; E3B has the following structures (IIIA), (IIIB), and (IIIC): is one selected from indicates that C may be in the R configuration, the S configuration, or a mixed RS configuration, and R １ indicates that H at any one of the other three positions on the benzene ring is replaced with F.

2. The L has the following structure (IIA) or (IIB): is one selected from X is CH ２ , O, NH, or CF ２ wherein n is an integer of 2-3, Y is NH or O, and Z is NH; E3B has the following structures (IIIB) and (IIIC): is one selected from 2. The compound according to claim 1, wherein C is in the R-configuration, the S-configuration or a mixed RS-configuration.

3. The L has the following structure (IIA) or (IIB): is one selected from X is CH ２ where n is the number 3, Y is NH, and Z is NH; E3B has the following structure (IIIB): is expressed as 3. The compound according to claim 2, wherein C may be in the R-configuration, the S-configuration or a mixed RS-configuration.

4. The compound has the following structure:

4. The compound according to claim 3, characterized in that it is selected from:

5. 10. Use of a compound according to any one of claims 1 to 4 in the preparation of a medicament for treating human tumor diseases.

6. The use according to claim 5, characterized in that the human tumor disease is prostate cancer, ovarian cancer, liver cancer, breast cancer, pancreatic cancer, glioma, cervical cancer, leukemia, gastric cancer or colon cancer.

7. The use according to claim 6, characterized in that the human tumor disease is prostate cancer.

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