Protac targeting bcl-xl protein and use thereof

Abstract

The present application relates to a PROTAC targeting a Bcl-xL protein and a use thereof. Specifically, the present invention provides a compound of formula I or a pharmaceutically acceptable salt, ester, prodrug, solvate, stereoisomer or deuterated compound thereof: A-L-E (I), wherein A is a small molecule ligand moiety targeting a Bcl-xL protein; E is a small molecule ligand moiety targeting an E3 ubiquitin ligase complex; and L is a linker. The compound or composition of the present invention can be used for selectively killing one or more senescent cells or treating senescence-related diseases or disorders in a subject.

Description

Protacs targeting bcl-xl protein and uses thereof TECHNICAL FIELD

[0001] The present invention relates to PROTACs targeting Bcl-xL protein and uses thereof. BACKGROUND

[0002] Members of the BCL-2 (B-cell lymphoma 2) gene family play a central role in regulating programmed cell death by controlling pro- and anti-apoptotic intracellular signals (Czabotar, P.E., et al., Nat Rev Mol Cell Biol 15, 49-63 (2014)). Anti-apoptotic BCL-2 family proteins such as BCL-2, BCL-XL, BCL-w, and Mcl-1 have been shown to be highly expressed in a variety of diseases, particularly BCL-xL protein is specifically highly expressed in a variety of senescent cells (He, et al., Nat Commun 11, 1996 (2020)). Currently, although the anti-aging effect of BCL-2 family inhibitors has been verified in a variety of preclinical models, it also has significant on-target hematotoxicity, including thrombocytopenia (Cang, et al., Journal of Hematology & Oncology 8, 129 (2015)). Therefore, it is necessary to develop more sophisticated and novel strategies to improve its efficacy and specificity for senescent cells and reduce toxicity.

[0003] The concept of PROTAC (Proteolysis targeting chimeras) was first proposed by Professor Craig Crews' team at Yale University in 2001. PROTAC molecules are composed of three parts: a target protein (POI) ligand, a linker, and an E3 ubiquitin ligase ligand. PROTAC is a heterobifunctional chimeric molecule that targets POI on one end and recruits E3 ubiquitin ligase on the other end, forming a ternary complex in the shape of a dumbbell. PROTAC molecules mediate transient interactions between E3 ligase and POI, and E3 ubiquitin ligase attaches ubiquitin to POI, marking it as a defective or damaged protein, which is then recognized and degraded by the proteasome. The main advantages of PROTAC technology include event-driven (rather than the usual occupancy-driven) activity, targeting undruggable proteins, overcoming drug resistance, and low dosage, which is expected to improve many limitations of traditional inhibitors. Because the earliest use of peptides to verify human-induced intracellular protein degradation required a molecular weight that was too large, cell-penetrating peptides were needed to improve cell permeability. Therefore, PROTAC technology developed slowly from 2001 to 2011. With the continuous discovery of small molecule E3 ligase ligands, PROTAC technology has developed rapidly. For example, mouse double minute 2 protein (MDM2), cellular inhibitor of apoptosis (cIAP), von Hippel-Lindau protein (VHL), and cereblon (CRBN) have been successfully used in small molecule PROTAC, and ARV-110 and ARV-471 based on CRBN have been used in clinical studies. However, due to the large molecular weight of PROTAC molecules, which do not meet the "drug-like five rules", leading to poor drugability, drug development based on this technology often has poor pharmacokinetic parameters in vivo, and there are challenges of low cell permeability and oral bioavailability, which require the screening of different E3 ligase ligands and careful optimization of the structure. At the same time, the overall degradation efficiency of PROTAC depends not only on the affinity of the target protein ligand and the type of E3 ligand, but also on the appropriate linker. A large amount of data shows that the length, composition, rigidity, and connection site of the linker are important for the formation of the PROTAC active ternary complex, degradation activity, and target selectivity. The design of the linker can further enhance potency by reducing free energy or limiting biologically active conformations. Optimizing the structure-activity relationship (SAR) within the linker will provide opportunities to adjust the pharmacokinetic properties of protein degraders. However, there is no universally applicable design strategy for the design and optimization of linkers, which often start with short and simple alkane or polyethylene glycol (PEG) chains, and then iteratively adjust their related properties.In summary, PROTAC is a powerful method to overcome the current obstacles in drug development, but further molecular design and optimization are needed in the future to gradually summarize the rules to improve the physicochemical properties and further explore the effectiveness and safety of PROTAC in the clinic. The target of PROTAC can be widely laid out, and the market is huge. With the continuous progress and improvement of this technology, PROTAC can become a successful therapy like small molecule inhibitors, mAbs and immunotherapy, so that more patients with diseases can benefit from it.

[0004] Senescence is a major risk factor for physiological deterioration, increased incidence of chronic diseases, and age-related mortality (Lopez-Otin et al., Cell 153, 1194-1217 (2013)). During senescence, senescent cells accumulate in multiple tissues and cause tissue dysfunction (van Deursen et al., Nature 509, 439-446 (2014); McHugh et al., J Cell Biol 217, 65-77 (2018)). Senescent cells also secrete a variety of pro-inflammatory factors, known as the senescence-associated secretory phenotype (SASP), which leads to age-related physical decline (Coppe et al., PLoS Biol 6, 2853-2868 (2008); Coppe et al., Annu Rev Pathol 5, 99-118 (2010)). Elimination of senescent cells has emerged as an attractive potential approach to ameliorate age-related diseases and improve health (Xu et al., Nat Med 24, 1246-1256 (2018); Baker et al., Nature 479, 232-236 (2011); Baker et al., Nature 530, 184-189 (2016)). However, due to the complexity and heterogeneity of senescent cells, it remains challenging to specifically and effectively eliminate multiple types of senescent cells (Kirkland et al. J Am Geriatr Soc 65, 2297-2301 (2017); Lozano-Torres et al., Nature Reviews Chemistry 3, 426-441 (2019)). Existing studies have shown that the Bcl-2 anti-apoptotic pathway plays a key role in maintaining the survival of senescent cells (Zhu et al., Aging Cell 14, 644-658 (2015), Chang et al., Nat Med 22, 78-83 (2016)). Compounds that selectively kill senescent cells, known as “senolytics”, have attracted considerable interest. BCL-xL inhibitors can effectively kill senescent cells as senolytics (Zhu et al., Aging Cell 15, 428-435 (2016)). Bcl-xL inhibitors induce senescent cells to undergo apoptosis by relieving the anti-apoptotic mechanism that is critical for senescent cells to maintain survival, achieving the effect of eliminating senescent cells in vitro and in vivo, which shows great potential in the treatment of multiple senescence-related diseases such as osteoarthritis, age-related macular degeneration, diabetic nephropathy, etc.

[0005] The structural modification of BCL-xL protein inhibitors combined with E3 ligase ligands to form PROTAC can effectively combine the advantages of both. On the one hand, it can specifically induce apoptosis of senescent cells, and on the other hand, it can effectively reduce the hematological toxicity of BCL-xL protein inhibitors, while optimizing the overall molecular structure to meet the effectiveness and safety of clinical application, providing a new idea for the treatment of a series of senile and aging-related diseases.

[0006] SUMMARY

[0007] It is an object of the present application to provide a compound of Formula I, or a pharmaceutically acceptable salt, ester, prodrug, solvate, stereoisomer, or deuterated compound thereof: A-L-E (I)

[0008] wherein A is a small molecule ligand targeting Bcl-xL protein; E is a small molecule ligand targeting E3 ubiquitin ligase complex; L is a linker group.

[0009] Small molecule ligand targeting Bcl-xL protein A

[0010] The small molecule ligand targeting Bcl-xL protein A has the structure of Formula 1 or 2:

[0011] wherein R0 is absent or selected from:

[0012] -COOR a , -CONR a R b , -SO3R a , -SO2NR a R b , -SO2NR a COR b , -CONR a SO2R b , -COSO2NR a R b ,

[0013] wherein X is selected from hydrogen, hydroxyl, amino or CH(OR a )2;

[0014] wherein R a and R b are independently selected from hydrogen, cyano, substituted or unsubstituted alkyl, or substituted or unsubstituted alkoxy.

[0015] Y is absent or selected from substituted or unsubstituted alkylene, substituted or unsubstituted alkyleneoxy, substituted or unsubstituted alkenylene, substituted or unsubstituted alkenyleneoxy, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene, substituted or unsubstituted cycloalkylene, and substituted or unsubstituted heterocyclylene;

[0016] Z1and Z2are independently absent or selected from substituted or unsubstituted -alkylene-cyclic group, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl.

[0017] In some embodiments, the small molecule ligand portion A targeting the Bcl-xL protein is a deuterated moiety having the structure of Formula 1’ or 2’:

[0018] wherein one or more hydrogens at position * are replaced with deuterium. In some embodiments, two hydrogens at position * are replaced with deuterium.

[0019] In some embodiments, R a and R b are independently selected from hydrogen, cyano, substituted or unsubstituted C1-C6alkyl, or substituted or unsubstituted C1-C6alkoxy.

[0020] In some embodiments, R a and R b are independently selected from hydrogen, cyano, alkyl (e.g., C1-C6alkyl), alkoxy (e.g., C1-C6alkoxy), halogen-substituted alkyl (e.g., C1-C6alkyl), or halogen-substituted alkoxy (e.g., C1-C6alkoxy).

[0021] In some embodiments, R0is absent or selected from: -COOR a , -CONHR b , -SO3R a , -SO2NHR b , -SO2NHCOR b , -CONHSO2R b , -COSO2NHR b , wherein R a and R b are independently selected from hydrogen, cyano, and alkyl (e.g., C1-C6alkyl).

[0022] In some embodiments, R0is selected from:

[0023] In some embodiments, Y is absent or selected from substituted or unsubstituted Ci-6alkylene, substituted or unsubstituted Ci-6alkylenoxy, substituted or unsubstituted C2-6 alkenylene, substituted or unsubstituted C2-6 alkenylenoxy, substituted or unsubstituted phenylene, substituted or unsubstituted 5- or 6-membered heteroarylene, substituted or unsubstituted C3-6 cycloalkylene, and substituted or unsubstituted 3-6 membered heterocyclylene.

[0024] In some embodiments, Y is selected from alkylene, alkylenoxy, alkenylene, alkenylenoxy, arylene, heteroarylene, cycloalkylene, and heterocyclylene optionally substituted with one or more substituents selected from halogen and alkyl (e.g., Ci-6alkyl).

[0025] In some embodiments, Y is selected from Ci-6alkylene, Ci-6alkylenoxy, C2-6 alkenylene, or C2-6 alkenylenoxy.

[0026] In some embodiments, Y is selected from the following structures:

[0027] wherein m is 0, 1, 2, or 3.

[0028] In some embodiments, Y is selected from phenylene and 5- or 6-membered heteroarylene optionally substituted with one or more substituents selected from halogen and alkyl.

[0029] In some embodiments, Y is selected from pyrazolylene optionally substituted with one or more substituents selected from halogen and alkyl.

[0030] In some embodiments, Y is selected from the following structures:

[0031] wherein the attachment to the Z1or Z2group is at a ring nitrogen atom.

[0032] In some embodiments, Y is selected from the following structures:

[0033] wherein the attachment to the Z1or Z2group is at a ring nitrogen atom.

[0034] In some embodiments, Z1and Z2are independently selected from substituted or unsubstituted -Ci-6alkylene-cyclic group, substituted or unsubstituted C5-8 cycloalkyl, substituted or unsubstituted 5-8 membered heterocyclyl, substituted or unsubstituted phenyl, substituted or unsubstituted 5-6 membered heteroaryl.

[0035] In some embodiments, Z1and Z2are independently selected from substituted or unsubstituted -alkylene-cyclic group, cycloalkyl-(R2’) 1’ , heterocyclyl-(R2’) p , heterocyclyl-(R2’)p , aryl-(R2') p , heteroaryl-(R2') p , aryl-propargylene, heteroaryl-propargylene, aryl-propargylene-N(R2')2, heteroaryl-propargylene-N(R2')2, aryl-propargylene-cycloalkyl-(R2') p , heteroaryl-propargylene-cycloalkyl-(R2') p , aryl-propargylene-heterocyclyl-(R2') p , and heteroaryl-propargylene-heterocyclyl-(R2') p .

[0036] In some embodiments, R 1’ is deuterium, halogen, amino, hydroxyl, nitro, thiol, cyano, isocyano, substituted or unsubstituted alkyl (e.g., alkyl substituted with halogen, amino, hydroxyl, nitro, carboxyl, etc.), substituted or unsubstituted alkoxy (e.g., alkoxy substituted with halogen, amino, hydroxyl, nitro, carboxyl, etc.), alkylamino, dialkylamino, carboxyl, carbonyl, amido, sulfonyl, sulfonic acid, phosphoryl, and phosphonyl.

[0037] In some embodiments, R2' is hydrogen, alkyl, aryl, heteroaryl, cycloalkyl, or heterocyclyl.

[0038] In some embodiments, p is 0, 1, 2, 3, or 4.

[0039] In some embodiments, the cyclic group is

[0040] In some embodiments, Z1and Z2are independently selected from the group consisting of C5-8cycloalkyl-(R2'), heterocyclyl-(R2'), phenyl-(R2'), heteroaryl-(R2'), aryl-(R2'), aryl-propargylene, heteroaryl-propargylene, aryl-propargylene-N(R2')2, heteroaryl-propargylene-N(R2')2, aryl-propargylene-cycloalkyl-(R2'), heteroaryl-propargylene-cycloalkyl-(R2'), aryl-propargylene-heterocyclyl-(R2'), and heteroaryl-propargylene-heterocyclyl-(R2'), optionally substituted with one or more R 1’ p , heterocyclyl-(R2') p , phenyl-(R2') p , heteroaryl-(R2') p , aryl-propargylene, heteroaryl-propargylene, aryl-propargylene-N(R2')2, heteroaryl-propargylene-N(R2')2, aryl-propargylene-cycloalkyl-(R2') p , heteroaryl-propargylene-cycloalkyl-(R2') p , aryl-propargylene-heterocyclyl-(R2') p , and heteroaryl-propargylene-heterocyclyl-(R2') p ​wherein the heterocyclyl is a 5- or 6-membered heterocyclyl containing one or two heteroatoms (e.g., piperidinyl, piperazinyl, or morpholinyl) or a 7-11 membered heterospirocyclyl (e.g., 3,9-diazaspiro[5.5]undecanyl), the heteroaryl is a 5- or 6-membered heteroaryl containing one or two heteroatoms (e.g., pyridinyl), and the heteroatoms are independently selected from nitrogen, oxygen, and sulfur.

[0041] In some embodiments, p is 0 or 1.

[0042] In some embodiments, R 1’ is selected from halogen (e.g., fluorine).

[0043] In some embodiments, R 2’ is hydrogen or alkyl (e.g., methyl).

[0044] In some embodiments, Z1is selected from the following groups:

[0045] In some embodiments, Z1may be selected from the following groups:

[0046] In some embodiments, Z1may be selected from the following groups:

[0047] In some embodiments, Z1is selected from the following groups:

[0048] In some embodiments, Z2is selected from the following groups:

[0049] In some embodiments, Z2is selected from:

[0050] In some embodiments, the small molecule ligand targeting a Bcl-xL protein Part A is selected from the following structures:

[0051] In some embodiments, the small molecule ligand targeting a Bcl-xL protein Part A is selected from the following structures:

[0052] In some embodiments, the small molecule ligand targeting a Bcl-xL protein Part A is selected from the following structures:

[0053] In some embodiments, the small molecule ligand portion A targeting Bcl-xL protein is selected from the following structures:

[0054] Small molecule ligand portion E targeting E3 ubiquitin ligase complex

[0055] In some embodiments, the small molecule ligand portion E targeting E3 ubiquitin ligase complex is selected from the following structures:

[0056] wherein R1 is O, NH, or absent; R2 is H or methyl; R3 is R c is F, methoxy, Cl, cyano.

[0057] In some embodiments, the small molecule ligand portion E targeting E3 ubiquitin ligase complex is selected from the following structures:

[0058] In some embodiments, the small molecule ligand portion E targeting E3 ubiquitin ligase complex is selected from the following structures:

[0059] Linker L

[0060] Linker L has the following structure:

[0061] Linker L is connected to small molecule ligand portion A targeting Bcl-xL protein at L1 and to small molecule ligand portion E targeting E3 ubiquitin ligase complex at L2,

[0062] wherein

[0063] L1 and L2 are independently absent or selected from:

[0064] R4 is independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted heterocyclyl;

[0065] R5 is independently selected from hydrogen, nitro, cyano, isocyano, amino, hydroxyl, thiol, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, alkylamino, dialkylamino, carboxyl, carbonyl, amido, sulfonyl, sulfonic acid, phosphoryl, and phosphinyl;

[0066] q and r are independently selected from integers from 0-10 (i.e., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10);

[0067] X is absent or has the following structure:

[0068] X1and X2are independently absent or selected from:

[0069] W is absent or selected from:

[0070] wherein m1, m2, n1, n2, s1, and s2are independently selected from an integer from 0-10 (i.e., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10).

[0071] In some embodiments, R4is independently selected from hydrogen, substituted or unsubstituted C1-6alkyl, substituted or unsubstituted C3-8cycloalkyl, and substituted or unsubstituted 3-8 membered heterocyclyl, wherein the alkyl, cycloalkyl, and heterocyclyl groups can be substituted with halogen, amino, hydroxyl, nitro, and / or carboxyl.

[0072] In some embodiments, R5is independently selected from hydrogen, nitro, cyano, isocyano, amino, hydroxyl, thiol, halogen, substituted or unsubstituted alkyl (e.g., alkyl substituted with halogen, amino, hydroxyl, nitro, and / or carboxyl), substituted or unsubstituted alkoxy (e.g., alkoxy substituted with halogen, amino, hydroxyl, nitro, and / or carboxyl), alkylamino, dialkylamino, carboxyl, substituted or unsubstituted carbonyl, amido, sulfonyl, sulfonic acid, phosphoryl, and phosphinyl.

[0073] In some embodiments, L1is absent or selected from

[0074] In some embodiments, L2is absent or selected from

[0075] In some embodiments, the X group can be selected from the following structures:

[0076] wherein u1, u2, u3, and v1are independently selected from an integer from 0-10 (i.e., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10).

[0077] In some embodiments, the linking group L is selected from the following structures:

[0078] In some embodiments, q, u1, u2, u3, and v1are independently selected from an integer from 0-10 (i.e., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10).

[0079] In some embodiments, L is absent.

[0080] In some embodiments, the linking group L is selected from the following structures:

[0081] In some embodiments, the linking group L is absent or selected from the following structures:

[0082] In some embodiments, the linking group L is absent or selected from the following structures:

[0083] In some embodiments, when the small molecule ligand portion A targeting the Bcl-xL protein has the structure of Formula 1, the linking group L is selected from the following structures:

[0084] and

[0085] The small molecule ligand portion E targeting the E3 ubiquitin ligase complex is selected from:

[0086] In some embodiments, when the small molecule ligand portion A targeting the Bcl-xL protein has the structure of Formula 2, the linking group L is absent or selected from the following structures:

[0087] and

[0088] The small molecule ligand portion E targeting the E3 ubiquitin ligase complex is selected from:

[0089] In some embodiments, the PROTAC molecule of Formula I has the following structure:

[0090] In one aspect, the application relates to a composition comprising an effective amount of a compound of Formula (I) and a pharmaceutically acceptable carrier.

[0091] In one aspect, the application relates to a method of selectively killing one or more senescent cells or treating a senescence-associated disease or disorder in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound or composition of the application.

[0092] In one aspect, the application relates to the use of a compound of Formula (I) of the application or a composition of the application in the manufacture of a medicament for selectively killing one or more senescent cells or treating a senescence-associated disease or disorder in a subject.

[0093] In some embodiments, the senescence-associated disease or disorder is a metabolic disease, an inflammatory disease or disorder, a pulmonary disease or disorder, a neurological disease or disorder, a proliferative disorder, a kidney disorder or disease, an ocular disease or disorder, or a dermatological disorder or disease.

[0094] In some embodiments, the senescence-associated disease or disorder is selected from:

[0095] (i) an inflammatory or autoimmune disease or disorder selected from oral mucositis, inflammatory bowel disease, kyphosis, and herniated intervertebral disc;

[0096] (ii) a neurological disease or disorder selected from Alzheimer's disease, Parkinson's disease, Huntington's disease, dementia, mild cognitive impairment, macular degeneration, and motor neuron dysfunction;

[0097] (iii) a metabolic disease selected from diabetic ulcer, metabolic syndrome, and obesity;

[0098] (iv) a pulmonary disease selected from pulmonary fibrosis, chronic obstructive pulmonary disease, asthma, cystic fibrosis, emphysema, bronchiectasis, and age-related loss of lung function;

[0099] (v) an ocular disease or disorder selected from macular degeneration, glaucoma, cataracts, presbyopia, and vision loss;

[0100] (vi) an age-related disorder selected from kidney failure, frailty, hearing loss, muscle fatigue, skin condition, skin wound healing, liver fibrosis, pancreatic fibrosis, oral submucosal fibrosis, and sarcopenia; and

[0101] (vii) a skin disease or disorder selected from eczema, psoriasis, hyperpigmentation, a nevus, a skin rash, atopic dermatitis, urticaria, diseases and disorders associated with photosensitivity or photoaging, wrinkles, pruritis, dysesthesia, eczematous skin rash, eosinophilic dermatosis, reactive neutrophilic dermatosis, pemphigus, pemphigoid, immunobullous dermatosis, cutaneous fibrohistiocytic proliferations, cutaneous lymphomas, and cutaneous lupus.

[0102] In some embodiments, the subject is diagnosed with a cancer and is optionally undergoing a cancer treatment selected from chemotherapy and radiation therapy.

[0103] In some embodiments, the compound of Formula (I) of the present application or the composition of the present application

[0104] (i) reduces a cell that has been pushed towards senescence or a cancer cell;

[0105] (ii) reduces one or more side effects produced by a senescent cell, wherein the side effects include inflammation, promotion of cancer growth, and promotion of metastasis;

[0106] (iii) reduces one or more side effects of chemotherapy; or

[0107] (iv) reduces one or more side effects of radiation therapy.

[0108] In some embodiments, the subject has a viral infection, optionally wherein the viral infection results in an overproduction of activated macrophages in the subject.

[0109] In some embodiments, the viral infection is a coronavirus infection selected from a coronavirus, a severe acute respiratory syndrome (SARS) coronavirus (CoV), SARS-CoV-2 (which causes COVID-19 (Coronavirus Disease) 2019), and Middle East respiratory syndrome (MERS)-CoV.

[0110] In some embodiments, the subject has a SARS-CoV-2 infection and has been diagnosed with Coronavirus Disease 2019.

[0111] In some embodiments, the compound or composition of the present application achieves the following structure after in vivo administration in an amount effective to kill one or more senescent cells:

[0112] wherein R°, Y, and Z1 are as described herein. BRIEF DESCRIPTION OF DRAWINGS

[0113] Figure 1 is a plot of the killing curves of test compounds on Molt-4 cells at different concentrations. Figure 1A is a plot of the killing curves of compound C1 and other known compounds on Molt-4 cells at different drug concentrations. Figure 1B is a plot of the killing curves of PROTAC molecules APQ2, APQ5b and APQ11 on Molt-4 cells at different drug concentrations. Figure 1C is a plot of the killing curves of PROTAC molecules APQ8, APQ9, APL-1 and APL-2 on Molt-4 cells at different drug concentrations. Figure 1D is a plot of the killing curves of PROTAC molecules APQ7a, APQ7b and APQ10 on Molt-4 cells at different drug concentrations. Figure 1E is a plot of the killing curves of PROTAC molecules APH1, APH7, APH10 and other known compounds on Molt-4 cells at different drug concentrations. Figure 1F is a plot of the killing curves of PROTAC molecules APH102, APH103, APH104, APH105 and other known compounds on Molt-4 cells at different drug concentrations. Figure 1G is a plot of the killing curves of PROTAC molecules APQ10-2, APQ902, APQ904, APH105 and other known compounds on Molt-4 cells at different drug concentrations. Figure 1H is a plot of the killing curves of PROTAC molecules APQ10-3, APQ902 and other known compounds on Molt-4 cells at different drug concentrations.

[0114] Figure 2 is the killing results of test compounds on Molt-4 cells at different concentrations in the presence or absence of pomalidomide. Figures 2A, 2B, 2C and 2D are plots of the killing curves of CRBN-E3 enzyme-based PROTAC molecules APQ2, APQ9, BF3 and BF4 on Molt-4 cells at different concentrations, respectively. Figures 2E, 2F, 2G and 2H are plots of the killing curves of compound C1, control compound ABT-263, known control PROTAC molecule DT-2216 based on VHL-E3 enzyme and known compound A1331852 on Molt-4 cells at different concentrations, respectively. Figures 2I, 2J, 2K and 2L are plots of the killing curves of CRBN-E3 enzyme-based PROTAC molecules APQ10, APQ10-2, APQ902 and APQ904 on Molt-4 cells at different concentrations, respectively. Veh: empty control group; +Poma: pomalidomide group.

[0115] Figure 3 is the killing results of test compounds on human embryonic fibroblast cells (HEF cells), etoposide-induced senescent human HEF cells, human embryonic lung fibroblast MRC5 cells, etoposide-induced senescent human embryonic lung fibroblast MRC5 cells, and human skin-derived BJ cells, bleomycin-induced senescent human skin-derived BJ cells at different concentrations. Figures 3A, 3B, 3C and 3D are the corresponding killing curves of PROTAC molecules APQ2, BF4, APQ9 and APQ10 on the HEF cell model, respectively; Figures 3E and 3F are the corresponding killing curves of commercially available control molecules A-1331852 and ABT-263 on the HEF cell model, respectively. Figures 3G, 3H, 3I, 3J, 3K and 3L are the corresponding killing curves of PROTAC molecules APQ9, APQ10, APQ10-2, APQ902, APQ904 and commercially available control molecule A-1331852 on the MRC5 cell model, respectively. Figures 3M, 3N and 3O are the corresponding killing curves of PROTAC molecules APQ10, APQ902 and APQ10-3 on the BJ cell model, respectively. HEF: normal culture HEF cell group; HEF-ETO: etoposide-induced senescent HEF cell group. MRC5: normal culture MRC5 cell group; MRC5-ETO: etoposide-induced senescent MRC5 cell group. BJ: normal culture BJ cell group; BJ-BLM: bleomycin-induced senescent BJ cell group.

[0116] Figure 4 is the degradation results of test compounds on target protein Bcl-xL. Figures 4A, 4B, 4C, 4D, 4E, 4F, 4G, 4H, 4I, 4J and 4K are the degradation results of PROTAC molecules APQ2, BF4, APQ9, APQ10, APQ10-2, APH1, APH7, APQ902, APQ904, APH105 and APH106 on Bcl-xL protein at each time point after drug addition. The test concentration of PROTAC molecules is 100 nM, the detection time points are 0 h, 2 h, 6 h and 16 h, and the control reference protein used is glyceraldehyde-3-phosphate dehydrogenase (GAPDH).

[0117] Figure 5 is the results of platelet toxicity of test compounds in vivo. Figure 5A is the results of comparing the effects of PROTAC molecule APQ2 and control molecule A-1331852 on the number of platelets in the peripheral blood of mice at different dosages. APQ2-0.1 mpk, APQ2-0.5 mpk, APQ-2.5 mpk are the dosage groups of APQ2 at 0.1, 0.5, 2.5 mg / kg, respectively; A133-0.5 mpk is the dosage group of A-1331852 at 0.5 mg / kg; Veh is the single solvent control group. The blood routine test time points are 0, 4, 24, 72 hours after administration. Figure 5B is the results of comparing the effects of PROTAC molecules APQ9, APQ10 and control molecules A-1331852, single solvent administration on the number of platelets in the peripheral blood of mice. Veh, APQ9-5 mpk, APQ10-5 mpk, A133-5 mpk are the single solvent administration group, the dosage group of APQ9 at 5 mg / kg, the dosage group of APQ10 at 5 mg / kg, and the dosage group of A-1331852 at 5 mg / kg, respectively. The blood routine test time points are 4, 24, 48 hours after administration. Figure 5C is the results of comparing the effects of APQ10 and A-1331852 on the number of platelets at different dosages. APQ10-2 mpk, APQ10-5 mpk, APQ10-20 mpk are the dosage groups of APQ10 at 2, 5, 20 mg / kg, respectively; A133-2 mpk, A133-5 mpk, A133-20 mpk are the dosage groups of A-1331852 at 2, 5, 20 mg / kg, respectively; Veh is the single solvent control group. The blood routine test time points are 24, 48, 72 hours after administration. The number of mice in each group in Figures 5A, 5B, 5C is 4. DETAILED DESCRIPTION

[0118] DEFINITIONS

[0119] As used herein, "substituted" refers to all permissible substituents of the compounds or functional groups described herein. In the broadest sense, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. Exemplary substituents include, but are not limited to, halogen, hydroxyl, or any other organic radical containing any number of carbon atoms, preferably 1-14 carbon atoms, and optionally including one or more heteroatoms, such as oxygen, sulfur, or nitrogen radicals, in linear, branched, or cyclic structural formats. Representative substituents include alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, phenyl, substituted phenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halogen, hydroxyl, alkoxy, substituted alkoxy, phenoxy, substituted phenoxy, aryloxy, substituted aryloxy, alkylthio, substituted alkylthio, phenylthio, substituted phenylthio, arylthio, substituted arylthio, cyano, isocyano, substituted isocyano, carbonyl, substituted carbonyl, carboxyl, substituted carboxyl, amino, substituted amino, acylamino, substituted acylamino, sulfonyl, substituted sulfonyl, sulfonic acid, phosphoryl, substituted phosphoryl, phosphonyl, substituted phosphonyl, polyaryl, substituted polyaryl, C3-C 20 cyclic groups, substituted C3-C 20 cyclic groups, heterocyclic, substituted heterocyclic, amino acids, poly(lactic-co-glycolic acid), peptides, and polypeptides. Such alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, phenyl, substituted phenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halogen, hydroxyl, alkoxy, substituted alkoxy, phenoxy, substituted phenoxy, aryloxy, substituted aryloxy, alkylthio, substituted alkylthio, phenylthio, substituted phenylthio, arylthio, substituted arylthio, cyano, isocyano, substituted isocyano, carbonyl, substituted carbonyl, carboxyl, substituted carboxyl, amino, substituted amino, acylamino, substituted acylamino, sulfonyl, substituted sulfonyl, sulfonic acid, phosphoryl, substituted phosphoryl, phosphonyl, substituted phosphonyl, polyaryl, substituted polyaryl, C3-C 20 cyclic groups, substituted C3-C 20 cyclic groups, heterocyclic, substituted heterocyclic, amino acids, poly(lactic-co-glycolic acid), poly(lactic-co-glycolic acid), peptides, and polypeptides can be further substituted.

[0120] Heteroatoms, such as nitrogen, can have hydrogen substituents and / or any permissible substituents of the organic compounds described herein that satisfy valence requirements. It should be understood that "substitution" or "substituted with" includes the implicit proviso that such substitution is in accordance with permitted valence of the atom and the substituent, and the substitution results in a stable compound, i.e., a compound that does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc.

[0121] Unless otherwise specifically and explicitly provided, the term "substituted" refers to a structure, e.g., a compound or a moiety on a larger compound, regardless of how the structure was formed. The structure is not limited to structures made by any particular method.

[0122] As used herein, "aryl" refers to a C5-C 26 Broadly defined, "aryl" as used herein includes 5-, 6-, 7-, 8-, 9-, 10-, 14-, 18-, and 24-membered monocyclic aromatic groups, which can include from zero to four heteroatoms, e.g., benzene, naphthalene, anthracene, phenanthrene, chrysene, pyrene, corannulene, coronene, and the like.

[0123] "Aryl" also includes polycyclic ring systems having two or more rings, wherein two or more carbons are shared by two adjacent rings (i.e., "fused rings"), wherein at least one ring is aromatic, e.g., other cyclic rings or multiple rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, and / or heterocycle.

[0124] The term "substituted aryl" refers to an aryl group in which one or more hydrogen atoms on the aromatic ring(s) are replaced by one or more substituents, including but not limited to halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, carbonyl (e.g., ketone, aldehyde, carboxyl, carboxyl ester, formyl, or acyl), silyl, ether, ester, thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxyl, phosphoryl, phosphonate, phosphinate, amino (or quaternized amino), amido, amidine, imine, cyano, nitro, azido, thiol, imino, sulfhydryl, sulfate, sulfonamide, sulfamoyl, sulfoxide, sulfonamido, sulfonyl, heterocyclic, alkylaryl, haloalkyl (e.g., CF3, -CH2-CF3, -CCl3), -CN, aryl, heteroaryl, and combinations thereof.

[0125] "Heterocycle," "heterocyclic," and "heterocyclyl" are used interchangeably and refer to a cyclic group of from 3 to 10 or 3 to 8 ring atoms, preferably 5 to 6 ring atoms, connected by ring carbon or nitrogen atoms of a monocyclic or bicyclic ring system, consisting of carbon and one to four heteroatoms selected from non-peroxidic oxygen, sulfur and N(Y), wherein Y is absent or is H, O, C1-C 10alkyl, phenyl or benzyl, and optionally contains 1-3 double bonds and is optionally substituted with one or more substituents. By definition, heterocyclyl is different from heteroaryl. Examples of heterocycles include, but are not limited to, piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, dihydrofuran[2,3-b]tetrahydrofuran, morpholinyl, piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pyranyl, 2H-pyrrolyl, 4H-quinolyl, quinuclidinyl, tetrahydrofuranyl, 6H-l,2,5-thiadiazinyl. The heterocyclyl group can be optionally substituted with one or more substituents as defined above for alkyl and aryl.

[0126] The term "heteroaryl" means a C5-C 26A heteroaryl group is a monocyclic aromatic, fused aromatic, diaromatic, or combination thereof in which one or more carbon atoms in one or more aromatic rings are replaced by heteroatoms. Suitable heteroatoms include, but are not limited to, oxygen, sulfur, and nitrogen. As used herein, “heteroaryl” in a broad sense includes 5-, 6-, 7-, 8-, 9-, 10-, 14-, 18-, and 24-membered monocyclic aromatic groups, which may include one to four heteroatoms, such as pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, tetraazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine. Heteroaryl groups may also be referred to as “aryl heterocycles” or “heteroaromatic compounds.” “Heteroaryl” also includes polycyclic systems having two or more rings, where two or more carbons are shared by two adjacent rings (i.e., “fused rings”), and where at least one ring is heteroaromatic; for example, the other rings or rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heterocyclic, or combinations thereof. Examples of heteroaromatic rings include, but are not limited to, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophene, benzooxazolyl, benzooxazolinyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzoisooxazolyl, benzoisothiazolyl, benzimidazolinyl, carbazole, 4aH-carbazole, carbolinyl, benzodihydropyranyl, benzopyranyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, furanyl, furazanyl, imidazoalkyl, imidazolinyl, imidazolyl, 1H-indazole, indolenyl, dihydroindoleyl, indolizinyl, indolyl, 3H-indoleyl. isatinoyl, isobenzofuranyl, isochromanyl, isoindazole, isodihydroindole, isoindole, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, naphthidyl, octahydroisoquinolinyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolylalkyl, oxazolyl, hydroxyindole, pyrimidinyl, phenanthidyl, phenanthrolinel, phenazinyl, phenthiazinyl, phenoxathinyl, phenoxazinyl, phthalazinyl, pteridinyl, purine, pyrazine The compounds include pyrazolyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridoxazolyl, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, pyrrolidinyl, quinazolinyl, quinolinyl, quinoxolinyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthyl, thiazolyl, thiazolyl, thienyl, thienothiazolyl, thienoxazolyl, thienoimidazole, phenylthio, and xanthonyl. One or more rings may be substituted, as defined below for "substituted heteroaryl".

[0127] The term "substituted heteroaryl" refers to a heteroaryl group in which one or more of the hydrogen atoms on one or more of the heteroaryl rings are replaced with one or more substituents including, but not limited to, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, carbonyl (e.g., ketone, aldehyde, carboxyl ester, formyl or acyl), silyl, ether, ester, thiocarbonyl (e.g., thioester, thioacetate or thioformate), alkoxyl, phosphoryl, phosphonate, phosphinate, amino (or quaternized amino), amido, amidine, imine, cyano, nitro, azido, thiol, imino, alkylthio, sulfate, sulfonate, sulfamido, sulfoxide, sulfonamido, sulfonyl, heterocyclic, aralkyl, haloalkyl (e.g., CF3, -CH2-CF3, -CC13), -CN, aryl, heteroaryl, and combinations thereof.

[0128] As used herein, "alkyl" refers to a group that is a saturated aliphatic group, including straight-chain alkyl or branched-chain alkyl, cycloalkyl (alicyclic), alkyl substituted cycloalkyl, and cycloalkyl substituted alkyl groups. In preferred embodiments, straight-chain or branched-chain alkyl groups have 30 or fewer carbon atoms in their backbone (e.g., C1-C30 alkyl), and more preferably 20 or fewer, more preferably 15 or fewer, and most preferably 10 or fewer. Likewise, preferred cycloalkyl groups have from 3-10 carbon atoms in their ring structure, and more preferably have 5, 6 or 7-carbon atoms in their ring structure. The term "alkyl" (or "lower alkyl") as used throughout the specification, examples and claims, is intended to include both "unsubstituted alkyls" and "substituted alkyls," the latter meaning alkyl moieties having one or more carbons on the hydrocarbon backbone replaced with a substituent. Such substituents include, but are not limited to, halogen, hydroxyl, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxyl, phosphoryl, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, thiol, alkylthio, sulfate, sulfonate, sulfamido, sulfoxide, sulfonamido, sulfonyl, heterocyclic, aralkyl, or aromatic or heteroaromatic moieties. 30 For straight-chain, C3-C 30 For branched-chain), preferably 20 or fewer, more preferably 15 or fewer, and most preferably 10 or fewer. Likewise, preferred cycloalkyl groups have from 3-10 carbon atoms in their ring structure, and more preferably have 5, 6 or 7-carbon atoms in their ring structure. The term "alkyl" (or "lower alkyl") as used throughout the specification, examples and claims, is intended to include both "unsubstituted alkyls" and "substituted alkyls," the latter meaning alkyl moieties having one or more carbons on the hydrocarbon backbone replaced with a substituent. Such substituents include, but are not limited to, halogen, hydroxyl, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxyl, phosphoryl, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, thiol, alkylthio, sulfate, sulfonate, sulfamido, sulfoxide, sulfonamido, sulfonyl, heterocyclic, aralkyl, or aromatic or heteroaromatic moieties.

[0129] "Lower alkyl" as used herein, unless otherwise specified, refers to alkyl groups as defined above, but having 1 to 10 carbon atoms in their backbone structure, more preferably 1 to 6 carbon atoms. Likewise, "lower alkenyl" and "lower alkynyl" have similar chain lengths. Throughout the application, preferred alkyl groups are lower alkyl groups. In preferred embodiments, substituents designated herein as alkyl groups are lower alkyl groups.

[0130] "Alkyl" includes one or more substitutions at one or more carbon atoms of the hydrocarbyl and heteroalkyl groups. Suitable substituents include, but are not limited to, halogen, e.g., fluorine, chlorine, bromine, or iodine; hydroxyl; -NRR', where R and R' are independently hydrogen, alkyl, or aryl, and where the nitrogen atom is optionally quaternized; -SR, where R is hydrogen, alkyl, or aryl; -CN; -NO2; -COOH; carboxylate; -COR, -COOR, or CON(R)2, where R is hydrogen, alkyl, or aryl; azide, aralkyl, alkoxy, imino, phosphonate, phosphinite, silyl, ether, sulfonyl, sulfonamido, heterocyclic, aromatic or heteroaromatic moieties, haloalkyl (e.g., -CF3, -CH2-CF3, -CC13); CN; -NCOCOCH2CH2; NCOCOCH; -NCS; and combinations thereof.

[0131] Those skilled in the art will appreciate that substituted moieties on hydrocarbon chains can themselves be substituted, if appropriate. For example, substituents of substituted alkyl groups can include halogen, hydroxyl, nitro, thiol, amino, azido, imino, amido, phosphoryl (including phosphonate and phosphinite), sulfonyl (including sulfate, sulfonamido, sulfamoyl, sulfoxide, and sulfonate ester), and silyl groups, as well as ether, alkylthio, carbonyl (including ketone, aldehyde, carboxylate, and ester), haloalkyl, -CN, and the like. Cycloalkyl groups can be substituted in the same manner.

[0132] The terms "alkenyl" and "alkynyl" refer to unsaturated aliphatic groups analogous in length and possible substitution to the alkyl radicals but that contain at least one double or triple bond, respectively.

[0133] The term "substituted alkenyl" refers to alkenyl moieties having one or more substituents replacing one or more hydrogens on one or more carbons of the hydrocarbon backbone. Such substituents include, but are not limited to, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, carbonyl (for example, carboxyl, alkoxy carbonyl, formyl, or acyl), silyl, ether, ester, thio carbonyl (for example, thioester, thioformate, or thioacetate), alkoxyl, phosphoryl, phosphinate, phosphonate, phosphinite, amino (or quaternized amino), amido, amidine, imine, cyano, nitro, azido, thiol, alkylthio, sulfate, sulfonamide, sulfamoyl, sulfoxide, sulfonamide, heterocyclyl, aralkyl, haloalkyl, -CN, -NCOCOCH2CH2; NCOCOCH; -NCS; and combinations thereof.

[0134] The term "substituted alkynyl" refers to an alkynyl moiety having one or more substituents replacing one or more hydrogens on one or more carbons of the hydrocarbon backbone. Such substituents include, but are not limited to, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, carbonyl (e.g., ketone, aldehyde, carboxyl ester, carboxyl, acyrl or anhydride), silyl, ether, ester, thio carbonyl (e.g., thioester, thioacetate or thiocarboxylate), alkoxyl, phosphoryl, phosphoester, phosphinoester, phosphine, amino (or quarternated amino), amido, amidine, imine, cyano, nitro, azido, thiol, alkylthio, sulfate, sulfonamide, sulfamoyl, sulffoxide, sulfonamido, sulfonyl, heterocyclyl, alkylaryl, haloalkyl, -CN, aryl, heteroaryl, and combinations thereof.

[0135] As used herein, "amino" and "amine" are art-recognized and refer to substituted and unsubstituted amines, for example, moieties that can be represented by the general structure:

[0136] wherein R, R' and R" each independently represent a hydrogen, a substituted or unsubstituted alkyl, a substituted or unsubstituted alkenyl, a substituted or unsubstituted alkynyl, a substituted or unsubstituted carbonyl, -(CH2) m R" or R and R' together with the N atom to which they are attached form a heterocyclic ring having from 3 to 14 atoms in the ring structure; R" represents a hydroxyl, a substituted or unsubstituted carbonyl, an aryl, a cycloalkyl ring, a cycloalkenyl ring, a heterocyclic ring, or a polycyclic ring; and m is 0 or an integer from 1 to 8. In preferred embodiments, only one of R and R' can be a carbonyl, for example, R and R' together with the nitrogen do not form a diacylimide. In preferred embodiments, R and R' (and optional R") each independently represent a hydrogen atom, a substituted or unsubstituted alkyl, a substituted or unsubstituted alkenyl, or -(CH2) m R". Thus, as used herein, the term "alkylamine" refers to an amine group as defined above having a substituted or unsubstituted alkyl group attached thereto (i.e., at least one of R, R' or R" is an alkyl group).

[0137] As used herein, "carbonyl" is art-recognized and includes moieties that can be represented by the general structure:

[0138] wherein X is a bond, or represents oxygen or sulfur, and R represents a hydrogen, a substituted or unsubstituted alkyl, a substituted or unsubstituted alkenyl, a substituted or unsubstituted alkynyl, a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted heterocyclyl, a substituted or unsubstituted alkylaryl, a substituted or unsubstituted arylalkyl, a substituted or unsubstituted aryl, or a substituted or unsubstituted heteroaryl, -(CH2) m-R" or a pharmaceutically acceptable salt, R' represents hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted alkylaryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl or (CH2) m -R"; R" represents hydroxyl, substituted or unsubstituted carbonyl, aryl, cycloalkyl ring, cycloalkenyl ring, heterocycle or polycycle; m is 0 or an integer from 1 to 8. Where X is oxygen and R is as defined above, the moiety is also referred to as a "carboxyl group." Where X is oxygen and R is hydrogen, the formula represents a "carboxylic acid." Where X is oxygen and R' is hydrogen, the formula represents a "formic acid." Where X is oxygen and R or R' is other than hydrogen, the formula represents an "ester." In general, where the oxygen atom of the above formula is replaced by a sulfur atom, the formula represents a 'thiocarbonyl' group. Where X is sulfur and R or R' is other than hydrogen, the formula represents a 'thioester.' Where X is sulfur and R is hydrogen, the formula represents a 'thiocarboxylic acid.' Where X is sulfur and R' is hydrogen, the formula represents a 'thioformate.' Where X is a bond and R is other than hydrogen, the above formula represents a 'ketone.' Where X is a bond and R is hydrogen, the above formula represents an 'aldehyde.'

[0139] The term "substituted carbonyl" refers to a carbonyl group as defined above, wherein R, R' or the moiety

[0140] one or more hydrogen atoms in the group to which it is attached are independently replaced with a substituent. Such substituents include, but are not limited to, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), silyl, ether, ester, thiocarbonyl (e.g., thioester, thioformate, or thioate), alkoxyl, phosphoryl, phosphoester, phosphinoester, phosphinothioester, amino (or quaternized amino), amido, amidine, imine, cyano, nitro, azido, thiol, alkylthio, sulfate, sulfonamide, sulfamoyl, sulffoxide, sulfonyl, heterocyclyl, alkylaryl, haloalkyl, -CN, aryl, heteroaryl, and combinations thereof.

[0141] The term "carboxyl" is defined above as

[0142] and more particularly by the formula -R iv COOH, wherein R iv is alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, alkylaryl, arylalkyl, aryl, or heteroaryl. In preferred embodiments, straight- chain or branched alkyl, alkenyl, and alkynyl groups have 30 or fewer carbon atoms in their main chain (e.g., C1-C30alkyl, C2-C30alkenyl, and C2-C30alkynyl). In more preferred embodiments, straight- chain or branched alkyl, alkenyl, and alkynyl groups have 20 or fewer carbon atoms in their main chain (e.g., C1-C20alkyl, C2-C20alkenyl, and C2-C20alkynyl). In more preferred embodiments, straight- chain or branched alkyl, alkenyl, and alkynyl groups have 10 or fewer carbon atoms in their main chain (e.g., C1-C10alkyl, C2-C10alkenyl, and C2-C10alkynyl). In more preferred embodiments, straight- chain or branched alkyl, alkenyl, and alkynyl groups have 5 or fewer carbon atoms in their main chain (e.g., C1-C5alkyl, C2-C5alkenyl, and C2-C5alkynyl). In more preferred embodiments, straight- chain or branched alkyl, alkenyl, and alkynyl groups have 4 or fewer carbon atoms in their main chain (e.g., C1-C4alkyl, C2-C4alkenyl, and C2-C4alkynyl). In more preferred embodiments, straight- chain or branched alkyl, alkenyl, and alkynyl groups have 3 or fewer carbon atoms in their main chain (e.g., C1-C3alkyl, C2-C3alkenyl, and C2-C3alkynyl). In more preferred embodiments, straight- chain or branched alkyl, alkenyl, and alkynyl groups have 2 or fewer carbon atoms in their main chain (e.g., C1-C2alkyl, C2-C2alkenyl, and C2-C2alkynyl). In more preferred embodiments, straight- chain or branched alkyl, alkenyl, and alkynyl groups have 1 carbon atom in their main chain (e.g., C1-C1alkyl, C2-C1alkenyl, and C2-C1alkynyl). 30 For straight-chain alkyl groups, C3-C 30 For branched alkyl groups, C2-C 30for straight chain alkenyl and alkynyl groups, C3-C 30 for branched chain alkenyl and alkynyl groups, preferably 20 or less, more preferably 15 or less, and most preferably 10 or less. Likewise, preferred cycloalkyl, heterocyclyl, aryl, and heteroaryl groups have 3-10 carbon atoms in their ring structure, and more preferably 5, 6, or 7 carbon atoms in the ring structure.

[0143] The term "substituted carboxyl" refers to a carboxyl group as defined above in which one or more of the hydrogen atoms is replaced by a substituent. Such substituents include, but are not limited to, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, carbonyl (for example, ketones, aldehydes, carboxyl, carboxyl esters, carboxyl amides), sulfanyl, silyl, ether, ester, thioether, alkoxyl, phosphoryl, phosphoester, phosphinoester, phosphine, amino, quaternary iv ammonium, acylamino, amidine, imine, cyano, nitro, azido, thiol, alkylthio, sulfate, sulfonamide, sulfamoyl, sulfoxide, sulfonamide, sulfonyl, heterocyclyl, alkylaryl, haloalkyl, -CN, aryl, heteroaryl, and combinations thereof.

[0144] As used herein, "heteroalkyl" refers to a straight chain or branched chain, or cyclic carbon-containing group, or combinations thereof, containing at least one heteroatom. Suitable heteroatoms include, but are not limited to, O, N, Si, P, and S, wherein the nitrogen, phosphorus and sulfur atoms are optionally oxidized and the nitrogen heteroatom is optionally quaternized.

[0145] Examples of saturated hydrocarbon groups include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, t-butyl, i-butyl, sec-butyl, cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl, and homologs and isomers thereof, for example, n-pentyl, n-hexyl, n-heptyl, n-octyl. Examples of unsaturated alkyl groups include, but are not limited to, ethenyl, 2-propenyl, crotyl, 2-iso-pentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, and 3-butynyl.

[0146] The terms "alkoxyl" or "alkoxy," "aroxy" or "aryloxy" generally describe compounds having the formula -OR v where R v includes, but is not limited to, substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, cycloalkenyl, heterocycloalkenyl, aryl, heteroaryl, aralkyl, heteroalkyl, alkylaryl, alkylheteroaryl.

[0147] As used herein, the term "alkoxy" or "alkoxy group" refers to an alkyl group as defined above, to which an oxygen group is attached. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy, etc. An "ether" is two hydrocarbons covalently linked by oxygen. Therefore, the substituent of the alkyl group that makes it an ether is or similar to an alkoxy group, and may be represented, for example, by one of -O-alkyl, -O-alkenyl, and -O-ynyl. Where valence permits, the term alkoxy also includes cycloalkyl, heterocyclic, cycloalkenyl, heterocyclic alkenyl, and aralkyl groups having an oxygen group attached to at least one carbon atom.

[0148] The term "substituted alkoxy" refers to an alkoxy group having one or more substituents that replace one or more hydrogen atoms on one or more carbons of the alkoxy backbone. Such substituents include, but are not limited to, halogens, azides, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), silyl, ether, ester, thiocarbonyl (e.g., thioester, thioacetate, or thiocarbamate), alkoxy, phosphoryl, phosphate, phosphonate, hypophosphonate, amino (or quaternized amino), amide, amidine, imine, cyano, nitro, azide, mercapto, alkylthio, sulfate, sulfonate, aminosulfonyl, sulfoxide, sulfonylamino, sulfonyl, heterocyclic, alkylaryl, haloalkyl, -CN, aryl, heteroaryl, and combinations thereof.

[0149] The term "alkathioyl" refers to an alkyl group as defined above, which is attached to a sulfur group. The "alkathioyl" part is represented by -S-alkyl. Representative alkathioyl groups include methylthioyl, ethylthioyl, etc. The term "alkathioyl" also includes cycloalkyl groups to which a sulfur group is attached.

[0150] The term "substituted alkathio" refers to an alkathio group having one or more substituents that replace one or more hydrogen atoms on one or more carbon atoms of the alkathio backbone. Such substituents include, but are not limited to, halogens, azides, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), silyl, ether, ester, thiocarbonyl (e.g., thioester, thioacetate, or thiocarbamate), alkoxy, phosphoryl, phosphate, phosphonate, hypophosphonate, amino (or quaternized amino), amide, amidine, imine, cyano, nitro, azide, mercapto, alkathio, sulfate, sulfonate, aminosulfonyl, sulfoxide, sulfonylamino, sulfonyl, heterocyclic, alkylaryl, haloalkyl, -CN, aryl, heteroaryl, and combinations thereof.

[0151] "Arylthio" refers to -S-aryl or -S-heteroaryl, where aryl and heteroaryl are as defined herein.

[0152] The term "substituted arylthio" represents -S-aryl or -S-heteroaryl groups having one or more substituents replacing one or more ring atoms of the aryl and heteroaryl rings defined herein. Such substituents include, but are not limited to, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxy, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), silyl, ether, ester, thiocarbonyl (e.g., thioester, thioformate, or thioate), alkoxyl, phosphoryl, phosphoester, phosphinoester, phosphinothioester, amino (or quaternary ammonium), amido, amidine, imine, cyano, nitro, azo, thio, sulfate, sulfyl, sulfonamide, sulfone, sulfoxide, heterocyclyl, alkylaryl, haloalkyl, -CN, -NO2, -N3, aryl, heteroaryl, and combinations thereof.

[0153] As used herein, "aralkyl" refers to an alkyl group substituted with a substituted or unsubstituted aryl or heteroaryl group.

[0154] As used herein, "alkaryl" refers to an aryl (e.g., aromatic or heteroaromatic group) substituted with a substituted or unsubstituted alkyl group.

[0155] The terms "amide" or "amido" are used interchangeably and refer to "unsubstituted amido" and "substituted amido" and are represented by the general formula:

[0156] wherein E is absent, or E is a substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aralkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocyclyl, wherein R and R' each independently represent, independently of E, a hydrogen, a substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbonyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted alkaryl, substituted or unsubstituted aralkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl, -(CH2) m R" or R and R' together with the N atom to which they are attached form a heterocyclic ring having from 3 to 14 atoms in the ring structure; R" represents a hydroxyl, substituted or unsubstituted carbonyl, aryl, cycloalkyl ring, cycloalkenyl ring, heterocyclic ring, or polycyclic ring; and m is 0 or an integer from 1 to 8. In preferred embodiments, only one of R and R' can be a carbonyl, e.g., R and R' do not form a diimide with the nitrogen. In preferred embodiments, R and R' each independently represent a hydrogen atom, a substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, or -(CH2) m- R'''. When E is oxygen, a carbamate is formed. As understood by one of ordinary skill in the art, a carbamate cannot be linked to another chemical species, such as to form an oxygen-oxygen bond or other unstable linkage.

[0157] The term "sulfonyl" is represented by the formula

[0158] where E is absent, or E is alkyl, alkenyl, alkynyl, aralkyl, alkaryl, cycloalkyl, aryl, heteroaryl, heterocyclyl, wherein independently of E, R represents hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted amine, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted alkaryl, substituted or unsubstituted aralkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl, -(CH2) m - R'''. When E is oxygen, a carbamate is formed. As understood by one of ordinary skill in the art, a carbamate cannot be linked to another chemical species, such as to form an oxygen-oxygen bond or other unstable linkage.

[0159] The term "substituted sulfonyl" represents a sulfonyl group in which E and R are independently substituted. Such substituents include, but are not limited to, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), silyl, ether, ester, thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxyl, phosphoryl, phosphoester, phosphinoester, phosphine, amino (or quarternized amino), amido, amidine, imine, cyano, nitro, azido, thiol, alkylthio, sulfate, sulfonamido, sulfamide, sulfone, sulfoxide, sulfonyl, heterocyclyl, alkaryl, haloalkyl, -CN, aryl, heteroaryl, and combinations thereof.

[0160] The term "sulfonic acid group" refers to a sulfonyl group as defined above, wherein R is hydroxyl, and E is absent, or E is substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted alkaryl, substituted or unsubstituted aralkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.

[0161] The term "sulfate" refers to a sulfonyl group as defined above, wherein E is absent, oxygen, alkoxy, aryloxy, substituted alkoxy, or substituted aryloxy, as defined above, and R is independently hydroxy, alkoxy, aryloxy, substituted alkoxy, or substituted aryloxy, as defined above. As understood by one of ordinary skill in the art, when E is oxygen, the sulfate cannot be linked to another chemical species, for example, to form an oxygen-oxygen bond or other unstable linkage.

[0162] The term "sulfonate" refers to a sulfonyl group as defined above, wherein E is oxygen, alkoxy, aryloxy, substituted alkoxy, or substituted aryloxy, as defined above, and R is independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted amine, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted alkylaryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl, -(CH2) m -R'" wherein R'" represents hydroxy, substituted or unsubstituted carbonyl, aryl, cycloalkyl ring, cycloalkenyl ring, heterocycle, or polycycle; and m is 0 or an integer from 1 to 8. As understood by one of ordinary skill in the art, when E is oxygen, the sulfonate cannot be linked to another chemical species, for example, to form an oxygen-oxygen bond or other unstable linkage.

[0163] The term "sulfonamide" refers to a sulfonamide or a sulfonamide represented by the formula

[0164] wherein E is absent, or E is substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aralkyl, substituted or unsubstituted alkylaryl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocyclyl, wherein each of E, R, and R' independently represents hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbonyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted alkylaryl, substituted or unsubstituted aralkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl, -(CH2) m -R'" or R and R' together with the N atom to which they are attached form a heterocyclic ring having from 3 to 14 atoms in the ring structure; R'" represents hydroxy, substituted or unsubstituted carbonyl, aryl, cycloalkyl ring, cycloalkenyl ring, heterocycle, or polycycle; and m is 0 or an integer from 1 to 8. In preferred embodiments, only one of R and R' can be carbonyl, for example, R and R' do not form a diimide together with the nitrogen.

[0165] The term "sulfoxide" is represented by the formula

[0166] wherein E is absent, or E is alkyl, alkenyl, alkynyl, aralkyl, alkaryl, cycloalkyl, aryl, heteroaryl, heterocyclyl, wherein, independently of E, R represents hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted amine, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted alkaryl, substituted or unsubstituted aralkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl, -(CH2) m -R'" or E and R together with the S atom to which they are attached form a heterocyclic ring having 3 to 14 atoms in the ring structure; R'" represents hydroxyl, substituted or unsubstituted carbonyl, aryl, cycloalkyl ring, cycloalkenyl ring, heterocyclic ring, or polycyclic ring; and m is 0 or an integer from 1 to 8.

[0167] The term "phosphono" is represented by the formula

[0168] wherein E is absent, or E is substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aralkyl, substituted or unsubstituted alkaryl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocyclyl, wherein, independently of E, R vi and R vii are independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbonyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted alkaryl, substituted or unsubstituted aralkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl, -(CH2) m -R'" or R and R' together with the P atom to which they are attached form a heterocyclic ring having 3 to 14 atoms in the ring structure; R'" represents hydroxyl, substituted or unsubstituted carbonyl, aryl, cycloalkyl ring, cycloalkenyl ring, heterocyclic ring, or polycyclic ring; and m is 0 or an integer from 1 to 8.

[0169] The term "substituted phosphono" represents wherein E, R vi and R viiphosphoryl, phosphonate, phosphinite, amino (or quaternized amino), amido, amidine, imine, cyano, nitro, azido, thiol, alkylthio, sulfate, sulfonate, sulfamoyl, sulfoxide, sulfonamido, sulfonyl, heterocyclyl, alkylaryl, haloalkyl, -CN, aryl, heteroaryl, and combinations thereof.

[0170] The term "phosphoryl" defines a phosphoryl group wherein E is absent, is oxygen, alkoxyl, aryloxyl, substituted alkoxyl, or substituted aryloxyl, as defined above, and R vi and R vii are independently hydroxyl, alkoxyl, aryloxyl, substituted alkoxyl, or substituted aryloxyl, as defined above. When E is oxygen, the phosphoryl group cannot be attached to another chemical species, for example, to form an oxygen-oxygen bond or other unstable bond, as understood by one of ordinary skill in the art. When E, R vi and R vii are substituted, the substituents include, but are not limited to, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, carbonyl (for example, carboxyl, alkoxycarbonyl, formyl, or acyl), silyl, ether, ester, thiocarbonyl (for example, thioester, thioacetate, or thioformate), alkoxyl, phosphoryl, phosphonate, phosphinite, amino (or quaternized amino), amido, amidine, imine, cyano, nitro, azido, thiol, alkylthio, sulfate, sulfonate, sulfamoyl, sulfoxide, sulfonamido, sulfonyl, heterocyclyl, alkylaryl, haloalkyl, -CN, aryl, heteroaryl, and combinations thereof.

[0171] The term "C3-C 20 cyclic group" refers to a substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted cycloalkynyl, substituted or unsubstituted heterocyclyl having from 3 to 20 carbon atoms, as geometric constraints allow. The cyclic structure is formed from a monocyclic or fused ring system. Substituted cycloalkyl, cycloalkenyl, cycloalkynyl, and heterocyclyl are substituted as defined above for alkyl, alkenyl, alkynyl, and heterocyclyl, respectively.

[0172] Example 1. Synthesis of compounds C1 and C4 as PROTAC targeting ligands

[0173] a) Synthesis of compound C1 as PROTAC targeting ligand

[0174] 1. Synthesis of compound C1-7-2a

[0175] Compound C1-7-1a (5.00 g, 21.0 mmol, 1.00 equiv), TBSCl (4.12 g, 27.3 mmol, 3.35 mL, 1.30 equiv) and imidazole (2.86 g, 42.0 mmol, 2.00 equiv) were mixed in DMF (30.0 mL), air was discharged and N2was filled, the mixture was stirred at 25 °C for 2 hours under N2environment. TLC (petroleum ether: ethyl acetate = 1:0, P1: R f = 0.86, R1: R f = 0.05) detection showed that C1-7-1a was completely reacted to generate a new main peak with low polarity. The reaction mixture was terminated by adding 300 mL of water, extracted with a total of 300 mL of DCM (100 mL x 3 times), and the obtained organic layer was washed with 300 mL of saturated brine, dried over Na2SO4, filtered and concentrated under reduced pressure to obtain the crude product, which was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 1:0) to obtain compound C1-7-2a (7.00 g, 19.9 mmol, 94.6% yield) as a colorless liquid.

[0176] HNMR: EC10607-21-P1A, CDCl3, 400MHz 1 H NMR (400 MHz, CDCl3) δ ppm 7.16 (dd, J = 10.0, 2.0 Hz, 1H), 7.07 (d, J = 8.4 Hz, 1H), 6.44 (t, J = 8.4 Hz, 1H), 0.78 (s, 10H), 0.03 (s, 6H).

[0177] 2. Synthesis of compound C1-7-3a

[0178] Compound C1-7-2a (2.00 g, 5.68 mmol, 1.00 equiv), C1-7-3 (1.91 g, 8.52 mmol, 1.50 equiv), CuI (54.1 mg, 284 μmol, 0.05 equiv) and Pd(PPh3)4(399 mg, 568 μmol, 0.10 equiv) were mixed in DMF (30 mL), stirred at 75 °C for 2 hours under N2environment. LCMS (EC10607-30-p1a1, P1: RT = 0.460 min) detection showed that about 58.1% of the target product was generated. TLC (petroleum ether: ethyl acetate = 3:1, P1: R f = 0.22, R1: R f= 0.98) showed that the reaction of compound C1-7-2a was complete, generating a new major peak with large polarity. The reaction mixture was terminated by adding 200 mL of water, extracted with 210 mL of DCM (70.0 mL x 3 times), and the organic layer was washed with 200 mL of saturated brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product, which was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 3: 1, Rf= 0.22) to obtain compound C1-7-3a (1.98 g, 4.41 mmol, 77.7% yield) as a yellow oil. f = 0.22) to obtain compound C1-7-3a (1.98 g, 4.41 mmol, 77.7% yield) as a yellow oil.

[0179] LCMS: EC EC10607-30-p1a1, RT = 0.460 min, M / Z (M+H+) = 449.9

[0180] 3. Synthesis of compound C1-7

[0181] To a mixture of compound C1-7-3a (1.98 g, 4.41 mmol, 1.00 equivalent) dissolved in THF (20.0 mL) was added pyridine hydrofluoric acid (875 mg, 8.83 mmol, 795 μL, 2.00 equivalents), and the mixture was stirred at 25°C under N2for 2 hours. LCMS (EC10607-35-p1a1, P1: RT = 0.241 min) detection showed that about 89.2% of the target product was generated. TLC (petroleum ether: ethyl acetate = 2: 1, Rf= 0.22, P1: Rf= 0.15) detection showed that the reaction of compound C1-7-3a was complete, generating a new major peak with large polarity. f = 0.22, P1: R f = 0.98) showed that the reaction of compound C1-7-3a was complete, generating a new major peak with large polarity. The reaction mixture was terminated by adding 300 mL of water, extracted with 300 mL of DCM (100 mL x 3 times), and the organic layer was washed with 300 mL of brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product, which was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 2: 1, Rf= 0.22, P1: Rf= 0.15) to obtain compound C1-7 (1.16 g, 3.47 mmol, 78.6% yield) as a red solid. f = 0.15) to obtain compound C1-7 (1.16 g, 3.47 mmol, 78.6% yield) as a red solid.

[0182] LCMS: EC EC10607-35-p1a1, RT = 0.241 min, M / Z (M+H+) = 335.4

[0183] HNMR: EC10607-21-P1A, CDCl3, 400 MHz

[0184] 1H NMR (400 MHz, CDC13) δ ppm 7.01-7.08 (m, 2 H), 6.90 (t, J = 8.8 Hz, 1 H), 3.49-3.57 (m, 6 H), 2.62 (br t, J = 4.5 Hz, 4 H), 1.47 (s, 9 H).

[0185] 4. Synthesis of compound C1-2

[0186] Compound C1-1 (1.50 g, 2.65 mmol, 1.00 equiv), compound 1a (2.37 g, 7.96 mmol, 3.00 equiv), Pd2(dba)3 (243 mg, 265 μmol, 0.10 equiv), K3PO4 (1.97 g, 9.28 mmol, 3.50 equiv) and (1S,3R,5R,7S)-1,3,5,7-tetramethyl-8-phenyl-2,4,6-trioxa-8- phosphortricyclo[3.3.1.13,7]decane (310 mg, 1.06 mmol, 0.40 equiv) were mixed in 1,4-dioxane (10.0 mL) and water (10.0 mL), air was purged by N2, the mixture was stirred at 90 °C for 2 hours under N2. LCMS (EC10607-26-p1a2, P1: RT = 0.649 min) detection showed about 67.5% of target product was generated. TLC (petroleum ether: ethyl acetate = 3:1, P1: Rf = 0.29, R1: Rf = 0.25) showed compound C1-1 was completely reacted and multiple new peaks were generated. The reaction mixture was quenched by adding 300 mL water, extracted with 300 mL DCM (100 mL x 3 times), the organic layer was washed with 300 mL brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give the product crude, which was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 3:1, Rf = 0.29) to give compound C1-2 (1.25 g, 1.90 mmol, 71.7% yield) as yellow oil. f f f = 0.29, R1: Rf = 0.25) showed compound C1-1 was completely reacted and multiple new peaks were generated. The reaction mixture was quenched by adding 300 mL water, extracted with 300 mL DCM (100 mL x 3 times), the organic layer was washed with 300 mL brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give the product crude, which was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 3:1, Rf = 0.29) to give compound C1-2 (1.25 g, 1.90 mmol, 71.7% yield) as yellow oil.

[0187] LCMS: EC10607-26-p1a2, RT = 0.650 min, M / Z (M+H + ) = 657.8

[0188] 5. Synthesis of compound C1-3

[0189] ​​To a solution of compound C1-2 (1.20 g, 1.83 mmol, 1.00 equiv) in THF (20.0 mL) and methanol (20.0 mL) was added Pd / C (1.00 g, 1.83 mmol, 10% purity, 1.00 equiv) and the reaction mixture was stirred at 25 °C under 50 psi H2(1.23 mg, 609 μmol) for 24 h. TLC (petroleum ether: ethyl acetate = 3:1, P1: Rf = 0.36, R1: Rf = 0.31) showed that the reaction of compound C1-2 was complete and a new peak was generated. The Pd / C was removed using celite filtration and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 3:1, Rf = 0.30) to give compound C1-3 (1.01 g, 1.53 mmol, 73.7% yield) as a yellow solid. f f f

[0190] 6. Synthesis of compound C1-4

[0191] To a mixture of compound C1-3 (1.00 g, 1.52 mmol, 1.00 equiv) in THF (15.0 mL) was added pyridine hydrofluoric acid (694 mg, 4.55 mmol, 631 μL, 65% purity, 3.00 equiv) and the mixture was stirred at 25 °C under N2for 2 h. LCMS (EC10607-37-p1a1, P1: RT = 0.410 min) showed that about 46.4% of the desired product was generated. TLC (petroleum ether: ethyl acetate = 1:1, R1: Rf = 0.60, P1: Rf = 0.16) showed that the reaction of C1-3 was complete and a new major peak of high polarity was generated. The reaction mixture was quenched by the addition of 300 mL of water and extracted with 300 mL of DCM (100 mL x 3 times). The organic layer was washed with 300 mL of brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product, which was purified by preparative TLC (SiO2, petroleum ether: ethyl acetate = 1:1, Rf = 0.16) to give compound C1-4 (700 mg, 1.29 mmol, 84.7% yield) as a white solid. f f f

[0192] LCMS: EC EC10607-37-p1a1, RT = 0.410 min, M / Z (M+H + ) = 545.4

[0193] 7. Synthesis of compound C1-5 ​​​​​​

[0194] Compound C1-4 (680 mg, 1.25 mmol, 1.00 equiv), TEA (133 mg, 1.31 mmol, 182 µL, 1.05 equiv), DMAP (1.53 mg, 12.5 µmol, 0.01 equiv) and 4-methylbenzenesulfonyl chloride (250 mg, 1.31 mmol, 1.05 equiv) were mixed in DCM (5.00 mL), air was purged with N2, and the mixture was stirred at 25 °C for 3 h under N2. LC-MS (EC10607-47-p1a1, P1: RT = 0.548 min) detection showed about 46.7% of the target product was generated. TLC (petroleum ether: ethyl acetate = 2:1, P1: R f = 0.36, R1: R f = 0.20) detection showed partial C1-4 reaction with multiple new peaks generated. The reaction mixture was quenched by adding 300 mL water, extracted with 300 mL DCM (100 mL x 3 times), and the organic layer was washed with 300 mL brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give the product crude. The crude was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 2:1, R f = 0.36) to give compound C1-5 (400 mg, 572 µmol, 45.8% yield) as a yellow solid.

[0195] LCMS: EC (EC10607-47-p1a1, RT = 0.548 min, M / Z (M+H + ) = 699.1

[0196] 8. Synthesis of compound C1-6

[0197] Compound C1-5 (100 mg, 143 µmol, 1.00 equiv), C1-7 (71.8 mg, 214 µmol, 1.50 equiv), Cs2CO3 (69.9 mg, 215 µmol, 1.50 equiv) were mixed in DMA (5.00 mL), air was purged with N2, and the mixture was stirred at 25 °C for 12 h under N2. LC-MS (EC10607-49-p1a1, P1: RT = 0.493 min) detection showed about 36.9% of the target product was generated. TLC (petroleum ether: ethyl acetate = 1:1, P1: R f = 0.24, R1: R f= 0.42) showed that C1-5 was consumed completely and multiple new peaks were generated. The reaction mixture was quenched by adding 30 mL of water and extracted with 30.0 mL of DCM (10.0 mL x 3 times). The organic layer was washed with 30.0 mL of brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product. The crude product was purified by preparative TLC (SiO2, Petroleum ether: ethyl acetate = 1:1, Rf = 0.24) to give compound C1-6 (106 mg, 123 pmol, 86.0% yield) as a yellow solid. f = 0.24) to give compound C1-6 (106 mg, 123 pmol, 86.0% yield) as a yellow solid.

[0198] LCMS: EC10607-49-p1a1, RT = 0.493 min, M / Z (M+H + ) = 861.2

[0199] 9. Synthesis of compound C1

[0200] Compound C1-6 (100 mg, 116 pmol, 1.00 equiv) was dissolved in HCl / dioxane (5.00 mL), air was purged and N2was filled, the mixture was stirred at 25 °C for 12 hours under N2environment. LC-MS (EC10607-51-p1a5, P1: RT = 0.412 min) detection showed that about 86.5% of the target product was generated. The reaction mixture was concentrated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC (HCl condition, column: Welch Xtimate C18 150*25mm*5pm; mobile phase: [water (HCl)-ACN]; B%: 26%-46%, 10 min) to give compound C1 (25.0 mg, 35.5 pmol, 89.3% yield) as a yellow solid. LCMS (EC10607-57-p1a1, P1: RT = 0.42 min) detection showed that the purity of the product was about 100%. HPLC (EC10607-57-p1a2, P1: RT = 2.027 min) detection showed that the purity of product C1 was about 96.1%.

[0201] LCMS: EC EC10607-51-p1a5, RT = 0.412, M / Z (M+H + ) = 705.2

[0202] 1H NMR (400 MHz, DMSO+D20) δ ppm 7.99 (d, J = 7.6 Hz, 1 H), 7.77 (d, J = 8.0 Hz, 1 H), 7.57 (d, J = 8.8 Hz, 1 H), 7.54 (d, J = 7.2 Hz, 1 H), 7.47 (t, J = 7.2 Hz, 1 H), 7.39-7.42 (m, 1 H), 7.32-7.37 (m, 3 H), 7.27 (d, J = 8.9 Hz, 1 H), 7.06-7.12 (m, 1 H), 7.01 (d, J = 8.8 Hz, 1 H), 4.91 (s, 2 H), 4.22 (s, 2 H), 4.08 (s, 2 H), 3.75-3.81 (m, 2 H), 3.33-3.43 (m, 8 H), 2.94-2.99 (m, 2 H), 2.77-2.83 (m, 2 H), 1.88-1.96 (m, 2 H).

[0203] b) Synthesis of compound C4 as PROTAC targeting ligand

[0204] 1. Synthesis of compound C4-2

[0205] To a solution of compound C4-1 (2.00 g, 17.7 mmol, 1.0 eq) in MeCN (20.0 mL) and TFA (3.34 mL) was added NIS (4.38 g, 19.5 mmol, 1.1 eq), and the reaction mixture was stirred at 25 °C for 12 h. TLC (petroleum ether: ethyl acetate = 0: 1, R1: R f = 0.4; P1: R f = 0.8) monitoring showed that C4-1 was consumed completely and a new compound peak was generated. The reaction mixture was filtered to give the crude product C4-2 (3.00 g, crude) as a white solid.

[0206] 2. Synthesis of compound C4-3

[0207] To a solution of compound C4-2 (1.00 g, 4.18 mmol, 1.0 eq) and C1-7-3 (1.41 g, 6.28 mmol, 1.5 eq) in DMF (20.0 mL) was added Cul (39.9 mg, 209 μmol, 0.05 eq), TEA (1.27 g, 12.6 mmol, 1.75 mL, 3.0 eq) and Pd(dppf)Cl2 (306 mg, 418 μmol, 0.1 eq), the reaction mixture was stirred at 90 °C for 2 h. LCMS (EC13784-5-P1A, P1: RT = 0.242 min) monitoring showed that the reaction of C4-2 was complete, and the main peak of the desired product was generated. The reaction mixture was extracted by adding 20.0 mL of water and 60.0 mL of ethyl acetate, and the resulting organic layer was washed with saturated brine (20.0 mL x 2 times), dried over Na2SO4, filtered and concentrated under reduced pressure to give the product crude C4-3 (500.0 mg) as a brown solid.

[0208] LC-MS: EC13784-5-P1A, product: RT = 0.242 min, m / z = 336.3 [M+H] +

[0209] 3. Synthesis of compound C4-4

[0210] To a solution of compound C4-3 (90.0 mg, 268 μmol, 1.0 eq) in DMA (3.00 mL) was added Cs2CO3 (175 mg, 537 μmol, 2.0 eq) and C1-3 (188 mg, 268 μmol, 1.0 eq), the reaction mixture was stirred at 25 °C for 12 h. LCMS (EC13784-6-P1A, P1: RT = 0.495 min) monitoring showed that the reaction of compound C4-3 was complete, and the main peak of the desired product with expected m / z was generated. TLC (petroleum ether: ethyl acetate = 1:2, R1: R f = 0.1; R2: R f = 0.85; P1: R f = 0.55) monitoring showed that C4-3 was completely consumed, and two new compound peaks were generated. The reaction mixture was extracted by adding 10.0 mL of water and 10.0 mL of ethyl acetate, and the resulting organic layer was washed with saturated brine (10.0 mL x 2 times), dried over Na2SO4, filtered and concentrated under reduced pressure to give the product crude, which was detected by TLC (petroleum ether: ethyl acetate = 1:2, P1: R f = 0.55; P2: R f = 0.30) and purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 1:2) to give compound C4-4 (55.0 mg) as a colorless oil.

[0211] LC-MS: EC13784-6-P1A, product: RT = 0.495 min, m / z = 862.6 [M+H] +

[0212] 4. Synthesis of compound C4

[0213] Compound C4-4 (50.0 mg, 58.0 μmol, 1.0 eq) was dissolved in HC1-dioxane (2.00 mL), the mixture was stirred at 25 °C for 1 h. LCMS (EC13784-11-P1A2, P1: RT = 0.418 min) monitoring showed that compound C4-4 was reacted completely, and the main peak of m / z was consistent with the expected product. The reaction mixture was concentrated under reduced pressure to obtain the crude product, which was purified by preparative HPLC (column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (HC1)-ACN]; gradient elution: 30%-60%B, 10 min) to obtain compound C4 (11.0 mg, 24.46% yield, 91.03% purity) as a colorless oil.

[0214] LC-MS: EC13784-11-P1A2, product: RT = 0.418 min, m / z = 706.3 [M+H] +

[0215] HPLC: EC13784-11-P1C; product: RT = 1.945 min

[0216] 1 H NMR: EC13784-11-P1C1 (400 MHz DMSO) δ ppm 8.17 (d, J = 1.6 Hz, 1H), 8.02 (d, J = 7.6 Hz, 1H), 7.90 (dd, J = 10.8, 1.8 Hz, 1H), 7.78 (d, J = 8.0 Hz, 1H), 7.59 (t, J = 8.8 Hz, 2H), 7.44-7.50 (m, 1H), 7.40-7.44 (m, 1H), 7.32-7.38 (m, 2H), 7.02 (d, J = 8.8 Hz, 1H), 4.92 (s, 2H), 4.37-4.42 (m, 2H), 4.29-4.37 (m, 2H), 3.83 (t, J = 6.0 Hz, 3H), 3.57 (d, J = 4.0 Hz, 5H), 3.42-3.49 (m, 4H), 2.98 (t, J = 5.6 Hz, 2H), 2.81 (t, J = 7.6 Hz, 2H), 1.92-2.01 (m, 2H).

[0217] Example 2. Synthesis of compound C1 as a targeting ligand and PROTAC molecule recruiting CRBN-E3 enzyme (AF3, AF4, BF3, BF4, APQ1, APQ2)

[0218] a) Synthesis of PROTAC molecules AF3 and AF4

[0219] Synthesis of AF3:

[0220] 1. Synthesis of compound AF3-2

[0221] Compound AF3-1 (4.02 g, 13.7 mmol, 1.00 equiv), compound 1 (3.80 g, 13.7 mmol, 1.00 equiv) and DIEA (3.56 g, 27.5 mmol, 4.79 mL, 2.00 equiv) were mixed in DMF (50.0 mL), N2was purged to remove air, and the mixture was stirred at 90 °C for 12 hours under N2. LC-MS (EC11272-85-P1A, P1: RT = 0.415 min) detection showed that compound 1 was completely reacted. The reaction mixture was terminated by adding 50.0 mL of water at 0 °C, diluted with 50.0 mL of EA, extracted with 150.0 mL of EA (50.0 mL x 3 times), and the organic layer was washed with 50.0 mL of saturated brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product, which was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 3: 1 to 0: 1; petroleum ether: ethyl acetate = 0: 1, Rf = 0.24). Then, HC1 / dioxane (4 M, 10.0 mL, 21.9 equiv) was added, N2was purged to remove air, and the mixture was stirred at 25 °C for 2 hours under N2. The crude product AF3-2 (500 mg, 1.11 mmol, 61.1% yield) was obtained by concentration under reduced pressure as a yellow solid. f

[0222] LCMS: EC11272-85-P1A, P1, RT = 0.415 min, M / Z (M+H + ) = 449.2

[0223] 2. Synthesis of compound AF3

[0224] ​Compound C1 (50.0 mg, 67.4 μmol, 1.00 equiv, HC1), AF3-2 (60.5 mg, 135 μmol, 2.00 equiv), EDCI (19.4 mg, 101 μmol, 1.50 equiv), HOBt (13.6 mg, 101 μmol, 1.50 equiv), DIEA (43.6 mg, 337 μmol, 58.7 μL, 5.00 equiv) were mixed in DMF (1.0 mL), N2 was bubbled to remove air, the mixture was stirred at 20 °C for 16 h under N2. LC-MS (EC13958-9-P1B1, p1: RT = 0.456 min) detection showed that the reaction of C1 was complete. The reaction mixture was concentrated under reduced pressure to obtain the crude product, which was purified by preparative HPLC (neutral conditions) to obtain compound AF3 (10 mg, 8.81 μmol, 13.0% yield) as a yellow solid.

[0225] LCMS: EC13958-9-P1B1, P1, RT = 0.456 min, M / Z (M+H + ) = 1135.8

[0226] HNMR: EC13958-9-P1R δ ppm 8.42 (t, J = 5.6 Hz, 1H), 7.98 (d, J = 7.6 Hz, 1H), 7.74 (d, J = 8.0 Hz, 1H), 7.60 (d, J = 6.8 Hz, 1H), 7.46-7.56 (m, 2H), 7.40-7.46 (m, 1H), 7.21-7.37 (m, 4H), 7.15 (s, 1H), 7.05-7.12 (m, 2H), 6.95-7.03 (m, 2H), 6.56 (s, 1H), 5.03 (dd, J = 5.2, 12.8 Hz, 1H), 4.98 (s, 2H), 4.01 (t, J = 6.8 Hz, 2H), 3.86 (t, J = 5.6 Hz, 2H), 3.54-3.57 (m, 2H), 3.45-3.50 (m, 6H), 3.43 (d, J = 4.4 Hz, 8H), 3.39 (dd, J = 8.4, 5.75 Hz, 8H), 2.93-2.99 (m, 4H), 2.78 (br t, J = 4.8 Hz, 4H), 2.47 (s, 2H), 1.88-2.09 (m, 4H).

[0227] Synthesis of AF4:

[0228] The synthesis of PROTAC molecule AF4 refers to that of AF3, but the starting material is changed to compound 1 and AF4-1. After synthesis, the target product AF4 is obtained by purification, which is a yellow solid with a yield of about 13%.

[0229] LCMS: EC13958-10-P1B1, P1, RT = 0.460 min, M / Z (M+H + ) = 1180.3

[0230] HNMR: EC13958-10-P1R, 400 MHz, DMSO-d6 δ ppm 8.39-8.46 (m, 1H), 7.97 (d, J = 8.0 Hz, 1H), 7.74 (d, J = 8.0 Hz, 1H), 7.58-7.62 (m, 1H), 7.52-7.58 (m, 1H), 7.49 (d, J = 8.8 Hz, 1H), 7.43 (t, J = 7.2 Hz, 1H), 7.23-7.37 (m, 4H), 7.14-7.19 (m, 1H), 7.06-7.13 (m, 2H), 6.94-7.03 (m, 2H), 6.57 (t, J = 5.6 Hz, 1H), 5.04 (dd, J = 12.8, 5.6 Hz, 1H), 4.99 (s, 2H), 4.01 (t, J = 6.4 Hz, 2H), 3.86 (t, J = 6.0 Hz, 2H), 3.55-3.59 (m, 2H), 3.49-3.51 (m, 2H), 3.45-3.48 (m, 4H), 3.43 (s, 8H), 3.42 (s, 4H), 3.37-3.41 (m, 6H), 2.93-2.99 (m, 4H), 2.76 (t, J = 4.8 Hz, 4H), 2.46 (d, J = 4.4 Hz, 4H), 1.90-2.06 (m, 4H)

[0231] b) Synthesis of PROTAC molecules BF3 and BF4

[0232] Synthesis of BF3:

[0233] 1. Synthesis of compound BF3-2

[0234] A mixture of compound BF3-1 (4.09 g, 21.1 mmol, 1.50 equiv), compound 1 (3.90 g, 14.1 mmol, 1.00 equiv) and DIEA (10.9 g, 84.7 mmol, 14.7 mL, 6.00 equiv) in DMSO (40.0 mL) was purged with N2to remove air and the mixture was stirred at 100 °C for 12 h under N2atmosphere. LC-MS (EC13798-1-p1a1, P1 RT = 0.312 min) detection showed the product of the desired molecular weight was generated. The reaction mixture was quenched by adding 300.0 mL of water at 25 °C, diluted with 100.0 mL of DCM, extracted with DCM (100.0 mL x 5 times), the organic layer was washed with 50.0 mL of saturated brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 5:1 to 0:1; petroleum ether: ethyl acetate = 0:1, Rf = 0.20) to give the crude product BF3-2 (5.00 g, 11.1 mmol, 78.7% yield) as a green oil. f

[0235] LCMS: EC13798-1-p1a1, P1 RT = 0.312 min, MS (ESI) m / z = 450.4 [M+H] +

[0236] 2. Synthesis of compound BF3-3

[0237] To a solution of compound BF3-2 (200 mg, 444 µmol, 1.00 equiv) in DCM (2.00 mL) was added DMP (377.47 mg, 889.97 µmol, 275.73 µL, 2.00 equiv) at 0 °C. The mixture was stirred at 25 °C for 2 h. LC-MS (EC13798-4-p1a1, P1 RT = 0.318 min) detection showed the product of the desired molecular weight was generated. The reaction mixture was quenched by adding saturated NaHCO3 (10.0 mL) and saturated Na2S2O3 (10.0 mL) at 25 °C, and stirred for another 30.0 min. The mixture was extracted with DCM (20.0 mL x 3 times), and the organic layer was washed with saturated brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product BF3-3 (200 mg) as a yellow oil.

[0238] LCMS: EC13798-4-p1a1, P1 RT = 0.318 min, MS (ESI) m / z = 448.3 [M+H] +

[0239] ​3. Synthesis of compound BF3

[0240] To a solution of compound BF3-3 (19.0 mg, 42.5 μmol, 1.00 equiv) in DMF (2.00 mL) was added compound Cl (30.0 mg, 42.5 μmol, 1.00 equiv) and NaBH(OAc)3 (27.0 mg, 127 μmol, 3.00 equiv), and the mixture was stirred at 25 °C for 2 h. LC-MS (EC13798-17-p1c8, P1 RT = 0.453 min) detection showed the product of the desired molecular weight was generated. The reaction mixture was concentrated under reduced pressure to give the crude product, which was purified by prep-HPLC (TFA condition; Column: Phenomenex luna C18 150*25.0mm*10.0μm; Mobile Phase: [water(TFA)-ACN]; Gradient: 36.0%-66.0%B in 15.0 min) to give compound BF3 (10.0 mg, 8.27 μmol, 19.4% yield, 94.0% purity) as a yellow solid.

[0241] LCMS: EC13798-17-p1c8, RT = 0.453 min, MS (ESI) m / z = 1136.4 [M+H] +

[0242] 1H NMR: EC13798-17-p1a2, 400 MHz, DMSO + D2O δ 8.02 (d, J = 7.6 Hz, 1H), 7.77 (d, J = 8.0 Hz, 1H), 7.52-7.61 (m, 3H), 7.47 (t, J = 8.0 Hz, 1H), 7.38-7.43 (m, 1H), 7.35 (m, 2H), 7.26 (dd, J = 12.0, 1.6 Hz, 1H), 7.18 (d, J = 8.8 Hz, 1H), 7.05-7.12 (m, 2H), 7.03 (d, J = 7.2 Hz, 1H), 6.96 (d, J = 8.4 Hz, 1H), 5.03 (dd, J = 13.2, 5.6 Hz, 1H), 4.87-4.95 (m, 2H), 4.01 (t, J = 6.0 Hz, 2H), 3.84 (t, J = 5.6 Hz, 2H), 3.69-3.73 (m, 2H), 3.64 (s, 2H), 3.60 (s, 4H), 3.43 (t, J = 5.2 Hz, 3H), 3.26 (s, 2H), 2.97 (t, J = 5.6 Hz, 2H), 2.84-2.91 (m, 2H), 2.77-2.83 (m, 3H), 2.55-2.63 (m, 2H), 1.88-2.06 (m, 4H).

[0243] Synthesis of BF4:

[0244] The synthesis of PROTAC molecule BF4 refers to that of BF3, but the starting material is changed to compound 1 and BF4-1. After synthesis, the target product BF4 is obtained by purification, which is a yellow solid with a purity of 94.1% and a yield of about 16%.

[0245] LCMS: EC13798-11-p1a2, RT = 0.456 min, MS (ESI) m / z = 1181.8 [M+H] +

[0246] 1H NMR: EC13798-11-p1a3, 400 MHz, DMSO + D2O δ 8.02 (d, J = 8.0 Hz, 1H), 7.78 (d, J = 7.6 Hz, 1H), 7.52-7.61 (m, 3H), 7.45-7.50 (m, 1H), 7.39-7.42 (m, 1H), 7.32-7.38 (m, 2H), 7.28 (dd, J = 11.6, 2.0 Hz, 1H), 7.19 (d, J = 9.2 Hz, 1H), 7.10 (d, J = 8.4 Hz, 2H), 7.03 (d, J = 6.8 Hz, 1H), 6.96 (d, J = 8.8 Hz, 1H), 5.03 (dd, J = 13.2, 6.0 Hz, 1H), 4.92 (s, 2H), 4.01 (t, J = 6.40 Hz, 2H), 3.83 (t, J = 6.00 Hz, 2H), 3.72 (s, 4H), 3.45-3.56 (m, 16H), 3.41-3.45 (m, 3H), 3.25-3.30 (m, 3H), 2.76-3.01 (m, 10H), 2.52-2.63 (m, 2H), 1.95 (m, 4H).

[0247] c) Synthesis of PROTAC molecule APQ1

[0248] 1. Synthesis of Q1-1

[0249] To a solution of compound 1A (451 mg, 2.84 mmol, 4.00 equiv) and CI (0.500 g, 709 µmol, 1.00 equiv) in DMF (5.00 mL) was added TBTU (455 mg, 1.42 mmol, 2.00 equiv) and DIEA (458 mg, 3.55 mmol, 617 mL, 5.00 equiv), the mixture was stirred at 25 °C for 2 h. After reaction, the mixture was quenched by water (10.0 mL), diluted with DCM (10.0 mL), extracted with DCM (30.0 mL, 10.0 mL x 3 times), the organic layer was washed with saturated brine (20.0 mL, 10.0 mL x 2 times), dried over Na2SO4, filtered and concentrated under reduced pressure to give the product as a crude, which was further purified by prep-HPLC (column: Phenomenex Luna C 18 150*25mm*10µm; mobile phase: [water(FA)-ACN]; gradient elution: 30%-60%B in 10 min) to give the product Q1-1 (90.0 mg, 106 µmol, 15.0% yield) as a white solid.

[0250] LCMS: EC13798-52-p1a2, P1 RT = 0.426 min, MS (ESI) m / z = 846.4 [M+H] +

[0251] 2. Synthesis of Q1-2

[0252] To compound Q1-1 (30.0 mg, 35.4 μmol, 1.00 equiv) was added formic acid (2.00 mL), the mixture was stirred at 25 °C for 1 h. After reaction, the mixture was concentrated under reduced pressure to give the product Q1-2 (25.0 mg, 31.2 μmol, 88.1% yield) as a yellow solid, which was used directly in the next step without purification.

[0253] LCMS: EC13798-58-p1a1, P1 RT = 0.394 min, MS (ESI) m / z = 800.5 [M+H] +

[0254] 3. Synthesis of Q1-4

[0255] To compound Q1-3 (0.500 g, 1.03 mmol, 1.00 equiv) was added HC1 / methanol (4.00 M, 12.5 mL, 48.5 equiv), the mixture was stirred at 25 °C for 0.3 h. LCMS (P1 RT = 0.162 min) monitoring showed the product with the desired molecular weight was generated. After reaction, the mixture was concentrated under reduced pressure to give the product Q1-4 (400 mg, crude product) as a yellow solid, which was used directly in the next step without purification.

[0256] LCMS: EC13798-46-p1a1, P1 RT = 0.162 min, MS (ESI) m / z = 386.3 [M+H] +

[0257] 4. Synthesis of APQ1

[0258] To a solution of compound Q1-2 (25.0 mg, 31.2 μmol, 1.00 equiv) and Q1-4 (12.0 mg, 31.2 μmol, 1.00 equiv) in DMA (0.50 mL) was added NaBH(OAc)3 (19.8 mg, 93.7 μmol, 3.00 equiv), the mixture was stirred at 25 °C for 1 h. LCMS (P1 RT = 0.375 min) monitoring showed the desired molecular weight product was generated. The reaction mixture was quenched with water (10.0 mL) at 25 °C, diluted with DCM (10.0 mL), extracted with DCM (30.0 mL, 10.0 mL x 3 times), the organic layer was washed with saturated brine (30.0 mL, 10.0 mL x 3 times), dried over Na2SO4, filtered and concentrated under reduced pressure to give the product crude, which was further purified by prep-HPLC (column: Phenomenex luna C 18 150*25mm*10μm; mobile phase: [water(HCl)-ACN]; gradient elution: 27%-47% B in 10.0 min) to give the product APQ1 (1.00 mg, crude) as a yellow solid.

[0259] LCMS: EC13798-59-p1c5, P1 RT = 0.375 min, MS (ESI) m / z = 585.7 [M / 2+H] +

[0260] HPLC: EC13798-59-p1c7, P1 RT = 2.385 min, 61.0% purity.

[0261] d) Synthesis of PROTAC molecule APQ2

[0262] 1. Synthesis of compound Q2-2

[0263] To a solution of compound 1A (2.00 g, 12.5 mmol, 1.00 equiv) in CH3CN (40.0 mL) was added K2CO3 (5.21 g, 37.7 mmol, 3.00 equiv) and compound Q2-1 (3.24 g, 12.5 mmol, 1.00 equiv), the mixture was stirred at 80 °C for 12 h. LC-MS (EC13784-1-p1a, P1 RT = 0.263 min) detection showed Q2-1 was reacted completely, product with target molecular weight was generated. To the reaction mixture was added 40.0 mL water and 40.0 mL ethyl acetate for extraction, the resulting organic layer was washed with saturated brine (30.0 mL x 2 times), dried over Na2SO4, filtered and concentrated under reduced pressure to give the product crude, which was purified using reverse phase HPLC (0.1% FA condition) to give Q2-2 (2.30 g, 5.91 mmol, 47.0% yield, 86.4% purity) as colorless oil.

[0264] LC-MS: EC13784-1-p1a, product: RT = 0.263 min, m / z = 337.5 [M+H] +

[0265] 1 H NMR: EC13784-1-p1d (400 MHz DMSO) δ 8.19 (s, 1H), 7.26-7.42 (m, 5H), 7.05-7.24 (m, 1H), 5.01 (s, 3H), 4.02 (d, J = 6.8 Hz, 1H), 3.24 (s, 6H), 3.05-3.18 (m, 2H), 2.90 (d, J = 10.8 Hz, 2H), 2.33-2.45 (m, 2H), 1.96 (t, J = 11.6 Hz, 2H), 1.44-1.66 (m, 3H), 1.12-1.32 (m, 2H).

[0266] 2. Synthesis of compound Q2-3

[0267] To a solution of compound Q2-2 (1.90 g, 5.65 mmol, 1.0 equiv) in MeOH (60.0 mL) was added Pd-C (190 mg, 10% purity), the mixture was hydrogenated at 50 Psi H2 at 20 °C for 12 h. LC-MS (EC13784-4-p1a, 0.446 min) detection showed Q2-2 was reacted completely, product with target molecular weight was generated. The reaction mixture was concentrated under reduced pressure to give the product crude, which was used without purification. Crude Q2-3 (1.00 g, 4.94 mmol, 87.53% yield) was colorless oil.

[0268] LC-MS: EC13784-4-p1a, Product: RT = 0.446 min, m / z = 203.6 [M+H] +

[0269] 3. Synthesis of compound Q2-4

[0270] Compound Q2-3 (1.23 g, 4.45 mmol, 1.0 eq), compound 1 (1.23 g, 4.45 mmol, 1.0 eq) and DIEA (1.15 g, 8.90 mmol, 1.55 mL, 2.0 eq) were mixed in DMF (20.0 mL), N2 was charged to discharge air, and the mixture was stirred at 90 °C for 3 hours under N2. LC-MS (EC13784-14-p1a1, P1: RT = 0.252 min) detection showed that Q2-3 was completely reacted to generate a product main peak with a target molecular weight. The reaction mixture was extracted by adding 30.0 mL of water and 90.0 mL of ethyl acetate, and the obtained organic layer was washed with saturated brine (40.0 mL x 2 times), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a crude product, which was detected by TLC (dichloromethane:methanol = 10:1, P1: RT = 0.46), and the crude product was purified by column chromatography (SiO2, DCM:MeOH = 10:1) to obtain Q2-4 (300 mg, crude) in the form of green oil.

[0271] LC-MS: EC13784-14-p1a1, Product: RT = 0.252 min, m / z = 459.2 [M+H] +

[0272] 4. Synthesis of compound Q2-5

[0273] To a solution of compound Q2-4 (80.0 mg, 174 μmol, 1.00 eq) dissolved in THF (5.00 mL) was added H2SO4 (34.2 mg, 348 μmol, 18.6 μL, 2.00 eq), and the mixture was stirred at 60 °C for 0.5 hours. LC-MS (EC13798-21-p1a1, P1 RT = 0.214 min) detection showed that a product with a target molecular weight was generated. The reaction mixture was concentrated under reduced pressure to obtain a crude product, which was not purified. The crude product Q2-5 (100 mg) was yellow oil.

[0274] LCMS: EC13798-21-p1a1, P1 RT = 0.214 min, MS (ESI) m / z = 413.3 [M+H] +

[0275] 5. Synthesis of compound APQ2

[0276] To a solution of compound Q2-5 (100 mg, 121 pmol, 1.00 equiv) and CI (85.4 mg, 121 pmol, 1.00 equiv) in DCM (5.00 mL), TEA (12.2 mg, 121 pmol, 16.8 pL, 1.00 equiv) was added dropwise at 25 °C for 5 min, followed by the addition of NaBH(OAc)3 (77.0 mg, 363 pmol, 3.00 equiv) at 25 °C, the mixture was stirred at 25 °C for 1 h. LC-MS (EC13798-22-p1c4, P1 RT = 1.825 min) detection showed the product of target molecular weight was generated. The reaction mixture was concentrated under reduced pressure to give the product crude, which was further purified by prep-HPLC (TFA condition; Column: Phenomenex luna C 18 150*25.0mm*10.0pm; mobile phase: [water(TFA)-ACN]; gradient: 32%-62%B in 10 min) to give APQ2 (12.2 mg, 10.2 pmol, 8.49% yield, 92.9% purity) as a yellow solid.

[0277] LCMS: EC13798-22-p1c4, P1 RT = 1.825 min, MS (ESI) m / z = 1102.8 [M+H] +

[0278] 1 H NMR: EC13798-22-p1c1, 400 MHz, DMSO d 12.83 (s, 1H), 11.10 (s, 1H), 9.40 (s, 1H), 8.04 (d, J = 8.0 Hz, 1H), 7.80 (d, J = 8.0 Hz, 1H), 7.55-7.65 (m, 3H), 7.42-7.49 (m, 1H), 7.37-7.42 (m, 1H), 7.29-7.37 (m, 3H), 7.16-7.24 (m, 2H), 7.09-7.16 (m, 2H), 6.97 (d, J = 8.8 Hz, 1H), 6.87 (t, J = 6.40 Hz, 1H), 5.07 (dd, J = 12.8, 5.2 Hz, 1H), 4.93 (s, 2H), 4.04 (t, J = 6.4 Hz, 2H), 3.86 (t, J = 6.4 Hz, 2H), 3.62-3.79 (m, 6H), 3.53 (s, 1H), 3.18-3.39 (m, 4H), 2.85-3.07 (m, 10H), 2.69-2.84 (m, 4H), 2.56 (m, 2H), 1.92-2.06 (m, 6H), 1.35-1.51 (m, 2H).

[0279] e) Synthesis of PROTAC molecule APQ4

[0280] 1. Synthesis of compound Q4-2

[0281] To a mixture of compound C1-3 (1.00 g, 1.52 mmol, 1.00 equiv) in THF (15.0 mL) was added pyridine-HF (694 mg, 4.55 mmol, 631 μL, 65% purity, 3.00 equiv) and the compound was stirred at 20 °C for 2 h. LC-MS (EC7111-187-p1a, P1: RT = 0.464 min) detection showed that the compound C1-3 reaction was complete. Multiple new compound peaks were generated, with the target m / z product accounting for about 89.8%. The mixture was concentrated under reduced pressure to obtain the product crude, which was further purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 1:0 to 1:2) to obtain compound Q4-2 (760 mg, 1.31 mmol, 86.4% yield, 94.0% purity) as a white solid.

[0282] LCMS: EC7111-187-p1a, P1: RT = 0.464 min, MS (M+H + ) = 545.5

[0283] 2. Synthesis of compound Q4-3

[0284] To a mixture of compound Q4-2 (300 mg, 551 μmol, 1.00 equiv) in DMF (3.00 mL) was added PPh3 (289 mg, 1.10 mmol, 2.00 equiv) and Br2 (132 mg, 826 μmol, 42.6 μL, 1.50 equiv), and the mixture was stirred at 20 °C for 24 h. LC-MS (EC7111-195-p1a, R1: RT = 0.425 min, P1: RT = 0.511 min) detection showed about 4.73% of compound Q4-2 remaining, and multiple new compound peaks were generated, with the target m / z product accounting for about 48.7%. After the reaction, the reaction mixture was terminated by adding NaHCO3 50.0 mL at 20 °C, and extracted with ethyl acetate 60.0 mL (20.0 mL x 3 times). The organic layer was dried over Na2SO4, and concentrated under reduced pressure to obtain the product crude, which was further purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 1:0 to 3:1) to obtain compound Q4-3 (220 mg, 347 μmol, 62.9% yield, 95.8% purity) as a white solid.

[0285] LCMS: EC7111-195-p1a, P1 : RT = 0.511 min, MS (M+H + ) = 609.3

[0286] 3. Synthesis of compound Q4-4

[0287] To a mixture of compound Q4-3 (210 mg, 345 pmol, 1.00 equiv) in ACN (3.00 mL) was added tert-butyl piperazine-1 -carboxylate hydrochloride (154 mg, 691 pmol, 2.00 equiv) and K2CO3 (239 mg, 1.73 mmol, 5.00 equiv), and the mixture was stirred at 20 °C for 24 h. LC-MS (EC7111-201-p1e, R1 : RT = 0.481 min, P1 : RT = 0.418 min) detection showed about 4.11% of compound Q4-3 remained, and the target m / z product accounted for about 69.4%. The mixture was concentrated under reduced pressure and filtered to give the product crude (200 mg) as a yellow solid, which was used directly in the next step without purification.

[0288] LCMS: EC7111-201-p1e, P1 : RT = 0.418 min, MS (M+H + ) = 713.3

[0289] 4. Synthesis of compound Q4-5

[0290] A mixture of compound Q4-4 (200 mg, 281 pmol, 1.00 equiv) in HC1 / EtOAc (3.00 mL) was stirred at 20 °C for 24 h under N2. LC-MS (EC15303-1-p1b, P1 : RT = 0.311 min) detection showed that compound Q4-4 was completely reacted, and several new compound peaks were generated, and the target m / z product accounted for about 58.7%. The mixture was concentrated under reduced pressure and filtered to give compound Q4-5 crude (250 mg) as a yellow solid, which was used directly in the next step without purification.

[0291] LCMS: EC15303-1-p1b, P1 : RT = 0.311 min, MS (M+H + ) = 557.2

[0292] 5. Synthesis of compound APQ4

[0293] A mixture of compound Q4-5 (130 mg, 233 μmol, 1.00 equiv), compound Q2-5 (96.3 mg, 233 μmol, 1.00 equiv) and TEA (189 mg, 1.87 mmol, 260 μL, 8.00 equiv) in DCM (2.00 mL) was stirred at 20 °C for 15 min, then NaBH(OAc)3 (148 mg, 700 μmol, 3.00 equiv) was added to the mixture, and the reaction was continued to stir at 20 °C for 1.75 h under N2. LC-MS (EC15303-3-P1A, P1: RT = 0.313 min) detection showed that the compound Q4-5 reaction was complete, and multiple new compound peaks were generated, with the target m / z product accounting for about 53.7%. The reaction mixture was concentrated under reduced pressure and filtered to obtain the product crude, which was further purified by preparative HPLC (FA condition: column: Welch Ultimate C18 150*25mm*5μm; mobile phase: [water (FA)-ACN]; gradient elution: 10%-40% B within 11 min) to obtain compound APQ4 (11.0 mg, 11.3 μmol, 4.86% yield, 98.3% purity) as a white solid.

[0294] LCMS: EC15303-3-P1A, P1: RT = 0.313 min, MS (M+H + ) = 477.7

[0295] 1H NMR: EC15303-3-p1a (400 MHz, DMSO-d6) δ 11.09 (s, 1H), 8.17 (s, 1H), 8.03 (d, J = 7.6 Hz, 1H), 7.79 (d, J = 8.2 Hz, 1H), 7.55-7.63 (m, 2H), 7.44-7.51 (m, 2H), 7.38-7.42 (m, 1H), 7.31-7.38 (m, 2H), 7.08 (d, J = 8.8 Hz, 1H), 7.02 (d, J = 7.2 Hz, 1H), 6.84 (d, J = 8.8 Hz, 1H), 6.75 (d, J = 3.2 Hz, 1H), 6.40-6.64 (m, 1H), 5.06 (dd, J = 12.8, 5.2 Hz, 1H), 4.89 (s, 2H), 3.84 (t, J = 6.0 Hz, 3H), 2.97 (t, J = 5.6 Hz, 3H), 2.86 (d, J = 11.6 Hz, 3H), 2.54-2.64 (m, 8H), 2.31-2.43 (m, 7H), 2.10 (d, J = 6.8 Hz, 2H), 1.88-2.06 (m, 4H), 1.72-1.79 (m, 2H), 1.61-1.68 (m, 2H), 1.40-1.50 (m, 1H), 1.03-1.15 (m, 2H)

[0296] Example 3. Synthesis of compound C1 as a targeting ligand and PROTAC molecule recruiting VHL-E3 enzyme or IAP-E3 enzyme (APQ3 and APL-1 to APL-6)

[0297] a) Synthesis of PROTAC molecule containing VHL-E3 enzyme recruiting ligand structure: APQ3

[0298] 1. Synthesis of compound Q3-2

[0299] To a mixture of compound 1A (1.00 g, 6.28 mmol, 1.00 equiv) and 3-bromomethyl methyl ester (1.05 g, 6.28 mmol, 687 μL, 1.00 equiv) in MeCN (15.0 mL) was added K2CO3 (2.60 g, 18.8 mmol, 3.00 equiv) and the mixture was stirred at 80 °C for 8 h. TLC (DCM / MeOH = 10:1, R1: Rf = 0.10, P1: Rf = 0.75) detection showed that compound 1A was completely reacted to generate several new products. The reaction mixture was concentrated by filtration under reduced pressure to obtain the product crude, which was purified by column chromatography (SiO2, DCM / MeOH = 100 / 1 to 0 / 1, P1: Rf = 0.75 to 0.10) to obtain compound 1B (1.00 g, 5.28 mmol, 83.6% yield) as a white solid. f f ​= 0.75), to give compound Q3-2 (1.15 g, 4.69 mmol, 74.6% yield) as a white solid.

[0300] 1 H NMR: EC12156-18-p1a1, DMSO, 400 MHZ δ 4.01-4.04 (m, 1H), 3.58 (s, 3H), 3.23 (s, 6H), 2.80 (d, J = 11.2 Hz, 2H), 2.50-2.54 (m, 2H), 2.44 (d, J = 6.0 Hz, 2H), 1.99 (s, 2H), 1.83 (td, J = 11.6, 2.0 Hz, 2H), 1.56 (d, J = 13.2 Hz, 2H), 1.50-1.46 (m, 1H), 1.13-1.18 (m, 3H)

[0301] 2. Synthesis of compound Q3-3

[0302] To a mixture of compound Q3-2 (1.10 g, 4.48 mmol, 1.00 equiv) in THF (4.00 mL) was added LiOH H2O (564 mg, 13.4 mmol, 3.00 equiv) and H2O (4.00 mL), and the mixture was stirred at 25 °C for 4 h. TLC (DCM:MeOH = 10:1, R1:R f = 0.63, P1:R f = 0.17) showed that the reaction of compound Q3-2 was complete to generate a single main peak compound. The reaction mixture was concentrated under reduced pressure to remove the solvent, and compound Q3-3 crude (2.20 g) was obtained as a white solid without purification.

[0303] 1 H NMR: EC12156-19-p1a1, D2O, 400 MHZ δ 4.16-4.26 (m, 1H), 3.36 (s, 6H), 2.89 (d, J = 11.6 Hz, 2H), 2.53-2.65 (m, 2H), 2.33 (dd, J = 8.8, 6.8 Hz, 2H), 2.01 (t, J = 11.2 Hz, 2H), 1.66-1.71 (m, 2H), 1.58-1.64 (m, 1H), 1.15-1.34 (m, 2H)

[0304] 3. Synthesis of compound Q3-4

[0305] To a mixture of compound 2 (384 mg, 864 pmol, 1.00 equiv) and compound Q3-3 (200 mg, 864.72 pmol, 1.00 equiv) in CH3CN (3.00 mL) was added HOBt (116 mg, 864 pmol, 1.00 equiv), Et3N (350 mg, 3.46 mmol, 481 pL, 4.00 equiv) and BTU (327 mg, 864 pmol, 1.00 equiv), and the compound was stirred at 25 °C for 8 h. LCMS (EC12156-24-p1a2, P1: RT = 0.299 min) detection showed that compound Q3-3 was completely reacted to generate a main peak of target m / z (m / z = 658.6 [M+H] + ), with a proportion of about 61.5%. The reaction mixture was concentrated under reduced pressure to remove the solvent, and further purified by preparative TLC (SiO2, DCM:MeOH = 7:1, P1: R f = 0.23) to obtain compound Q3-4 (179 mg, 272 pmol, 31.4% yield) as a white solid.

[0306] LCMS: EC12156-24-p1a2, P1: RT = 0.299 min, MS (ESI) m / z = 658.6 [M+H] +

[0307] 4. Synthesis of compound Q3-5

[0308] To compound Q3-4 (50.0 mg, 76.0 pmol, 1.00 equiv) was added H2SO4 (14.9 mg, 152 pmol, 8.10 pL, 2.00 equiv) and THF (2.00 mL), and the mixture was stirred at 60 °C for 4 h. LCMS (EC12156-30-p1a1, P1: RT = 0.298 min) detection showed that compound Q3-4 was completely reacted to generate a new main peak, with a proportion of about 89.4%, m / z consistent with the target product (m / z = 612.5 [M+H] + ). The reaction mixture was diluted with 15.0 mL of water, extracted with DCM 300 mL (100 mL x 3 times), and the obtained organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to obtain compound Q3-5 product crude (61.0 mg) as a yellow solid.

[0309] LCMS: EC12156-30-p1a1, P1: RT = 0.298 min, MS (ESI) m / z = 612.5 [M+H] +

[0310] 1H NMR: EC12156-30-p1a1, CDC13, 400 MHz δ 9.68 (s, 1H), 9.17 (d, J = 7.4 Hz, 1H), 8.65-8.69 (m, 1H), 7.64 (d, J = 7.8 Hz, 1H), 7.39 (q, J = 8.2 Hz, 4H), 5.30 (s, 1H), 5.07 (m, J = 7.2 Hz, 1H), 4.77 (t, J = 7.6 Hz, 1H), 4.48 (s, 1H), 4.41 (d, J = 7.6 Hz, 1H), 4.16 (d, J = 10.4 Hz, 1H), 3.56 (dd, J = 11.2, 3.6 Hz, 1H), 2.87-3.00 (m, 2H), 2.59-2.63 (m, 2H), 2.53 (s, 3H), 2.37-2.42 (m, 2H), 2.27-2.37 (m, 2H), 2.14-2.24 (m, 2H), 1.94-2.10 (m, 4H), 1.87-1.92 (m, 1H), 1.66-1.86 (m, 5H), 1.46 (d, J = 6.8 Hz, 3H), 1.06 (s, 7H), 0.87 (d, J = 10.4 Hz, 5H)

[0311] 5. Synthesis of compound APQ3

[0312] To a mixture of compound C1 (50.6 mg, 71.9 pmol, 1.00 equiv) in DCM (3.00 mL) was added NaBH(OAc)3 (45.7 mg, 215 pmol, 3.00 equiv), compound Q3-5 (44.0 mg, 71.9 pmol, 1.00 equiv) and TEA (21.8 mg, 215 pmol, 30.0 pL, 3.00 equiv), the mixture was stirred at 25 °C for 4 h. LCMS (EC12156-36-p1a3, P1: RT = 0.418 min) detection showed that the reaction of compound Q3-5 was complete, a new main peak was generated, accounting for about 81.2%, m / z consistent with the target product (m / z = 1300.7 [M+H] + ). The reaction mixture was terminated by adding water 5.0 mL at 25 °C, filtered and concentrated under reduced pressure to obtain the product crude, which was further purified by preparative HPLC to obtain compound APQ3 (6.12 mg, 4.29 pmol, 5.96% yield, 91.1% purity) as a white solid.

[0313] LCMS: EC12156-36-p1a3, P1: RT = 0.418 min, MS (ESI) m / z = 1300.7 [M+H] +

[0314] HPLC: EC12156-36-p1a10, RT = 1.899 min, 91.1% purity.

[0315] 1 H NMR: EC12156-36-p1a2, DMSO, 400 MHZ δ 8.97 (s, 1H), 8.57-8.67 (m, 1H), 8.38 (d, J = 8.0 Hz, 1H), 8.03 (d, J = 7.4 Hz, 1H), 7.78 (d, J = 8.4 Hz, 1H), 7.60 (d, J = 7.2 Hz, 1H), 7.31-7.51 (m, 10H), 7.26 (dd, J = 12.0, 2.0 Hz, 1H), 7.15-7.20 (m, 1H), 7.09 (t, J = 8.8 Hz, 1H), 6.89 (s, 1H), 4.86-4.96 (m, 3H), 4.47-4.56 (m, 1H), 4.36-4.47 (m, 1H), 4.27 (d, J = 1.6 Hz, 1H), 4.01 (t, J = 6.8 Hz, 2H), 3.8 (t, J = 5.6 Hz, 2H), 3.55-3.65 (m, 3H), 3.44 (s, 4H), 2.97 (t, J = 5.4 Hz, 3H), 2.72-2.81 (m, 2H), 2.64-2.70 (m, 2H), 2.45 (s, 6H), 2.29-2.36 (m, 6H), 2.09 (dd, J = 7.2, 2.8 Hz, 2H), 1.88-2.03 (m, 6H), 1.73-1.85 (m, 3H), 1.60-1.70 (m, 3H), 1.43-1.52 (m, 2H), 1.37 (d, J = 6.8 Hz, 3H), 1.23 (s, 1H), 1.04-1.19 (m, 3H), 0.94 (s, 9H).

[0316] b) Synthesis of PROTAC molecules containing IAP-E3 enzyme recruiting structures: APL-1, APL-2, APL-3, APL-4, APL-5, APL-6

[0317] Synthesis of APL-1:

[0318] 1. Synthesis of L1-2

[0319] A mixture of 2-iodoethanol (137 mg, 801 μmol, 62.6 μL, 2.4 eq), compound 3 (200 mg, 334 μmol, 1.0 eq) and K2CO3(46.1 mg, 334 μmol, 1.0 eq) in DMF (2.0 mL) was stirred at 70 °C for 48 h under N2. LC-MS (EC16364-17-P1A, P1, RT = 0.432 min) monitoring showed compound 3 was consumed completely. The reaction mixture was quenched by the addition of water (20.0 mL) at 0 °C, diluted with ethyl acetate (20.0 mL), and extracted with ethyl acetate (60.0 mL, 20.0 mL x 3 times). The resulting organic layer was washed with saturated brine (20.0 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the L1-2 product crude (140 mg, 217 μmol, 65.2% yield) as a white oil, which was used directly in the next step without further purification.

[0320] LC-MS: EC16364-17-P1A, product: RT = 0.432 min, m / z = 643.3 [M+H]+

[0321] 2. Synthesis of L1-3

[0322] To a solution of compound L1-2 (140 mg, 217 μmol, 1.0 eq) in DCM (3.0 mL) was added TsCl (83.0 mg, 435 μmol, 2.0 eq) and Et3N (66.1 mg, 653 μmol, 90.9 μL, 3.0 eq), and the mixture was stirred at 25 °C for 12 h. LC-MS (EC16364-18-P1A1, P1, RT = 0.495 min) monitoring showed compound L1-2 was consumed completely. The reaction mixture was quenched by the addition of water (20.0 mL) at 0 °C, diluted with DCM (20.0 mL), and extracted with DCM (60.0 mL, 20.0 mL x 3 times). The resulting organic layer was washed with saturated brine (20.0 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the L1-3 product crude (170 mg, 213 μmol, 97.9% yield) as a white oil, which was used directly in the next step without further purification.

[0323] LC-MS: EC16364-18-P1A1, product: RT = 0.495 min, m / z = 797.5 [M+H] +

[0324] 3. Synthesis of L1-4

[0325] To a solution of compound L1-3 (100 mg, 125 pmol, 1.0 eq) in DCM (2.0 mL) was added Et3N (63.5 mg, 627 pmol, 87.3 pL, 5.0 eq) and compound 1A (99.9 mg, 627 pmol, 5.0 eq), the mixture was stirred at 85 °C for 24 h. LCMS (EC16364-42-p1a, P1, RT = 0.303 min) monitoring showed compound L1-3 was completely reacted. After reaction, the mixture was quenched by water (20.0 mL) at 0 °C, diluted with ethyl acetate (20.0 mL), extracted with ethyl acetate (60.0 mL, 20.0 mL x 3 times). The resulting organic layer was washed with saturated brine (20.0 mL), dried over Na2S04, filtered and concentrated under reduced pressure to give L1-4 product crude, which was further purified by preparative TLC (Si02, petroleum ether: ethyl acetate = 0: 1, P1, Rf = 0.3) to give product L1-4 (20.0 mg, 25.5 pmol, 20.3% yield) as a white solid. f

[0326] LC-MS: EC16364-42-p1a, product: RT = 0.303 min, m / z = 684.4 [M+H] +

[0327] 4. Synthesis of L1-5

[0328] Compound L1-4 (20 mg, 25.51 pmol, 1.0 eq) was added to formic acid (2 mL), the mixture was stirred at 25 °C for 2 h under N2. TLC (petroleum ether: ethyl acetate = 0: 1) monitoring showed L1-4 was completely reacted, a more polar main peak was generated. After reaction, the mixture was concentrated under reduced pressure to give L1-5 product crude (15 mg, 20.3 pmol, 79.6% yield) as a yellow oil, the product was used directly in the next step without purification.

[0329] 5. Synthesis of APL-1

[0330] ​To a solution of compound L1-5 (15.0 mg, 20.3 pmol, 1.0 eq) in DCM (1 mL) was added TEA (20.6 mg, 203 pmol, 28.3 pL, 10.0 eq) and compound C1 (14.3 mg, 20.3 pmol, 1.0 eq), the mixture was stirred at 25 °C for 0.5 h, NaBH(OAc)3 (12.9 mg, 61.0 pmol, 3.0 eq) was added, stirred at 25 °C for 3.5 h. LC-MS (EC16364-45-P1A, P1, RT = 0.381 min) monitoring showed that L1-5 reaction was complete. The reaction mixture was concentrated under reduced pressure to give the product crude, which was further purified by preparative HPLC column (FA condition) to give compound APL-1 (10 mg, 7.34 pmol, 36.12% yield, 97.4 purity) as a white solid.

[0331] LC-MS: EC16364-45-P1A, product: RT = 0.381 min, m / z = 1326.8 [M+H] +

[0332] 1 H NMR: EC16364-45-P1D (400 MHz DMSO) d ppm 8.51 (s, 1H) 8.22 (s, 1H) 8.09 (d, J = 8.8 Hz, 1H) 8.00 - 8.06 (m, 1H) 7.79 (d, J = 8.0 Hz, 1H) 7.71 (s, 1H) 7.57 - 7.66 (m, 2H) 7.42 - 7.50 (m, 3H) 7.38 - 7.42 (m, 1H) 7.34 (td, J = 7.2, 3.38 Hz, 2H) 7.22 - 7.29 (m, 2H) 7.18 (d, J = 9.2 Hz, 1H) 7.05 - 7.14 (m, 1H) 6.84 (d, J = 9.2 Hz, 1H) 5.39 (dd, J = 7.6, 2.81 Hz, 1H) 4.90 (s, 2H) 4.47 (t, J = 7.6 Hz, 1H) 4.13 t, J = 5.2 Hz, 2H) 4.01 (t, J = 6.4 Hz, 2H) 3.78 - 3.85 (m, 4H) 3.46 (s, 2H) 3.17 (d, J = 6.8 Hz, 2H) 2.97 (t, J = 5.2 Hz, 2H) 2.91 (d, J = 10.0 Hz, 2H) 2.69 - 2.77 (m, 4H) 2.23 - 2.43 (m, 6H) 2.22 (s, 3H) 1.93 - 2.13 (m, 10H) 1.58 - 1.69 (m, 6H) 1.51 (d, J = 7.6 Hz, 2H) 1.46 (s, 1H) 1.15 (d, J = 6.8 Hz, 3H) 0.89 - 1.11 (m, 8H).

[0333] Synthesis of APL-2:

[0334] 1. Synthesis of compound L2-1

[0335] To a solution of compound 3 (280 mg, 468 pmol, 1.0 eq) in DCM (10.0 mL) was added PhNTf2 (267 mg, 748 pmol, 1.6 eq), TEA (94.6 mg, 935 pmol, 130 pL, 2.0 eq) and DMAP (5.71 mg, 46.8 pmol, 0.1 eq), the mixture was stirred at 25 °C for 16 h. After reaction, the mixture was concentrated under reduced pressure to remove the solvent to give the product crude, which was further purified by reverse phase HPLC (0.1% FA condition) to give compound L2-1 (300 mg, about 98% purity) as a white solid.

[0336] LC-MS: EC16331-27-P1A, product: RT = 0.568 min, m / z = 731.3 [M+H] +

[0337] 2. Synthesis of compound L2-2

[0338] Compound L2-1 (160 mg, 219 pmol, 1.0 eq), compound 1A (174 mg, 1.09 mmol, 5.0 eq), Cs2CO3 (214 mg, 657 pmol, 3.0 eq) and XPhos PdG4 (18.8 mg, 21.9 pmol, 0.1 eq) were mixed in dioxane (3.00 mL), stirred at 90 °C for 2 h under N2. After reaction, the mixture was liquid-liquid partitioned by adding water (10.0 mL) and DCM (60.0 mL, 10.0 mL x 6 times), the obtained organic layer was washed with saturated brine (60.0 mL, 30.0 mL x 2 times), dried over Na2SO4, filtered, concentrated under reduced pressure to give the product crude. The crude was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 5: 1) to give compound L2-2 (120 mg, 129.57 pmol, 59.18% yield, 79.9% purity) as a yellow oil.

[0339] LC-MS: EC16331-11-P1A, product: RT = 0.501 min, m / z = 740.3 [M+H] +

[0340] 1H NMR: EC16331-11-P1A (400 MHz CDC13) δ ppm 8.10 (s, 1H), 7.74 (d, J = 1.6 Hz, 1H), 7.56-7.66 (m, 1H), 7.34 (d, J = 1.2 Hz, 1H), 7.13-7.21 (m, 1H), 5.56 (dd, J = 7.6, 2.4 Hz, 1H), 4.64 (dd, J = 8.8, 6.4 Hz, 2H), 3.86-3.91 (m, 1H), 3.78 (d, J = 11.6 Hz, 2H), 3.39 (s, 6H), 2.81 (s, 3H), 2.75 (d, J = 8.0 Hz, 2H), 2.50 (dt, J = 6.4, 3.2 Hz, 1H), 2.27-2.32 (m, 1H), 2.18 (d, J = 7.2 Hz, 1H), 2.09-2.13 (m, 1H), 1.85-1.90 (m, 2H), 1.71 (s, 6H), 1.58 (s, 9H), 1.49 (s, 11H), 1.34 (d, J = 7.2 Hz, 3H).

[0341] 3. Synthesis of compound L2-3

[0342] Compound L2-2 (50.0 mg, 67.6 μmol, 1.0 eq) was mixed with formic acid (0.5 mL) and stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure to remove the solvent to give the crude product of compound L2-3 (30.0 mg) as a yellow oil, which was used directly in the next step without further purification.

[0343] LC-MS: EC16331-15-P1A, product: RT = 0.322 min, m / z = 594.3 [M+H] +

[0344] 4. Synthesis of compound APL-2

[0345] Compound L2-3 (30.0 mg, 50.5 μmol, 1.0 eq), CI (35.6 mg, 50.5 μmol, 1.0 eq) and NaBH(OAc)3 (32.1 mg, 152 μmol, 3.0 eq) were mixed in DMF (0.5 mL) and stirred at 25 °C for 2 h under N2 protection. The reaction mixture was filtered and concentrated under reduced pressure to give the crude product, which was further purified by reverse phase HPLC (0.1% FA condition) to give compound APL-2 (10.0 mg, 7.61 μmol, 15.06% yield, 97.61% purity) as a yellow solid.

[0346] LC-MS: EC16331-16-P1A, Product: RT = 0.420 min, m / z = 1283.5 [M+H] +

[0347] 1 H NMR: EC16331-16-P1C (400 MHz DMSO) δ ppm 8.44 (s, 1 H), 8.11 (d, J = 8.4 Hz, 1 H), 8.02 (d, J = 7.6 Hz, 1 H), 7.75-7.82 (m, 1 H), 7.69 (s, 1 H), 7.60 (d, J = 7.13 Hz, 1 H), 7.44-7.50 (m, 2 H), 7.39 (d, J = 5.2 Hz, 2 H), 7.35 (dt, J = 7.2, 3.6 Hz, 3 H), 7.14-7.30 (m, 4 H), 7.04-7.14 (m, 1 H), 6.77-6.85 (m, 1 H), 5.38 (dd, J = 6.6, 3.6 Hz, 1 H), 4.90 (s, 2 H), 4.43-4.50 (m, 2 H), 3.98-4.05 (m, 4 H), 3.83 (d, J = 5.2 Hz, 6 H), 3.70-3.79 (m, 10 H), 3.20 (s, 2 H), 2.97 (d, J = 4.8 Hz, 2 H), 2.71-2.76 (m, 4 H), 2.37-2.42 (m, 2 H), 2.23 (s, 4 H), 2.15 (d, J = 6.13 Hz, 2 H), 2.00 (dd, J = 16.8, 6.8 Hz, 5 H), 1.78 (d, J = 10.4 Hz, 2 H), 1.68 (d, J = 3.2 Hz, 2 H), 1.61 (s, 2 H), 1.54 (d, J = 9.6 Hz, 2 H), 1.24 (s, 2 H), 1.15 (d, J = 6.8 Hz, 2 H), 1.06 (t, J = 6.8 Hz, 3 H).

[0348] Synthesis of APL-3 and APL-4:

[0349] Synthesis of APL-3:

[0350] 1. Synthesis of L3-2

[0351] To a solution of compound 3 (100 mg, 167 μmol, 1.0 eq) in MeCN (1 mL) was added K2CO3 (69.2 mg, 501 μmol, 3.0 eq) at 25 °C, stirred for 30 min at 25 °C, then 1,4-dibromobutane (180 mg, 835 μmol, 100 μL, 5.0 eq) was added dropwise at 25 °C, the mixture was stirred at 80 °C for 12 h. After reaction, the mixture was quenched by water (20.0 mL) at 0 °C, diluted with ethyl acetate (20.0 mL), extracted with ethyl acetate (60.0 mL, 20.0 mL x 3 times). The organic layer was washed with saturated brine (20.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product L3-2, which was further purified by preparative TLC (SiO2, petroleum ether: ethyl acetate = 1:1, P1, Rf = 0.3) to give the product L3-2 (100 mg, 136 μmol, 40.8% yield, 100% purity) as a white solid. f

[0352] LC-MS: EC16364-66-P1A, product: RT = 0.591 min, m / z = 733.4 [M+H] +

[0353] 2. Synthesis of L3-3

[0354] Compound C1 (76.8 mg, 109 μmol, 1.0 eq), L3-2 (80.0 mg, 109 μmol, 1.0 eq) and DIEA (42.3 mg, 327 μmol, 56.9 μL, 3.0 eq) were mixed in DMF (1.0 mL), stirred at 80 °C for 6 h under N2. LC-MS (EC16364-70-P1A1, P1, RT = 0.504 min) monitoring showed that L3-2 was completely reacted. After reaction, the mixture was concentrated under reduced pressure to give the crude product, which was further purified by preparative HPLC column chromatography (FA condition) to give the product L3-3 (30.0 mg, 22.1 μmol, 20.2% yield) as a white solid.

[0355] LC-MS: EC16364-70-P1A1, product: RT = 0.504 min, m / z = 1357.6 [M+H] +

[0356] 3. Synthesis of APL-3

[0357] ​Compound L3-3 (30.0 mg, 22.1 μmol, 1.0 equiv) and HC1 / dioxane (4 M, 5.0 mL, 905 equiv) were mixed and stirred at 25 °C for 1 h under N2. LC-MS (EC16364-80-P1B2, P1, RT = 0.428 min) monitoring showed the reaction of L3-3 was complete. The reaction mixture was concentrated under reduced pressure to give the product crude, which was further purified by preparative HPLC column (HC1 condition) to give product APL-3 (5 mg, 3.81 μmol, 17.2% yield, 95.7% purity) as a brown solid.

[0358] LC-MS: EC16364-80-P1B2, product: RT = 0.428 min, m / z = 1279.6 [M+H] +

[0359] 1 H NMR: EC16364-80-P1D (400 MHz DMSO) δ ppm 9.30 (s, 1H) 9.07 (s, 1H) 8.87 (s, 1H) 8.75 (d, J = 7.6 Hz, 1H) 8.49 (s, 1H) 8.04 (d, J = 8.0 Hz, 1H) 7.79 (d, J = 8.0 Hz, 1H) 7.68 (d, J = 7.6 Hz, 1H) 7.53 - 7.63 (m, 4H) 7.47 (br t, J = 7.6 Hz, 3H) 7.39 - 7.43 (m, 1H) 7.30 - 7.39 (m, 4H) 7.22 - 7.27 (m, 2H) 7.09 - 7.15 (m, 1H) 6.95 - 7.00 (m, 1H) 5.35 - 5.41 (m, 1H) 4.92 (s, 2H) 4.47 (t, J = 7.6 Hz, 1H) 4.18 (s, 1H) 4.01 - 4.10 (m, 6H) 3.86 (t, J = 5.6 Hz, 4H) 3.76 - 3.82 (m, 4H) 2.98 (t, J = 5.2 Hz, 2H) 2.81 (d, J = 7.2 Hz, 2H) 2.32 (t, J = 4.8 Hz, 1H) 2.13 - 2.29 (m, 3H) 1.94 - 2.06 (m, 5H) 1.87 - 1.92 (m, 2H) 1.82 (d, J = 6.4 Hz, 2H) 1.59 - 1.72 (m, 5H) 1.52 - 1.58 (m, 3H) 1.35 (d, J = 6.8 Hz, 3H) 0.99 - 1.15 (m, 6H).

[0360] Synthesis of APL-4:

[0361] The synthesis of PROTAC molecule APL-4 was referred to APL-3, but the starting material was changed to compound 3 and L4-1. The product was purified by preparative HPLC column (HCl condition) to give the target product APL-4 (5 mg, 3.59 pmol, 32.8% yield, 91.3% purity) as a white solid.

[0362] LC-MS: EC16364-88-P1B2, product: RT = 0.427 min, m / z = 1293.6 [M+Na] +

[0363] 1 H NMR: EC16364-88-p1c (400 MHz DMSO) δ ppm 9.18 (s, 1H) 8.84 (s, 1H) 8.73-8.73 (m, 1H) 8.74 (d, J = 8.0 Hz, 1H) 8.49 (s, 1H) 8.04 (d, J = 8.0 Hz, 1H) 7.79 (d, J = 8.0 Hz, 1H) 7.68 (d, J = 7.6 Hz, 1H) 7.54-7.62 (m, 4H) 7.47 (td, J = 7.6, 4.13 Hz, 3H) 7.41 (d, J = 7.2 Hz, 2H) 7.36 (d, J = 6.0 Hz, 2H) 7.32 (d, J = 9.6 Hz, 1H) 7.23-7.29 (m, 2H) 7.17-7.22 (m, 1H) 7.13 (t, J = 8.8 Hz, 1H) 6.97 (d, J = 8.8 Hz, 1H) 5.39 (dd, J = 7.6, 2.69 Hz, 1H) 4.92 (s, 2H) 4.47 (t, J = 7.6 Hz, 1H) 4.05 (d, J = 9.2 Hz, 4H) 3.86 (d, J = 5.2 Hz, 4H) 3.81 (dd, J = 6.4, 2.25 Hz, 2H) 3.10-3.13 (m, 2H) 2.98 (t, J = 5.2 Hz, 2H) 2.78-2.83 (m, 2H) 2.54 (s, 2H) 2.17-2.25 (m, 2H) 2.02-2.08 (m, 2H) 1.95-2.01 (m, 2H) 1.75-1.80 (m, 4H) 1.61-1.70 (m, 4H) 1.53-1.58 (m, 2H) 1.45-1.51 (m, 2H) 1.34 (d, J = 6.8 Hz, 3H) 1.16-1.25 (m, 2H) 1.12 (d, J = 7.2 Hz, 2H) 1.07 (s, 1H) 1.05 (s, 2H) 1.03-1.04 (m, 1H).

[0364] Synthesis of APL-5 and APL-6:

[0365] Synthesis of APL-5:

[0366] 1. Synthesis of L5-2

[0367] To a solution of compound 3 (100 mg, 167 pmol, 1.0 eq) in MeCN (3 mL) was added K2CO3 (69.2 mg, 501 pmol, 3.0 eq) at 25 °C, stirred for 30 min, then L5-1 (230 mg, 835 pmol, 5.0 eq) was added dropwise at 25 °C, the mixture was stirred at 80 °C for 12 h. LC-MS (EC16364-78-P1A, P1, RT = 0.495 min) monitoring showed compound 3 was completely reacted. After reaction, the mixture was quenched by water (20.0 mL) at 0 °C, diluted with ethyl acetate (20.0 mL), extracted with ethyl acetate (60.0 mL, 20.0 mL x 3 times). The resulting organic layer was washed with saturated brine (20.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product of L5-2, which was further purified by preparative TLC (SiO2, petroleum ether: ethyl acetate = 1:2, P1, Rf = 0.3) to give the product L5-2 (90.0 mg, 113 pmol, 67.9% yield, 100% purity) as a white solid. f

[0368] LC-MS: EC16364-78-P1A, product: RT = 0.495 min, m / z = 793.3 [M+H] +

[0369] 2. Synthesis of L5-3

[0370] Compound L5-2 (50.0 mg, 63.0 pmol, 1.0 eq), CI (35.5 mg, 50.4 pmol, 0.8 eq) and DIEA (24.4 mg, 188 pmol, 32.9 pL, 3.0 eq) were mixed in DMF (1.0 mL), stirred at 80 °C under N2for 6 h. LC-MS (EC16364-91-P1A1, P1, RT = 0.500 min) monitoring showed L5-2 was completely reacted. After reaction, the mixture was concentrated under reduced pressure to give the crude product, which was further purified by preparative HPLC column chromatography (FA condition) to give the product L5-3 (20.0 mg, 14.1 pmol, 22.4% yield) as a white solid.

[0371] ​LC-MS: EC16364-91-P1A1, product: RT = 0.500 min, m / z = 1418.6 [M+H] +

[0372] 3. Synthesis of APL-5

[0373] Compound L5-3 (20.0 mg, 14.1 pmol, 1.0 eq) and HCl / ethyl acetate (4 M, 3.53 pL) were mixed and stirred at 25 °C for 1 h under N2. LC-MS (EC16364-100-P1A1, P1, RT = 0.413 min) monitoring showed the reaction of L5-3 was complete. The reaction mixture was concentrated under reduced pressure to give the crude product, which was further purified by preparative HPLC column (HCl condition) to give the product APL-5 (13.1 mg, 9.07 pmol, 64.2% yield, 91.9% purity) as a brown solid.

[0374] LC-MS: EC16364-100-P1A1, product: RT = 0.413 min, m / z = 659.7 [M / 2+H] +

[0375] 1H NMR: EC16364-100-P1C (400 MHz DMSO) δ ppm 9.26 (s, 1 H) 8.80-8.93 (m, 1 H) 8.74 (d, J=8.0 Hz, 1 H) 8.46-8.51 (m, 1 H) 8.04 (d, J=7.6 Hz, 1 H) 7.79 (d, J=7.6 Hz, 1 H) 7.68 (d, J=7.6 Hz, 1 H) 7.54-7.62 (m, 3 H) 7.39-7.50 (m, 4 H) 7.31-7.39 (m, 3 H) 7.26 (dd, J=8.0, 2.63 Hz, 2 H) 7.12 (t, J=8.8 Hz, 1 H) 6.97 (d, J=9.2 Hz, 1 H) 5.38 (dd, J=7.6, 3.31 Hz, 1 H) 5.32 (t, J=4.4 Hz, 1 H) 4.92 (s, 2 H) 4.47 (t, J=7.6 Hz, 1 H) 4.15-4.19 (m, 2 H) 4.03 (t, J=6.4 Hz, 2 H) 3.85 (dd, J=11.6, 5.50 Hz, 6 H) 3.77-3.80 (m, 4 H) 3.33 (s, 4 H) 2.98 (t, J=5.6 Hz, 2 H) 2.78-2.83 (m, 2 H) 2.47 (s, 2 H) 2.46 (s, 1 H) 2.15-2.24 (m, 2 H) 2.00 (dd, J=15.2, 7.63 Hz, 8 H) 1.60-1.70 (m, 4 H) 1.56 (d, J=8.4 Hz, 2 H) 1.41-1.49 (m, 2 H) 1.34 (d, J=6.8 Hz, 3 H) 1.03-1.16 (m, 6 H) 0.83-0.88 (m, 2 H).

[0376] Synthesis of APL-6:

[0377] The synthesis of PROTAC molecule APL-6 followed the procedure of APL-5, but with compound 3 and L6-1 as starting materials. The desired product APL-6 (11.0 mg, 7.32 pmol, 53.5% yield, 90.6% purity) was obtained as a white solid after purification.

[0378] LC-MS: EC16364-101-P1A1, product: RT = 0.418 min, m / z = 1384.5 [M+Na] +

[0379] 1H NMR: EC16364-101-P1C (400 MHz DMSO) δ ppm 9.24-9.36 (m, 1 H) 8.80-8.94 (m, 1 H) 8.48 (s, 1 H) 8.04 (d, J=7.6 Hz, 1 H) 7.79 (d, J=7.6 Hz, 1 H) 7.68 (d, J=7.6 Hz, 1 H) 7.54-7.62 (m, 3 H) 7.40-7.50 (m, 4 H) 7.33-7.38 (m, 2 H) 7.23-7.31 (m, 2 H) 7.13 (t, J=8.8 Hz, 1 H) 6.97 (d, J=8.8 Hz, 1 H) 5.38 (dd, J=7.6, 3.06 Hz, 1 H) 5.32 (t, J=4.8 Hz, 1 H) 4.92 (s, 2 H) 4.47 (t, J=7.6 Hz, 1 H) 4.14-4.17 (m, 2 H) 4.01-4.04 (m, 2 H) 3.87 (d, J=5.6 Hz, 4 H) 3.82 (d, J=4.4 Hz, 4 H) 3.76 (br s, 4 H) 3.59-3.61 (m, 4 H) 3.57 (s, 4 H) 3.34 (d, J=1.2 Hz, 2 H) 2.98 (t, J=5.6 Hz, 2 H) 2.80 (t, J=7.2 Hz, 2 H) 2.46 (s, 1 H) 2.33 (s, 3 H) 2.19-2.23 (m, 1 H) 2.02 (d, J=7.2 Hz, 2 H) 1.98 (d, J=7.6 Hz, 2 H) 1.64 (d, J=8.0 Hz, 2 H) 1.55 (dd, J=8.0, 1.69 Hz, 2 H) 1.43-1.48 (m, 1 H) 1.35 (d, J=6.8 Hz, 3 H) 1.23 (s, 8 H) 1.08 (d, J=6.4 Hz, 2 H) 0.83-0.86 (m, 1 H).

[0380] Example 4. Synthesis of PROTAC molecules based on compound A1331852 as targeting ligand and CRBN-E3 enzyme (APF1, APF2, APF3, APF4, APF1A, APR1, APR2, APR3, APR4)

[0381] a) Synthesis of PROTAC molecules APF1, APF2, APF3, APF4, APF1A

[0382] Synthesis of APF1:

[0383] 1. Synthesis of compound APF1-2

[0384] A solution of compound 1 (200 mg, 724 pmol, 1.00 equiv) and compound APF1-1 (177 mg, 869 pmol, 1.20 equiv) in DMSO (2.00 mL) was added with DIEA (281 mg, 2.17 mmol, 378 pL, 3.00 equiv), the mixture was stirred at 75 °C for 12 h. LC-MS (EC11255-27-P1A, P1 RT = 0.357 min) detection showed compound 1 reacted completely, produced multiple new compounds, about 56.0% target compound was detected. The mixture was extracted with ethyl acetate 45 mL (15.0 mL x 3 times), the resulting organic layer was washed with brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (SiO2, PE:EA = 1 / 0 to 1 / 5), the resulting product was dissolved in 5 mL hydrochloric acid / dioxane, N2 was filled to expel air, stirred at 25 °C for 2 h. The reaction solution was concentrated under vacuum to give the crude product, compound APF1-2 (160 mg, 347 pmol, 48.0% yield) was a light yellow solid.

[0385] LCMS: EC11255-27-P1A, P1 RT = 0.357 min, MS (ESI) m / z = 361.2 [M+H] +

[0386] 2. Synthesis of compound APF1

[0387] A mixture of compound A1331852 (60.0 mg, 91.0 pmol, 1.00 equiv), APF1-2 (49.2 mg, 136 pmol, 1.50 equiv), HATU (51.9 mg, 136 pmol, 1.50 equiv) and DIEA (58.8 mg, 455 pmol, 79.3 pL, 5.00 equiv) in DCM (1 mL) was filled with N2 to expel air, stirred at 25 °C for 2 h. LC-MS (EC11272-23-P1B) detection showed that compound A1331852 reacted completely, the reaction solution was concentrated under vacuum to give the crude product, which was further purified by preparative HPLC (basic conditions) to give compound APF1 (21.6 mg, 20.0 pmol, 22.0% yield, 93.0% purity) as a yellow solid.

[0388] LCMS: EC11272-23-P1B, RT = 0.569 min, M / Z (M+H + ) = 1001.3

[0389] LCMS: EC11272-23-P1C, RT = 0.563 min, M / Z (M+H+ ) = 1001.3

[0390] HNMR: EC11272-23-P1C, DMSO + D20, 400 MHz δ 11.08 (s, 1H), 8.14 (t, J = 5.69 Hz, 1H), 7.99 (d, J = 7.6 Hz, 1H), 7.77 (d, J = 8.0 Hz, 1H), 7.58 (d, J = 7.6 Hz, 1H), 7.51 (t, J = 8.0 Hz, 1H), 7.42-7.48 (m, 2H), 7.29-7.41 (m, 3H), 7.25 (s, 1H), 7.06 (d, J = 8.4 Hz, 1H), 6.99 (d, J = 7.2 Hz, 1H), 6.94 (d, J = 8.8 Hz, 1H), 6.50-6.57 (m, 1H), 4.92-5.05 (m, 3H), 3.86 (t, J = 5.2 Hz, 2H), 3.64 (s, 2H), 3.38 (d, J = 6.4 Hz, 4H), 3.24 (d, J = 5.2 Hz, 2H), 2.96-3.01 (m, 2H), 2.70-2.92 (m, 2H), 2.02 (s, 3H), 1.91-2.00 (m, 2H), 1.88 (s, 3H), 1.57-1.64 (m, 3H), 1.46-1.54 (m, 9H).

[0391] Synthesis of APF2:

[0392] The synthesis steps of the PROTAC molecule APF2 refer to APF1, but the starting material is changed to compound 1 and APF2-1, and the target product APF2 is obtained after purification, which is a yellow solid with a purity of 93.7% and a yield of about 23.9%.

[0393] LCMS: EC11272-24-P1B, RT = 0.572 min, M / Z (M+H + ) = 1045.3

[0394] LCMS: EC11272-24-P1C, RT = 0.567 min, M / Z (M+H + ) = 1045.3

[0395] HNMR: EC11272-24-P1A, DMSO, 400MHz δ 11.07-11.11 (m, 1H), 8.18 (t, J = 5.6 Hz, 1H), 7.97-8.02 (m, 1H), 7.73-7.79 (m, 1H), 7.58-7.64 (m, 1H), 7.54 (t, J = 7.2 Hz, 1H), 7.41-7.48 (m, 2H), 7.29-7.41 (m, 3H), 7.25 (s, 1H), 7.08 (d, J = 8.4 Hz, 1H), 7.01 (d, J = 7.2 Hz, 1H), 6.94 (d, J = 8.8 Hz, 1H), 6.57 (t, J = 6.0 Hz, 1H), 4.94-5.08 (m, 3H), 3.89 (t, J = 6.0 Hz, 2H), 3.66 (s, 2H), 3.55-3.58 (m, 2H), 3.46-3.50 (m, 2H), 3.39-3.41 (m, 4H), 3.23-3.26 (m, 4H), 2.97-3.01 (m, 2H), 2.79-2.92 (m, 2H), 2.04 (s, 3H), 1.94-2.02 (m, 2H), 1.90 (s, 3H), 1.59-1.65 (m, 3H), 1.48-1.55 (m, 9H).

[0396] Synthesis of APF3:

[0397] The synthesis of the PROTAC molecule APF3 refers to that of APF1, but the starting material is changed to compound 1 and APF3-1. The target product APF3 is obtained after purification, which is a yellow solid with a purity of 97.4% and a yield of about 11.4%.

[0398] LCMS: EC12165-1-P1A, P1, RT = 0.513 min, M / Z (M+H + ) = 1089.7

[0399] LCMS: EC12165-1-P1C, RT = 0.570 min, M / Z (M+H + ) = 1089.2

[0400] HNMR: EC12165-1-P1C, DMSO, 400 MHz δ 12.85 (s, 1H), 11.09 (s, 1H), 8.16 (t, J = 6.0 Hz, 1H), 8.01 (d, J = 7.6 Hz, 1H), 7.78 (d, J = 8.0 Hz, 1H), 7.52-7.63 (m, 2H), 7.43-7.49 (m, 2H), 7.39-7.43 (m, 1H), 7.31-7.37 (m, 2H), 7.25 (s, 1H), 7.10 (d, J = 8.8 Hz, 1H), 7.02 (d, J = 7.2 Hz, 1H), 6.94 (d, J = 8.8 Hz, 1H), 6.58 (t, J = 5.6 Hz, 1H), 5.04 (dd, J = 12.8, 5.4 Hz, 1H), 4.97 (s, 2H), 3.89 (t, J = 6.0 Hz, 2H), 3.67 (s, 2H), 3.55-3.60 (m, 2H), 3.47-3.53 (m, 4H), 3.41-3.46 (m, 4H), 3.38 (dd, J = 6.0, 3.6 Hz, 4H), 3.22-3.26 (m, 2H), 3.00 (t, J = 5.6 Hz, 2H), 2.81-2.92 (m, 1H), 2.05 (s, 3H), 1.96-2.03 (m, 1H), 1.90 (s, 3H), 1.60-1.66 (m, 3H), 1.49-1.57 (m, 9H).

[0401] Synthesis of APF4:

[0402] The synthesis of the PROTAC molecule APF4 refers to that of APF1, but the starting material is changed to compound 1 and APF4-1. The target product APF4 is obtained after purification, which is a yellow solid with a purity of 93.4% and a yield of about 20.7%.

[0403] LCMS: EC12165-2-P1C, RT = 0.572 min, M / Z (M+Na + ) = 1155.2

[0404] HNMR: EC12165-2-P1C, DMSO, 400 MHz δ 12.84 (s, 1H), 11.09 (s, 1H), 8.17 (t, J = 5.6 Hz, 1H), 8.01 (d, J = 8.0 Hz, 1H), 7.78 (d, J = 8.0 Hz, 1H), 7.53 - 7.64 (m, 2H), 7.43 - 7.50 (m, 2H), 7.39 - 7.43 (m, 1H), 7.31 - 7.38 (m, 2H), 7.25 (s, 1H), 7.11 (d, J = 8.4 Hz, 1H), 7.02 (d, J = 7.2 Hz, 1H), 6.95 (d, J = 8.8 Hz, 1H), 6.58 (t, J = 5.6 Hz, 1H), 5.04 (dd, J = 12.8, 5.2 Hz, 1H), 4.98 (s, 2H), 3.89 (t, J = 6.0 Hz, 2H), 3.67 (s, 2H), 3.56 - 3.61 (m, 2H), 3.50 - 3.54 (m, 2H), 3.47 - 3.50 (m, 2H), 3.45 (s, 4H), 3.39 - 3.44 (m, 4H), 3.35 - 3.39 (m, 4H), 3.22 - 3.26 (m, 2H), 3.00 (t, J = 5.6 Hz, 2H), 2.87 (ddd, J = 17.2, 14.0, 5.2 Hz, 1H), 2.05 (s, 3H), 1.97 - 2.03 (m, 1H), 1.91 (s, 3H), 1.60 - 1.66 (m, 3H), 1.49 - 1.58 (m, 9H).

[0405] Synthesis of APF1A:

[0406] 1. Synthesis of compound APF1A-2

[0407] To a solution of compound 1 (500 mg, 1.81 mmol, 1.00 equiv) and compound APF1A-1 (405 mg, 2.72 mmol, 1.50 equiv) in dioxane (10.0 mL) was added DIEA (1.40 g, 10.9 mmol, 1.89 mL, 6.00 equiv), and the mixture was stirred at 100 °C for 20 h. LCMS (EC11255-19-P1a, P1 RT = 0.283 min) detection showed compound 1 was fully reacted, and multiple new compound peaks appeared, with the target compound detected at about 55.5%. The mixture was reduced in pressure to remove dioxane, diluted with 10.0 mL of water, extracted with 30 mL (10.0 mL x 3 times) of ethyl acetate, washed with 10 mL of brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (SiO2, PE:EA = 1 / 0 to 9 / 1) to give compound APF1A-2 (140 mg, 345 µmol, 19.1% yield) as a light yellow oil. LC-MS (EC11255-19-P1z, P1 RT = 0.272 min) detection showed a purity of about 97.2%.

[0408] LCMS: EC11255-19-P1a, P1 RT = 0.283 min, MS (ESI) m / z = 406.3 [M+H] +

[0409] LCMS: EC11255-19-P1z, P1 RT = 0.272 min, MS (ESI) m / z = 406.3 [M+H] +

[0410] 2. Synthesis of compound APF1A

[0411] Compound A1331852 (100 mg, 151 µmol, 1.00 equiv), APF1A-2 (73.8 mg, 182 µmol, 1.20 equiv), DMAP (37.1 mg, 303 µmol, 2.00 equiv), and EDCI (116 mg, 607 µmol, 4.00 equiv) were mixed in DCM (1 mL), N2 was filled to expel air, and the mixture was stirred at 25 °C for 16 h. LC-MS (EC11272-31-P1A, P1, RT = 0.594 min) detection showed compound A1331852 was fully reacted, and the reaction solution was concentrated under vacuum to give the crude product, which was further purified by preparative HPLC (basic conditions) to give compound APF1A (20.2 mg, 18.8 µmol, 12.4% yield, 97.3% purity) as a yellow solid.

[0412] LCMS: EC11272-31-P1A, RT = 0.569 min, M / Z (M+H + ) = 524.2

[0413] LCMS: EC11272-31-P1C, RT = 0.593 min, M / Z (M+H + ) = 1446.3

[0414] HNMR: EC11272-31-P1C, DMSO, 400 MHz δ 12.85 (s, 1H), 11.08 (s, 1H), 8.02 (d, J = 7.6 Hz, 1H), 7.78 (d, J = 8.0 Hz, 1H), 7.60 (d, J = 8.0 Hz, 1H), 7.50-7.56 (m, 2H), 7.40-7.48 (m, 2H), 7.35 (q, J = 7.6 Hz, 2H), 7.24 (s, 1H), 7.08 (d, J = 8.2 Hz, 1H), 6.94-7.03 (m, 2H), 6.56 (t, J = 5.6 Hz, 1H), 5.00-5.07 (m, 1H), 4.95 (s, 2H), 4.02-4.09 (m, 2H), 3.84 (t, J = 5.6 Hz, 2H), 3.68 (s, 2H), 3.56 (t, J = 5.2 Hz, 2H), 3.45-3.49 (m, 2H), 3.40 (d, J = 5.2 Hz, 4H), 3.29 (s, 2H), 3.00 (t, J = 5.6 Hz, 2H), 2.77-2.92 (m, 2H), 2.07 (s, 3H), 1.95-2.04 (m, 2H), 1.89 (s, 3H), 1.58-1.64 (m, 3H), 1.47-1.55 (m, 9H).

[0415] b) Synthesis of PROTAC molecules APR1, APR2, APR3, APR4

[0416] Synthesis of APR1:

[0417] 1. Synthesis of compound APR1-2

[0418] To a solution of compound 1 (1.50 g, 5.43 mmol, 1.00 eq) and compound APR1-1 (538 mg, 5.43 mmol, 1.00 eq) in DMSO (15.0 mL) was added DIPEA (1.40 g, 10.8 mmol, 1.89 mL, 2.00 eq), the mixture was stirred at 70 °C for 16 h. LC-MS (EC7111-89-p1a1, rt [P1] = 0.362 min) detection showed compound 1 was completely reacted, multiple new compound peaks appeared, the target compound was detected about 35.3%. The reaction mixture was diluted with 30.0 mL water, extracted with 60 mL ethyl acetate (20.0 mL x 3 times), the organic layer was washed with 30 mL brine, dried over Na2S04, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography (Si02, PE:EA = 5 / 1 to 0 / 1) and further purified by preparative HPLC (neutral condition, column: Phenomenex C18 250*50mm*10μm; mobile phase: [water(NH4HCO3)-ACN]; B%: 22%-52%, 9 min) to give compound APR1-2 (0.522 g, crude) as a yellow solid.

[0419] LCMS: EC7111-89-p1a1, RT = 0.362 min. M / Z (M+H + ) = 356.3

[0420] 2. Synthesis of compound APR1

[0421] Intermediate compound 2A (100 mg, 137 μmol, 1.00 equiv), APR1-2 (53.8 mg, 151 μmol, 1.10 equiv), CuSO4(548 μg, 3.44 μmol, 0.025 equiv) and (2R)-2-[(1S)-1,2-dihydroxyethyl]-4-hydroxy-5-oxo-2H-furan-3-olate sodium salt (1.36 mg, 6.88 μmol, 0.05 equiv) were mixed in THF (4.0 mL), water (1.0 mL) and methanol (6.0 mL), purged with N2to remove air, and stirred at 25 °C for 3 h. LC-MS (EC12165-6-P1A, P1: RT = 0.508 min) detection showed compound 2A was fully reacted. The reaction mixture was quenched with water 20.0 mL at 0 °C, diluted with DCM 20 mL, extracted with ethyl acetate 60 mL (20.0 mL x 3 times), the organic layer was washed with 20 mL saturated brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product, which was purified by prep-HPLC (neutral condition) to give compound APR1 (60.0 mg, 55.4 μmol, 40.3% yield) as a yellow solid. LC-MS (EC12165-6-P1C, P1 RT = 0.515 min) detection purity was 96.0%.

[0422] LCMS: EC12165-6-P1a, RT = 0.508 min, M / Z (M+H + ) = 1082.7

[0423] LCMS: EC12165-6-P1C, RT = 0.515 min, M / Z (M+H + ) = 1083.6

[0424] HNMR: EC12165-6-P1C, DMSO, 400 MHz δ 12.85 (s, 1H), 11.09 (s, 1H), 8.48 (t, J = 6.0 Hz, 1H), 7.99 (d, J = 7.8 Hz, 1H), 7.93 (s, 1H), 7.76 (d, J = 8.0 Hz, 1H), 7.52-7.64 (m, 2H), 7.40-7.50 (m, 3H), 7.30-7.38 (m, 2H), 7.23 (s, 1H), 7.08 (d, J = 8.4 Hz, 1H), 7.02 (d, J = 7.2 Hz, 1H), 6.97 (d, J = 8.8 Hz, 1H), 6.57 (t, J = 6.0 Hz, 1H), 4.96-5.09 (m, 3H), 4.55 (s, 2H), 4.41 (t, J = 6.4 Hz, 2H), 3.90 (t, J = 6.0 Hz, 2H), 3.67 (s, 2H), 3.59-3.63 (m, 2H), 3.53-3.57 (m, 2H), 3.43 (q, J = 5.6 Hz, 2H), 3.01 (t, J = 5.6 Hz, 2H), 2.82-2.92 (m, 1H), 2.02 (s, 4H), 1.90 (s, 3H), 1.58-1.65 (m, 3H), 1.49-1.56 (m, 9H).

[0425] Synthesis of APR2:

[0426] The synthesis of PROTAC molecule APR2 refers to that of APR1, but the starting material is changed to compound 1 and APR2-1. The target product APR2 is obtained by purification after synthesis, which is a yellow solid with a purity of 96.4% and a yield of about 24.4%.

[0427] LCMS: EC12165-5-P1C, RT = 0.503 min, M / Z (M+H + ) = 1126.7

[0428] HNMR: EC12165-5-P1C, DMSO, 400 MHz δ 12.85 (s, 1H), 11.09 (s, 1H), 8.48 (t, J = 6.0 Hz, 1H), 7.99 (d, J = 8.0 Hz, 1H), 7.90 (s, 1H), 7.75 (d, J = 8.4 Hz, 1H), 7.52-7.62 (m, 2H), 7.40-7.49 (m, 3H), 7.29-7.38 (m, 2H), 7.22 (s, 1H), 7.10 (d, J = 8.8 Hz, 1H), 7.02 (d, J = 7.2 Hz, 1H), 6.96 (d, J = 8.8 Hz, 1H), 6.58 (t, J = 5.6 Hz, 1H), 5.04 (dd, J = 12.8, 5.2 Hz, 1H), 4.99 (s, 2H), 4.49 (s, 2H), 4.39 (t, J = 6.0 Hz, 2H), 3.90 (t, J = 6.0 Hz, 2H), 3.67 (s, 2H), 3.54-3.59 (m, 4H), 3.52 (s, 4H), 3.42 (q, J = 5.6 Hz, 2H), 3.00 (t, J = 5.6 Hz, 2H), 2.80-2.92 (m, 1H), 2.01 (s, 3H), 1.99 (s, 1H), 1.90 (s, 3H), 1.59-1.65 (m, 3H), 1.49-1.55 (m, 9H).

[0429] Synthesis of APR3:

[0430] The synthesis of PROTAC molecule APR3 refers to that of APR1, but the starting material is changed to compound 1 and APR3-1. The target product APR3 is obtained by purification after synthesis, which is a yellow solid with a purity of 97.7% and a yield of about 18.2%.

[0431] LCMS: EC12286-2-p1A, RT = 0.511 min, MS (M+H + ) = 1171.7

[0432] 1H NMR: EC12886-2-P1C (400 MHz, DMSO) δ 12.01 (s, 1H), 10.25 (s, 1H), 7.58 - 7.67 (m, 1H), 7.14 (d, J = 7.6 Hz, 1H), 7.05 (s, 1H), 6.91 (d, J = 8.0 Hz, 1H), 6.67 - 6.78 (m, 2H), 6.59 (dd, J = 18.0, 8.4 Hz, 3H), 6.45 - 6.54 (m, 2H), 6.37 (s, 1H), 6.26 (d, J = 8.4 Hz, 1H), 6.17 (d, J = 7.2 Hz, 1H), 6.12 (d, J = 8.8 Hz, 1H), 5.73 (t, J = 5.6 Hz, 1H), 4.08 - 4.25 (m, 3H), 3.62 (s, 2H), 3.50 - 3.58 (m, 2H), 3.05 (t, J = 5.2 Hz, 2H), 2.82 (s, 2H), 2.54 - 2.77 (m, 16H), 2.13 - 2.20 (m, 2H), 1.96 - 2.08 (m, 1H), 1.16 (s, 4H), 1.05 (s, 3H), 0.74 - 0.81 (m, 3H), 0.60 - 0.69 (m, 9H).

[0433] Synthesis of APR4:

[0434] The synthesis of PROTAC molecule APR4 refers to that of APR1, but the starting material is changed to compound 1 and APR4-1. The target product APR4 is obtained by purification after synthesis, which is a light yellow solid with a purity of 92.8% and a yield of about 27.4%.

[0435] LCMS: EC12886-1-p1a1, P1: RT = 0.500 min, MS (M+H + ) = 1215.7

[0436] 1 H NMR: 1HNMR (EC12886-1-P1B4) (400 MHz, DMSO) δ 12.00 (s, 1H), 10.26 (s, 1H), 7.65 (t, J = 5.6 Hz, 1H), 7.15 (d, J = 5.2 Hz, 1H), 7.07 (s, 1H), 6.86-6.98 (m, 1H), 6.77 (d, J = 7.2 Hz, 1H), 6.71-6.75 (m, 1H), 6.56-6.65 (m, 3H), 6.47-6.54 (m, 2H), 6.39 (s, 1H), 6.29 (d, J = 8.8 Hz, 1H), 6.19 (d, J = 7.00 Hz, 1H), 6.05-6.17 (m, 1H), 5.76 (t, J = 5.6 Hz, 1H), 4.22 (dd, J = 12.8, 5.2 Hz, 1H), 4.16 (s, 2H), 3.64 (s, 2H), 3.57 (d, J = 6.6 Hz, 2H), 3.01-3.14 (m, 2H), 2.83 (s, 2H), 2.74-2.78 (m, 3H), 2.69-2.71 (m, 2H), 2.65-2.67 (m, 4H), 2.59-2.64 (m, 10H), 2.18 (d, J = 5.6 Hz, 2H), 1.99-2.09 (m, 1H), 1.18 (s, 4H), 1.07 (s, 3H), 0.76-0.81 (m, 3H), 0.67-0.72 (m, 9H).

[0437] Synthesis of intermediate compound 2A:

[0438] 1. Synthesis of compound 2A-2

[0439] To a solution of compound 2A-1 (10.0 g, 44.6 mmol, 1.00 equiv) in DMF (100 mL) was added NaN3(4.35 g, 66.9 mmol, 1.50 equiv), and the mixture was stirred at 75 °C for 16 h. TLC (PE:EA = 5:1, P1, Rf = 0.4) detection showed that compound 2A-1 was completely reacted to generate one major peak with large polarity. After the reaction was completed, it was restored to room temperature, and 16.5 g of KOH was added gradually in small amounts. After being restored to room temperature, 200 mL of ethyl acetate was used for extraction (40 mL x 5 times), and the obtained organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (SiO2, PE / EA = 10 / 1 to 5 / 1, PE:EA = 5:1, P1, Rf = 0.4) to obtain compound 2A-2 (7.50 g, 40.3 mmol, 90.2% yield) in the form of white oil. f f Compound 2A-2 (7.50 g, 40.3 mmol, 1.00 equiv) was dissolved in THF (100 mL), and the solution was cooled to -78 °C. Then, 2.5 mL of n-BuLi (2.5 M, 6.25 mmol, 0.15 equiv) was added dropwise, and the mixture was stirred at -78 °C for 1 h. Then, 2.5 mL of DMF (25.0 mmol, 0.62 equiv) was added dropwise, and the mixture was stirred at -78 °C for 1 h. After the reaction was completed, the mixture was warmed to room temperature, and 100 mL of saturated NH4Cl was added. The mixture was extracted with 200 mL of ethyl acetate (40 mL x 5 times), and the obtained organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (SiO2, PE / EA = 10 / 1 to 5 / 1, PE:EA = 5:1, P1, Rf = 0.4) to obtain compound 2A-3 (5.00 g, 27.9 mmol, 69.2% yield) in the form of white oil.​

[0440] 2. Synthesis of compound 2A-3

[0441] Compound 2A-2 (2.00 g, 10.7 mmol, 1.00 equiv) was added to hydrochloric acid / dioxane (4 M, 26.8 mL, 10.0 equiv), N2 was purged to remove air, and the mixture was stirred at 25 °C for 1 h under N2. TLC (PE:EA = 5:1, R1, R f = 0.4) detection showed that compound 2A-2 was fully reacted. The reaction was concentrated under vacuum to give the crude product, which was used in the next step without further purification to give compound 2A-3 (1.20 g, 9.79 mmol, 91.16% yield, HC1) as a white solid.

[0442] 3. Synthesis of compound 2A

[0443] Compound A1331852 (200 mg, 303.58 pmol, 1 equiv), 2-azidoethanamine (186 mg, 1.52 mmol, 5.00 equiv, HC1), HATU (173 mg, 455 pmol, 1.5.0 equiv), and DIEA (196 mg, 1.52 mmol, 264 pL, 5.00 equiv) were mixed in DCM (5 mL), N2 was purged to remove air, and the mixture was stirred at 25 °C for 1 h under N2. LC-MS (EC11272-32-P1A, P1, RT = 0.590 min) detection showed that compound A1331852 was fully reacted. The reaction was concentrated under vacuum to give the crude product, which was purified by column chromatography (SiO2, PE:EA = 3:1 to 2:1, PE:EA = 2:1, P1:R f = 0.3) to give compound 2A (90 mg, 112 pmol, 36.9% yield, 90.6% purity) as a beige solid.

[0444] LCMS: EC11272-32-P1A, RT = 0.590 min. M / Z (M+H + ) = 727.4

[0445] Example 5. Synthesis of deuterium-substituted compounds of A1331852 as PROTAC targeting ligands and corresponding deuterated PROTAC molecules (ligands A13D-1, A13D-2, A13D-5, A13D-6 and deuterated PROTAC molecules A13D6-PF3)

[0446] Synthesis of A13D-1:

[0447] 1. Synthesis of compound D1-2

[0448] To compound D1-1 (4.00 g, 12.8 mmol, 1.00 equiv) was added methanol (32.0 mL), DMF (8.00 mL), TEA (3.89 g, 38.4 mmol, 5.35 mL, 3.00 equiv) and Pd(dppf)Cl2(937 mg, 1.28 mmol, 0.10 equiv) at 25 °C, and the reaction was stirred at 110 °C under CO (2.00 Mpa) for 24 h. TLC (petroleum ether: ethyl acetate = 2:1) detection showed that compound D1-1 was completely reacted to generate a new compound main peak (R f = 0.51). Water (50.0 mL) and ethyl acetate (100 mL) were added to the reaction mixture, and extraction was performed with ethyl acetate (50.0 mL x 2 times). The organic layer was washed with saturated brine (50.0 mL x 5 times), dried over Na2SO4, filtered, and concentrated to obtain a crude product. The crude product (TLC: petroleum ether: ethyl acetate = 2:1, R f = 0.51) was purified by column chromatography (ethyl acetate: petroleum ether = 0-5%) to obtain compound D1-2 (3.61 g, 12.4 mmol, 96.7% yield) in the form of yellow oil.

[0449] 1 H NMR: EB8953-5-P1N2 (400 MHz, DMSO-d6) δ ppm 1.41 (s, 9H), 2.85 (t, J = 6.40 Hz, 2H), 3.55 (br t, J = 5.60 Hz, 2H), 3.83 (s, 3H), 4.73-4.90 (m, 2H), 7.26-7.36 (m, 1H), 7.41 (d, J = 7.20 Hz, 1H), 7.76 (d, J = 7.60 Hz, 1H)

[0450] 2. Synthesis of compound D1-3

[0451] To compound D1-2 (1.02 g, 3.50 mmol, 1.00 equiv) was added ethyl acetate (2.00 mL) and HCl / ethyl acetate (7.00 mL) at 25 °C, and the reaction was stirred at 25 °C for 2 h. TLC (petroleum ether: ethyl acetate = 3:1) detection showed that compound D1-2 was completely reacted to generate a new compound main peak (R f = 0.00). The reaction mixture was concentrated to obtain a crude product of compound D1-3 (0.80 g).

[0452] 3. Synthesis of compound D1-4

[0453] To compound 3A (500 mg, 1.13 mmol, 1.00 eq) was added dioxane (5.00 mL), compound D1-3 (216 mg, 1.13 mmol, 1.00 eq), Cs2CO3 (1.47 g, 4.52 mmol, 4.00 eq), x-antphos (131 mg, 226 µmol, 0.20 eq) and Pd2(dba)3 (104 mg, 113 µmol, 0.10 eq) at 25 °C successively, and stirred at 100 °C for 12 h. LCMS (EB9483-5-IPCL1) detection showed that the reaction of compound D1-3 was complete, and the product with the target molecular weight was generated (R t = 1.96 min). Water (100 mL) was added to the reaction mixture, extracted with ethyl acetate (100 mL x 2 times), and the organic layer was washed with saturated brine (20.0 mL), dried over Na2SO4, filtered, and concentrated to give the crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 100:1 to 10:1) to give compound D1-4 (400 mg, 670 µmol, 59.25% yield) as a yellow solid.

[0454] LCMS: EB9483-5-IPCL1, product: R t = 1.964 min, MS (ESI) m / z = 597.4 [M+H] +

[0455] 1 H NMR: EB9483-5-P1N1 (400 MHz, DMSO-d6) δ ppm 1.32 (s, 10H) 1.51-1.70 (m, 13H) 1.92 (br s, 3H) 2.11 (s, 3H) 2.98 (br t, J = 5.60 Hz, 2H) 3.72 (s, 2H) 3.80-3.95 (m, 5H) 5.03 (s, 2H) 6.94 (d, J = 8.80 Hz, 1H) 7.23 (s, 1H) 7.29-7.37 (m, 1H) 7.41-7.52 (m, 2H) 7.76 (dd, J = 7.60, 1.20 Hz, 1H)

[0456] 4. Synthesis of compound D1-5

[0457] To compound D1-4 (320 mg, 536 μmol, 1.00 equiv) was added methanol (2.50 mL), NaOH (42.9 mg, 1.07 mmol, 2.00 equiv) and H2O (0.50 mL) sequentially at 25 °C, and the reaction was stirred at 25 °C for 12 h. LCMS (EB9483-12-IPCL2) detection showed that the reaction of compound D1-4 was complete, and the product with the target molecular weight (R t = 1.038 min) was generated. Water (3.00 mL) was added to the reaction mixture, the pH was adjusted to about 5 with acetic acid, and extraction was performed with ethyl acetate (20.0 mL x 3 times), the obtained organic layer was washed with saturated brine (50.0 mL), dried over Na2SO4, filtered, and concentrated to obtain a crude product. The crude product was purified by preparative HPLC (column: Xtimate C18 150*40mm*10μm; mobile phase: [water (NH4HCO3)-ACN]; B%: 20%-60%, 25 min) to obtain compound D1-5 (108 mg, 185 μmol, 34.56% yield) as a yellow solid.

[0458] LCMS: EB9483-12-IPCL2, product: R t = 1.038 min, MS (ESI) m / z = 583.3 [M+H] +

[0459] 1 H NMR: EB9483-12-P1N1 (400 MHz, DMSO-d6) δ ppm 1.29 (s, 9 H) 1.50 - 1.73 (m, 12 H) 1.88 - 1.96 (m, 3 H) 2.07 (br s, 2 H) 2.88 (br s, 2 H) 3.72 (s, 2 H) 3.82 (br t, J = 5.6 Hz, 2 H) 4.99 (s, 2 H) 6.88 (br d, J = 8.8 Hz, 1 H) 7.01 - 7.65 (m, 5 H)

[0460] 5. Synthesis of compound D1-6

[0461] To compound D1-5 (150 mg, 257 μmol, 1.00 equiv) was added CH2Cl2(2.00 mL), EDCl (148 mg, 772 μmol, 3.00 equiv), DMAP (94.4 mg, 772 μmol, 3.00 equiv) and compound D1-0 (59.6 mg, 386 μmol, 1.50 equiv) sequentially at 0 °C, and the reaction was stirred at 25 °C for 12 h. LCMS (EB9483-13-IPCL2) detection showed that the reaction of compound D1-5 was complete, and the product with the target molecular weight (Rt = 2.057 min). The reaction mixture was adjusted to pH ~ 6 with acetic acid, extracted with CH2Cl2(20.0 mL x 3 times), the organic layer was washed with saturated brine (30.0 mL), dried over Na2SO4, filtered, concentrated to get the crude product. The crude product was purified by prep-HPLC (column: Xtimate C18 150*40mm*10um; mobile phase: [water (NH4HCO3) - ACN]; B%: 58%-98%, 25 min) to get compound D1-6 (110 mg, 139 pmol, 54.04% yield) as yellow oil.

[0462] LCMS: EB9483-13-IPCL2, product: R t = 2.057 min, MS (ESI) m / z = 719.3 [M+H] +

[0463] 1 H NMR: EB9483-13-P1N1, (400 MHz, DMSO-d6) δ ppm 1.21 (s, 9 H) 1.48 - 1.70 (m, 13 H) 1.92 (br s, 3 H) 2.08 (s, 3 H) 3.01 (br t, J=5.60 Hz, 2 H) 3.71 (s, 2 H) 3.81 (br t, J=6.40 Hz, 2 H) 4.89 (s, 2 H) 6.90 (d, J=8.80 Hz, 1 H) 7.21 (s, 1 H) 7.36 (quin, J=7.60 Hz, 2 H) 7.42 - 7.48 (m, 2 H) 10.68 (s, 1 H)

[0464] 6. Synthesis of compound A13D-1

[0465] To compound D1-6 (100 mg, 139 pmol, 1.00 equiv) was added CH2Cl2(1.00 mL) and TFA (5.12 g, 44.9 mmol, 3.33 mL, 323 equiv) sequentially at 25 °C, stirred for 12 h at 25 °C. LCMS (EB9484-16-IPCL1) detection showed that compound D1-6 was completely reacted to generate the target molecular weight product (R t = 1.833 min). The reaction mixture was concentrated to get the product crude, which was purified by prep-HPLC (column: Welch Xtimate C18 150*30mm*5um; mobile phase: [water (FA) - ACN]; B%: 56%-96%, 25 min) to get compound A13D-1 (30.0 mg, 43.4 pmol, 34.64% yield, 95.8% purity) as white solid.

[0466] LCMS: EB9483-16-IPCL1, Product: R t = 1.833 min, MS (ESI) m / z = 663.3 [M+H] +

[0467] HPLC: EB9483-16-P1H5, Product: R t = 4.492 min, Purity: 95.8%

[0468] HRMS: X00470-2-P1S2, D2: 0.35%; D3: 3.12%; D4: 94.30%;

[0469] 1 H NMR: EB9483-16-P1N1 (400 MHz, DMSO-d6) δ ppm 1.50-1.60 (m, 10 H) 1.62-1.70 (m, 3 H) 1.93 (br s, 3 H) 2.10 (s, 3 H) 2.99 (br t, J=5.60 Hz, 2 H) 3.71 (s, 2 H) 3.86 (t, J=6.00 Hz, 2 H) 4.88 (s, 2 H) 6.91 (d, J=8.80 Hz, 1 H) 7.20 (s, 1 H) 7.32-7.41 (m, 2 H) 7.44-7.52 (m, 2 H) 10.69 (s, 1 H) 12.80 (br s, 1 H)

[0470] Synthesis of A13D-2:

[0471] The synthesis of deuterated molecule A13D-2 was performed according to the procedure of A13D-1, but using intermediate compounds D2-1 and D0-0 as starting materials. The desired product A13D-2 was obtained as a yellow solid with a purity of 99.1% and a yield of about 18.8%.

[0472] LCMS: EB9776-40-IPCL2, Product: R T = 1.793 min, MS (ESI) m / z = 665.2 [M+H] +

[0473] HPLC: EB9776-40-P1H2, Purity 99.1% (area%)

[0474] 1HNMR: EB9776-40-P1N5 (400 MHz, DMSO-d6) δ 8.21 (br s, 1H), 8.03 (br d, J = 7.6 Hz, 1H), 7.79 (br d, J = 8.0 Hz, 1H), 7.62 (br d, J = 7.6 Hz, 1H), 7.51-7.32 (m, 5H), 7.28 (s, 1H), 6.89 (br d, J = 8.8 Hz, 1H), 3.69 (br s, 2H), 2.10 (s, 3H), 1.92 (br s, 3H), 1.72-1.35 (m, 13H)

[0475] Synthesis of A13D-5:

[0476] The synthesis of deuterated molecule A13D-5 was performed according to the procedure of A13D-1, but using intermediate compounds D5-1 and D0-0 as starting materials. The desired product A13D-5 was obtained as a yellow solid with a purity of 98.9% and a yield of about 42.9% after purification.

[0477] LCMS: EB9776-26-IPCL2, product: R T = 0.82 min, MS (ESI) m / z = 661.4 [M+H] +

[0478] HPLC: EB9776-26-P1H1, purity 98.9% (area%)

[0479] 1 HNMR: EB9776-26-P1N3 (400 MHz, DMSO-d6) δ 13.27-12.28 (m, 1H), 8.04 (d, J = 7.6 Hz, 1H), 7.79 (d, J = 8.0 Hz, 1H), 7.62 (d, J = 7.6 Hz, 1H), 7.52-7.32 (m, 5H), 7.29 (s, 1H), 6.92 (br d, J = 8.8 Hz, 1H), 4.95 (s, 2H), 3.87 (s, 2H), 3.70 (s, 2H), 2.11 (s, 3H), 1.93 (br s, 3H), 1.69-1.49 (m, 13H)

[0480] Synthesis of A13D-6:

[0481] The synthesis of deuterated molecule A13D-6 was performed according to the procedure of A13D-1, but using intermediate compounds D6-1 and D0-0 as starting materials. The desired product A13D-6 was obtained as a white solid with a purity of 98.4% and a yield of about 18.8% after purification.

[0482] LCMS: EB9776-48-IPCL1, Product: R T = 0.815 min, MS (ESI) m / z = 661.4 [M+H] +

[0483] HPLC: EB9776-48-P1H3, purity 98.4% (area%)

[0484] 1 HNMR: EB9776-48-P1N1 (400 MHz, DMSO-d6) 8.15 (s, 1H), 8.04 (d, J = 7.6 Hz, 1H), 7.79 (d, J = 8.0 Hz, 1H), 7.61 (d, J = 7.2 Hz, 1H), 7.52-7.32 (m, 5H), 7.27 (s, 1H), 6.93 (d, J = 8.8 Hz, 1H), 4.95 (s, 2H), 3.70 (s, 2H), 2.99 (s, 2H), 2.10 (s, 3H), 1.92 (br s, 3H), 1.69-1.49 (m, 13H)

[0485] Synthesis of intermediate compound 3A:

[0486] 1. Synthesis of compound 3A-3

[0487] To compound 3A-1 (11.0 g, 66.2 mmol, 1.00 equiv) was added THF (300 mL), compound 3A-2 (19.3 g, 99.2 mmol, 1.50 equiv), PPh3 (26.0 g, 99.2 mmol, 1.50 equiv) and DIAD (20.1 g, 99.2 mmol, 19.3 mL, 1.50 equiv) at 25 °C under N2 atmosphere. The reaction was stirred at 25 °C for 36 h. LCMS (EB8953-61-IPCL4) detection showed that compound 3A-1 was completely consumed and the product with the desired molecular weight was generated (R t = 1.344 min). The reaction mixture was concentrated to give the product as a crude product, which was purified by column chromatography (ethyl acetate: petroleum ether = 0-5%) to give compound 3A-3 as a white solid.

[0488] LCMS: EB8953-61-IPCL4, Product: R t = 1.344 min, MS (ESI) m / z = 557.3 [M+H] +

[0489] 1 H NMR: EB8953-61-P1N1 (400 MHz, DMSO-d6) δ ppm 1.42 (d, J = 2.00 Hz, 6 H), 1.48 - 1.56 (m, 3 H), 1.59 - 1.67 (m, 3 H), 1.91 (br s, 3 H), 3.78 (s, 2 H), 7.48 (s, 1 H), 7.80 (s, 1 H)

[0490] 2. Synthesis of compound 3A-4

[0491] To THF (160 mL) was added DIPA (6.24 g, 61.7 mmol, 8.72 mL, 1.10 eq) at 25 °C under N2, n-BuLi (2.50 M, 29.2 mL, 1.30 eq) was added at -78 °C and stirred for 30 min, compound 3A-3 (19.2 g, 56.1 mmol, 1.00 eq) dissolved in THF (80.0 mL) was added at -78 °C and stirred for 1 h, Mel (9.08 g, 64.0 mmol, 3.98 mL, 1.14 eq) was added at -78 °C and stirred for 4 h, LCMS (EB8953-61-IPCL4) showed compound 3A-3 was consumed completely and the product of interest (R t = 1.130 min) was formed. To the reaction mixture was added saturated NH4Cl solution at 0 °C to terminate the reaction, water (100 mL) and ethyl acetate (200 mL) were added, extracted with ethyl acetate (200 mL x 2 times), the organic layer was washed with saturated brine (100 mL), dried over Na2SO4, filtered and concentrated to give the crude product. The crude product was triturated in MeOH (100 mL) at 25 °C to give compound 3A-4 as a white solid.

[0492] LCMS: EB8953-68-IPCL1, product: R t = 1.130 min, MS (ESI) m / z = 357.0 [M+H] +

[0493] 1 H NMR: EB8953-68-P1N1 (400 MHz, CDCl3) δ ppm 1.55 - 1.76 (m, 12 H), 1.99 (br s, 3 H), 2.29 (s, 3 H), 3.78 (s, 2 H), 7.44 (s, 1 H)

[0494] 3. Synthesis of compound 3A-6

[0495] To THF (100 mL) was added toluene (100 mL), compound 3A-4 (15.6 g, 43.8 mmol, 1.00 equiv), and triisopropyl borate (14.8 g, 78.8 mmol, 18.1 mL, 1.80 equiv) at 25 °C under N2, n-BuLi (2.50 M, 26.3 mL, 1.50 equiv) was added at -78 °C and stirred for 4 h, 2,3-dimethylbutane-2,3-dione (7.76 g, 65.7 mmol, 1.50 equiv) dissolved in toluene (80.0 mL) was added at -78 °C, the mixture was slowly warmed to 20 °C for 1 h, and stirred at 25 °C for 12 h. LCMS (EB8953-69-IPCL1) showed compound 3A-4 was completely reacted and the product with the desired molecular weight was generated (R t = 1.145 min). To the reaction mixture was added saturated NH4Cl solution at 0 °C to quench the reaction, water (100 mL) and THF (200 mL) were added, and the mixture was extracted with THF (300 mL), the organic layer was washed with saturated brine (200 mL), dried over Na2SO4, filtered, and concentrated to give the crude product. The crude product was recrystallized in DMSO (180 mL) at 25 °C to give compound 3A-6 as a white solid.

[0496] LCMS: EB8953-69-IPCL1, product: R t = 1.145 min, MS (ESI) m / z = 357.2 [M+H] +

[0497] 1 H NMR: EB8953-69-P1N2 (400 MHz, CDC13) δ ppm 1.31 (s, 12 H), 1.56 - 1.72 (m, 12 H), 1.97 (br s, 3 H), 2.42 (s, 3 H), 3.71 (s, 2 H), 7.69 (s, 1 H)

[0498] 4. Synthesis of compound 3A

[0499] To compound 3A-7 (2.76 g, 9.43 mmol, 1.20 equiv) was added dioxane (30.0 mL), water (6.00 mL), compound 3A-6 (2.80 g, 7.86 mmol, 1.00 equiv), Cs2CO3(7.68 g, 23.6 mmol, 3.00 equiv) and 4-di-tert-butylphosphoryl-N,N-dimethylamine dichloropalladium (556 mg, 786 μmol, 556 μL, 0.10 equiv) at 25 °C under N2. The mixture was stirred at 80 °C for 12 h. TLC (petroleum ether: ethyl acetate = 3: 1) detection showed that compound 3A-6 was completely reacted to generate a new compound main peak (R f = 0.56). Water (200.0 mL) was added to the reaction mixture, which was extracted with ethyl acetate (200.0 mL x 2 times), and the obtained organic layer was washed with saturated brine (200 mL), dried over Na2SO4, filtered and concentrated to obtain a crude product. The crude product (TLC: petroleum ether: ethyl acetate = 3: 1, R f = 0.51) was purified by column chromatography (petroleum ether: ethyl acetate = 100: 1 to 10: 1) to obtain compound 3A (2.70 g, 6.11 mmol, 77.7% yield) as a white solid.

[0500] 1 H NMR: EB9483-4-P1N1 (400 MHz, DMSO-d6) δ ppm 1.35 (s, 9H) 1.49-1.70 (m, 12H) 1.88-2.02 (m, 3H) 2.17 (s, 3H) 3.77 (s, 2H) 7.36 (s, 1H) 7.66 (d, J = 8.40 Hz, 1H) 7.85 (d, J = 8.40 Hz, 1H)

[0501] Synthesis of intermediate D2-1:

[0502] 1. Synthesis of compound D2-1-2

[0503] Compound D2-1-1 (19.4 g, 97.0 mmol, 1.00 equiv), dimethyl malonate (19.2 g, 145 mmol, 16.7 mL, 1.50 equiv) and K2CO3(33.5 g, 242 mmol, 2.50 equiv) were mixed in DMF (100 mL) at 25 °C, and the mixture was stirred at 60 °C for 12 h. TLC (petroleum ether: ethyl acetate = 3: 1) detection showed that compound D2-1-1 was completely reacted to generate a new compound main peak (R f= 0.39). Water (200 mL) and ethyl acetate (200 mL) were added to the post-reaction mixture, the organic layer was separated, extracted with ethyl acetate (100 mL x 2 times), the obtained organic layer was washed with saturated brine (200 mL), dried over Na2SO4, filtered, and concentrated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 100:1 to 10:1) to obtain compound D2-1-2 (19.1 g, 61.1 mmol, 63.0% yield) as a white solid.

[0504] 1 H NMR: EB9915-3-P1N1 (400 MHz, DMSO-d6) δ 7.92 (dd, J = 1.2, 8.1 Hz, 1H), 7.72-7.66 (m, 1H), 7.64-7.60 (m, 1H), 5.36 (s, 1H), 3.74 (s, 6H)

[0505] 2. Synthesis of compound D2-1-3

[0506] To compound D2-1-2 (19.1 g, 61.1 mmol, 1.00 equiv) was added ethyl acetate (190 mL) and Lil (81.8 g, 611 mmol, 23.4 mL, 10.0 equiv) at 25 °C, and the reaction was stirred at 80 °C for 4 h. TLC (petroleum ether: ethyl acetate = 2:1) detection showed that compound D2-1-2 was completely reacted to generate a new compound main peak (R f = 0.19). The post-reaction mixture was terminated by adding citric acid (100 mL), extracted with ethyl acetate (200 mL x 2 times), the obtained organic layer was washed with saturated brine (200 mL), dried over Na2SO4, filtered, and concentrated to obtain the crude product. The crude product (TLC: petroleum ether: ethyl acetate = 2:1, R f = 0.19) was purified by column chromatography (ethyl acetate / petroleum ether = 0-10%) to obtain compound D2-1-3 (5.60 g, 23.3 mmol, 38.2% yield) as a white solid.

[0507] 1 H NMR: EB9915-4-P1N1 (400 MHz, DMSO-d6) δ 12.82 (br s, 1H), 7.79 (dd, J = 1.2, 7.9 Hz, 1H), 7.64-7.50 (m, 2H), 3.90 (s, 2H)

[0508] 3. Synthesis of compound D2-1-4

[0509] To compound D2-1-3 (1.00 g, 4.17 mmol, 1.00 equiv) and K2CO3 (691 mg, 5.00 mmol, 1.20 equiv) in DMF at 25 °C, add Mel (887 mg, 6.25 mmol, 389 μL, 1.50 equiv) at 0 °C, stir the reaction at 25 °C for 1 h. TLC (petroleum ether: ethyl acetate = 1:1) detection shows that compound D2-1-3 is completely reacted to generate a new compound main peak (Rf = 0.40). The reaction mixture is terminated by adding water (10.0 mL), extracted with ethyl acetate (20.0 mL x 2 times), the obtained organic layer is washed with saturated brine (20.0 mL x 3 times), dried over Na2SO4, filtered, concentrated to obtain the crude product. The crude product (TLC: petroleum ether: ethyl acetate = 1:1, Rf = 0.40) is purified by column chromatography (ethyl acetate / petroleum ether = 0-20%) to obtain compound D2-1-4 (640 mg, 2.52 mmol, 60.5% yield) as a white solid. f

[0510] 1 H NMR: EB9915-10-P1N1 (400 MHz, DMSO-d6) δ 7.81 (dd, J = 1.2, 8.0 Hz, 1H), 7.64-7.54 (m, 2H), 4.05-3.96 (m, 2H), 3.66 (s, 3H)

[0511] 4. Synthesis of compound D2-1-5

[0512] To compound D2-1-4 (640 mg, 2.52 mmol, 1.00 equiv) in THF (1.00 mL) at 25 °C, add K2CO3 (17.4 mg, 126 μmol, 0.05 equiv) dissolved in D2O (1.01 g, 50.4 mmol, 20.0 equiv) at 0 °C, stir the reaction at 25 °C for 12 h. The reaction mixture is terminated by adding D2O (5.00 mL), extracted with ethyl acetate (20.0 mL x 2 times), the obtained organic layer is dried over Na2SO4, filtered, concentrated to obtain compound D2-1-5 crude product (590 mg, 2.30 mmol, 91.5% yield) as a white solid.

[0513] 1 H NMR: EB9915-11-P1N1 (400 MHz, DMSO-d6) δ 7.81 (dd, J = 1.2, 7.9 Hz, 1H), 7.66-7.50 (m, 2H), 4.02-3.97 (m, 1H), 3.66 (s, 3H)

[0514] ​5. Synthesis of compound D2-1-6

[0515] To compound D2-1-5 (490 mg, 1.91 mmol, 1.00 equiv) was added BD3.THF (0.75 M, 7.65 mL, 3.00 equiv) at 0 °C under N2environment, the mixture was stirred at 25 °C for 12 h. LCMS (EB9776-22-IPCL1) detection showed that compound D2-1-5 was fully reacted to generate the product of the desired molecular weight (R t = 0.335 min). To the reaction mixture was added CD3OD (15.0 mL), and stirred at 80 °C for 16 h. The mixture was concentrated to give compound D2-1-6 (2.30 g) as a yellow oil.

[0516] LCMS: EB9776-22-IPCL1, product: R T = 0.335 min, MS (ESI) m / z = 237.5 [M+H] +

[0517] 6. Synthesis of compound D2-1-7

[0518] To compound D2-1-6 (2.46 g, 10.4 mmol, 1.00 equiv) was added methanol (15.0 mL), DIEA (1.62 g, 12.5 mmol, 2.18 mL, 1.20 equiv) and (Boc)20 (2.27 g, 10.4 mmol, 2.39 mL, 1.00 equiv), and stirred at 25 °C for 16 h. LCMS (EB9776-30-IPCL1) detection showed that compound D2-1-6 was fully reacted to generate the product of the desired molecular weight (R t = 0.657 min). The reaction mixture was concentrated to give the product crude, which was purified by column chromatography (ethyl acetate: petroleum ether = 0-20%) to give compound D2-1-7 (250 mg, 743 µmol, 7.14% yield). f = 0.25) to give compound D2-1-7 (250 mg, 743 µmol, 7.14% yield).

[0519] LCMS: EB9776-30-IPCL1, product: R T = 0.657 min, MS (ESI) m / z = 412.3 [M+H] +

[0520] 1H NMR: EB9776-30-P1N1 (400 MHz, DMSO-d6) δ 7.46 (dd, J = 1.2, 7.9 Hz, 1H), 7.20 (dd, J = 1.2, 7.6 Hz, 1H), 7.14 - 7.09 (m, 1H), 5.20 (br s, 1H), 4.52 (br d, J = 5.6 Hz, 1H), 3.80 (s, 1H), 3.12 - 3.03 (m, 1H), 1.45 (s, 9H)

[0521] 7. Synthesis of compound D2-1-8

[0522] To compound D2-1-7 (830 mg, 2.47 mmol, 1.00 equiv) was added DCM (15.0 mL) and 4-methylmorpholine (749 mg, 7.41 mmol, 814 μL, 3.00 equiv) at 0 °C under N2environment, stirred for 0.5 h at 0 °C, added methylsulfonyl chloride (650 mg, 5.67 mmol, 439 μL, 2.30 equiv), stirred for 2 h at 0 °C. LCMS (EB9776-32-IPCL1) detection showed compound D2-1-7 was fully reacted to generate the product of interest (R t = 0.613 min). To the reaction mixture was added water (10.0 mL), extracted with CH2Cl2(10.0 mL x 3 times), the obtained organic layer was washed with saturated brine (10.0 mL), dried over Na2SO4, filtered, concentrated to give the crude product, which was purified by column chromatography (ethyl acetate: petroleum ether = 1:1, R f = 0.34) to give compound D2-1-8 (780 mg, 1.88 mmol, 76.3% yield) as yellow oil.

[0523] LCMS: EB9776-32-IPCL1, product: R T = 0.613 min, MS (ESI) m / z = 357.9 [M-t-Bu+H] +

[0524] 1 H NMR: EB9776-32-P1N1 (400 MHz, DMSO-d6) δ 7.51 (dd, J = 1.2, 8.0 Hz, 1H), 7.24 - 7.19 (m, 1H), 7.17 - 7.08 (m, 1H), 5.08 (br s, 1H), 2.94 (s, 3H), 1.45 (s, 9H)

[0525] 8. Synthesis of compound D2-1

[0526] To compound D2-1-8 (680 mg, 1.64 mmol, 1.00 eq) was added THF (10.0 mL) and NaH (72.2 mg, 1.81 mmol, 60.0% purity, 1.10 eq) at 0 °C under N2, and stirred at 25 °C for 3 h. LCMS (EB9776-34-IPCL1) detection showed the product with the desired molecular weight was generated (R t = 0.813 min). To the reaction mixture was added water (10.0 mL), extracted with ethyl acetate (10.0 mL x 3 times), the organic layer was washed with saturated brine (10.0 mL), dried over Na2SO4, filtered, and concentrated to give the product D2-1 (590 mg) as a yellow oil.

[0527] LCMS: EB9776-34-IPCL1, product: R T = 0.813 min, MS (ESI) m / z = 217.9 [M-Boc+H] +

[0528] 1 H NMR: EB9776-34-P1N1 (400 MHz, CHLOROFORM-d) δ 7.42 (dd, J = 1.2, 7.6 Hz, 1H), 7.13 - 6.98 (m, 2H), 4.51 (br s, 1H), 3.02 (s, 1H), 1.51 (s, 9H)

[0529] Synthesis of intermediate D5-1:

[0530] 1. Synthesis of compound D5-1-2

[0531] To a mixture of compound D5-1-1 (45.0 g, 196 mmol, 1.00 eq) dissolved in CCl4(500 mL) was added benzoyl peroxide (4.76 g, 19.6 mmol, 0.10 eq) and NBS (35.0 g, 196 mmol, 1.00 eq) at 25 °C under N2, and stirred at 80 °C for 12 h. TLC (petroleum ether: ethyl acetate = 5:1) detection showed that compound D5-1-1 was completely reacted and a new compound main peak (R f= 0.48). The reaction mixture was filtered after it was returned to room temperature, the filtrate was washed with saturated brine (600 mL), dried over Na2S04, filtered, and concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 100: 1 to 10: 1) to give compound D5-1-2 (39.0 g, 127 mmol, 64.5% yield) as a colorless oil.

[0532] 1 H NMR: EB9495-2-P1N1 (400 MHz, DMSO-d6) δ 7.69 (dd, J = 0.8, 8.1 Hz, 1H), 7.59 (d, J = 7.6 Hz, 1H), 7.47 - 7.39 (m, 1H), 4.64 (s, 2H), 3.91 (s, 3H)

[0533] 2. Synthesis of compound D5-1-3

[0534] Compound D5-1-2 (34.0 g, 110 mmol, 1.00 equiv) and NaCN (6.43 g, 131 mmol, 1.19 equiv) were mixed in DMF (170 mL) at 25 °C, and the reaction was stirred at 60 °C for 6 h. TLC (petroleum ether: ethyl acetate = 10: 1) detection showed that compound D5-1-2 was completely reacted to generate a new compound main peak. The reaction mixture was terminated by adding water (300 mL), extracted with ethyl acetate (200 mL x 2 times), and the obtained organic layer was washed with saturated brine (200 mL), dried over Na2S04, filtered, and concentrated under reduced pressure to give the crude product. The crude product was detected by TLC (petroleum ether: ethyl acetate = 10: 1, Rf = 0.33), and purified by column chromatography (petroleum ether: ethyl acetate = 10: 1 to 10: 0) to give compound D5-1-3 (20.2 g, 79.5 mmol, 72.0% yield) as a light yellow oil. f = 0.33). The reaction mixture was filtered after it was returned to room temperature, the filtrate was washed with saturated brine (600 mL), dried over Na2S04, filtered, and concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 100: 1 to 10: 1) to give compound D5-1-2 (39.0 g, 127 mmol, 64.5% yield) as a colorless oil.

[0535] 1 H NMR: EB9495-2-P1N1 (400 MHz, DMSO-d6) δ 7.69 (dd, J = 0.8, 8.1 Hz, 1H), 7.59 (d, J = 7.6 Hz, 1H), 7.47 - 7.39 (m, 1H), 4.64 (s, 2H), 3.91 (s, 3H)

[0536] 3. Synthesis of compound D5-1-4

[0537] To a mixture of compound K2CO3(136 mg, 984 pmol, 0.05 eq) in D2O (7.88 g, 394 mmol, 20.0 eq) was added compound D5-1-3 (5.00 g, 19.7 mmol, 1.00 eq) and THF (10.0 mL) at 25 °C under N2. The reaction was stirred at 25 °C for 12 h. HNMR detection showed that compound D5-1-3 was completely reacted to generate the target product. The reaction mixture was transferred to a separatory funnel to obtain an aqueous phase, which was extracted with MTBE (10.0 mL x 2 times). The obtained organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to obtain the crude compound D5-1-4 (4.80 g, 10.0 mmol, 50.9% yield, 53.4% purity) in the form of colorless oil.

[0538] 1 H NMR: EB9495-10-P1N1 (400 MHz, DMSO-d6) δ 7.73 (dd, J = 1.2, 8.0 Hz, 1H), 7.56-7.52 (m, 1H), 7.50-7.45 (m, 1H), 3.91 (s, 3H)

[0539] EB9495-10-P1N2 (400 MHz, DMSO-d6) δ 7.73 (dd, J = 1.2, 7.9 Hz, 1H), 7.56-7.52 (m, 1H), 7.50-7.45 (m, 1H), 3.91 (s, 3H)

[0540] 4. Synthesis of compound D5-1-5

[0541] To compound D5-1-4 (3.30 g, 12.9 mmol, 1.00 eq) was added BH3.THF (1.00 M, 77.3 mL, 6.00 eq) at 0 °C under N2, and the reaction was stirred at 25 °C for 16 h. LCMS (EB9776-9-IPCL2) detection showed that compound D5-1-4 was completely reacted to generate a new compound with the target molecular weight (R t = 1.070 min). The reaction was transferred to a new bottle, methanol (155 mL) was added thereto, and the reaction was stirred at 80 °C for 16 h. The mixture was concentrated under reduced pressure to obtain the product crude (dichloromethane:methanol = 10:1, R f = 0.20), which was purified by column chromatography (petroleum ether: ethyl acetate = 1:0 to 1:1) to obtain compound D5-1-5 (1.69 g, 7.41 mmol, 57.5% yield) in the form of yellow oil.

[0542] 1H NMR: EB9776-9-P1N1 (400 MHz CHLOROFORM-d) δ 7.61 (dd, J = 1.6, 7.8 Hz, 1H), 7.25 - 7.16 (m, 2H), 6.54 (br s, 1H), 3.48 (d, J = 3.6 Hz, 2H)

[0543] LCMS: EB9776-9-IPCL2, product: R T = 1.070 min, MS (ESI) m / z = 227.9 [M+H] +

[0544] 5. Synthesis of compound D5-1-6

[0545] To compound D5-1-5 (1.69 g, 7.41 mmol, 1.00 equiv) was added BH3.THF (1.00 M, 51.9 mL, 7.00 equiv) at 0 °C under N2, stirred at 25 °C for 16 h, then BH3.THF (1.00 M, 22.2 mL, 3.00 equiv) was added at 0 °C, stirred at 25 °C for 16 h, then BH3.THF (1.00 M, 14.8 mL, 2.00 equiv) was added at 0 °C, stirred at 25 °C for 4 h, then BH3.THF (1.00 M, 22.2 mL, 3.00 equiv) was added at 0 °C, stirred at 25 °C for 4 h, then BH3.THF (1.00 M, 14.8 mL, 2.00 equiv) was added at 0 °C, stirred at 25 °C for 4 h. LCMS (EB9776-11-IPCL15) detection showed the product of target molecular weight was generated (R t = 0.441 min). The reaction was transferred to a new flask, to which was added methanol (220 mL), and the reaction was stirred at 80 °C for 16 h. The mixture was concentrated under reduced pressure to give the product of crude compound D5-1-6 (2.62 g) as a yellow oil.

[0546] LCMS: EB9776-11-IPCL15, product: R T = 0.441 min, MS (ESI) m / z = 214.05 [M+H] +

[0547] 6. Synthesis of compound D5-1

[0548] To a mixture of compound D5-1-6 (2.62 g, 12.2 mmol, 1.00 equiv) in methanol (25.0 mL) was added DIEA (1.90 g, 14.7 mmol, 2.56 mL, 1.20 equiv) and (Boc)20 (2.67 g, 12.2 mmol, 2.81 mL, 1.00 equiv) and stirred at 25 °C for 16 h. LCMS (EB9776-14-IPCL2) detection showed that compound D5-1-6 was completely reacted to generate the product with the target molecular weight (R t = 1.803 min). The mixture was concentrated under reduced pressure to give the crude product, which was purified by column chromatography (petroleum ether: ethyl acetate = 5:1, R f = 0.62) to give compound D5-1 (870 mg, 2.77 mmol, 22.6% yield) as a yellow oil.

[0549] LCMS: EB9776-14-IPCL2, product: R T = 1.803 min, MS (ESI) m / z = 214.0 [M-Boc+H] +

[0550] 1 H NMR: EB9776-14-P1N1 (400 MHz CHLOROFORM-d) δ 7.42 (dd, J = 0.8, 7.7 Hz, 1H), 7.13-6.98 (m, 2H), 4.54 (br s, 2H), 3.63 (br s, 2H), 1.51 (s, 9H)

[0551] Synthesis of intermediate D6-1:

[0552] 1. Synthesis of compound D6-1-2

[0553] A mixture of compound D6-1-1 (10.0 g, 50.0 mmol, 1.00 equiv), dimethyl malonate (9.91 g, 75.0 mmol, 8.59 mL, 1.50 equiv) and K2CO3 (17.3 g, 125 mmol, 2.50 equiv) in DMF (50.0 mL) was stirred at 60 °C for 16 h. TLC (petroleum ether: ethyl acetate = 3:1) detection showed that compound D6-1-1 was completely reacted to generate a new compound with the main peak (R f= 0.53). Water (200 mL) and ethyl acetate (200 mL) were added to the post-reaction mixture, the organic layer was separated, extracted using ethyl acetate (100 mL x 2 times), the obtained organic layer was washed with saturated brine (200 mL), dried over Na2SO4, filtered, concentrated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 100:1 to 10:1) to obtain compound D6-1-2 (7.70 g, 24.7 mmol, 49.3% yield) as a white solid.

[0554] 1 H NMR: EB9495-16-P1N1 (400 MHz, DMSO-d6) δ 7.91 (d, J = 7.6 Hz, 1H), 7.71-7.65 (m, 1H), 7.64-7.59 (m, 1H), 5.36 (s, 1H), 3.74 (s, 6H)

[0555] 2. Synthesis of compound D6-1-3

[0556] To compound D6-1-2 (6.70 g, 21.5 mmol, 1.00 eq) was added ethyl acetate (70 mL) and Lil (28.7 g, 215 mmol, 8.23 mL, 10.0 eq) at 25 °C, and the reaction was stirred at 80 °C for 4 h. TLC (petroleum ether: ethyl acetate = 3:1) detection showed that compound D6-1-2 was completely reacted to generate a new compound main peak (R f = 0.34). The post-reaction mixture was terminated by adding citric acid (3.0 mL), extracted using ethyl acetate (20 mL x 2 times), the obtained organic layer was washed with saturated brine (10 mL), dried over Na2SO4, filtered, concentrated to obtain the crude product. The crude product (TLC: petroleum ether: ethyl acetate = 3:1, R f = 0.34) was purified by column chromatography (ethyl acetate / petroleum ether = 0-10%) to obtain compound D6-1-3 (3.90 g, 16.3 mmol, 75.7% yield) as a white solid.

[0557] 1 H NMR: EB9495-20-P1N1 (400 MHz, DMSO-d6) δ 7.79 (dd, J = 0.8, 7.9 Hz, 1H), 7.62-7.57 (m, 1H), 7.56-7.52 (m, 1H), 3.89 (s, 2H)

[0558] 3. Synthesis of compound D6-1-4

[0559] To compound D6-1-3 (1.70 g, 7.08 mmol, 1.00 equiv) was added THF (10.0 mL) at 0 °C, and tri-deuterium borane (0.75 M, 13.2 mL, 1.40 equiv) was added at 25 °C. The reaction was stirred at 0 °C for 4 h. TLC (petroleum ether: ethyl acetate = 1 : 1) detection showed that the reaction of compound D6-1-3 was complete, and a new compound main peak (R f = 0.41) was generated. The reaction mixture was concentrated to obtain the product crude, and the crude (TLC: petroleum ether: ethyl acetate = 1 : 1, R f = 0.41) was purified by column chromatography (petroleum ether: ethyl acetate = 100: 1 to 10: 1) to obtain compound D6-1-4 (1.40 g, 4.91 mmol, 69.3% yield, 80% purity) as a colorless oil.

[0560] LCMS: EB9495-21-P1L1, product: R T = 1.108 min, MS (ESI) m / z = 227.8 [M+H] +

[0561] 1 H NMR: EB9495-21-P1N1 (400 MHz, DMSO-d6) δ 7.72 (dd, J = 0.8, 7.8 Hz, 1H), 7.62 - 7.47 (m, 2H), 4.81 (s, 1H), 2.95 (s, 2H)

[0562] 4. Synthesis of compound D6-1-5

[0563] To compound D6-1-4 (400 mg, 1.75 mmol, 1.00 equiv) was added THF (3.00 mL) at 25 °C, and BH3.THF (1.00 M, 5.26 mL, 3.00 equiv) was added at 0 °C. The reaction was stirred at 25 °C for 10 h, and BH3.THF (1.00 M, 3.51 mL, 2.00 equiv) was added at 0 °C. The reaction was stirred at 25 °C for 10 h, and BH3-Me2S (10.0 M, 5.00 mL, 28.5 equiv) was added at 0 °C. The reaction was stirred at 25 °C for 10 h. LCMS (EB9495-38-IPCL10) detection showed that the reaction of compound D6-1-4 was complete, and a product of the target molecular weight was generated (R T = 0.965 min). The mixture was quenched by adding methanol (10.0 mL) at 80 °C, and the reaction was terminated after 12 h. The mixture was concentrated to obtain compound D6-1-5 (820 mg, crude) as a yellow oil.

[0564] LCMS: EB9495-38-IPCL10, Product: R T = 0.965 min, MS (ESI) m / z = 231.8 [M+H] +

[0565] 5. Synthesis of compound D6-1-6

[0566] To compound D6-1-5 (820 mg, 3.53 mmol, 1.00 equiv) was added THF (5.00 mL), DIEA (548 mg, 4.24 mmol, 738 μL, 1.20 equiv) and (Boc)20 (771 mg, 3.53 mmol, 812 μL, 1.00 equiv) at 25 °C under N2environment. The reaction was stirred at 25 °C for 4 h. TLC (petroleum ether: ethyl acetate = 1:1) detection showed that compound D6-1-5 was completely reacted to generate a new compound main peak (R f = 0.30). The reaction mixture was concentrated to obtain the product crude, which was purified by column chromatography (ethyl acetate: petroleum ether = 0-20%) to obtain compound D6-1-6 (150 mg, 452 μmol, 12.8% yield) in the form of yellow oil. f

[0567] 1 H NMR: EB9495-42-P1N4 (400 MHz, DMSO-d6) δ 7.45 (d, J = 7.6 Hz, 1H), 7.23 (d, J = 7.6 Hz, 1H), 7.17-7.11 (m, 1H), 6.90 (br s, 1H), 4.74-4.67 (m, 1H), 4.33 (br d, J = 4.8 Hz, 2H), 2.85 (s, 2H), 1.44-1.36 (m, 11H)

[0568] 6. Synthesis of compound D6-1-7

[0569] To compound D6-1-6 (400 mg, 1.20 mmol, 1.00 equiv) was added DCM (10.0 mL) and 4-methylmorpholine (365 mg, 3.61 mmol, 397 μL, 3.00 equiv) at 0 °C under N2environment, and stirred at 0 °C for 0.5 h. Methylsulfonyl chloride (340 mg, 2.97 mmol, 230 μL, 2.47 equiv) was added at 0 °C, and stirred at 0 °C for 2 h. LCMS (EB9776-41-IPCL1) detection showed that compound D6-1-6 was completely reacted to generate the product with the target molecular weight (R T ​= 0.702 min). To the reaction mixture, ice water (10.0 mL) was added, extracted with CH2Cl2(10.0 mL x 3 times), the organic layer was washed with saturated brine (10 mL), dried over Na2SO4, filtered, concentrated to give the crude product. The crude product (TLC: Petroleum ether: ethyl acetate = 1:1, Rf = 0.34) was purified by column chromatography (ethyl acetate / petroleum ether = 0-20%) to give compound D6-1-7 (450 mg, 1.10 mmol, 91.1% yield) as a yellow oil. f = 0.34) was purified by column chromatography (ethyl acetate / petroleum ether = 0-20%) to give compound D6-1-7 (450 mg, 1.10 mmol, 91.1% yield) as a yellow oil.

[0570] LCMS: EB9776-41-IPCL1, product: R T = 0.702 min, MS (ESI) m / z = 354.0 [M-t-Bu+H] +

[0571] 1 H NMR: EB9776-41-P1N1 (400 MHz, CHLOROFORM-d) δ 7.51 (dd, J = 0.8, 7.9 Hz, 1H), 7.23 (d, J = 7.2 Hz, 1H), 7.16 - 7.10 (m, 1H), 5.10 (br s, 1H), 4.51 (d, J = 6.0 Hz, 2H), 3.31 (s, 2H), 2.94 (s, 3H), 1.47 - 1.42 (m, 9H)

[0572] 7. Synthesis of compound D6-1

[0573] To compound D6-1-7 (450 mg, 1.10 mmol, 1.00 eq) was added THF (8.00 mL) and NaH (48.3 mg, 1.21 mmol, 60.0% purity, 1.10 eq) at 0 °C under N2environment, stirred at 25 °C for 3 hours. LCMS (EB9776-42-IPCL2) detection showed that compound D6-1-7 was completely reacted to generate the product of target molecular weight (R T = 0.804 min). To the mixture, water (10.0 mL) was added, extracted with ethyl acetate (10.0 mL x 3 times), the organic layer was washed with saturated brine (10 mL), dried over Na2SO4, filtered, concentrated to give the crude compound D6-1 (330 mg, 1.05 mmol, 95.8%) as a yellow oil.

[0574] LCMS: EB9776-42-IPCL2, product: R T = 0.804 min, MS (ESI) m / z = 215.3 [M-Boc+H] +

[0575] 1 H NMR: EB9776-42-P1N1 (400 MHz, CHLOROFORM-d) δ 7.42 (d, J = 7.6 Hz, 1H), 7.13 - 6.96 (m, 2H), 4.55 (br s, 2H), 2.83 (s, 2H), 1.51 (s, 9H)

[0576] Synthesis of intermediate D1-0:

[0577] 1. Synthesis of compound D1-0-2

[0578] To ethanol (180 mL) was added water (60.0 mL), compound D1-0-1 (10.0 g, 78.0 mmol, 1.00 equiv), Fe (21.8 g, 390 mmol, 5.00 equiv) and NH4CI (20.9 g, 390 mmol, 5.00 equiv) at 25 °C under N2environment, the mixture was stirred at 60 °C for 12 h. TLC (petroleum ether: ethyl acetate = 10: 1) detection showed that compound D1-0-1 was completely reacted to generate a new compound main peak (R f = 0.15). After the reaction mixture was returned to room temperature, it was filtered, the filter residue was washed with ethyl acetate (500 mL x 2 times), extracted with ethyl acetate (300 mL x 2 times), the obtained organic layer was washed with saturated brine (500 mL), dried over Na2S04, filtered, concentrated to give the crude product. The crude product (TLC: petroleum ether: ethyl acetate = 10: 1, R f = 0.15) was purified by column chromatography (ethyl acetate / petroleum ether = 0-10%) to give compound D1-0-2 (83.4% yield) as a red liquid.

[0579] 1 H NMR: EB8953-2-P1N2 (400 MHz, DMSO-d6) δ ppm 4.98 (br s, 2H).

[0580] 2. Synthesis of compound D1-0

[0581] To acetic acid (30.0 mL) at 10 °C was added KSCN (4.98 g, 51.3 mmol, 4.98 mL, 3.00 equiv) and compound D1-0-2 (2.00 g, 17.1 mmol, 1.96 mL, 1.00 equiv) under N2, stirred for 30 min, then Br2(2.73 g, 17.1 mmol, 881 μL, 1.00 equiv) dissolved in acetic acid (10.0 mL) was added at 10 °C, the mixture was stirred at 25 °C for 6 h. LCMS (EB8953-44-IPCL2) detection showed that compound D1-0-2 was completely reacted to generate the product of the target molecular weight (R t = 1.068 min). The reaction mixture was adjusted to pH ~ 8 with ammonia gas (50%), water (100 mL) was added, extracted with ethyl acetate (100 mL x 2 times), the obtained organic layer was washed with saturated brine (100 mL), dried over Na2SO4, filtered and concentrated to obtain the crude product. The crude product (TLC: petroleum ether: ethyl acetate = 2:1, R f = 0.54) was purified by column chromatography (ethyl acetate / petroleum ether = 0-10%) to obtain compound D1-0 (74.7% yield) as a yellow solid.

[0582] LCMS: EB8953-44-IPCL2, product: Rt = 1.068 min, MS (ESI) m / z = 331.8 [2M+Na] +

[0583] 1 H NMR: EB8953-44-P1N1 (400 MHz, DMSO-d6) δ ppm 5.78 (s, 2H).

[0584] Synthesis of deuterated PROTAC molecule A13D6-PF3:

[0585] The synthesis of deuterated PROTAC molecule A13D6-PF3 followed the synthesis of non-deuterated PROTAC molecule APF3, but the starting material A-1331852 was replaced by deuterated molecule A13D-6, and the final product was purified by reverse phase HPLC (neutral condition) to obtain the target product A13D6-PF3 (12.6 mg, 11.2 μmol, 36.9% yield, 96.9% purity) as a yellow solid.

[0586] LCMS: EC15866-4-P1C2, RT = 0.567 min, M / Z (M+H + ) = 1091.4

[0587] 1HNMR: EC15866-4-P1A (400 MHz, DMSO-d6) δ ppm 12.84 (s, 1 H) 11.08 (s, 1 H) 8.17 (t, J=5.6 Hz, 1 H) 8.01 (d, J=7.6 Hz, 1 H) 7.77 (d, J=8.4 Hz, 1 H) 7.52 - 7.62 (m, 2 H) 7.43 - 7.48 (m, 2 H) 7.38 - 7.42 (m, 1 H) 7.31 - 7.37 (m, 2 H) 7.25 (s, 1 H) 7.10 (d, J=8.4 Hz, 1 H) 6.99 - 7.05 (m, 1 H) 6.91 - 6.98 (m, 1 H) 6.58 (t, J=5.6 Hz, 1 H) 5.04 (dd, J=12.8, 5.32 Hz, 1 H) 4.93 - 5.01 (m, 2 H) 3.67 (s, 2 H) 3.55 - 3.62 (m, 2 H) 3.47 - 3.53 (m, 4 H) 3.41 - 3.46 (m, 4 H) 3.37 - 3.40 (m, 2 H) 3.33 - 3.36 (m, 2 H) 3.22 - 3.26 (m, 2 H) 2.95 - 3.01 (m, 3 H) 2.81 - 2.92 (m, 1 H) 2.58 (s, 2 H) 2.05 (s, 3 H) 1.98 - 2.02 (m, 1 H) 1.90 (s, 3 H) 1.60 - 1.65 (m, 3 H) 1.49 - 1.56 (m, 9 H).

[0588] Example 6. Synthesis of PROTAC molecules recruiting CRBN-E3 enzymes based on novel ligand structures

[0589] a) Synthesis of APQ5a and APQ5b

[0590] Synthesis of APQ5a:

[0591] 1. Synthesis of compound Q5a-2

[0592] To a solution of compound 5A (1.00 g, 3.65 mmol, 1.0 eq) in DMF (20.0 mL) was added DMAP (22.3 mg, 182 pmol, 0.05 eq), EDCI (769 mg, 4.01 mmol, 1.1 eq) and acrylic acid (525 mg, 7.29 mmol, 500 pL, 2.0 eq), the mixture was stirred at 25 °C for 8 h. LC-MS (EC16331-35-P1G, P1: RT = 0.390 min, R1: RT = 0.341 min) monitoring showed that compound 5A was completely reacted to generate the main peak of m / z consistent with the desired product. After reaction, the mixture was concentrated under reduced pressure to remove the solvent to obtain the crude product, which was further purified by reverse phase HPLC (0.1% NH3-H2O) column chromatography to obtain compound Q5a-2 (200 mg, 609 pmol, 16.7% yield) as a brown solid.

[0593] LC-MS: EC16331-35-P1G, product: RT = 0.390 min, m / z = 329.1 [M+H]+

[0594] 2. Synthesis of compound APQ5a

[0595] To a solution of compound C1 (150 mg, 213 pmol, 1.0 eq) in THF (2.00 mL) was added TEA (216 mg, 2.13 mmol, 296 pL, 10 eq) and compound Q5a-2 (70.0 mg, 213 pmol, 1.0 eq), the mixture was stirred at 25 °C for 16 h. LC-MS (EC16331-41-P1E, P1: RT = 0.413 min) monitoring showed that compound Q5a-2 was completely reacted to generate the main peak of m / z consistent with the desired product. After reaction, the mixture was concentrated under reduced pressure to remove the solvent to obtain the crude product, which was further purified by preparative HPLC (column: Phenomenex luna C18 150*25mm*10pm; mobile phase: [water(FA)-ACN]; gradient elution: 31%-51%B in 15 min) column chromatography to obtain compound APQ5a (20.0 mg, 17.9 pmol, 8.39% yield, 92.41% purity) as a white solid.

[0596] LC-MS: EC16331-41-P1E, product: RT = 0.413 min, m / z = 1033.5 [M+H] +

[0597] 1H NMR: EC16331-41-P1K (400 MHz DMSO) δ ppm 10.27 (s, 1H), 8.20 (s, 1H), 8.03 (d, J = 7.8 Hz, 1H), 7.79 (d, J = 7.8 Hz, 1H), 7.59 (d, J = 6.6 Hz, 1H), 7.44-7.52 (m, 2H), 7.38-7.42 (m, 1H), 7.32-7.37 (m, 2H), 7.26 (dd, J = 12.0, 2.0 Hz, 1H), 7.14-7.19 (m, 3H), 7.05-7.11 (m, 1H), 6.94 (d, J = 9.2 Hz, 2H), 6.89 (d, J = 8.8 Hz, 1H), 4.91 (s, 2H), 4.00 (t, J = 6.4 Hz, 2H), 3.85 (t, J = 5.6 Hz, 2H), 3.69 (t, J = 6.6 Hz, 3H), 3.59 (d, J = 3.6 Hz, 5H), 3.45 (s, 3H), 3.13 (s, 3H), 3.07 (s, 3H), 2.97 (t, J = 5.6 Hz, 3H), 2.77 (t, J = 7.6 Hz, 3H), 2.67 (t, J = 6.6 Hz, 3H), 2.54 (s, 4H), 1.92-2.00 (m, 2H).

[0598] Synthesis of APQ5b:

[0599] 1. Synthesis of compound Q5b-2

[0600] To a solution of compound Q5b-1 (1.00 g, 6.17 mmol, 1.0 eq) in methanol (10.0 mL) was added NaOH (1 M, 6.17 mL, 1.0 eq) and the reaction was stirred at 25 °C for 2 h. After reaction, the mixture was adjusted to pH less than 6 using HC1 (1 M), and separated by extraction using H2O (20.0 mL) and DCM (30.0 mL, 3 times), the organic layer was washed with saturated brine (30.0 mL, 2 times), dried over Na2S04, and concentrated under reduced pressure to give compound Q5b-2 product crude (500 mg, 3.37 mmol, 54.73% yield) as a colorless oil.

[0601] 2. Synthesis of compound Q5b-3

[0602] Compound Q5b-2 (500 mg, 3.37 mmol, 1.0 eq), compound 5A (925 mg, 3.37 mmol, 1.0 eq), DMAP (20.6 mg, 168 µmol, 0.05 eq) and EDCI (711 mg, 3.71 mmol, 1.1 eq) were mixed in DCM (10.0 mL) and stirred at 25 °C for 8 h under N2. After reaction, the mixture was separated by extraction with H2O (10.0 mL) and DCM (10.0 mL, 3 times), the organic layer was washed with saturated brine (20.0 mL, 2 times), dried over Na2SO4, and concentrated under reduced pressure to give compound Q5b-3 (400 mg) as a colorless oil.

[0603] 3. Synthesis of compound Q5b-4

[0604] Compound Q5b-3 (500 mg, 1.24 mmol, 1.0 eq) was dissolved in formic acid (5.00 mL) and stirred at 25 °C for 2 h. After reaction, the mixture was concentrated under reduced pressure to give a crude product, which was further purified by reverse phase HPLC (0.1% FA condition) to give compound Q5b-4 (30.0 mg, about 75% purity) as a brown solid.

[0605] 4. Synthesis of APQ5b

[0606] Compound Q5b-4 (20.0 mg, 55.8 µmol, 1.0 eq), compound C1 (39.3 mg, 55.8 µmol, 1.0 eq) and NaBH(OAc)3 (35.5 mg, 167 µmol, 3.0 eq) were mixed in DMF (1.00 mL) and stirred at 25 °C for 2 h under N2. After reaction, the mixture was concentrated under reduced pressure to remove the solvent to give a crude product, which was purified by preparative HPLC (column: Phenomenex luna C18 150*25mm*10µm; mobile phase: [water(FA)-ACN]; gradient: 32%-52%B in 10 min) to give compound APQ5b (6.3 mg, 9.47 µmol, 16.97% yield, 99.2% purity) as a white solid.

[0607] LC-MS: EC16331-37-P1C1, Product: RT = 0.418 min, m / z = 1047.5 [M+H] +

[0608] 1H NMR: EC16331-37-P1A (400 MHz DMSO) δ ppm 10.26 (s, 1H), 8.18 (s, 1H), 8.03 (d, J = 7.6 Hz, 1H), 7.79 (d, J = 8.4 Hz, 1H) 7.60 (d, J = 7.2 Hz, 1H), 7.51 (d, J = 8.8 Hz, 1H), 7.45-7.48 (m, 1H), 7.39-7.42 (m, 1H), 7.32-7.38 (m, 2H), 7.26 (dd, J = 11.6, 1.6 Hz, 1H), 7.13-7.19 (m, 3H), 7.08 (d, J = 8.8 Hz, 1H), 6.89-6.94 (m, 3H), 4.91 (s, 2H), 4.00 (t, J = 6.4 Hz, 2H), 3.85 (t, J = 5.6 Hz, 2H), 3.69 (t, J = 6.8 Hz, 3H), 3.57-3.60 (m, 4H), 3.45 (br s, 3H), 3.12 (s, 3H), 3.06 (s, 2H), 2.97 (t, J = 5.6 Hz, 3H), 2.78 (t, J = 7.38 Hz, 2H) 2.67 (t, J = 6.8 Hz, 3H), 2.32-2.37 (m, 3H), 2.28 (t, J = 7.2 Hz, 2H), 1.91-2.00 (m, 3H), 1.67 (q, J = 7.2 Hz, 3H).

[0609] b) Synthesis of APQ6a and APQ6b

[0610] Synthesis of APQ6a:

[0611] 1. Synthesis of Q6a-2

[0612] To compound Q6a-1 (2.00 g, 11.6 mmol, 1.00 equiv) was added acrylic acid (3.35 g, 46.5 mmol, 3.19 mL, 4.00 equiv) and stirred at 110 °C for 3 hours, then CO(NH2)2 (4.48 g, 74.6 mmol, 4.01 mL, 6.40 equiv) and AcOH (20 mL) were added, the mixture was stirred at 120 °C for 12 hours. The reaction mixture was added to water (50.0 mL) at 0 °C, the residue was obtained by filtration, washed with water (20.0 mL), and concentrated under reduced pressure to obtain compound Q6a-2 product crude (2.20 g, 8.18 mmol, 70.3% yield) as a brown solid.

[0613] LCMS: EC16416-3-p1a, P1 RT = 0.265 min, m / z = 269.0 [M+H] +

[0614] 2. Synthesis of Q6a-4

[0615] Compound Q6a-3 (2.44 g, 22.3 mmol, 3.00 equiv, HC1), Q6a-2 (2.00 g, 7.43 mmol, 1.00 equiv), Cs2C03(7.26 g, 22.3 mmol, 3.00 equiv), RuPhos (693 mg, 1.49 mmol, 0.200 equiv) and Pd2(dba)3(681 mg, 743 pmol, 0.100 equiv) were mixed in dioxane (20.0 mL), the mixture was stirred at 100 °C for 16 h under N2. The reaction mixture was concentrated under reduced pressure to give the crude product, which was further purified by prep-HPLC (neutral condition) to give product Q6a-4 (300 mg, 1.15 mmol, 15.4% yield) as a white solid.

[0616] LCMS: EC16364-67-P1A1, P1 RT = 0.465 min, m / z = 262.1 [M+H] +

[0617] 3. Synthesis of Q6a-5

[0618] To a solution of Py-S03(122 mg, 765 pmol, 4.00 equiv) in DMSO (1.00 mL) was added TEA (96.8 mg, 957 pmol, 133 pL, 5.00 equiv) dropwise at 0 °C, stirred at 0 °C for 15 min, then compound Q6a-4 (50.0 mg, 191 pmol, 1.00 equiv) dissolved in DMSO (1.00 mL) was added dropwise at 0 °C, the mixture was stirred at 25 °C for 3.75 h. LCMS (P1, RT = 0.331 min) monitoring showed Q6a-4 was completely reacted. The reaction mixture was concentrated under reduced pressure to give the crude product, which was purified by prep-HPLC (FA condition) to give product Q6a-5 (25.0 mg, 96.4 pmol, 50.4% yield) as a white solid.

[0619] LCMS: EEC16364-119-P1A1, P1 RT = 0.331 min, m / z = 260.0 [M+H] +

[0620] 4. Synthesis of Q6a

[0621] To a solution of compound C1 (27.2 mg, 38.6 pmol, 1.00 equiv) in DCM (1 mL) was added TEA (39.0 mg, 386 pmol, 53.7 pL, 10.0 equiv) and compound Q6a-5 (10.0 mg, 38.6 pmol, 1.00 equiv), stirred at 25 °C for 0.5 h, then added NaBH(OAc)3 (24.5 mg, 116 pmol, 3.00 equiv), the mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure to give the product crude, purified by prep-HPLC (neutral condition) to give product APQ6a (8.05 mg, 8.44 pmol, 21.9% yield) as a white solid.

[0622] LCMS: EC16364-125-P1X, P1 RT = 0.470 min, m / z = 948.1 [M+H] +

[0623] 1 H NMR: EC16364-125-P1C (400 MHz DMSO-d6) d 10.22 (s, 1H) 8.03 (d, J = 7.6 Hz, 1H) 7.79 (d, J = 8.0 Hz, 1H) 7.59 (d, J = 7.2 Hz, 1H) 7.44 - 7.54 (m, 2H) 7.38 - 7.42 (m, 1H) 7.32 - 7.38 (m, 2H) 7.26 (dd, J = 12.0, 1.88 Hz, 1H) 7.14 - 7.20 (m, 1H) 7.09 (d, J = 8.8 Hz, 3H) 6.90 (d, J = 8.8 Hz, 1H) 6.41 (d, J = 8.8 Hz, 2H) 4.91 (s, 2H) 4.01 (t, J = 6.4 Hz, 2H) 3.85 (t, J = 6.0 Hz, 2H) 3.90 (t, J = 7.2 Hz, 2H) 3.66 (t, J = 6.8 Hz, 2H) 3.54 - 3.58 (m, 2H) 3.48 (s, 2H) 3.21 - 3.26 (m, 5H) 2.97 (t, J = 5.6 Hz, 2H) 2.77 (t, J = 7.2 Hz, 2H) 2.66 (t, J = 6.8 Hz, 2H) 2.52 - 2.57 (m, 2H) 2.38 (d, J = 2.4 Hz, 2H) 1.92 - 2.00 (m, 2H).

[0624] Synthesis of APQ6b:

[0625] 1. Synthesis of Q6b-2

[0626] A mixture of compound Q6a-2 (2.00 g, 7.43 mmol, 1.00 equiv), Q6b-1 (1.01 g, 8.18 mmol, 1.01 equiv, HC1), Cs2C03(7.26 g, 22.3 mmol, 3.00 equiv) and RuPhos Pd G3 (1.24 g, 1.49 mmol, 0.200 equiv) in dioxane (20.0 mL) was stirred at 100 °C for 16 h under N2. The reaction mixture was diluted with water (30.0 mL), extracted with ethyl acetate (120.0 mL, 40.0 mL x 3 times), the organic layer was washed with saturated brine (120.0 mL, 60.0 mL x 2 times), dried over Na2S04, filtered and concentrated under reduced pressure to give the product crude, which was further purified by reverse phase HPLC (FA condition) to give product Q6b-2 (300 mg, crude) as a brown solid.

[0627] LCMS: EC16331-20-P1B, product: RT = 0.297 min, m / z = 275.9 [M+H] +

[0628] 2. Synthesis of Q6b-3

[0629] To a solution of compound Q6b-2 (100 mg, 363 pmol, 1.00 equiv) in DCM (1.00 mL) was added TEA (110 mg, 1.09 mmol, 152 pL, 3.00 equiv), TsCl (208 mg, 1.09 mmol, 3.00 equiv) and DMAP (178 mg, 1.45 mmol, 4.00 equiv), the mixture was stirred at 50 °C for 4 h. The reaction mixture was quenched by adding water (20.0 mL) at 0 °C, diluted with ethyl acetate (20.0 mL), extracted with ethyl acetate (60.0 mL, 20.0 mL x 3 times), the organic layer was washed with saturated brine (20.0 mL), dried over Na2S04, filtered and concentrated under reduced pressure to give the product crude, which was further purified by column chromatography (Si02, petroleum ether: ethyl acetate = 10: 1 to 5: 1) to give product Q6b-3 (70.0 mg, 163 pmol, 44.9% yield) as a white solid.

[0630] LCMS: EC16364-56-P1A, product: RT = 0.395 min, m / z = 430.1 [M+H] +

[0631] 3. Synthesis of APQ6b

[0632] To a solution of compound Q6b-3 (50.0 mg, 116 pmol, 1.00 equiv) and CI (49.2 mg, 69.8 pmol, 0.600 equiv) in DMF (0.500 mL) was added DIEA (60.2 mg, 465 pmol, 81.1 pL, 4.00 equiv) and KI (48.3 mg, 291 pmol, 2.50 equiv), the mixture was stirred at 60 °C for 2 h. After reaction, the mixture was filtered, concentrated under reduced pressure to give the product crude, which was further purified by prep-HPLC (FA condition and neutral condition) to give product APQ6b (2.40 mg, 2.49 pmol, 2.14% yield) as a white solid.

[0633] LCMS: EC16331-43-P1B, P1 RT = 0.427 min, m / z = 962.3 [M+H] +

[0634] 1 H NMR: EC16364-64-P1D (400 MHz DMSO-d6) d 10.23 (s, 1H) 7.87 (d, J = 7.2 Hz, 1H) 7.68 (d, J = 8.8 Hz, 1H) 7.57 - 7.63 (m, 1H) 7.54 (d, J = 8.8 Hz, 1H) 7.23 - 7.50 (m, 3H) 7.22 - 7.28 (m, 2H) 7.18 (d, J = 8.8 Hz, 1H) 7.08 - 7.12 (m, 1H) 7.04 (d, J = 7.2 Hz, 2H) 6.94 - 6.98 (m, 1H) 6.31 (d, J = 7.6 Hz, 1H) 4.95 (s, 2H) 4.28 (d, J = 6.0 Hz, 2H) 3.98 - 4.03 (m, 2H) 3.75 (d, J = 6.0 Hz, 2H) 3.70 - 3.74 (m, 2H) 3.59 - 3.65 (m, 4H) 3.48 - 3.52 (m, 4H) 3.42 (s, 2H) 2.93 (d, J = 5.6 Hz, 2H) 2.73 (s, 4H) 2.66 (s, 2H) 2.44 (s, 2H) 2.33 (s, 1H) 1.89 - 2.01 (m, 4H).

[0635] c) Synthesis of APQ7a and APQ7b

[0636] Synthesis of APQ7a:

[0637] 1. Synthesis of compound Q7a-2

[0638] To a solution of compound 4A (6.58 g, 55.3 mmol, 4.77 mL, 1.5 eq) in a mixture of H2O (50.0 mL) and CHCl3(50.0 mL) was added K2CO3(15.3 g, 110 mmol, 3 eq) and compound Q7a-1 (5.00 g, 36.9 mmol, 1.0 eq), and the mixture was stirred at 20 °C for 8 h. After reaction, the mixture was separated and the organic layer was washed with saturated brine (30.0 mL, 2 times), dried over Na2SO4, and concentrated under reduced pressure to give the product as a crude, which was further purified by column chromatography (SiO2, dichloromethane:methanol = 97:3) to give compound Q7a-2 as a yellow oil.

[0639] 1 H NMR: EC9174-266-P1C (400 MHz CDCl3) δ ppm 3.45 (d, J = 2.4 Hz, 2H), 2.87 (t, J = 6.0 Hz, 4H), 2.50 (t, J = 6.0 Hz, 4H), 2.29 (t, J = 2.4 Hz, 1H).

[0640] 2. Synthesis of compound Q7a-3

[0641] Compound 6A (867 mg, 3.64 mmol, 1.0 eq), Pd(PPh3)2Cl2(51.2 mg, 72.9 μmol, 0.02 eq), CuI (27.8 mg, 146 μmol, 0.04 eq) and TEA (10.0 mL) were mixed, and compound Q7a-2 (500 mg, 3.64 mmol, 1.0 eq) was added under N2environment, and the mixture was stirred at 25 °C for 8 h. After reaction, the mixture was extracted with H2O (30.0 mL) and ethyl acetate (40.0 mL), and filtered under reduced pressure to give Q7a-3 product as a crude (1.00 g, crude) as a brown solid.

[0642] LC-MS: EC9174-317-P1E7, product: RT = 0.302 min, m / z = 246.3 [M+H] +

[0643] 1 H NMR: EC9174-317-P1A (400 MHz CDCl3) δ ppm 7.09-7.15 (m, 2H), 6.93 (t, J = 8.8 Hz, 1H), 3.64 (s, 2H), 2.95 (t, J = 6.0 Hz, 4H), 2.55 (t, J = 6.0 Hz, 4H).

[0644] 3. Synthesis of compound Q7a-4

[0645] To a mixture of compound Q7a-3 (400 mg, 1.62 mmol, 1.0 eq), compound 5A (466 mg, 1.70 mmol, 1.05 eq) and NaBH(OAc)3 (686 mg, 3.24 mmol, 2.0 eq) in DCE (10.0 mL), HOAc (117 mg, 1.94 mmol, 111 μL, 1.2 eq) was added dropwise at 25 °C within 30 min, and the mixture was stirred at 25 °C for 1.5 h. After reaction, the mixture was concentrated under reduced pressure to give the crude product, which was further purified by column chromatography (0.1% NH3·H2O) to give compound Q7a-4 (100 mg) as a brown solid.

[0646] LC-MS: EC16331-24-P1A, Product: RT = 0.392 min, m / z = 506.2 [M+H] +

[0647] 4. Synthesis of compound Q7a-5

[0648] A mixture of compound Q7a-4 (80.0 mg, 158 μmol, 1.0 eq), compound Q4-3 (106 mg, 174 μmol, 1.1 eq) and Cs2CO3 (103 mg, 316 μmol, 2.0 eq) in DMA (2.00 mL) was stirred at 25 °C for 2 h under N2. After reaction, the mixture was concentrated under reduced pressure to give the crude product, which was further purified by column chromatography (0.1% NH3·H2O) to give compound Q7a-5 (30.0 mg) as a white solid.

[0649] 5. Synthesis of compound APQ7a

[0650] Compound Q7a-5 (30.0 mg, 29.1 μmol, 1.0 eq) was dissolved in HC1-dioxane (2.00 mL), the reaction was stirred at 25 °C for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the product crude, which was further purified by reverse phase HPLC column (column: Daisogel SP ODS RPS 150*25mm*5μm; mobile phase: [water(NH4HCO3)-ACN]; gradient elution: 28%-58%B in 10 min) to obtain compound APQ7a (8.60 mg, 10.2 μmol, 35.25% yield, 95.76% purity) as a white solid.

[0651] LC-MS: EC16331-38-P1D2, Product: RT = 1.578 min, m / z = 976.2 [M+H] +

[0652] 1 H NMR: EC16331-38-P1A (400MHz DMSO) δ ppm 10.25 (s, 1H), 8.03 (d, J = 7.50 Hz, 1H), 7.78 (d, J = 7.6 Hz, 1H), 7.60 (d, J = 7.2 Hz, 1H), 7.48-7.51 (m, 1H), 7.47 (s, 1H), 7.38-7.42 (m, 1H), 7.35 (dd, J = 7.2, 2.8 Hz, 2H), 7.27 (dd, J = 11.6, 2.0 Hz, 1H), 7.16-7.19 (m, 1H), 7.14 (d, J = 9.2 Hz, 2H), 7.10 (d, J = 8.8 Hz, 1H), 6.92 (d, J = 9.2 Hz, 2H), 6.89 (s, 1H), 4.91 (s, 2H), 4.01 (t, J = 6.4 Hz, 2H), 3.85 (t, J = 6.0 Hz, 2H), 3.69 (t, J = 6.4 Hz, 3H), 3.45 (s, 3H), 3.11 (s, 4H), 2.97 (t, J = 5.6 Hz, 3H), 2.89 (d, J = 10.8 Hz, 3H), 2.77 (t, J = 7.2 Hz, 3H), 2.67 (t, J = 6.8 Hz, 3H), 2.62 (s, 5H), 2.13-2.23 (m, 4H), 1.93-2.00 (m, 2H), 1.80 (d, J = 11.6 Hz, 2H), 1.38-1.52 (m, 3H).

[0653] Synthesis of APQ7b:

[0654] The synthesis of the PROTAC molecule APQ7b followed the procedure of APQ7a, but the starting material compound Q7a-1 was replaced by compound 1A, and the final product was purified by preparative HPLC (column: Welch Xtimate C18 150*25mm*5um; mobile phase: [water(HCl)-ACN]; gradient elution: 18%-48%B in 8 min) column chromatography to give compound APQ7b (13.7 mg, 13.4 pmol, 82.4% yield, 96.8% purity) as a brown solid.

[0655] LC-MS: EC11255-211-P1A1, product: RT = 0.372 min, m / z = 496.0 [M / 2+H] +

[0656] 1 H NMR: EC11255-211-P1B (400 MHz DMSO) d ppm 8.04 (d, J = 7.8 Hz, 1H), 7.79 (d, J = 8.0 Hz, 1H), 7.54-7.65 (m, 2H), 7.29-7.50 (m, 6H), 7.13-7.24 (m, 3H), 6.97-7.05 (m, 3H), 4.93 (s, 2H), 4.30 (s, 2H), 4.06 (t, J = 6.4 Hz, 2H), 3.86 (t, J = 5.6 Hz, 2H), 3.79 (d, J = 12.0 Hz, 2H), 3.70 (t, J = 6.6 Hz, 2H), 3.64 (s, 4H), 3.22-3.30 (m, 2H), 3.04-3.18 (m, 6H), 2.98 (t, J = 5.8 Hz, 2H), 2.78-2.86 (m, 2H), 2.69 (t, J = 6.6 Hz, 2H), 2.13 (d, J = 13.8 Hz, 3H), 1.92-2.02 (m, 2H), 1.52-1.72 (m, 2H).

[0657] d) Synthesis of APQ8, APQ9, APQ10 and APQ11

[0658] Synthesis of APQ8:

[0659] 1. Synthesis of compound Q8-3

[0660] To a solution of compound Q8-2 (2.95 g, 17.4 mmol, 2.05 mL, 1.2 eq) in DMF (35.0 mL) was added compound Q8-1 (3.00 g, 14.5 mmol, 1.0 eq), KI (2.66 g, 16.00 mmol, 1.1 eq) and K2CO3 (4.02 g, 29.1 mmol, 2.0 eq), the mixture was stirred at 115 °C for 16 h, additional Q8-1 (1.50 g, 8.87 mmol, 1.04 mL, 0.6 eq) was added, the reaction was continued to stir at 115 °C for 3 h. After reaction, the mixture was filtered, concentrated under reduced pressure to give the product crude, which was purified by preparative HPLC column (TFA condition) to give compound Q8-3 (1.60 g, 5.44 mmol, 37.37% yield) as a white solid.

[0661] LC-MS: EC16416-5-P1da, product: RT = 0.326 min, m / z = 231.1 [M+H] +

[0662] 2. Synthesis of compound Q8-4

[0663] Compound Q8-3 (300 mg, 1.02 mmol, 1.0 eq) was dissolved in formic acid (5.00 mL), the mixture was stirred at 25 °C for 2 h. After reaction, the mixture was concentrated under reduced pressure to give compound Q8-4 product crude (385 mg) as a yellow oil.

[0664] LC-MS: EC11532-166-p1a1, product: RT = 0.183 min, m / z = 231.1 [M+H] +

[0665] 3. Synthesis of compound Q8-5

[0666] To a solution of compound Q8-4 (288 mg, 1.16 mmol, 1.0 eq) in DCM (10.0 mL) was added TEA (1.17 g, 11.6 mmol, 1.61 mL, 10.0 eq) and compound 1A (184 mg, 1.16 mmol, 1.0 eq), the mixture was stirred at 25 °C for 0.5 h, then NaBH(OAc)3 (737 mg, 3.48 mmol, 3.0 eq) was added and stirred at 25 °C for 1 h. The mixture was quenched by adding water (5.00 mL) at 25 °C, extracted with DCM (20.0 mL, 5.00 mL x 4 times), the organic layer was washed with saturated brine (10.0 mL, 5.00 mL x 2 times), dried over Na2SO4, filtered, concentrated under reduced pressure to give compound Q8-5 product crude (388 mg, 991 µmol, 85.43% yield) as a yellow solid, which was used directly in the next reaction without purification.

[0667] LC-MS: EC11532-169-p1a1, Product: RT = 0.188 min, m / z = 392.1 [M+H] +

[0668] 4. Synthesis of compound Q8-6

[0669] Compound Q8-5 (100 mg, 255 µmol, 1.0 eq) was dissolved in formic acid (0.5 mL), the mixture was stirred at 25 °C for 2 h. After reaction, the mixture was concentrated under reduced pressure to remove the solvent to give compound Q8-6 product crude (80.0 mg) as a colorless oil.

[0670] LC-MS: EC16331-29-P1A, Product: RT = 0.172 min, m / z = 346.1 [M+H] +

[0671] 5. Synthesis of compound APQ8

[0672] To a solution of compound Q8-6 (80.0 mg, 231 pmol, 1.0 eq) and CI (163 mg, 231 pmol, 1 eq) in DMF (2.00 mL) was added TEA (234 mg, 2.32 mmol, 322 pL, 10.0 eq) and NaBH(OAc)3 (147 mg, 695 pmol, 3.0 eq), the mixture was stirred at 25 °C for 2 hours. After reaction, the mixture was concentrated under reduced pressure to give the product crude, which was purified by preparative HPLC column chromatography (column: Phenomenex luna C18 150*25mm*10pm; mobile phase: [water(FA)-ACN]; gradient elution: 20%-50%B in 10 min) to give compound APQ8 (30.0 mg, 28.6 pmol, 12.33% yield, 98.45% purity) as a white solid.

[0673] LC-MS: EC16331-30-P1A1, Product: RT = 0.379 min, m / z = 1034.5 [M+H] +

[0674] 1H NMR: EC16331-30-P1A (400 MHz DMSO) δ ppm 10.29 (s, 1 H), 8.18 (s, 1 H), 8.03 (d, J = 7.6 Hz, 1 H), 7.79 (d, J = 8.0 Hz, 1 H), 7.59 (d, J = 7.6 Hz, 1 H), 7.51 (d, J = 8.8 Hz, 1 H), 7.45-7.49 (m, 1 H), 7.39-7.42 (m, 1 H), 7.35 (s, 1 H), 7.34 (s, 1 H), 7.26 (dd, J = 11.6, 1.6 Hz, 1 H), 7.21 (d, J = 9.2 Hz, 2 H), 7.17 (d, J = 10.4 Hz, 1 H), 7.06-7.12 (m, 1 H), 6.94 (s, 1 H), 6.92 (s, 1 H), 6.89-6.92 (m, 1 H), 4.91 (s, 2 H), 4.03 (dt, J = 16.8, 6.4 Hz, 5 H), 3.85 (t, J = 6.0 Hz, 2 H), 3.70 (t, J = 6.8 Hz, 3 H), 3.44 (s, 3 H), 2.97 (t, J = 5.6 Hz, 2 H), 2.90 (d, J = 11.6 Hz, 2 H), 2.78 (t, J = 7.6 Hz, 3 H), 2.65-2.71 (m, 5 H), 2.29-2.40 (m, 4 H), 2.09 (d, J = 7.2 Hz, 2 H), 1.92-2.04 (m, 5 H), 1.60-1.67 (m, 2 H), 1.41-1.49 (m, 1 H), 1.02-1.14 (m, 2 H).

[0675] Synthesis of APQ9:

[0676] 1. Synthesis of compound Q9-1

[0677] To a solution of compound Q4-3 (100 mg, 164 μmol, 1.0 eq) in DMA (2 mL) was added Cs2CO3 (134 mg, 411 μmol, 2.5 eq) and compound 6A (58.7 mg, 246 μmol, 1.5 eq), stirred at 25 °C for 16 h. The mixture was quenched by adding water (10.0 mL) at 0 °C, extracted with ethyl acetate (30.0 mL, 10.0 mL x 3 times), the organic layer was washed with saturated brine (10.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give the product as a crude, which was further purified by column chromatography (SiO2, ethyl acetate: petroleum ether = 3: 1 to 1: 1) to give compound Q9-1 (60.0 mg, 72.4 μmol, 43.9% yield, 92.2% purity) as a yellow solid.

[0678] LC-MS: EC16364-30-P1A, Product: RT = 0.619 min, m / z = 765.1 [M+H] +

[0679] 2. Synthesis of compound Q9-2

[0680] A mixture of 3-bromo-l-propyne (104 mg, 874 μmol, 75.4 μL, 1.2 eq), compound 5A (200 mg, 729 μmol, 1.0 eq) and DIEA (565 mg, 4.37 mmol, 762 μL, 6.0 eq) in DMF (5 mL) was stirred at 60 °C for 16 h under N2. The reaction mixture was concentrated under reduced pressure to give the product crude, which was further purified by preparative HPLC column (basic condition) to give compound Q9-2 (140 mg, 448 μmol, 61.5% yield) as a yellow oil.

[0681] LC-MS: EC16364-15-P1A, Product: RT = 0.411 min, m / z = 313.3 [M+H] +

[0682] 3. Synthesis of compound Q9-5

[0683] A mixture of compound Q9-1 (70.0 mg, 91.5 μmol, 1.0 eq), Pd(PPh3)2Cl2 (1.29 mg, 1.83 μmol, 0.02 eq), CuI (697 μg, 3.66 μmol, 0.04 eq) and TEA (1 mL) was protected by N2, then compound Q9-2 (42.9 mg, 137.3 μmol, 1.5 eq) was added. The mixture was stirred at 25 °C for 8 h. The mixture was concentrated under reduced pressure, and triturated with water to remove impurities to give compound Q9-3 product crude (80.0 mg, 84.3 μmol, 92.1%) as a yellow solid.

[0684] LC-MS: EC16364-32-P1A, Product: RT = 0.484 min, m / z = 949.3 [M+H] +

[0685] 1H NMR: EC17015-67-P1B (400 MHz DMSO) δ ppm 12.54-13.15 (m, 1 H), 10.26 (s, 1 H), 8.01 (d, J = 7.8 Hz, 1 H), 7.76 (d, J = 7.8 Hz, 1 H), 7.56 (d, J = 7.4 Hz, 1 H), 7.50 (d, J = 8.8 Hz, 1 H), 7.42-7.48 (m, 1 H), 7.37-7.42 (m, 1 H), 7.31-7.36 (m, 2 H), 7.29 (dd, J = 12.0, 1.8 Hz, 1 H), 7.17-7.22 (m, 1 H), 7.15 (d, J = 8.8 Hz, 2 H), 7.07-7.13 (m, 1 H), 6.94 (d, J = 9.0 Hz, 2 H), 6.91 (d, J = 8.8 Hz, 1 H), 4.93 (s, 2 H), 4.02 (t, J = 6.0 Hz, 2 H), 3.79 (t, J = 6.0 Hz, 2 H), 3.69 (t, J = 6.6 Hz, 2 H), 3.55 (s, 2 H) 3.17 (d, J = 3.8 Hz, 4 H), 2.98 (t, J = 6.0 Hz, 2 H), 2.63-2.70 (m, 8 H), 1.88-1.98 (m, 2 H), 1.30 (s, 9 H).

[0686] 4. Synthesis of compound APQ9

[0687] Compound Q9-3 (50.0 mg, 52.7 μmol, 1.0 eq) and TFA (1.54 g, 13.5 mmol, 1 mL, 255 eq) were mixed in DCM (1 mL) and stirred at 25 °C for 2 h under N2. The mixture was concentrated under reduced pressure to give the product crude, which was further purified by preparative HPLC column (HCl condition and neutral condition) to give compound APQ9 (6 mg, 6.24 μmol, 11.85% yield, 92.9% purity) as a white solid.

[0688] LC-MS: EC16364-35-P1R; Product: RT = 0.416 min

[0689] 1H NMR: EC16364-35-P1R (400 MHz DMSO) δ ppm 10.26 (s, 1 H) 8.03 (d, J=8.0 Hz, 1 H) 7.78 (d, J=8.0 Hz, 1 H) 7.60 (d, J=7.2 Hz, 1 H) 7.43 - 7.55 (m, 2 H) 7.38 - 7.42 (m, 1 H) 7.32 - 7.37 (m, 2 H) 7.28 (dd, J=12.0, 1.75 Hz, 1 H) 7.13 - 7.21 (m, 3 H) 7.07 - 7.12 (m, 1 H) 6.88 - 6.96 (m, 3 H) 4.92 (s, 2 H) 4.02 (t, J=6.4 Hz, 2 H) 3.85 (t, J=6.0 Hz, 2 H) 3.69 (t, J=6.8 Hz, 2 H) 3.55 (s, 2 H) 3.16 (s, 4 H) 2.97 (t, J=5.6 Hz, 2 H) 2.78 (t, J=7.2 Hz, 2 H) 2.65 - 2.70 (m, 6 H) 1.91 - 2.02 (m, 2 H).

[0690] Synthesis of APQ10:

[0691] 1. Synthesis of compound Q10-2

[0692] Compound 1A (218 mg, 1.11 mmol, 3.0 eq, HC1), 1-(4-bromophenyl)hexahydropyrimidine-2,4-dione (100 mg, 371.62 pmol, 1 eq), Cs2CO3 (363 mg, 1.11 mmol, 3.0 eq) and RuPhos Pd G3 (31.1 mg, 37.1 pmol, 0.1 eq) were mixed in dioxane (5 mL), protected by N2inflation, and stirred at 90 °C for 2 hours. The mixture was concentrated under reduced pressure to obtain the product crude, which was further purified by preparative HPLC column (FA condition) to obtain compound Q10-2 (60.0 mg, 172 pmol, 46.5% yield) as a white oil.

[0693] LC-MS: EC16364-40-P1A, product: RT = 0.206 min, m / z = 348.1 [M+H] +

[0694] 2. Synthesis of compound Q10-3

[0695] Compound Q10-2 (60 mg, 172 pmol, 1 eq) was mixed with formic acid (1.0 mL) and stirred at 25 °C for 1 h under N2. The mixture was concentrated under reduced pressure to give the crude product, which was used in the next step without purification to give compound Q10-3 (35.0 mg, 116.1 pmol, 67.2% yield) as a yellow oil.

[0696] 3. Synthesis of compound APQ10

[0697] To a solution of compound Q10-3 (35 mg, 116 pmol, 1.0 eq) in DCM (5.0 mL) was added TEA (117 mg, 1.16 mmol, 161 pL, 10.0 eq) and compound Cl (90 mg, 127.69 pmol, 1.1 eq), and the mixture was stirred at 25 °C for 0.5 h, followed by the addition of NaBH(OAc)3 (73.81 mg, 348.26 pmol, 3.0 eq) and stirred at 25 °C for 1 h. The mixture was concentrated under reduced pressure to give the crude product, which was further purified by preparative HPLC column (FA condition) to give compound APQ10 (13 mg, 12.8 pmol, 11.1% yield, 98.0% purity) as a yellow solid.

[0698] LC-MS: 16364-47-P1A, Product: RT = 0.386 min, m / z = 990.5 [M+H] +

[0699] 1H NMR: EC16364-47-P1C (400 MHz DMSO) δ ppm 12.85 (s, 1H), 11.69 (s, 1H), 10.44 (s, 1H), 8.04 (d, J = 8.0 Hz, 1H), 7.79 (d, J = 8.0 Hz, 1H), 7.69-7.77 (m, 1H), 7.52-7.61 (m, 3H), 7.44-7.51 (m, 3H), 7.40-7.43 (m, 1H), 7.42 (d, J = 8.0 Hz, 1H), 7.29-7.39 (m, 3H), 7.10 -7.19 (m, 1H), 6.98 (d, J = 8.8 Hz, 1H), 4.93 (s, 2H), 4.30 (s, 2H), 4.02-4.08 (m, 2H), 3.86 (t, J = 6.0 Hz, 2H), 3.80 (t, J = 6.4 Hz, 4H), 3.60-3.64 (m, 2H), 3.33-3.53 (m, 4H), 3.14-3.27 (m, 2H), 2.98 (t, J = 5.6 Hz, 2H), 2.78-2.86 (m, 2H), 2.71 (t, J = 6.4 Hz, 2H), 2.16-2.27 (m, 2H), 1.97 (dt, J = 14.4, 7.04 Hz, 2H), 1.88 (s, 1H).

[0700] Synthesis of APQ11:

[0701] The synthesis of the PROTAC molecule APQ11 refers to APQ2, but the intermediate compound 1 is replaced by compound 4. After synthesis, compound APQ11 (30.0 mg, 26.5 μmol, 18.6% yield, 97.1% purity) is obtained by column chromatography (FA condition) purification, which is a white solid.

[0702] LC-MS: EC16364-14-P1A1, product: RT = 1.33 min, m / z = 1101.7 [M+H] +

[0703] 1H NMR: EC16364-14-P1C (400 MHz DMSO) δ ppm 11.12 (s, 1 H) 8.19 (s, 1 H) 8.03 (d, J=8.0 Hz, 1 H) 7.89 (d, J=9.2 Hz, 1 H) 7.78 (d, J=8.0 Hz, 1 H) 7.59 (d, J=7.2 Hz, 1 H) 7.44 - 7.53 (m, 2 H) 7.38 - 7.42 (m, 1 H) 7.30 - 7.38 (m, 3 H) 7.24 - 7.29 (m, 1 H) 7.13 - 7.20 (m, 2 H) 7.04 - 7.13 (m, 2 H) 6.90 (d, J=8.8 Hz, 1 H) 5.84 (dd, J=12.4, 5.25 Hz, 1 H) 4.91 (s, 2 H) 4.01 (t, J=6.4 Hz, 2 H) 3.85 (t, J=6.0 Hz, 2 H) 3.44 (s, 2 H) 3.30 (dd, J=12.0, 5.57 Hz, 4 H) 2.86 - 3.00 (m, 6 H) 2.77 (t, J=7.6 Hz, 2 H) 2.64 - 2.71 (m, 2 H) 2.29 - 2.41 (m, 4 H) 2.21 (d, J=5.2 Hz, 2 H) 2.10 (d, J=6.8 Hz, 2 H) 1.92 - 2.01 (m, 4 H) 1.65 (d, J=12.0 Hz, 2 H) 1.40 - 1.54 (m, 2 H) 1.04 - 1.18 (m, 2 H).

[0704] e) Synthesis of APQ9-1, APQ9-2, APQ10-1, APQ10-2, APQ10-3

[0705] Synthesis of APQ9-1:

[0706] To a solution of compound APQ9 (20.0 mg, 22.40 pmol, 1.00 equiv) in methanol (5.0 mL) was added SOCl2(13.3 mg, 112 pmol, 8.13 pL, 5.00 equiv), the mixture was stirred at 80 °C for 3 hours. LCMS (EC17015-82-P1D, P1: RT = 0.449 min) monitoring showed 47.6% of the desired compound was generated. The reaction mixture was extracted with CH2Cl2(10.0 mL, 2 times), the resulting organic layer was washed with saturated brine (10.0 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the product crude, which was further purified by prep-HPLC (column: Phenomenex luna C18 150*25mm*10 pm; mobile phase: [water (FA)-ACN]; gradient elution: 37%-57% B in 13 min) to give compound APQ9-1 (6.20 mg, 12.2% yield, 96.3% purity) as a white solid.

[0707] LC-MS: EC17015-82-P1D, product: RT = 0.449 min, m / z = 454.4 [M / 2+H] +

[0708] 1 H NMR: EC19038-3-P1A1 (400 MHz DMSO) d ppm 10.26 (s, 1H), 8.03 (d, J = 7.6 Hz, 1H), 7.78 (d, J = 8.0 Hz, 1H), 7.52-7.61 (m, 2H), 7.43-7.50 (m, 1H), 7.37-7.42 (m, 1H), 7.32-7.37 (m, 2H), 7.29 (dd, J = 12.0, 1.8 Hz, 1H), 7.20 (d, J = 9.0 Hz, 1H), 7.15 (d, J = 8.8 Hz, 2H), 7.05-7.12 (m, 1H), 6.95 (dd, J = 8.8, 5.0 Hz, 3H), 4.90 (s, 2H), 4.00 (t, J = 6.4 Hz, 2H), 3.77-3.86 (m, 2H), 3.69 (t, J = 6.8 Hz, 2H), 3.62 (s, 3H), 3.55 (s, 2H), 3.17 (s, 4H), 2.98 (t, J = 5.8 Hz, 2H), 2.62-2.77 (m, 8H), 1.86-1.99 (m, 2H).

[0709] Synthesis of APQ9-2:

[0710] Compound APQ9 (50.0 mg, 56.0 μmol, 1.00 equiv), methyl sulfonamide (7.99 mg, 84.0 μmol, 1.50 equiv), EDCI (17.2 mg, 89.6 μmol, 1.60 equiv) and DMAP (13.7 mg, 112 μmol, 2.0 equiv) were mixed in DCM (1.00 mL) and stirred at 25 °C for 3 h under N2 atmosphere. LCMS (EC17015-45-P1A, P1: RT = 0.435 min) monitoring showed 75.6% of the target compound was generated. The reaction mixture was concentrated under reduced pressure to give the product crude, which was further purified by prep-HPLC (column: Phenomenex luna C18 150*25mm*10μm; mobile phase: [water(FA)-ACN]; gradient elution: 30%-60%B in 10 min) to give compound APQ9-2 (19.9 mg, 18.5 μmol, 33.0% yield, 90.1% purity) as a white solid.

[0711] LC-MS: EC17015-45-P1A, product: RT = 0.435 min, m / z = 970.3 [M+H] +

[0712] 1 H NMR: EC17015-45-P1A (400MHz DMSO) δ ppm 10.25 (s, 1H), 8.14 (s, 1H), 8.02 (d, J = 7.2 Hz, 1H), 7.78 (d, J = 8.2 Hz, 1H), 7.59 (d, J = 7.6 Hz, 1H), 7.44-7.52 (m, 2H), 7.39-7.43 (m, 1H), 7.32-7.37 (m, 2H), 7.25-7.30 (m, 1H), 7.07-7.22 (m, 5H), 6.94 (d, J = 9.0 Hz, 3H), 4.86 (s, 2H), 4.02 (t, J = 6.0 Hz, 2H), 3.84-3.93 (m, 2H), 3.69 (t, J = 6.6 Hz, 2H), 3.54 (s, 2H), 3.16 (s, 4H), 2.99 (d, J = 5.6 Hz, 2H), 2.72-2.78 (m, 2H), 2.67 (dd, J = 3.8, 1.8 Hz, 9H), 1.92-2.01 (m, 2H).

[0713] Synthesis of APQ10-1:

[0714] 1. Synthesis of Q10-1-2

[0715] To a solution of compound Q10-1-1 (334 mg, 1.43 mmol, 1.00 equiv) in DCM (10.0 mL) was added TEA (1.44 g, 14.3 mmol, 1.99 mL, 10.0 equiv) and compound Q10-3 (430 mg, 1.43 mmol, 1.00 equiv), the mixture was stirred at 25 °C for 0.5 h, then NaBH(OAc)3 (907 mg, 4.28 mmol, 3.00 equiv) was added and the reaction was continued to stir at 25 °C for 1 h. After reaction, the mixture was diluted with water (5.0 mL), extracted with DCM (5.0 mL*4), the organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to give the product crude. The crude was purified by reverse phase HPLC (0.1% FA condition) to give compound Q10-1-2 (160 mg, 308 µmol, 21.6% yield) as a yellow solid.

[0716] LCMS: EC19038-2-P1A1, RT = 0.238 min, MS (ESI) m / z = 520.3 [M+H] +

[0717] 2. Synthesis of APQ10-1

[0718] Compound Q4-3 (117 mg, 192 µmol, 1.00 equiv), Q10-1-2 (100 mg, 192 µmol, 1.00 equiv) and Cs2CO3 (157 mg, 481 µmol, 2.50 equiv) were mixed in DMA (2.00 mL), stirred at 25 °C for 16 h under N2. LCMS (EC17015-91-P1A, P1: RT = 0.433 min) monitoring showed 57.6% of the desired compound was generated. After reaction, the mixture was concentrated under reduced pressure to give the product crude, which was further purified by prep-HPLC (column: Phenomenex luna C18 150*25mm*10µm; mobile phase: [water(FA)-ACN]; gradient elution: 36%-66%B in 10 min) to give compound APQ10-1 (63.8 mg, 56.0 µmol, 31.7% yield, 95.8% purity) as a white solid.

[0719] LCMS: EC17015-91-P1A, product: RT = 0.433 min, m / z = 1046.4 [M+H] +

[0720] 1H NMR: EC17015-93-P1A (400 MHz DMSO)

[0721] δ ppm 10.25 (s, 1H), 8.20 (s, 1H), 8.02 (d, J = 7.8 Hz, 1H), 7.77 (d, J = 8.2 Hz, 1H), 7.56 (d, J = 7.4 Hz, 1H), 7.43 - 7.52 (m, 2H), 7.38 - 7.42 (m, 1H), 7.34 (td, J = 7.2, 2.2 Hz, 2H), 7.24 - 7.30 (m, 1H), 7.15 - 7.23 (m, 1H), 7.05 - 7.14 (m, 3H), 6.91 (dd, J = 8.8, 4.2 Hz, 3H), 4.93 (s, 2H), 4.02 (t, J = 6.2 Hz, 2H), 3.79 (t, J = 5.94 Hz, 2H), 3.58 - 3.72 (m, 5H), 3.46 (s, 3H), 2.98 (t, J = 5.6 Hz, 2H), 2.57 - 2.73 (m, 7H), 2.27 - 2.45 (m, 4H), 2.15 (d, J = 6.8 Hz, 2H), 1.87 - 1.98 (m, 2H), 1.76 (d, J = 11.0 Hz, 2H), 1.56 - 1.70 (m, 1H), 1.30 (s, 9H) 1.10 - 1.25 (m, 3H).

[0722] Synthesis of APQ10-2:

[0723] A mixture of compound APQ10 (50.0 mg, 50.5 μmol, 1.00 equiv), methyl sulfonamide (7.21 mg, 75.8 μmol, 1.50 equiv), EDCI (15.5 mg, 80.8 μmol, 1.60 equiv) and DMAP (12.3 mg, 101 μmol, 2.00 equiv) in DCM (2.00 mL) was stirred at 25 °C for 1 h under N2. LCMS (EC17015-89-P1B, P1: RT = 0.416 min) monitoring showed about 76.1% of the target compound was generated. The reaction mixture was diluted with water (20.0 mL), extracted with DCM (20.0 mL*3), the organic layer was washed with saturated brine (30.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give the product as a crude. The crude was purified by prep-HPLC (column: Phenomenex luna C18 150*25mm*10μm; mobile phase: [water(FA)-ACN]; gradient elution: 28%-58%B in 10 min) to give compound APQ10-2 (17.0 mg, 15.1 μmol, 29.8% yield, 94.6% purity) as a white solid.

[0724] LCMS: EC17015-89-P1B, product: RT = 0.416 min, m / z = 1067.5 [M+H] +

[0725] 1 H NMR: EC17015-89-P1A (400MHz DMSO)

[0726] δ ppm 10.24 (s, 1H), 8.16 (s, 1H), 8.03 (d, J = 8.4 Hz, 1H), 7.78 (d, J = 8.2 Hz, 1H), 7.58 (d, J = 7.8 Hz, 1H), 7.44-7.50 (m, 1H), 7.40 (d, J = 7.8 Hz, 2H), 7.31-7.37 (m, 2H), 7.26 (d, J = 12.6 Hz, 1H), 7.09-7.19 (m, 4H), 6.91 (d, J = 9.0 Hz, 2H), 6.67-6.75 (m, 1H), 4.83 (s, 2H), 4.01 (t, J = 6.8 Hz, 2H), 3.86 (t, J = 5.8 Hz, 2H), 3.63-3.73 (m, 4H), 3.46 (s, 2H), 2.97 (t, J = 5.2 Hz, 2H), 2.91 (s, 3H), 2.58-2.73 (m, 8H), 2.44 (s, 4H), 2.17 (d, J = 6.4 Hz, 2H), 1.91-2.03 (m, 2H), 1.76 (d, J = 12.6 Hz, 2H), 1.65 (s, 2H), 1.08-1.29 (m, 3H).

[0727] Synthesis of APQ10-3

[0728] 1. Synthesis of Q10-3-2

[0729] To compound Q10-3-1 (5.00 g, 26.3 mmol, 1.00 equiv) was added acrylic acid (3.79 g, 52.6 mmol, 3.61 mL, 2.00 equiv) at 20 °C, the mixture was stirred at 110 °C for 5 h, then acetic acid (50.0 mL) and urea (7.11 g, 118 mmol, 6.37 mL, 4.50 equiv) were added, the mixture was stirred at 120 °C for 12 h. LC-MS (EC21184-306-P1B, P1: RT = 0.598 min) detected about 76.5% of the desired product. The reaction mixture was diluted with water (200 mL), filtered, concentrated under reduced pressure to give compound Q10-3-2 product (2.50 g, 8.11 mmol, 30.8% yield, 93.1% purity) as a white solid.

[0730] LC-MS: EC21184-306-P1B, P1: RT = 0.598 min, m / z = 288.9 [M+H] +

[0731] 2. Synthesis of Q10-3-3

[0732] A mixture of compound Q10-3-2 (2.20 g, 7.66 mmol, 1.00 equiv), compound 1A (6.10 g, 38.3 mmol, 5.00 equiv), RuPhos Pd G3 (641 mg, 766 µmol, 0.10 equiv) and Cs2CO3 (4.99 g, 15.3 mmol, 2.00 equiv) in dioxane (25.0 mL) was stirred at 90 °C for 12 h under N2. LC-MS (EC21184-315-P1, P1: RT = 0.272 min) indicated about 12.0% of desired product was formed. The reaction mixture was filtered, concentrated under reduced pressure and purified by preparative HPLC (neutral condition) column chromatography to give compound Q10-3-3 (200 mg, 321 µmol, 4.20% yield, 58.7% purity) as a brown solid.

[0733] LC-MS: EC21184-315-P1, P1: RT = 0.272 min, m / z = 366.2 [M+H] +

[0734] 3. Synthesis of Q10-3-4

[0735] To compound Q10-3-3 (50.0 mg, 137 µmol, 1.00 equiv) was added formic acid (0.50 mL) and stirred at 25 °C for 0.5 h under N2. LC-MS (EC21184-321-P1A, P1: RT = 0.221 min) indicated about 60.9% of desired product was formed. The reaction mixture was filtered, concentrated under reduced pressure to give compound Q10-3-4 (50.0 mg, crude) as a white solid.

[0736] LC-MS: EC21184-321-P1A, P1: RT = 0.221 min, m / z = 320.2 [M+H] +

[0737] 4. Synthesis of APQ10-3

[0738] To a solution of compound Q10-3-4 (50.0 mg, 157 pmol, 1.00 equiv) and compound Cl (66.2 mg, 93.9 pmol, 0.60 equiv) in DCM (1.00 mL) was added TEA (158 mg, 1.57 mmol, 218 pL, 10.0 equiv) and NaBH(OAc)3 (100 mg, 470 pmol, 3.00 equiv), and the mixture was stirred at 25 °C for 4 h. LC-MS (EC21184-322-P1B, P1: RT = 1.447 min) detected about 44.1% of the desired product. The reaction mixture was diluted with water (2.0 mL) after reaction, filtered, concentrated under reduced pressure to get the crude product, which was further purified by prep-HPLC (column: CD02-Waters Xbidge BEH C18 150*25*10um; mobile phase: [water(NH4HCO3)-ACN]; gradient: 28%-58%B in 10 min) to give compound APQ10-3 product (8.00 mg, 7.94 pmol, 4.57% yield, 90.3% purity) as a white solid.

[0739] LC-MS: EC17015-651-P1B1, P1: RT = 0.424 min, m / z = 1008.5 [M+H] +

[0740] 1H NMR: EC17015-651-P1A (400 MHz DMSO) δ 10.36 (s, 1H), 8.03 (d, J = 7.6 Hz, 1H), 7.78 (d, J = 8.0 Hz, 1H), 7.60 (d, J = 6.8 Hz, 1H), 7.52 (d, J = 8.8 Hz, 1H), 7.47 (t, J = 7.2 Hz, 1H), 7.38-7.43 (m, 1H), 7.31-7.38 (m, 2H), 7.27 (dd, J = 11.6, 1.6 Hz, 1H), 7.13-7.22 (m, 2H), 7.05-7.12 (m, 1H), 6.92 (d, J = 8.4 Hz, 1H), 6.70-6.82 (m, 2H), 4.92 (s, 2H), 4.02 (t, J = 6.4 Hz, 2H), 3.86 (t, J = 6.0 Hz, 2H), 3.68-3.77 (m, 2H), 3.61 (t, J = 6.8 Hz, 2H), 3.43-3.48 (m, 4H), 2.94-3.01 (m, 2H), 2.79 (t, J = 7.2 Hz, 2H), 2.63-2.74 (m, 6H), 2.30-2.42 (m, 4H), 2.14 (d, J = 7.2 Hz, 2H), 1.91-2.02 (m, 2H), 1.63-1.81 (m, 3H), 1.05-1.21 (m, 2H).

[0741] f) Synthesis of APQ901, APQ902, APQ902a, APQ903, APQ904

[0742] Synthesis of APQ901:

[0743] The synthesis of the PROTAC molecule APQ901 refers to APQ902, and the raw material Q901-1 can be directly purchased. The starting material is changed to Q901-1 and acrylic acid, and the target product APQ901 (17.0 mg, 16.0 μmol, 19.3% yield, 85.6% purity) is obtained after synthesis and purification. It is a white solid.

[0744] LC-MS: EC21184-138-p1a, P1: RT = 1.820 min, m / z = 911.6 [M+H] +

[0745] 1 H NMR: EC21184-138-P1CC (400 MHz DMSO)

[0746] δ 12.40-13.11 (m, 2H), 10.37 (s, 1H), 8.04 (d, J = 8.0 Hz, 1H), 7.79 (d, J = 8.0 Hz, 1H), 7.59 (d, J = 7.2 Hz, 1H), 7.54 (d, J = 8.8 Hz, 1H), 7.44-7.50 (m, 1H), 7.39-7.43 (m, 1H), 7.32-7.38 (m, 2H), 7.30 (dd, J = 12.0, 2.0 Hz, 1H), 7.15-7.22 (m, 2H), 7.11 (d, J = 8.8 Hz, 1H), 7.03-7.09 (m, 2H), 6.95 (d, J = 8.8 Hz, 1H), 4.92 (s, 2H), 4.02 (t, J = 6.4 Hz, 2H), 3.86 (t, J = 6.0 Hz, 2H), 3.74 (t, J = 6.8 Hz, 2H), 3.55 (s, 2H), 3.04 (s, 4H), 2.98 (t, J = 5.6 Hz, 2H), 2.80 (t, J = 7.6 Hz, 2H), 2.68 (t, J = 6.4 Hz, 6H), 1.92-2.02 (m, 2H).

[0747] Synthesis of APQ902:

[0748] 1. Synthesis of compound Q902-2

[0749] Compound Q902-1 (20.0 g, 126 mmol, 13.8 mL, 1.00 equiv), compound 9A (23.4 g, 126 mmol, 1.00 equiv) and TEA (38.2 g, 377 mmol, 52.5 mL, 3.00 equiv) were mixed in DMF (200 mL), the mixture was stirred at 90 °C for 12 h under N2. After reaction, the mixture was added water (600 mL), extracted with ethyl acetate (200 mL*3), the organic layer was washed with saturated brine (200 mL), dried over Na2S04, filtered and concentrated under reduced pressure to give the product crude, which was further purified by column chromatography (Si02, petroleum ether / ethyl acetate = 1 / 0 to 9 / 1) to give compound Q902-2 product (13.1 g, crude) as a yellow solid.

[0750] LC-MS: EC17015-352-P1A, Product: RT = 0.445 min

[0751] 1H NMR: EC17015-352-P1A (400 MHz CDCh) δ ppm 8.01 (t, J=9.2 Hz, 1 H), 6.38 - 6.64 (m, 2 H), 3.53 - 3.71 (m, 4 H), 3.36 - 3.47 (m, 4 H), 1.48 (s, 9 H).

[0752] 2. Synthesis of compound Q902-3

[0753] To a solution of compound Q902-2 (2.00 g, 6.15 mmol, 1.00 equiv) in methanol (20.0 mL) was added Pd / C (1.00 g, 10% purity), the reaction was shaken under H2(30 psi) at 25 °C for 12 hours. After reaction, the mixture was filtered, washed with methanol, the resulting organic layer was collected and concentrated, dried to give compound Q902-3 product crude (1.70 g, 5.76 mmol, 93.6% yield) as a brown solid.

[0754] 1 H NMR: EC17015-352-P1A (400 MHz CDCh) δ ppm 8.01 (t, J=9.2 Hz, 1 H), 6.38 - 6.64 (m, 2 H), 3.53 - 3.71 (m, 4 H), 3.36 - 3.47 (m, 4 H), 1.48 (s, 9 H).

[0755] 3. Synthesis of compound Q902-4

[0756] Compound Q902-3 (5.00 g, 16.9 mmol, 1.00 equiv) and acrylic acid (4.88 g, 67.7 mmol, 4.64 mL, 4.00 equiv) were mixed, the reaction was stirred at 110 °C for 3 hours, then urea (6.53 g, 109 mmol, 5.84 mL, 6.42 equiv) and AcOH (50.0 mL) were added, the mixture was stirred at 120 °C for 12 hours. After reaction, the mixture was concentrated under reduced pressure to give compound Q902-4 product (6.60 g, crude) as a black brown oil, which was used directly in the next step without purification.

[0757] 4. Synthesis of compound Q902-5

[0758] Compound Q902-4 (6.60 g, 16.8 mmol, 1.00 equiv) was mixed in HCl (33.0 mL) and H2O (33.0 mL), and the mixture was stirred at 50 °C for 12 h. LCMS (EC17015-374-P1A, P1: RT = 0.509 min) detection showed about 49.1% of the desired product was generated. The reaction mixture was concentrated under reduced pressure to give the product crude, which was purified by prep-HPLC (column: CD19-Daisogel SP-100-8-ODS-PK 200*50*10 pm; mobile phase: [water(NH3H2O)-ACN]; gradient elution: 8%-36% B in 10 min) to give compound Q902-5 product (160 mg, 547 pmol, 8.00% yield) as a brown solid.

[0759] LC-MS: EC17015-374-P1A, product: RT = 0.509 min, m / z = 293.1 [M+H] +

[0760] 5. Synthesis of compound Q902-6

[0761] Compound Q902-5 (140 mg, 479 pmol, 1.00 equiv), compound 4A (57.0 mg, 479 pmol, 41.3 pL, 1.00 equiv), and NaHC03(120.7 mg, 1.44 mmol, 55.9 pL, 3.00 equiv) were mixed in DMF (2.00 mL), and the mixture was stirred at 25 °C for 12 h. LCMS (EC17015-400-P1W, P1: RT = 0.566 min) detection showed about 81.3% of the desired product was generated. The reaction mixture was filtered and concentrated under reduced pressure to give the product crude, which was purified by prep-HPLC (column: CD02-Waters Xbidge BEH C18 150*25*10 pm; mobile phase: [water(NH3H2O)-ACN]; gradient elution: 12%-42% B in 10 min) to give compound Q902-6 product (90.0 mg, 272 pmol, 56.9% yield) as a brown solid.

[0762] LC-MS: EC17015-400-P1W, product: RT = 0.566 min, m / z = 331.2 [M+H] +

[0763] 1 H NMR: EC17015-400-P1A (400 MHz DMSO)

[0764] δ ppm 10.24-10.44 (m, 1H), 7.20 (t, J = 9.0 Hz, 1H), 6.72-6.90 (m, 2H), 3.62 (t, J = 6.6 Hz, 2H), 3.33-3.34 (m, 2H), 3.15-3.24 (m, 5H), 2.68 (t, J = 6.6 Hz, 2H), 2.54-2.60 (m, 4H).

[0765] 6. Synthesis of compound Q902-7

[0766] Q9-1 (108 mg, 141 μmol, 1.00 equiv), compound Q902-6 (70.0 mg, 212 μmol, 1.50 equiv), CuI (1.35 mg, 7.06 μmol, 0.05 equiv) and Pd(PPh3)2Cl2 (9.92 mg, 14.1 μmol, 0.10 equiv) were mixed in TEA (1.50 mL) and DMF (0.50 mL), the reaction was stirred at 25 °C for 12 h under N2 atmosphere. LCMS (EC17015-412-P1A, P1: RT = 0.449 min) detection showed about 31.6% of the desired product was generated. The reaction mixture was diluted with ice water (20.0 mL), filtered under reduced pressure, concentrated to give the crude product, which was further purified by column chromatography (SiO2, dichloromethane / ethyl acetate = 1 / 4 to 0 / 1) to give compound Q902-7 product (60.0 mg, 62.0 μmol, 43.9% yield) as a yellow solid.

[0767] LC-MS: EC17015-412-P1A, product: RT = 0.449 min, m / z = 967.4 [M+H] +

[0768] 7. Synthesis of compound APQ902

[0769] Compound Q902-7 (60.0 mg, 62.0 μmol, 1.00 equiv) was mixed in HC1 / dioxane (5.00 mL), the mixture was stirred at 20 °C for 12 h. LCMS (EC17015-419-P1B1, P1: RT = 0.422 min) detection showed about 71.4% of the desired product was generated. The reaction mixture was concentrated under reduced pressure to give the product crude, which was further purified by prep-HPLC (column: CD03-Welch Xtimate C18 150*25*5 μm; mobile phase: [water (FA)-ACN]; gradient elution: 28%-58% B in 10 min) to give compound APQ902 product (15.0 mg, 14.6 μmol, 23.6% yield, 88.8% purity) as a white solid.

[0770] LC-MS: EC17015-419-P1B1, product: RT = 0.422 min, m / z = 911.3 [M+H] +

[0771] 1 H NMR: EC17015-419-P1A (400 MHz DMSO)

[0772] δ ppm 10.38 (s, 1H), 8.17 (s, 1H), 8.03 (d, J = 7.8 Hz, 1H), 7.79 (d, J = 8.0 Hz, 1H), 7.59 (d, J = 7.2 Hz, 1H), 7.52 (d, J = 8.8 Hz, 1H), 7.47 (t, J = 7.6 Hz, 1H), 7.39-7.42 (m, 1H), 7.32-7.38 (m, 2H), 7.28 (dd, J = 12.0, 1.6 Hz, 1H), 7.16-7.24 (m, 2H), 7.07-7.14 (m, 1H), 6.92 (d, J = 8.6 Hz, 1H), 6.81-6.87 (m, 1H), 6.74-6.80 (m, 1H), 4.91 (s, 2H), 4.02 (t, J = 6.4 Hz, 2H), 3.85 (t, J = 5.6 Hz, 2H), 3.60-3.64 (m, 2H), 3.55 (s, 2H), 3.21 (s, 4H), 2.97 (t, J = 5.4 Hz, 2H), 2.75-2.82 (m, 2H), 2.63-2.71 (m, 6H), 1.89-2.07 (m, 2H).

[0773] Synthesis of APQ902a:

[0774] The synthesis of PROTAC molecule APQ902a followed the procedure of APQ902, but changed the starting material to compound Q902a-1 and 9A. The desired product APQ902a (8.00 mg, 8.46 µmol, 6.90% yield, 97.6% purity) was obtained as a white solid after purification.

[0775] LC-MS: EC17015-420-P1B1, Product: RT = 0.413 min, m / z = 923.3 [M+H] +

[0776] 1 H NMR: EC17015-420-P1A (400 MHz DMSO) δ ppm 10.20 (s, 1H), 8.03 (d, J = 8.0 Hz, 1H), 7.78 (d, J = 7.6 Hz, 1H), 7.60 (d, J = 7.6 Hz, 1H), 7.48-7.53 (m, 1H), 7.42-7.48 (m, 1H), 7.38-7.42 (m, 1H), 7.31-7.37 (m, 2H), 7.25-7.31 (m, 1H), 7.19 (d, J = 8.8 Hz, 1H), 7.10 (t, J = 8.8 Hz, 1H), 7.03 (d, J = 8.4 Hz, 1H), 6.90 (d, J = 9.2 Hz, 1H), 6.62 (d, J = 1.6 Hz, 1H), 6.49 (dd, J = 8.8, 2.4 Hz, 1H), 4.91 (s, 2H), 4.02 (t, J = 6.4 Hz, 2H), 3.85 (t, J = 6.0 Hz, 2H), 3.77 (s, 3H), 3.55 (s, 2H), 3.49 (d, J = 6.4 Hz, 2H), 3.21 (s, 4H), 2.94-3.01 (m, 2H), 2.74-2.80 (m, 2H), 2.61-2.68 (m, 6H), 1.93-2.02 (m, 2H).

[0777] Synthesis of APQ903:

[0778] The synthesis of PROTAC molecule APQ903 followed the procedure of APQ902, but changed the starting material to compound Q903-1 and 9A. The desired product APQ903 (12.0 mg, 12.0 µmol, 29.5% yield, 92.6% purity) was obtained as a white solid after purification.

[0779] LC-MS: EC17015-421-P1B1, Product: RT = 0.418 min, m / z = 929.3 [M+H] +

[0780] 1 H NMR: EC17015-421-P1A (400 MHz DMSO) δ ppm 10.47 (s, 1 H), 8.03 (d, J = 8.4 Hz, 1 H), 7.78 (d, J = 8.0 Hz, 1 H), 7.59 (d, J = 7.2 Hz, 1 H), 7.52 (d, J = 8.8 Hz, 1 H), 7.47 (t, J = 7.2 Hz, 1 H), 7.38 - 7.43 (m, 1 H), 7.27 - 7.37 (m, 3 H), 7.20 (d, J = 8.8 Hz, 1 H), 7.06 - 7.13 (m, 3 H), 6.92 (d, J = 8.0 Hz, 1 H), 4.85 - 4.99 (m, 2 H), 4.02 (t, J = 6.4 Hz, 2 H), 3.86 (t, J = 5.6 Hz, 2 H), 3.76 (t, J = 6.8 Hz, 2 H), 3.54 (s, 2 H), 3.13 (s, 4 H), 2.95 - 3.00 (m, 2 H), 2.76 - 2.81 (m, 2 H), 2.62 - 2.68 (m, 6 H), 1.92 - 2.02 (m, 2 H).

[0781] Synthesis of APQ904:

[0782] The synthesis of the PROTAC molecule APQ904 followed the procedure of APQ902, the intermediate APQ904-3 was synthesized by a different method, and the synthesis from APQ904-3 to APQ904 followed the procedure of APQ902. The target product APQ904 (18.7 mg, 19.5 pmol, 32.0% yield, 96.9% purity) was obtained as a white solid after purification.

[0783] LC-MS: EC21184-159-p1a, P1: RT = 0.409 min, m / z = 929.2 [M+H] +

[0784] 1H NMR: EC21184-159-p1dg1 (400 MHz DMSO) δ 12.41-13.04 (m, 2H), 10.47 (s, 1H), 8.04 (d, J = 8.0 Hz, 1H), 7.79 (d, J = 8.0 Hz, 1H), 7.60 (d, J = 7.2 Hz, 1H), 7.55 (d, J = 8.8 Hz, 1H), 7.47 (t, J = 7.2 Hz, 1H), 7.39-7.43 (m, 1H), 7.27-7.38 (m, 4H), 7.17-7.22 (m, 1H), 7.07-7.13 (m, 1H), 6.92-7.01 (m, 2H), 4.92 (s, 2H), 4.02 (t, J = 6.4 Hz, 2H), 3.86 (t, J = 6.0 Hz, 2H), 3.66 (t, J = 6.8 Hz, 2H), 3.55 (s, 2H), 3.08 (s, 4H), 2.98 (t, J = 5.6 Hz, 2H), 2.80 (t, J = 7.6 Hz, 2H), 2.63-2.71 (m, 6H), 1.93-2.00 (m, 2).

[0785] Synthesis of intermediate APQ904-3:

[0786] 1. Synthesis of compound Q904-1

[0787] To a solution of compound 10A (24.0 g, 115 mmol, 1.00 equiv) in ACN (240 mL) was added BnBr (59.2 g, 346 mmol, 41.1 mL, 3.0 equiv) and K2CO3 (47.8 g, 346 mmol, 3.00 equiv), and the mixture was stirred at 85 °C for 16 h. LC-MS (EC21184-81-p1d, P1: RT = 0.577 min) detection showed about 65.7% of the desired product was generated. The reaction mixture was concentrated under reduced pressure to give the product crude, which was further purified by column chromatography (SiO2, Petroleum ether / Ethyl acetate = 100 / 0) to give compound Q904-1 product (40.0 g, crude) as a white solid.

[0788] LC-MS: EC21184-81-p1d, P1: RT = 0.577 min, m / z = 387.9 [M+H] +

[0789] 1H NMR: EC21184-81-p1gz (400 MHz DMSO) δ 7.54 (dd, J = 12.4, 6.8 Hz, 1H), 7.16 - 7.40 (m, 10H), 6.91 (dd, J = 11.2, 7.6 Hz, 1H), 4.40 (s, 4H).

[0790] 2. Synthesis of compound Q904-2

[0791] Compound Q904-1 (34.0 g, 87.6 mmol, 1.00 equiv), compound 9A (17.9 g, 96.3 mmol, 1.10 equiv), Cs2C03(57.1 g, 175.2 mmol, 2.00 equiv), BINAP (10.9 g, 17.5 mmol, 0.20 equiv) and Pd(OAc)2(1.97 g, 8.76 mmol, 0.10 equiv) were mixed in toluene (340 mL), purged with N2and stirred at 100 °C for 16 h. LC-MS (EC21184-91-p1a, P1 : RT = 0.602 min) detection showed about 52.7% of the desired product was generated. The reaction mixture was concentrated under reduced pressure to give the crude product, which was further purified by column chromatography (Si02, petroleum ether / ethyl acetate = 100 / 0 to 85 / 15) to give compound Q904-2 product (36.0 g, 67.8 mmol, 77.5% yield, 93.0% purity) as a white solid.

[0792] LC-MS: EC21184-91-p1a, P1 : RT = 0.602 min, m / z = 494.2 [M+H] +

[0793] 3. Synthesis of compound Q904-3

[0794] To a solution of compound Q904-2 (34.0 g, 68.9 mmol, 1.00 equiv) in methanol (700 mL) was added Pd / C (3.67 g, 3.44 mmol, 10% purity, 0.05 equiv) under N2environment, followed by H2charge, and the reaction was stirred under H2(40 Psi) at 20 °C for 16 h. LC-MS (EC21184-96-p1b2, R1: RT = 0.391 min, P1: RT = 0.391 min) detection showed about 81.3% of the desired product was generated. The reaction mixture was concentrated under reduced pressure to give the crude product, which was further purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 0 to 88 / 12) to give compound Q904-3 product (18.0 g, 57.5 mmol, 83.4% yield, 100% purity) as a white solid.

[0795] LC-MS: EC21184-96-p1b2, P1: RT = 0.391 min, m / z = 258.0 [M-55+H] +

[0796] 1 H NMR: EC21184-96-p1gz (400 MHz CHLOROFORM) δ 6.62-6.88 (m, 1H), 6.52 (dd, J = 12.8, 8.0 Hz, 1H), 3.56-3.62 (m, 4H), 2.91 (s, 4H), 1.48 (s, 9H).

[0797] Example 7. Synthesis of PROTAC molecules recruiting VHL-E3 enzyme (APH1-APH20 and APH101-APH106)

[0798] a) Synthesis of APH1, APH2, APH3, APH4, APH5

[0799] Synthesis of APH1:

[0800] 1. Synthesis of compound H1-2

[0801] To a solution of compound 2 (200 mg, 450 pmol, 1.0 eq) in DMF (2.00 mL) was added HATU (257 mg, 674 pmol, 1.5 eq), DIEA (174 mg, 1.35 mmol, 235 pL, 3.0 eq) and H1-1 (200 mg, 1.35 mmol, 3.0 eq), the mixture was stirred at 25 °C for 3 h. LCMS (EC16331-51-P1A6, P1: RT = 0.359 min) detection showed that compound 2 was completely reacted to generate the main peak of the desired product with m / z consistent with the expectation. After reaction, the mixture was extracted by water (5.00 mL) and DCM (5.00 mL*3) to separate the layers, the organic layer was washed with saturated brine (5.00 mL*2), dried over Na2S04, filtered and concentrated under reduced pressure to give compound H1-2 product crude (120 mg, 186.97 pmol, 13.85% yield, 78% purity) as a white solid.

[0802] LC-MS: EC16331-51-P1C; Product: RT = 0.361 min

[0803] 1 H NMR: EC16331-51-P1C (400 MHz DMSO) d ppm 8.99 (s, 1H), 8.46 (br d, J = 7.6 Hz, 1H), 7.32-7.47 (m, 5H), 4.97-5.00 (m, 1H), 4.86-4.96 (m, 1H), 4.40-4.51 (m, 2H), 4.28 (br s, 1H), 3.56 (br s, 5H), 2.45 (s, 3H), 2.01-2.10 (m, 1H), 1.77 (ddd, J = 12.8, 8.8, 4.4 Hz, 1H), 1.37 (br d, J = 6.8 Hz, 3H), 0.93 (s, 9H).

[0804] 2. Synthesis of compound APH1

[0805] To a solution of compound H1-2 (100 mg, 200 pmol, 1.0 eq) in DMF (2.00 mL) was added TEA (202 mg, 2.00 mmol, 278 pL, 10.0 eq), NaBH(OAc)3 (127 mg, 600 pmol, 3.0 eq) and compound C1 (70.4 mg, 99.9 pmol, 0.5 eq), the mixture was stirred at 25 °C for 2 hours. LCMS (EC16331-55-P1A, P1: RT = 0.462 min; R2: RT = 0.409 min) detection showed that compound C1 reacted completely to generate the main peak of m / z consistent with the expected product. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain compound APH1 product crude, which was purified by preparative-HPLC (FA condition; column: Phenomenex luna C18 150*25mm*10 pm; mobile phase: [water(FA)-ACN]; gradient: 23%-53%B in 10 min) column chromatography to obtain product APH1 (7.00 mg, 5.31 pmol, 2.66% yield, 90.23% purity) as a white solid.

[0806] LC-MS: EC16331-55-P1E; product: RT = 0.456 min

[0807] 1H NMR: EC16331-55-P1T (400 MHz DMSO) δ ppm 8.94-8.99 (m, 1 H), 8.39-8.46 (m, 1 H), 8.21 (s, 1 H), 8.03 (br d, J = 7.6 Hz, 1 H), 7.73-7.81 (m, 2 H), 7.59 (br d, J = 6.8 Hz, 1 H), 7.47 (br dd, J = 14.0, 8.2 Hz, 2 H), 7.40 (br t, J = 7.2 Hz, 3 H), 7.32-7.36 (m, 3 H), 7.26 (br d, J = 12.0 Hz, 1 H), 7.19 (br d, J = 8.2 Hz, 1 H), 7.06-7.12 (m, 1 H), 6.88 (br d, J = 8.4 Hz, 1 H), 4.83-4.93 (m, 3 H), 4.37-4.53 (m, 3 H), 4.28 (br s, 1 H), 4.01 (br t, J = 6.0 Hz, 3 H), 3.84 (br s, 3 H), 3.58 (br s, 3 H), 3.48 (br s, 4 H), 2.90-3.08 (m, 6 H), 2.72-2.82 (m, 3 H), 2.42-2.46 (m, 3 H), 2.00-2.08 (m, 1 H), 1.97 (br d, J = 6.0 Hz, 2 H), 1.72-1.80 (m, 1 H), 1.38 (br s, 1 H), 1.28-1.37 (m, 3 H), 1.23 (br s, 1 H), 0.93 (br s, 9 H).

[0808] Synthesis of APH2:

[0809] 1. Synthesis of compound H2-3

[0810] To a solution of compound H2-2 (10.3 g, 76.9 mmol, 1.5 eq) in THF (120 mL) was added NaH (3.08 g, 76.9 mmol, 60% purity, 1.5 eq) at 0 °C under N2 atmosphere, the mixture was stirred at 0 °C for 0.5 h, then compound H2-1 (10.0 g, 51.3 mmol, 7.57 mL, 1.0 eq) dissolved in THF (20.0 mL) was added dropwise at 0 °C, the mixture was stirred at 15 °C for 2 h under N2 atmosphere. TLC (PE:EA = 15:1, R1:R f = 0.92; R2:R f = 0.1; P1:R f= 0.44) showed that compound H2-1 reacted completely and a new product peak was generated. The reaction mixture was terminated by adding NH4CI (200 mL) at 0 °C, diluted with water (200 mL), and extracted with ethyl acetate (160 mL*3). The obtained organic layer was washed with saturated brine (200 mL*2), dried over Na2S04, filtered and concentrated under reduced pressure to obtain the crude product, which was purified by column chromatography (Si02, petroleum ether: ethyl acetate = 98:1 to 95:1) to obtain compound H2-3 product (1.30 g, 5.24 mmol, 10.21% yield) as a colorless liquid.

[0811] 1 H NMR: EC9174-340-P1A (400 MHz DMSO) δ ppm 4.58 (t, J = 5.2 Hz, 1H), 4.00 (s, 2H), 3.56-3.65 (m, 2H), 3.45-3.52 (m, 2H), 3.43 (d, J = 5.2 Hz, 2H), 1.42 (s, 9H), 1.11 (t, J = 7.2 Hz, 6H).

[0812] 2. Synthesis of compound H2-4

[0813] To a solution of compound H2-3 (210 mg, 846 μmol, 1.0 eq) in THF (1.00 mL), methanol (1.00 mL) and H20 (2.00 mL) was added LiOH-H20 (355 mg, 8.46 mmol, 10.0 eq), and the mixture was stirred at 25 °C for 2 h. TLC (PE:EA = 15:1, R1:R2= 1:2) detection showed that compound H2-3 reacted completely and a new compound peak was generated. The reaction mixture was adjusted to pH 6 with hydrochloric acid (1 M) after the reaction, extracted with ethyl acetate (4.00 mL*3), and the obtained organic layer was washed with saturated brine (10.0 mL*2), dried over Na2S04, filtered and concentrated under reduced pressure to obtain the crude product of compound H2-4 (110 mg, 572 μmol, 67.67% yield), which was a yellow oil and used directly in the next step without purification. f = 0.38) showed that compound H2-3 reacted completely and a new compound peak was generated. The reaction mixture was adjusted to pH 6 with hydrochloric acid (1 M) after the reaction, extracted with ethyl acetate (4.00 mL*3), and the obtained organic layer was washed with saturated brine (10.0 mL*2), dried over Na2S04, filtered and concentrated under reduced pressure to obtain the crude product of compound H2-4 (110 mg, 572 μmol, 67.67% yield), which was a yellow oil and used directly in the next step without purification.

[0814] 1 H NMR: EC16331-59-P1A1 (400 MHz DMSO) δ ppm 4.58 (t, J = 5.2 Hz, 1H), 4.00 (s, 2H), 3.56-3.65 (m, 2H), 3.45-3.52 (m, 2H), 3.43 (d, J = 5.2 Hz, 2H), 1.42 (s, 9H), 1.11 (t, J = 7.2 Hz, 6H).

[0815] 3. Synthesis of compound 4

[0816] Compound H2-4 (80.0 mg, 416 pmol, 1.0 eq), compound 2 (185 mg, 416 pmol, 1.0 eq), HATU (237 mg, 624 pmol, 1.5 eq) and TEA (126 mg, 1.25 mmol, 174 pL, 3.0 eq) were mixed in DMF (2.00 mL) and the reaction was stirred at 25 °C for 3 h under N2 atmosphere. LC-MS (EC16331-60-P1A1, P1: RT = 0.440 min; R2: RT = 0.313 min) detection showed that compound H2-4 reacted completely to generate the main peak of m / z consistent with the expected product. The reaction mixture was filtered under reduced pressure and concentrated to give the crude product, which was purified by preparative HPLC (FA condition; column: Phenomenex luna C 18 150*25mm*10 pm; mobile phase: [water(FA)-ACN]; gradient: 30%-60%B in 10 min) to give compound H2-5 product (80.0 mg, 114 pmol, 27.33% yield, 88% purity).

[0817] LC-MS: EC16331-60-P1C; product: RT = 0.405 min

[0818] 1 H NMR: EC16331-60-P1S (400 MHz DMSO) d ppm 8.99 (s, 1H), 8.45 (br d, J = 7.6 Hz, 1H), 7.41-7.45 (m, 2H), 7.35-7.40 (m, 3H), 4.91 (quin, J = 7.2 Hz, 1H), 4.63 (t, J = 5.2 Hz, 1H), 4.55 (br d, J = 9.6 Hz, 1H), 4.41-4.47 (m, 1H), 4.28 (br s, 1H), 3.98 (s, 2H), 3.44-3.67 (m, 9H), 2.45 (s, 3H), 1.99-2.10 (m, 1H), 1.77 (ddd, J = 12.8, 8.8, 4.4 Hz, 1H), 1.37 (br d, J = 7.2 Hz, 3H), 1.09-1.16 (m, 6H), 0.94 (s, 9H).

[0819] 4. Synthesis of compound H2-6

[0820] To a solution of compound H2-5 (70.0 mg, 113 μmol, 1.0 eq) in DMF (1.00 mL) was added formic acid (1.00 mL), the mixture was stirred at 25 °C for 0.5 h. LC-MS (EC16331-68-P1A2, P1: RT = 0.325 min) detection showed that the reaction of compound H2-5 was complete, and the main peak of m / z of the desired product was generated. The reaction mixture was filtered and concentrated under reduced pressure to obtain compound H2-6 product (80.0 mg) as a yellow oil, which was directly used in the next step without purification.

[0821] LC-MS: EC16331-68-P1A2, product: RT = 0.440 min, m / z = 545.2 [M+H] +

[0822] 5. Synthesis of compound APH2

[0823] To a solution of compound H2-6 (80.0 mg, 147 μmol, 1.0 eq) in DMF (1.00 mL) was added TEA (149 mg, 1.47 mmol, 204.44 μL, 10.0 eq), NaBH(OAc)3 (93.4 mg, 441 μmol, 3.0 eq) and compound C1 (72.5 mg, 103 μmol, 0.7 eq), the mixture was stirred at 25 °C for 1 h. LC-MS (EC16331-69-P1A, P1: RT = 0.448 min) detection showed that the reaction of compound H2-6 was complete, and the main peak of m / z of the desired product was generated. The reaction mixture was filtered and concentrated under reduced pressure to obtain the product crude, which was purified by preparative HPLC (column: Waters Xbridge 150*25mm*5μm; mobile phase: [water(NH3H2O)-ACN]; gradient: 15%-45%B in 10 min) to obtain compound APH2 product (20.0 mg, 14.6 μmol, 9.96% yield, 90.18% purity) as a white solid.

[0824] LC-MS: EC16331-69-P1E; product: RT = 0.497 min

[0825] 1H NMR: EC16331-69-P1W (400 MHz DMSO) δ ppm 8.95-9.00 (m, 1 H), 8.45 (br d, J = 7.6 Hz, 1 H), 8.26 (s, 1 H), 8.02 (d, J = 7.6 Hz, 1 H), 7.78 (d, J = 8.0 Hz, 1 H), 7.59 (br d, J = 7.6 Hz, 1 H), 7.40-7.51 (m, 6 H), 7.31-7.39 (m, 6 H), 7.26 (br d, J = 12.0 Hz, 1 H), 7.15-7.19 (m, 1 H), 7.09-7.09 (m, 1 H), 7.05-7.12 (m, 1 H), 6.83 (br d, J = 8.8 Hz, 1 H), 4.90 (s, 3 H), 4.49-4.57 (m, 2 H), 4.44 (br t, J = 8.0 Hz, 2 H), 4.28 (br s, 2 H), 3.97-4.02 (m, 3 H), 3.92-3.96 (m, 2 H), 3.83 (br t, J = 5.6 Hz, 3 H) 3.56-3.61 (m, 5 H), 3.45 (s, 3 H), 2.95-2.95 (m, 1 H), 2.96 (br t, J = 5.2 Hz, 1 H), 2.74 (br t, J = 6.8 Hz, 2 H), 2.43-2.46 (m, 4 H), 2.01-2.08 (m, 1 H), 1.91-2.01 (m, 2 H), 1.76 (ddd, J = 12.8, 8.8, 4.4 Hz, 1 H), 1.34-1.38 (m, 3 H), 0.90-0.97 (m, 12 H).

[0826] Synthesis of APH3:

[0827] 1. Synthesis of compound H3-2

[0828] To a solution of compound C1-8 (150 mg, 197 pmol, 1.00 equiv) and compound H3-1 (141 mg, 789 pmol, 4.00 equiv) in DMF (2.00 mL) was added HATU (113 mg, 296 pmol, 1.50 equiv) and TEA (59.8 mg, 591 pmol, 82.3 pL, 3.00 equiv), and the mixture was stirred at 20 °C for 1 h. LC-MS (EC21184-30-p1a, P1: RT = 0.439 min) detection showed that compound C1-8 was completely reacted to generate the target product with a proportion of about 84.7%. After reaction, the mixture was extracted with water (50.0 mL) and ethyl acetate (30.0 mL*2), and the organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to give compound H3-2 product (180 mg, crude product) as a white solid.

[0829] LC-MS: EC21184-30-p1a, P1: RT = 0.439 min, m / z = 921.4 [M+H] +

[0830] 2. Synthesis of compound APH3-3

[0831] To a solution of compound H3-2 (180 mg, 195 pmol, 1.00 equiv) and compound 2 (86.9 mg, 195 pmol, 1.00 equiv) in DMF (2.00 mL) was added HATU (111 mg, 293 pmol, 1.50 equiv) and TEA (59.3 mg, 586 pmol, 81.6 pL, 3.00 equiv), and the mixture was stirred at 20 °C for 1 h. LC-MS (EC21184-31-p1a1, P1: RT = 0.810 min) detection showed that compound H3-2 was completely reacted to generate the target product with a proportion of about 8.41%. After reaction, the mixture was extracted with water (100 mL) and ethyl acetate (25.0 mL*3), and the organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to give the product crude, which was further purified by preparative HPLC (neutral condition; column: CD07-Daisogel SP-100-8-ODS-PK 150*25*10 pm; mobile phase: [water (NH4HCO3)-ACN]; gradient: 57%-87% B in 10 min) to give compound H3-3 (50.0 mg, 36.1 pmol, 18.5% yield, 97.3% purity) as a white solid.

[0832] LC-MS: EC21184-31-p1a1, P1: RT = 0.810 min, m / z = 1347.5 [M+H]+

[0833] 1 H NMR: EC21184-31-p1qc (400 MHz DMSO) δ ppm 12.76-12.93 (m, 1 H), 8.97 (s, 1 H), 8.43 (d, J = 7.6 Hz, 1 H), 8.02 (d, J = 8.0 Hz, 1 H), 7.77 (d, J = 8.4 Hz, 1 H), 7.56 (d, J = 8.0 Hz, 1 H), 7.30-7.53 (m, 10 H), 7.27 (dd, J = 12.0, 2.0 Hz, 1 H), 7.17 (d, J = 8.8 Hz, 1 H), 7.05-7.12 (m, 1 H), 6.91 (d, J = 8.8 Hz, 1 H), 5.13 (d, J = 3.2 Hz, 1 H), 4.86-4.96 (m, 3 H), 4.54 (d, J = 9.2 Hz, 1 H), 4.44 (t, J = 8.0 Hz, 1 H), 4.27 (s, 1 H), 4.20 (d, J = 2.4 Hz, 2 H), 4.01 (t, J = 6.0 Hz, 2 H), 3.96 (s, 2 H), 3.78 (br t, J = 6.0 Hz, 2 H), 3.54-3.65 (m, 6 H), 3.39-3.53 (m, 6 H), 2.98 (t, J = 5.6 Hz, 2 H), 2.62-2.71 (m, 2 H), 2.44 (s, 5 H), 2.00-2.09 (m, 1 H), 1.87-1.97 (m, 2 H), 1.71-1.82 (m, 1 H), 1.36 (d, J = 7.2 Hz, 3 H), 1.30 (s, 9 H), 0.93 (s, 9 H).

[0834] 3. Synthesis of compound APH3

[0835] Compound H3-3 (45.0 g, 33 μmol, 1.00 equiv) was mixed in HC1 / dioxane (25.0 mL) and stirred at 20 °C for 4 h under N2 atmosphere. LC-MS (EC21184-35-p1a1, P1 : RT = 0.450 min) detection showed that the desired product was generated with about 88.5% yield. The reaction mixture was filtered and concentrated under reduced pressure to give the product crude, which was further purified by prep-HPLC (neutral condition; column: CD02-Waters Xbidge BEH C18 150*25*10 μm; mobile phase: [water (NH4HC03)-ACN]; gradient: 25%-55% B in 10 min) to give compound APH3 (13.0 mg, 9.70 μmol, 29.0% yield, 96.4% purity) as a white solid.

[0836] LC-MS: EC21184-35-p1a1, P1 : RT = 0.450 min, m / z = 1313.3 [M+Na] +

[0837] 1 H NMR: EC21184-35-p1qc (400 MHz DMSO) δ 8.97 (s, 1H), 8.42 (d, J = 7.6 Hz, 1H), 8.03 (d, J = 8.0 Hz, 1H), 7.78 (d, J = 8.0 Hz, 1H), 7.60 (d, J = 8.0 Hz, 1H), 7.30 - 7.54 (m, 10H), 7.26 (dd, J = 12.0, 2.0 Hz, 1H), 7.14 - 7.19 (m, 1H), 7.05 - 7.12 (m, 1H), 6.89 (d, J = 8.4 Hz, 1H), 4.85 - 4.95 (m, 3H), 4.54 (d, J = 9.6 Hz, 1H), 4.44 (t, J = 8.0 Hz, 1H), 4.28 (s, 1H), 4.20 (d, J = 1.6 Hz, 2H), 4.01 (t, J = 6.4 Hz, 2H), 3.96 (s, 2H), 3.85 (t, J = 5.6 Hz, 2H), 3.55 - 3.67 (m, 7H), 3.42 - 3.55 (m, 10H), 2.97 (t, J = 5.6 Hz, 2H), 2.77 (t, J = 7.2 Hz, 2H), 2.44 (s, 5H), 2.01 - 2.10 (m, 1H), 1.91 - 2.00 (m, 2H), 1.77 (ddd, J = 13.2, 8.8, 4.4 Hz, 1H), 1.37 (d, J = 6.8 Hz, 3H), 0.93 (s, 9H).

[0838] Synthesis of APH4:

[0839] 1. Synthesis of compound H4-2

[0840] To a solution of compound 2 (200 mg, 450 pmol, 1.0 eq) in methanol (3.00 mL) was added NaHC03(189 mg, 2.25 mmol, 87.52 pL, 5.0 eq) and compound H4-1 (105 mg, 675 pmol, 1.5 eq), the mixture was stirred at 60 °C for 0.5 h. LC-MS (EC16331-49-P1A, P1: RT = 0.497 min) detection showed compound 2 was completely reacted, and the main peak of m / z was consistent with the expected product. After reaction, the mixture was filtered and concentrated under reduced pressure to give the crude product, which was purified by preparative HPLC (neutral condition; column: Waters Xbridge BEH C18 150*25mm*5pm; mobile phase: [water (NH4HC03)-ACN]; gradient: 25%-55% B in 10 min) to give compound H4-2 product (160 mg, 292 pmol, 64.83% yield, 96% purity) as a white solid.

[0841] LC-MS: EC16331-49-P1C; product: RT = 0.488 min

[0842] 1 H NMR: EC16331-49-P1C (400 MHz DMSO) d ppm 8.98 (s, 1H), 8.34 (d, J = 7.6 Hz, 1H), 7.34-7.45 (m, 4H), 5.07 (d, J = 3.6 Hz, 1H), 4.85-4.94 (m, 1H), 4.44 (t, J = 7.6 Hz, 1H), 4.27 (br d, J = 2.0 Hz, 1H), 3.51-3.62 (m, 3H), 3.29 (s, 1H), 3.17 (dt, J = 11.6, 5.6 Hz, 2H), 2.83 (dt, J = 11.6, 6.0 Hz, 2H), 2.45 (s, 3H), 2.26-2.35 (m, 4H), 1.99-2.07 (m, 1H), 1.79 (ddd, J = 12.8, 7.6, 4.8 Hz, 1H), 1.37 (d, J = 7.2 Hz, 3H), 0.99 (s, 9H).

[0843] 2. Synthesis of compound APH4

[0844] To a solution of compound H4-2 (100 mg, 190 pmol, 1.0 eq) in DMF (2.00 mL) was added TEA (192 mg, 1.90 mmol, 264 pL, 10.0 eq), NaBH(OAc)3 (121 mg, 570 pmol, 3.0 eq) and compound C1 (66.9 mg, 94.9 pmol, 0.5 eq), the mixture was stirred at 25 °C for 2 h. LC-MS (EC16331-54-P1A1, P1: RT = 0.769 min; R1: RT = 0.700 min) detection showed the generation of the desired product with m / z consistent with expectations. After reaction, the mixture was filtered, concentrated under reduced pressure to give the product crude, which was purified by prep-HPLC (neutral condition; column: Daisogel SP ODS RPS 150*25mm*5pm; mobile phase: [water(NH4HCO3)-ACN]; gradient: 37%-67%B in 10 min) to give compound APH4 product (20.0 mg, 16.5 pmol, 8.67% yield, 100% purity) as a white solid.

[0845] LC-MS: EC16331-54-P1T; product: RT = 0.417 min

[0846] 1H NMR: EC16331-54-P1M (400 MHz DMSO) δ ppm 8.98 (s, 1 H), 8.31 (br d, J = 7.6 Hz, 1 H), 8.02 (d, J = 7.6 Hz, 1 H), 7.78 (d, J = 8.0 Hz, 1 H), 7.59 (br d, J = 7.2 Hz, 1 H), 7.31 - 7.50 (m, 9 H), 7.25 (dd, J = 12.0, 1.2 Hz, 1 H), 7.16 (br d, J = 8.6 Hz, 1 H), 7.04 - 7.11 (m, 1 H), 6.85 (br d, J = 8.8 Hz, 1 H), 4.86 - 4.93 (m, 3 H), 4.43 (br t, J = 7.6 Hz, 1 H), 4.26 (br s, 2 H), 4.00 (br t, J = 6.4 Hz, 2 H), 3.84 (br t, J = 5.6 Hz, 3 H), 3.57 (br s, 4 H), 3.16 - 3.23 (m, 4 H), 3.07 (br s, 2 H), 2.97 (br d, J = 5.2 Hz, 3 H), 2.85 - 2.92 (m, 2 H), 2.70 - 2.79 (m, 3 H), 2.45 (s, 3 H), 1.94 (br s, 6 H), 1.76 - 1.85 (m, 1 H), 1.62 - 1.72 (m, 2 H), 1.37 (br d, J = 6.8 Hz, 2 H), 1.28 (br d, J = 9.2 Hz, 1 H), 1.23 (br s, 1 H), 0.92 (s, 9 H).

[0847] Synthesis of APH5:

[0848] 1. Synthesis of compound H5-2

[0849] To a solution of compound H5-1 (5.00 g, 39.63 mmol, 1.00 equiv) in THF (238 mL) was added LDBBA (0.5 M, 118 mL, 1.50 equiv) dropwise at -40 °C, the mixture was stirred at -40 °C for 0.5 h. TLC (PE:EA = 7:1, R1:R f = 0.80; P1:R f = 0.75) detection showed that compound H5-1 was completely reacted, two new compound peaks were generated. The reaction mixture was terminated by adding HC1 (1 M, 30.0 mL) at -60 °C after the reaction, dried with Na2S04, filtered and concentrated under reduced pressure to obtain H5-2 product crude (5.20 g) as a brown liquid, which was directly used in the next step reaction without purification.

[0850] 1H NMR: EC16331-78-P1A (400 MHz DMSO) δ ppm 9.59 (d, J = 1.2 Hz, 1H), 5.63 (s, 2H), 3.11-3.15 (m, 1H), 2.55-2.61 (m, 4H).

[0851] 2. Synthesis of compound H5-3

[0852] To a solution of compound H5-2 (5.00 g, 52.0 mmol, 1.00 equiv) in toluene (60.0 mL) was added HOCH2CH2OH (32.3 g, 520 mmol, 29.01 mL, 10.0 equiv) and TsOH (89.6 mg, 520 μmol, 0.01 equiv) under N2environment, the mixture was stirred at 110 °C for 12 h. TLC (PE:EA = 7:1, R1:R f = 0.80; P1:R f = 0.75) detection showed that the reaction of compound H5-2 was complete, a new compound peak was generated. The reaction mixture was extracted by adding water (100.0 mL) and ethyl acetate (80.0 mL*3), the organic layer was washed with saturated brine (100 mL*2), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain the crude product, which was further purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 99:1) to obtain compound H5-3 product (700 mg, 4.99 mmol, 9.60% yield) as a yellow oil.

[0853] 1 H NMR: EC16331-78-P1A (400 MHz DMSO) δ ppm 9.59 (d, J = 1.2 Hz, 1H), 5.63 (s, 2H), 3.11-3.15 (m, 1H), 2.55-2.61 (m, 4H).

[0854] 3. Synthesis of compound H5-4

[0855] To a solution of compound H5-3 (400 mg, 2.85 mmol, 1.00 equiv) in DCM (20.0 mL) at -78 °C, O3 (1.37 g, 2.85 mmol, 1.00 equiv) was bubbled for 1 min, PPh3 (898.11 mg, 3.42 mmol, 1.20 equiv) was added, stirred at -78 °C for 0.5 h, then stirred at 25 °C for 1 h. The reaction was quenched by adding HC1 (1 M, 30.0 mL) at -60 °C, dried over Na2S04, filtered, concentrated under reduced pressure to give the product as a crude, which was further purified by preparative TLC (Si02, PE:EA = 0:1; P1 :R f = 0.30) to give compound H5-4 product (100 mg, crude) as a colorless oil.

[0856] 4. Synthesis of compound H5-5

[0857] To a solution of compound H5-4 (50.0 mg, 290 pmol, 1.00 equiv) in DCM (1.00 mL), TEA (2.93 g, 2.90 mmol, 404 pL, 10.0 equiv), NaBH(OAc)3 (185 mg, 871 pmol, 3.0 equiv) and compound 2 (103 mg, 232 pmol, 0.8 equiv) were added, the mixture was stirred at 25 °C for 2 h. The reaction mixture was extracted by water (5.00 mL) and ethyl acetate (5.00 mL*3), the organic layer was washed by saturated brine (30.0 mL), dried over Na2S04, filtered, concentrated under reduced pressure to give the product as a crude, which was further purified by preparative HPLC (FA condition; column: Phenomenex luna C18 150*25 mm*10 pm; mobile phase: [water (FA)-ACN]; gradient: 9%-39% B in 10 min) to give compound H5-5 product (60.0 mg, 99.19 pmol, 34.16% yield, 96.67% purity) as a yellow oil.

[0858] LC-MS: EC16331-100-P1C; Product: RT = 0.307 min

[0859] 1H NMR: EC16331-100-P1A (400 MHz DMSO) δ ppm 8.70 (s, 1 H), 8.36 (s, 1 H), 7.66 (br d, J = 7.6 Hz, 1 H), 7.40 (q, J = 8.2 Hz, 4 H), 7.28 (s, 1 H), 5.06 (quin, J = 7.2 Hz, 1 H), 4.78 - 4.95 (m, 2 H), 4.65 (d, J = 4.0 Hz, 1 H), 4.60 (br s, 1 H), 3.84 - 3.97 (m, 4 H), 3.77 - 3.83 (m, 1 H), 3.66 (br dd, J = 10.8, 4.8 Hz, 1 H), 3.52 (s, 1 H), 3.45 (br d, J = 12.4 Hz, 1 H), 3.18 (br d, J = 9.6 Hz, 1 H), 2.96 (br s, 9 H), 2.75 - 2.84 (m, 1 H), 2.64 - 2.72 (m, 1 H), 2.57 (br s, 1 H), 2.51 - 2.53 (m, 1 H), 2.03 (ddd, J = 12.8, 8.4, 4.0 Hz, 1 H), 1.70 - 1.82 (m, 2 H), 1.60 (br d, J = 7.2 Hz, 2 H), 1.49 (br d, J = 6.8 Hz, 3 H), 1.10 (s, 9 H).

[0860] 5. Synthesis of compound H5-6

[0861] Compound H5-5 (30.0 mg, 51.3 μmol, 1.00 equiv) was dissolved in formic acid (1.00 mL), and the mixture was stirred at 25 °C for 12 hours. LCMS (EC17015-215-P1A, P1: RT = 1.353 min) detection showed that the desired product was generated. After reaction, the mixture was filtered, concentrated under reduced pressure to obtain compound H5-6 product (28.0 mg, crude), which was used directly in the next step without purification.

[0862] LC-MS: EC17015-215-P1A, product: RT = 1.353 min, m / z = 541.3 [M+H] +

[0863] 6. Synthesis of compound APH5

[0864] To a solution of compound H5-6 (28.0 mg, 51.8 μmol, 1.00 eq) in DCM (1.00 mL) was added TEA (52.4 mg, 518 μmol, 72.1 μL, 10.0 eq) and compound C1 (18.2 mg, 25.9 μmol, 0.50 eq), the mixture was stirred at 25 °C for 0.5 h, then NaBH(OAc)3 (32.9 mg, 155 μmol, 3.00 eq) was added, stirred at 25 °C for 12 h. LCMS (EC17015-217-P1B, P1: RT = 1.388 min) detection showed the generation of the desired product. The reaction mixture was filtered, concentrated under reduced pressure to give the product crude, which was further purified by prep-HPLC (column: CD02-Waters Xbidge BEH C18 150*25*10 μm; mobile phase: [water(NH3H2O)-ACN]; gradient: 24%-54% B in 10 min) to give compound APH5 product (5.00 mg, 3.69 μmol, 7.13% yield, 90.8% purity) as a white solid.

[0865] LC-MS: EC17015-217-P1B, product: RT = 1.388 min, m / z = 1230.5 [M+H] +

[0866] 1H NMR: EC17015-217-P1A (400 MHz DMSO) δ 8.98 (s, 1H), 8.29 (d, J = 7.0 Hz, 1H), 8.01 (d, J = 8.2 Hz, 1H), 7.77 (d, J = 7.6 Hz, 1H), 7.60 (d, J = 7.6 Hz, 1H), 7.41-7.47 (m, 3H), 7.38 (s, 3H), 7.30-7.35 (m, 2H), 7.26 (d, J = 12.6 Hz, 1H), 7.17 (d, J = 7.6 Hz, 1H), 7.05-7.12 (m, 1H), 6.79 (dd, J = 7.0, 2.0 Hz, 1H), 5.30 (s, 1H), 4.90 (s, 3H), 4.40-4.47 (m, 1H), 4.23-4.30 (m, 1H), 4.00 (s, 2H), 3.84 (d, J = 4.4 Hz, 2H), 3.56-3.63 (m, 3H), 3.05 (s, 2H), 2.96 (s, 2H), 2.78-2.86 (m, 2H), 2.72 (s, 2H), 2.45 (s, 6H), 2.29-2.37 (m, 3H), 2.05-2.15 (m, 3H), 1.91-2.04 (m, 4H), 1.75-1.84 (m, 1H), 1.50-1.68 (m, 3H), 1.37 (d, J = 7.0 Hz, 4H), 1.20-1.28 (m, 4H), 0.92 (s, 9H).

[0867] b) Synthesis of APH6, APH7, APH8, APH9

[0868] Synthesis of APH6:

[0869] 1. Synthesis of compound H6-2

[0870] To a solution of compound H6-1 (100 mg, 291 pmol, 1.0 eq) in DMSO (1.00 mL) was added EDCI (83.7 mg, 436 pmol, 1.5 eq), 1-hydroxy-7-azabenzotriazole (59.5 mg, 436 pmol, 61.10 pL, 1.5 eq), NMM (88.3 mg, 873 pmol, 96.0 pL, 3.0 eq) and compound 7A (43.1 mg, 145 pmol, 0.5 eq), the mixture was stirred at 25 °C for 2 h. After reaction, the mixture was filtered, concentrated under reduced pressure to give the product crude, which was further purified by prep-HPLC (FA condition; column: Phenomenex luna C18 150*25mm*10pm; mobile phase: [water(FA)-ACN]; gradient: 16%-46%B in 10 min) to give product H6-2 (80.0 mg, 129 pmol, 44.19% yield) as a white solid.

[0871] LC-MS: EC16331-96-P1A, Product: RT = 0.328 min

[0872] 1 H NMR: EC16331-96-P1A1 (400 MHz DMSO) d ppm 8.95-9.03 (m, 1H), 8.43-8.53 (m, 1H), 7.29-7.58 (m, 4H), 6.09-6.28 (m, 1H), 5.21-5.35 (m, 1H), 5.09 (br s, 1H), 4.19-4.41 (m, 2H), 3.70-3.79 (m, 4H), 3.65 (br dd, J = 12.0, 5.6 Hz, 1H), 3.55 (br d, J = 10.0 Hz, 1H), 2.89-3.02 (m, 2H), 2.42-2.47 (m, 3H), 2.34 (br d, J = 5.6 Hz, 3H), 2.20 (s, 4H), 1.94-2.06 (m, 1H), 1.71-1.85 (m, 1H), 1.34-1.57 (m, 1H), 1.06-1.32 (m, 1H), 0.95 (br d, J = 5.6 Hz, 3H), 0.73-0.82 (m, 3H).

[0873] 2. Synthesis of compound APH6

[0874] To a solution of compound H6-2 (40.0 mg, 64.3 μmol, 1.0 eq) in DCM (1.00 mL) was added TEA (65.1 mg, 643 μmol, 89.5 μL, 10.0 eq), NaBH(OAc)3 (40.9 mg, 193 μmol, 3.0 eq) and compound C1 (22.67 mg, 32.17 μmol, 0.5 eq), the mixture was stirred at 25 °C for 2 h. After reaction, the mixture was extracted with water (5.00 mL) and ethyl acetate (5.00 mL*3), the organic layer was washed with saturated brine (10.0 mL*2), dried over Na2SO4, filtered and concentrated under reduced pressure to give the product as a white solid. Further purification by prep-HPLC (FA condition; Column: Phenomenex luna C18 150*25mm*10μm; mobile phase: [water(FA)-ACN]; gradient: 22%-52% B in 8 min) to give compound APH6 (10.0 mg, 7.05 μmol, 10.96% yield, 92.44% purity) as a white solid.

[0875] LC-MS: EC16331-105-P1Y; Product: RT = 0.445 min

[0876] 1H NMR: EC16331-105-P1E (400 MHz DMSO) δ ppm 8.92-8.99 (m, 1, H), 8.46 (br t, J = 8.4 Hz, 1 H), 8.20 (s, 1 H), 8.03 (d, J = 7.6 Hz, 1 H), 7.79 (d, J = 8.0 Hz, 1 H), 7.59 (br d, J = 7.6 Hz, 1 H), 7.31 -7.53 (m, 9 H), 7.23-7.30 (m, 1 H), 7.17 (br d, J = 8.0 Hz, 1 H), 7.05-7.13 (m, 1 H), 6.90 (br d, J = 8.8 Hz, 1 H), 6.21 (s, 1 H), 5.13-5.24 (m, 1 H), 4.91 (s, 2 H), 4.61 -4.72 (m, 1 H), 4.22-4.44 (m, 5 H), 4.01 (br t, J = 6.2 Hz, 2 H), 3.85 (br t, J = 5.6 Hz, 3 H), 3.73 (br dd, J = 9.6, 4.8 Hz, 3 H), 3.41 (br s, 1 H), 3.38-3.38 (m, 1 H), 3.38 (br s, 1 H), 3.17 (s, 1 H), 2.97 (br t, J = 5.6 Hz, 3 H), 2.85-2.93 (m, 3 H), 2.74-2.81 (m, 3 H), 2.46 (br d, J = 2.8 Hz, 5 H), 2.36 (br s, 3 H), 2.19 (s, 2 H), 2.17 (s, 1 H), 1.90-2.06 (m, 4 H), 1.73-1.83 (m, 1 H), 1.61 -1.72 (m, 2 H), 1.23 (s, 2 H), 0.96 (br d, J = 6.4 Hz, 3 H), 0.78 (br s, 1 H), 0.76 (d, J = 6.8 Hz, 2 H).

[0877] Synthesis of APH7:

[0878] 1. Synthesis of compound H7-2

[0879] To a solution of compound H7-1 (200 mg, 724 pmol, 1.00 equiv) and compound 7A (193 mg, 651 pmol, 0.90 equiv) in DMSO (2.00 mL) was added EDCI (208 mg, 1.09 mmol, 1.50 equiv), HOAt (148 mg, 1.09 mmol, 152 pL, 1.50 equiv) and NMM (220 mg, 2.17 mmol, 239 pL, 3.00 equiv), and the mixture was stirred at 25 °C for 1 h. After reaction, the mixture was diluted with water (10.0 mL), extracted with DCM (10.0 mL*6), and the organic layer was dried over Na2S04, filtered and concentrated under reduced pressure to give the product crude, which was further purified by preparative TLC (petroleum ether: ethyl acetate = 0: 1, P1 : Rf = 0.50) to give compound H7-2 product (240 mg, 433 pmol, 59.8% yield) as a yellow oil. f

[0880] LC-MS: EC17015-204-P1A, product: RT = 0.363 min, m / z = 555.2 [M+H] +

[0881] 2. Synthesis of compound H7-3

[0882] To a solution of compound H7-2 (240 mg, 433 pmol, 1.00 equiv) in methanol (3.00 mL) and water (1.00 mL) was added LiOH.H20 (54.5 mg, 1.30 mmol, 3.00 equiv), and the mixture was stirred at 25 °C for 1 h. LCMS (EC17015-206-P1A, P1 : RT = 0.328 min) detection showed that the desired product was generated with 96.0% ratio. After reaction, the mixture was filtered and concentrated under reduced pressure to give compound H7-3 crude (230 mg) as a yellow solid, which was used directly in the next reaction without purification.

[0883] LC-MS: EC17015-206-P1A, product: RT = 0.328 min, m / z = 541.2 [M+H] +

[0884] 1 H NMR: EC17015-206-P1A (400 MHz DMSO)

[0885] ​δ ppm 8.93 (s, 1H), 7.17-7.47 (m, 4H), 6.12-6.35 (m, 1H), 4.78-5.00 (m, 1H), 4.13-4.41 (m, 1H), 3.65-3.88 (m, 2H), 3.50 (s, 2H), 2.41 (s, 3H), 2.25 (dt, J = 14.2, 6.8 Hz, 3H), 2.10-2.19 (m, 3H), 1.91-2.03 (m, 1H), 1.72-1.85 (m, 1H), 1.24 (d, J = 12.2 Hz, 1H), 0.93 (dd, J = 6.2, 2.8 Hz, 3H), 0.68-0.83 (m, 3H).

[0886] 3. Synthesis of compound H7-4

[0887] To a solution of compound H7-3 (230 mg, 425 μmol, 1.00 equiv) and compound 1A (102 mg, 638 μmol, 1.50 equiv) in DMF (2.00 mL) was added HATU (243 mg, 638 μmol, 1.50 equiv) and DIEA (330 mg, 2.55 mmol, 445 μL, 6.00 equiv), and the mixture was stirred at 25 °C for 1 h. LCMS (EC17015-213-P1A, P1: RT = 0.356 min) detected that the target product accounted for 42.4%. The reaction mixture was extracted with ethyl acetate (15.0 mL*3) after water (20.0 mL) was added, and the organic layer was dried over Na2S04, filtered and concentrated under reduced pressure to obtain the product crude, which was further purified by preparative TLC (dichloromethane:methanol = 10 / 1, P1: Rf = 0.45) to obtain compound H7-4 product (100 mg, 147 μmol, 34.5% yield) as a colorless oil. f

[0888] LC-MS: EC17015-213-P1A, product: RT = 0.356 min, m / z = 682.2 [M+H] +

[0889] 4. Synthesis of compound H7-5

[0890] ​To a solution of compound H7-4 (45.0 mg, 77.0 μmol, 1.00 equiv) in formic acid (1.00 mL) was added, and the mixture was stirred at 25 °C for 0.5 h. LCMS (EC17015-224-P1A, P1: RT = 0.341 min) detected the desired product with 91.5% yield. The reaction mixture was filtered, concentrated under reduced pressure to give compound H7-5 (42.0 mg, crude) as a yellow oil, which was used directly in the next step without further purification.

[0891] LC-MS: EC17015-224-P1A, product: RT = 0.341 min, m / z = 636.2 [M+H] +

[0892] 5. Synthesis of compound APH7

[0893] To a solution of compound H7-5 (42.0 mg, 66.1 μmol, 1.00 equiv) in DCM (1.00 mL) was added TEA (66.8 mg, 661 μmol, 92.0 μL, 10.0 equiv) and compound C1 (46.6 mg, 66.1 μmol, 1.00 equiv), and the mixture was stirred at 25 °C for 0.5 h, followed by the addition of NaBH(OAc)3 (42.0 mg, 198 μmol, 3.00 equiv) and continued stirring at 25 °C for 1 h. LCMS (EC17015-225-P1B, P1: RT = 0.448 min) detected the desired product with 52.0% yield. The reaction mixture was filtered, concentrated under reduced pressure to give the product crude, which was further purified by prep-HPLC (column: CD01-Phenomenex luna C18 150*25*10 μm; mobile phase: [water (FA)-ACN]; gradient: 30%-50% B in 10 min) to give compound APH7 (12.0 mg, 8.71 μmol, 13.2% yield, 96.1% purity) as a white solid.

[0894] LC-MS: EC17015-225-P1B, product: RT = 0.448 min, m / z = 1324.6 [M+H] +

[0895] 1 H NMR: EC17015-225-P1A (400 MHz DMSO)

[0896] 8.90-9.05 (m, 1H), 8.18 (s, 1H), 8.03 (d, J = 7.8 Hz, 1H), 7.78 (d, J = 8.0 Hz, 1H), 7.59 (d, J = 7.4 Hz, 1H), 7.48-7.54 (m, 1H), 7.46 (d, J = 8.2 Hz, 1H), 7.30-7.44 (m, 7H), 7.26 (d, J = 12.4 Hz, 1H), 7.14-7.21 (m, 1H), 7.05-7.13 (m, 1H), 6.91 (d, J = 8.8 Hz, 1H), 6.20 (s, 1H), 5.12-5.24 (m, 1H), 4.91 (s, 2H), 4.20-4.43 (m, 4H), 4.01 (t, J = 6.4 Hz, 3H), 3.85 (t, J = 5.6 Hz, 4H), 3.69-3.77 (m, 4H), 2.97 (t, J = 5.4 Hz, 4H), 2.74-2.81 (m, 3H), 2.45 (s, 6H), 2.23-2.34 (m, 4H), 2.14-2.21 (m, 4H), 1.86-2.07 (m, 6H), 1.50-1.70 (m, 4H), 0.95 (d, J = 6.4 Hz, 3H), 0.78 (dd, J = 14.2, 6.8 Hz, 5H).

[0897] Synthesis of APH8:

[0898] 1. Synthesis of compound H8-2

[0899] To a solution of compound H8-1 (200 mg, 812 pmol, 1.0 eq) in DMSO (2.00 mL) was added EDCI (233 mg, 1.22 mmol, 1.5 eq), HOAt (166 mg, 1.22 mmol, 170 pL, 1.5 eq), compound 7A (120 mg, 406.00 pmol, 0.5 eq) and NMM (246 mg, 2.44 mmol, 267.82 pL, 3.0 eq), the mixture was stirred at 25 °C for 1 h. After reaction, the mixture was extracted with ethyl acetate (15.0 mL*3) after adding water (15.0 mL), the organic layer was washed with saturated brine (25.0 mL*2), dried over Na2S04, filtered and concentrated under reduced pressure to give the product crude, which was further purified by preparative TLC (Si02, PE:EA = 0:1, P1 :R f = 0.1) to give compound H8-2 (100 mg, 177 pmol, 21.74% yield, 92.6% purity) as colorless oil.

[0900] LC-MS: EC16331-79-P1C2, Product: RT = 0.317 min

[0901] 2. Synthesis of compound APH8

[0902] To a solution of compound H8-2 (70.0 mg, 133 pmol, 1.0 eq) in DCM (2.00 mL) was added TEA (135 mg, 1.33 mmol, 185.72 pL, 10.0 eq), NaBH(OAc)3 (84.8 mg, 400 pmol, 3.0 eq) and compound Cl (47.0 mg, 66.7 pmol, 0.5 eq), the mixture was stirred at 25 °C for 2 h. After reaction, the mixture was filtered, concentrated under reduced pressure to give the product as a crude, which was further purified by prep-HPLC (FA condition; column: Phenomenex luna C18 150*25mm*10pm; mobile phase: [water(FA)-ACN]; gradient: 29%-59%B in 10 min) to give compound APH8 (10.0 mg, 7.57 pmol, 5.67% yield, 91.86% purity) as a white solid.

[0903] LC-MS: EC16331-94-P1B1, Product: RT = 0.440 min, ½ m / z = 607.5 [M+H] + ;

[0904] 1H NMR: EC16331-94-P1H (400 MHz DMSO) δ ppm 8.39 (br d, J=8.4 Hz, 1 H), 8.32 (s, 1 H), 7.98-8.05 (m, 1 H), 7.77 (d, J=8.0 Hz, 1 H), 7.59 (br d, J=6.8 Hz, 1 H), 7.45 (br t, J=7.2 Hz, 1 H), 7.34-7.42 (m, 3 H), 7.33 (br dd, J=7.2, 3.75 Hz, 2 H), 7.25-7.30 (m, 4 H), 7.24 (br s, 1 H), 7.21 (br d, J=4.4 Hz, 1 H), 7.18 (br d, J=4.0 Hz, 1 H), 7.15 (br s, 1 H), 7.06-7.12 (m, 1 H), 6.71-6.78 (m, 1 H), 6.21 (s, 1 H), 5.10-5.21 (m, 1 H), 4.88 (br s, 2 H), 4.21-4.36 (m, 5 H), 3.95-4.02 (m, 4 H), 3.81 (br d, J=5.2 Hz, 5 H), 3.71 (br d, J=2.4 Hz, 6 H), 2.95 (br s, 3 H), 2.80-2.88 (m, 2 H), 2.65-2.73 (m, 3 H), 2.30-2.37 (m, 1 H), 2.15-2.24 (m, 4 H), 1.91-2.03 (m, 3 H), 1.52-1.79 (m, 3 H), 1.23 (br s, 2 H), 1.05 (br t, J=7.2 Hz, 1 H), 0.92-1.01 (m, 3 H), 0.71-0.82 (m, 3 H).

[0905] Synthesis of APH9:

[0906] 1. Synthesis of compound H9-2

[0907] A mixture of compound H9-1 (500 mg, 2.79 mmol, 1.00 equiv), compound 7A (413 mg, 1.39 mmol, 0.50 equiv), EDCI (802 mg, 4.18 mmol, 1.50 equiv), HOAt (570 mg, 4.18 mmol, 585 μL. 1.50 equiv) and NMM (846 mg, 8.37 mmol, 920 μL, 3.00 equiv) in DMSO (5.00 mL) was stirred at 25 °C for 1 h. LCMS (EC17015-207-PA, P1: RT = 0.357 min) detected the desired product with 99.4% ratio. The reaction mixture was diluted with water (10.0 mL), extracted with DCM (10.0 mL*6), the organic layer was washed with saturated brine (10.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product, which was further purified by preparative TLC (petroleum ether: ethyl acetate = 0 / 1, P1: Rf = 0.50) to give compound H9-2 (700 mg, 1.53 mmol, 54.8% yield) as colorless oil. f

[0908] LC-MS: EC17015-207-P1A, product: RT = 0.357 min, m / z = 458.2 [M+H] +

[0909] 1 H NMR: EC17015-207-P1A (400 MHz CDCl3)

[0910] δ ppm 7.16-7.31 (m, 5H), 5.96-6.12 (m, 1H), 5.31 (d, J = 8.2 Hz, 1H), 4.39-4.56 (m, 2H), 3.72 (dd, J = 10.4, 4.8 Hz, 1H), 3.44-3.64 (m, 5H), 2.63-2.93 (m, 3H), 2.25-2.41 (m, 2H), 2.10-2.22 (m, 3H), 1.85-2.04 (m, 2H), 0.97 (d, J = 6.4 Hz, 3H), 0.81 (d, J = 6.6 Hz, 3H).

[0911] 2. Synthesis of compound H9-3

[0912] ​To a solution of compound H9-2 (700 mg, 1.53 mmol, 1.00 equiv) in methanol (6.00 mL) and water (2.00 mL) was added LiOH.H2O (193 mg, 4.59 mmol, 3.00 equiv), the mixture was stirred at 25 °C for 1 h. LCMS (EC17015-210-P1A, P1: RT = 0.327 min) detection showed that the target product was generated with 100% ratio. After reaction, the mixture was filtered, concentrated under reduced pressure to give the product H9-3 crude (700 mg) as a white solid, which was used directly in the next step without purification.

[0913] LC-MS: EC17015-210-P1A, product: RT = 0.327 min, m / z = 444.2 [M+H] +

[0914] 3. Synthesis of compound H9-4

[0915] To a solution of compound H9-3 (350 mg, 790 µmol, 1.00 equiv) and compound 1A (188 mg, 1.18 mmol, 1.50 equiv) in DMF (4.00 mL) was added HATU (360 mg, 947 µmol, 1.20 equiv) and DIEA (612 mg, 4.74 mmol, 825 µL, 6.00 equiv), the mixture was stirred at 25 °C for 1 h. LCMS (EC17015-216-P1B, P1: RT = 0.343 min) detection showed that the target product was generated with 18.8% ratio. After reaction, the mixture was filtered, concentrated under reduced pressure to give the product crude, which was further purified by reverse phase HPLC (neutral condition) to give compound H9-4 product (140 mg, 239 µmol, 30.3% yield) as a yellow solid.

[0916] LC-MS: EC17015-216-P1B, product: RT = 0.343 min, m / z = 585.0 [M+H] +

[0917] 4. Synthesis of compound H9-5

[0918] To a solution of compound H9-4 (60.0 mg, 103 µmol, 1.00 equiv) in DCM (2.00 mL) was added formic acid (1.00 mL), the mixture was stirred at 25 °C for 0.5 h. LCMS (EC17015-230-P1A, P1: RT = 0.303 min) detection showed that the target product was generated with 87.8% yield. After reaction, the mixture was filtered, concentrated under reduced pressure to give H9-5 product (52.0 mg, 96.5 µmol, 94.1% yield) as a yellow oil.

[0919] LC-MS: EC17015-230-P1A, product: RT = 0.303 min, m / z = 539.1 [M+H] +

[0920] 5. Synthesis of compound APH9

[0921] To a solution of compound H9-5 (52.0 mg, 96.5 µmol, 1.00 equiv) in DCM (2.00 mL) was added TEA (97.7 mg, 965 µmol, 134 µL, 10.0 equiv) and compound C1 (54.4 mg, 77.2 µmol, 0.80 equiv), the mixture was stirred at 25 °C for 0.5 h, then NaBH(OAc)3 (61.4 mg, 290 µmol, 3.00 equiv) was added, and the reaction was continued to stir at 25 °C for 12 h. LCMS (EC17015-231-P1A, P1: RT = 0.447 min) detection showed that the target product was generated with 63.3% yield. After reaction, the mixture was filtered, concentrated under reduced pressure to give the crude product, which was further purified by prep-HPLC (column: CD02-Waters Xbidge BEH C18 150*25*10 µm; mobile phase: [water(NH4HCO3)-ACN]; gradient: 38%-58% B in 10 min) to give compound APH9 (16.0 mg, 12.1 µmol, 12.5% yield, 92.8% purity) as a white solid.

[0922] LC-MS: EC17015-231-P1A, product: RT = 0.447 min, m / z = 1227.5 [M+H] +

[0923] 1H NMR: EC17015-231-P1A (400 MHz DMSO) δ ppm 8.03 (d, J = 7.8 Hz, 1 H), 7.78 (d, J = 8.2 Hz, 1 H), 7.60 (d, J = 7.2 Hz, 1 H), 7.52 (d, J = 8.4 Hz, 1 H), 7.47 (t, J = 7.6 Hz, 1 H), 7.38-7.43 (m, 1 H), 7.35 (d, J = 7.4 Hz, 2 H), 7.30-7.33 (m, 1 H), 7.29 (s, 4 H), 7.15-7.22 (m, 2 H), 7.05-7.13 (m, 1 H), 6.91 (d, J = 9.0 Hz, 1 H), 6.14-6.23 (m, 1 H), 5.07-5.23 (m, 1 H), 4.92 (s, 2 H), 4.14-4.46 (m, 3 H), 4.01 (t, J = 6.4 Hz, 2 H), 3.85 (t, J = 5.6 Hz, 3 H), 3.72 (d, J = 9.8 Hz, 2 H), 3.43-3.58 (m, 6 H), 2.97 (t, J = 5.6 Hz, 2 H), 2.74-2.87 (m, 5 H), 2.64-2.73 (m, 2 H), 2.28-2.36 (m, 3 H), 2.14-2.21 (m, 4 H), 2.07 (d, J = 5.2 Hz, 2 H), 1.90-2.03 (m, 5 H), 1.50-1.80 (m, 5 H), 0.95 (d, J = 6.6 Hz, 3 H), 0.68-0.86 (m, 5 H).

[0924] c) Synthesis of APH10, APH11, APH12

[0925] Synthesis of APH10:

[0926] 1. Synthesis of compound H10-1

[0927] A mixture of compound 8A (200 mg, 418 pmol, 1.00 equiv), compound 1A (100 mg, 628 pmol, 1.50 equiv), HATU (239 mg, 628 pmol, 1.50 equiv) and DIEA (324 mg, 2.51 mmol, 437 pL, 6.00 equiv) in DMF (3.00 mL) was stirred at 25 °C for 2 h. LCMS (EC17015-293-P1B, P1: RT = 0.370 min) detection showed that the target product was generated with 20.1% yield. The reaction mixture was extracted with water (20.0 mL) and ethyl acetate (15.0 mL*5), the organic layer was washed with saturated brine (10.0 mL), dried over Na2S04, filtered and concentrated under reduced pressure to give the product crude, which was further purified by prep-HPLC (column: CD07-Daisogel SP-100-8-ODS-PK 150*25*10 pm; mobile phase: [water(NH4HCO3)-ACN]; gradient: 24%-54% B in 12 min) to give compound H10-1 product (165 mg, 266 pmol, 63.7% yield) as colorless oil.

[0928] LC-MS: EC17015-293-P1B, product: RT = 0.370 min, m / z = 619.1 [M+H] +

[0929] 1 H NMR: EC17015-293-P1A (400 MHz CDCl3)

[0930] ppm 7.31-7.47 (m, 4H), 6.14-6.32 (m, 1H), 5.23-5.52 (m, 1H), 4.51-4.71 (m, 2H), 3.89-4.00 (m, 1H), 3.62-3.83 (m, 3H), 3.34-3.52 (m, 6H), 2.73-3.16 (m, 3H), 2.45-2.60 (m, 2H), 2.35 (d, J = 9.6 Hz, 4H), 2.16-2.26 (m, 1H), 2.15-2.15 (m, 1H), 1.92-2.13 (m, 2H), 1.71-1.87 (m, 3H), 1.70-1.71 (m, 1H), 1.09-1.22 (m, 3H), 0.84-0.89 (m, 1H), 0.98 (t, J = 6.2 Hz, 3H)

[0931] 2. Synthesis of compound H10-2

[0932] To a solution of compound H10-2 (71.0 mg, 124 pmol, 1.00 eq) in DCM (3.00 mL) was added TEA (125 mg, 1.24 mmol, 172 pL, 10.0 eq) and compound C1 (43.7 mg, 62.0 pmol, 0.50 eq), the mixture was stirred at 25 °C for 0.5 h, then NaBH(OAc)3 (78.8 mg, 372 pmol, 3.00 eq) and TEA (125 mg, 1.24 mmol, 172 pL, 10.0 eq) were added, the mixture was stirred at 25 °C for 1 h. LCMS (EC17015-299-P1A, P1: RT = 0.610 min) detection showed that the target product was generated with a proportion of 68.0%. After reaction, the mixture was filtered and concentrated under reduced pressure to obtain the product crude, which was further purified by preparative HPLC (column: CD02-Waters Xbidge BEH C18 150*25*10 pm; mobile phase: [water(NH4HCO3)-ACN]; gradient: 32%-62%B within 13 min) to obtain compound APH10 (26.0 mg, 19.4 pmol, 15.7% yield, 94.2% purity) as a white solid.

[0933] LC-MS: EC17015-299-P1Q1, product: RT = 0.618 min, m / z = 1261.2 [M+H] +

[0934] 3. Synthesis of compound APH10

[0935] To a solution of compound H10-2 (71.0 mg, 124 pmol, 1.00 eq) in DCM (3.00 mL) was added TEA (125 mg, 1.24 mmol, 172 pL, 10.0 eq) and compound C1 (43.7 mg, 62.0 pmol, 0.50 eq), the mixture was stirred at 25 °C for 0.5 h, then NaBH(OAc)3 (78.8 mg, 372 pmol, 3.00 eq) and TEA (125 mg, 1.24 mmol, 172 pL, 10.0 eq) were added, the mixture was stirred at 25 °C for 1 h. LCMS (EC17015-299-P1A, P1: RT = 0.610 min) detection showed that the target product was generated with a proportion of 68.0%. After reaction, the mixture was filtered and concentrated under reduced pressure to obtain the product crude, which was further purified by preparative HPLC (column: CD02-Waters Xbidge BEH C18 150*25*10 pm; mobile phase: [water(NH4HCO3)-ACN]; gradient: 32%-62%B within 13 min) to obtain compound APH10 (26.0 mg, 19.4 pmol, 15.7% yield, 94.2% purity) as a white solid.

[0936] LC-MS: EC17015-299-P1Q1, product: RT = 0.618 min, m / z = 1261.2 [M+H] +

[0937] 1H NMR: EC17015-299-P1A (400 MHz DMSO) δ ppm 8.04 (d, J = 7.6 Hz, 1 H), 7.79 (d, J = 8.0 Hz, 1 H), 7.60 (d, J = 7.6 Hz, 1 H), 7.53 (d, J = 8.8 Hz, 1 H), 7.48 (t, J = 7.6 Hz, 1 H), 7.39-7.43 (m, 1 H), 7.27-7.39 (m, 6 H), 7.21-7.26 (m, 1 H), 7.06-7.13 (m, 1 H), 7.18 (d, J = 8.8 Hz, 1 H), 6.93 (d, J = 8.4 Hz, 1 H), 6.14-6.25 (m, 1 H), 5.06-5.22 (m, 1 H), 4.92 (s, 2 H), 4.20-4.45 (m, 3 H), 4.02 (t, J = 6.2 Hz, 2 H), 3.86 (t, J = 5.6 Hz, 2 H), 3.66-3.77 (m, 2 H), 3.43-3.61 (m, 7 H), 3.10-3.17 (m, 2 H), 2.98 (t, J = 5.4 Hz, 2 H), 2.86-2.92 (m, 1 H), 2.79 (t, J = 7.4 Hz, 2 H), 2.61-2.75 (m, 2 H), 2.27-2.43 (m, 5 H), 2.16-2.21 (m, 3 H), 2.08 (s, 2 H), 1.91-2.03 (m, 4 H), 1.51-1.79 (m, 4 H), 1.19-1.29 (m, 1 H), 0.95 (d, J = 6.4 Hz, 3 H), 0.68-0.83 (m, 4 H).

[0938] Synthesis of APH11:

[0939] 1. Synthesis of compound H11-2

[0940] To a solution of compound H11-1 (8.00 g, 26.7 mmol, 1.00 equiv) and K2CO3 (7.38 g, 53.4 mmol, 2.00 equiv) in DMF (120 mL) was added Mel (19.0 g, 133 mmol, 8.31 mL, 5.00 equiv) dropwise at 25 °C under N2 atmosphere for 30 min, the mixture was stirred at 80 °C for 2.5 h. After reaction, the mixture was diluted with water (200 mL), extracted with ethyl acetate (120 mL*3), the organic layer was washed with saturated brine (120 mL*2), dried over Na2SO4, filtered and concentrated under reduced pressure to give compound H11-2 product crude (9.00 g) as a yellow solid, which was used directly in the next step without purification.

[0941] LC-MS: EC16331-109-P1A, Product: RT = 0.439 min

[0942] 1 H NMR: EC16331-109-P1B (400 MHz DMSO) δ ppm 7.51 (d, J = 8.4 Hz, 1 H), 7.30 - 7.40 (m, 4 H), 4.84 - 4.97 (m, 1 H), 3.55 (s, 3 H), 2.76 - 2.81 (m, 1 H), 2.63 - 2.69 (m, 1 H), 1.34 (s, 8 H).

[0943] 2. Synthesis of compound H11-3

[0944] To compound H11-2 (9.00 g, 28.7 mmol, 1.00 equiv) was added HC1-dioxane (2 M, 71.7 mL, 5.00 equiv), the mixture was stirred at 25 °C for 2 h. After reaction, the mixture was filtered, concentrated under reduced pressure to give compound H11-3 product (7.70 g, crude, HC1 salt) as a yellow solid, the crude was used in the next step without purification.

[0945] 1 H NMR: EC16331-109-P1B (400 MHz DMSO) δ ppm 8.86 (s, 3 H), 7.60 (d, J = 8.4 Hz, 2 H), 7.48 (d, J = 8.4 Hz, 2 H), 4.52 - 4.68 (m, 1 H), 3.54 (s, 3 H), 3.24 (dd, J = 16.4, 5.6 Hz, 1 H), 3.02 (dd, J = 16.4, 8.8 Hz, 1 H).

[0946] 3. Synthesis of compound H11-4

[0947] Compound H11-3 (1.80 g, 7.20 mmol, 1.00 eq, HC1), compound 7A (1.07 g, 3.60 mmol, 0.50 eq), EDCI (2.07 g, 10.8 mmol, 1.50 eq), NMM (2.18 g, 21.6 mmol, 2.37 mL, 3.00 eq) and HOAt (1.47 g, 10.8 mmol, 1.51 mL, 1.50 eq) were mixed in DMSO (20.0 mL) and the reaction was stirred at 25 °C for 1 h. LCMS (EC17015-237-P1A1, P1: RT = 0.353 min) detection showed that the target product was generated with 84.0% yield. The reaction mixture was filtered, concentrated under reduced pressure to give the product crude, which was further purified by reverse phase HPLC (0.1% FA condition) to give compound H11-4 (1.60 g, 3.25 mmol, 45.2% yield) as a yellow solid.

[0948] LC-MS: EC17015-237-P1A1, product: RT = 0.353 min, m / z = 492.0 [M+H] +

[0949] 4. Synthesis of compound 8A

[0950] To a solution of compound H11-4 (1.60 g, 3.25 mmol, 1.00 eq) in methanol (12.0 mL) and water (4.00 mL) was added LiOH.H2O (409 mg, 9.76 mmol, 3.00 eq) and the mixture was stirred at 25 °C for 1 h. LCMS (EC17015-249-P1A, P1: RT = 0.326 min) detection showed that the target product was generated with 100% yield. The reaction mixture was filtered, concentrated under reduced pressure to give compound 8A product (1.80 g, crude) as a yellow solid, which was used in the next step without purification.

[0951] LC-MS: EC17015-249-P1A, product: RT = 0.326 min, m / z = 478.0 [M+H] +

[0952] 5. Synthesis of compound APH11

[0953] Compound 8A (10.0 mg, 20.9 pmol, 1.00 equiv), compound C1 (10.3 mg, 14.6 pmol, 0.70 equiv), PyBOP (13.1 mg, 25.1 pmol, 1.20 equiv) and TEA (6.35 mg, 62.8 pmol, 8.74 pL, 3.00 equiv) were mixed in DMA (0.50 mL) and stirred at 25 °C for 1 h. LCMS (EC17015-294-P1E1, P1: RT = 0.423 min) detected 18.8% of the desired product. The reaction mixture was filtered under reduced pressure, concentrated to give the product crude, which was further purified by prep-HPLC (column: CD02-Waters Xbidge BEH C18 150*25*10 pm; mobile phase: [water (NH4HCO3)-ACN]; gradient: 22%-52% B in 10 min) to give compound APH11 (8.30 mg, 5.61 pmol, 2.68% yield, 93.4% purity) as a white solid.

[0954] LC-MS: EC17015-294-P1E1, product: RT = 0.423 min, m / z = 1164.0 [M+H] +

[0955] 1 H NMR: EC17015-294-P1A (400 MHz DMSO) d ppm 8.02 (d, J = 8.2 Hz, 1H), 7.78 (d, J = 8.4 Hz, 1H), 7.60 (d, J = 7.2 Hz, 1H), 7.43-7.51 (m, 2H), 7.38-7.42 (m, 1H), 7.23-7.37 (m, 7H), 7.17 (d, J = 8.4 Hz, 1H), 7.05-7.13 (m, 1H), 6.85 (d, J = 7.8 Hz, 1H), 6.09-6.28 (m, 1H), 5.07-5.22 (m, 1H), 4.91 (s, 2H), 4.18-4.40 (m, 2H), 4.01 (s, 2H), 3.80-3.87 (m, 2H), 3.68-3.75 (m, 2H), 3.47 (s, 4H), 2.95-2.99 (m, 2H), 2.72-2.79 (m, 4H), 2.30-2.45 (m, 5H), 2.18 (s, 3H), 1.88-2.05 (m, 4H), 1.67-1.78 (m, 1H), 1.23 (s, 2H), 0.94 (d, J = 6.0 Hz, 3H), 0.68-0.86 (m, 4H).

[0956] Synthesis of APH12:

[0957] 1. Synthesis of compound H12-2

[0958] Compound 8A (200 mg, 418 pmol, 1.00 equiv), compound H12-1 (66.0 mg, 628 pmol, 68.4 pL, 1.50 equiv), HATU (239 mg, 628 pmol, 1.50 equiv) and DIEA (324 mg, 2.51 mmol, 437 pL, 6.00 equiv) were mixed in DMF (3.00 mL) and stirred at 25 °C for 1 h. LCMS (EC17015-279-P1B, P1: RT = 0.337 min) detected the desired product with 99.7% ratio. The reaction mixture was concentrated under reduced pressure to give the product crude, which was further purified by prep-HPLC (neutral condition; column: CD07-Daisogel SP-100-8-ODS-PK 150*25*10 pm; mobile phase: [water(NH4HCO3)-ACN]; gradient: 22%-52%B, 10 min) to give compound H12-2 product (160 mg, crude) as a white solid.

[0959] LC-MS: EC17015-279-P1B, product: RT = 0.337 min

[0960] 2. Synthesis of compound H12-3

[0961] To compound H12-2 (70.0 mg, 124 pmol, 1.00 equiv) was added formic acid (1.00 mL) and the mixture was stirred at 25 °C for 0.5 h. The reaction mixture was filtered and concentrated under reduced pressure to give compound H12-3 product (60.0 mg, crude) as a yellow oil, which was used in the next step without further purification.

[0962] LC-MS: EC16331-182-P1A, product: RT = 0.332 min;

[0963] 3. Synthesis of compound APH12

[0964] To a solution of compound H12-3 (60.0 mg, 116 pmol, 1.00 equiv) and compound Cl (40.7 mg, 57.8 pmol, 0.50 equiv) in DCM (2.00 mL) was added TEA (117 mg, 1.16 mmol, 161 pL, 10.0 equiv) and NaBH(OAc)3 (73.5 mg, 347 pmol, 3.00 equiv), the mixture was stirred at 25 °C for 2 h. After reaction, the mixture was filtered, concentrated under reduced pressure to give the product crude (60.0 mg), which was further purified by prep-HPLC (column: Phenomenex luna C18 150*25mm*10pm; mobile phase: [water(HCl)-ACN]; gradient: 37%-67% B in 14 min) to give compound APH12 (25.0 mg, 19.3 pmol, 16.7% yield, 96.2% purity, HC1) as a white solid.

[0965] LC-MS: EC16331-183-P1A1, Product: RT = 1.853 min

[0966] 1H NMR: EC16331-183-P1R (400 MHz DMSO) δ ppm 8.26-8.34 (m, 1 H), 8.04 (d, J = 7.6 Hz, 1 H), 7.79 (d, J = 8.0 Hz, 1 H), 7.55-7.63 (m, 2 H), 7.44-7.51 (m, 2 H), 7.40-7.43 (m, 1 H), 7.28-7.39 (m, 7 H), 7.24 (br d, J = 8.4 Hz, 1 H), 7.15 (br t, J = 8.8 Hz, 1 H), 6.98 (d, J = 8.8 Hz, 1 H), 6.20-6.24 (m, 1 H), 5.14 (dt, J = 14.8, 7.6 Hz, 1 H), 4.93 (s, 2 H), 4.41 (br d, J = 7.6 Hz, 1 H), 4.33-4.38 (m, 2 H), 4.23 (br d, J = 3.2 Hz, 2 H), 4.16 (br s, 2 H), 4.04 (br t, J = 6.4 Hz, 3 H), 3.86 (br t, J = 5.6 Hz, 2 H), 3.70-3.75 (m, 2 H), 3.44-3.55 (m, 5 H), 3.34-3.43 (m, 4 H), 3.11-3.21 (m, 2 H), 2.98 (br t, J = 5.2 Hz, 2 H), 2.81 (br t, J = 7.6 Hz, 2 H), 2.65 (br dd, J = 14.8, 7.6 Hz, 1 H), 2.56 (br d, J = 6.8 Hz, 1 H), 2.25 (br d, J = 7.6 Hz, 1 H), 2.18 (s, 3 H), 1.92-2.02 (m, 3 H), 1.64-1.73 (m, 1 H), 1.20-1.27 (m, 1 H), 0.91-1.00 (m, 3 H), 0.70-0.82 (m, 4 H).

[0967] d) Synthesis of APH14, APH15, APH16

[0968] Synthesis of APH14:

[0969] 1. Synthesis of compound H14-2

[0970] To a solution of compound H14-1 (3.00 g, 21.5 mmol, 1.00 equiv, HC1) in ACN (15.0 mL) was added DIEA (5.06 g, 39.1 mmol, 6.81 mL, 1.82 equiv) and compound 4A (2.33 g, 19.6 mmol, 1.69 mL, 0.91 equiv), the mixture was stirred at 25 °C for 12 h. After reaction, the mixture was filtered, concentrated under reduced pressure to give the product crude, which was further purified by column chromatography (Si02, Petroleum ether / Ethyl acetate = 4 / 1 to 1 / 1) to give the product H14-2 (2.25 g, 15.9 mmol, 74.2% yield) as yellow oil.

[0971] 1 H NMR: EC17015-223-P1A (400 MHz CDCl3) δ ppm 3.71 (s, 3H), 3.47 (d, J = 2.0 Hz, 2H), 3.31 (s, 2H), 2.39 (s, 3H), 2.20-2.28 (m, 1H).

[0972] 2. Synthesis of compound H14-3

[0973] To a solution of compound 6A (200 mg, 840 pmol, 1.00 equiv), compound H14-2 (178 mg, 1.26 mmol, 1.50 equiv), Pd(PPh3)2Cl2 (59.0 mg, 84.0 pmol, 0.10 equiv) and Cul (8.00 mg, 42.0 pmol, 0.05 equiv) in TEA (2.00 mL) was stirred at 75 °C for 2 h under N2. LCMS (EC17015-226-P1A, P1: RT = 0.341 min) detection showed that the target product was generated with 49.3% ratio. After reaction, the mixture was concentrated under reduced pressure to give the product crude, which was further purified by column chromatography (Si02, Petroleum ether / Ethyl acetate = 7 / 3 to 1 / 1) to give compound H14-3 product (170 mg, 677 pmol, 80.5% yield) as yellow oil.

[0974] LC-MS: EC17015-226-P1A, product: RT = 0.341 min, m / z = 250.0 [M+H] +

[0975] 1H NMR: EC17015-226-P1A (400 MHz CDC13) δ ppm 7.08 - 7.19 (m, 2 H), 6.89 - 6.97 (m, 1 H), 3.75 (s, 3 H), 3.67 (s, 2 H), 3.40 (s, 2 H), 2.48 (s, 3 H).

[0976] 3. Synthesis of compound H14-4

[0977] To a solution of compound Q4-3 (200 mg, 329 pmol, 1.00 equiv), compound H14-3 (91.0 mg, 362 pmol, 1.10 equiv) and Cs2C03(322 mg, 988 pmol, 3.00 equiv) in DMA (4.00 mL) was stirred at 25 °C for 4 h under N2. LCMS (EC17015-229-P1A, P1: RT = 0.469 min) detection showed that the target product was generated with 48.0% yield. The reaction mixture was extracted with water (20.0 mL) and ethyl acetate (15.0 mL*3), the organic layer was washed with saturated brine (10.0 mL), dried over Na2S04, filtered and concentrated under reduced pressure to give the product crude, which was further purified by column chromatography (Si02, petroleum ether / ethyl acetate = 3 / 2 to 1 / 1) to give compound H14-4 (228 mg, 293 pmol, 89.0% yield) as a yellow oil.

[0978] LC-MS: EC17015-229-P1A, product: RT = 0.469 min, m / z = 778.3 [M+H] +

[0979] 1 H NMR: EC17015-229-P1A (400 MHz DMSO) δ ppm 12.84 (s, 1 H), 8.03 (d, J = 7.8 Hz, 1 H), 7.78 (d, J = 8.0 Hz, 1 H), 7.55 (d, J = 7.2 Hz, 1 H), 7.43 - 7.52 (m, 2 H), 7.27 - 7.42 (m, 4 H), 7.18 (d, J = 8.2 Hz, 1 H), 7.05 - 7.13 (m, 1 H), 6.91 (d, J = 8.8 Hz, 1 H), 4.93 (s, 2 H), 4.02 (t, J = 6.07 Hz, 2 H), 3.78 (t, J = 6.0 Hz, 2 H), 3.56 - 3.64 (m, 5 H), 3.33 (s, 4 H), 2.96 - 3.00 (m, 2 H), 2.66 (t, J = 7.6 Hz, 2 H), 2.33 (s, 3 H), 1.29 (s, 9 H).

[0980] 4. Synthesis of compound H14-5

[0981] To a solution of compound H14-4 (228 mg, 293 mΐ, 1.00 equiv) in methanol (3.00 mL) and H20 (1.00 mL) was added LiOH.H20 (36.9 mg, 880 mΐ, 3.00 equiv), and the mixture was stirred at 25 °C for 0.5 h. LCMS (EC17015-232-P1W, P1: RT = 0.464 min) detection showed that the desired product was generated with 68.5% yield. The reaction mixture was adjusted to pH 3 with 1 M hydrochloric acid, diluted with water (10.0 mL), extracted with DCM and methanol solution (DCM:methanol = 10:1, 15.0 mL*3), and the organic layer was dried over Na2S04, filtered and concentrated under reduced pressure to give H14-5 product crude (200 mg, 262 mΐ, 89.3% yield) as a yellow solid, which was used directly in the next step without purification.

[0982] LC-MS: EC17015-232-P1W, product: RT = 0.464 min, m / z = 764.2 [M+H] +

[0983] 5. Synthesis of compound H14-6

[0984] Compound H14-5 (120 mg, 157 mΐ, 1.00 equiv), compound 2 (69.8 mg, 157 mΐ, 1.00 equiv), PyBOP (98.1 mg, 188 mΐ, 1.20 equiv) and TEA (47.7 mg, 471 mΐ, 65.6 pL, 3.00 equiv) were mixed in DMA (2.00 mL), and the mixture was stirred at 25 °C for 1 h. LCMS (EC17015-242-P1C, P1: RT = 0.859 min) detection showed that the desired product was generated with 92.3% yield. The reaction mixture was diluted with water (20.0 mL), extracted with DCM (15.0 mL*3), and the organic layer was washed with saturated brine (10.0 mL), dried over Na2S04, filtered and concentrated under reduced pressure to give H14-6 product crude (160 mg, crude product) as a yellow oil, which was used directly in the next step without purification.

[0985] LC-MS: EC17015-242-P1C, product: RT = 0.859 min, m / z = 1190.4 [M+H] +

[0986] 6. Synthetic compound APH14

[0987] To compound H14-6 (150 mg, 126 pmol, 1.00 equiv) was added HC1 / dioxane (5.00 mL), the mixture was stirred at 25 °C for 6 h. LCMS (EC17015-250-P1B2, P1: RT = 0.607 min) detected the desired product with 35.2% yield. The reaction mixture was concentrated under reduced pressure to give the product crude, which was further purified by prep-HPLC (column: CD02-Waters Xbidge BEH C18 150*25*10 pm; mobile phase: [water(NH4HCO3)-ACN]; gradient: 36%-66% B in 10 min) to give compound APH14 (20.0 mg, 17.6 pmol, 14.0% yield, 96.1% purity) as a white solid.

[0988] LC-MS: EC17015-250-P1B2, product: RT = 0.607 min, m / z = 1134.4 [M+H] +

[0989] 1 H NMR: EC21184-4-P1A (400 MHz DMSO) d ppm 8.93-9.02 (m, 1H), 8.44 (d, J = 7.6 Hz, 1H), 8.03 (d, J = 8.0 Hz, 1H), 7.76-7.81 (m, 1H), 7.69 (d, J = 9.6 Hz, 1H), 7.60 (d, J = 7.6 Hz, 1H), 7.51-7.55 (m, 1H), 7.27-7.50 (m, 9H), 7.18-7.23 (m, 1H), 7.08-7.15 (m, 1H), 6.89-6.96 (m, 1H), 4.85-4.98 (m, 3H), 4.52 (d, J = 9.6 Hz, 1H), 4.39-4.48 (m, 1H), 4.29 (d, J = 2.8 Hz, 1H), 4.02 (t, J = 6.4 Hz, 2H), 3.85 (t, J = 5.8 Hz, 2H), 3.54-3.65 (m, 4H), 3.11 (d, J = 2.8 Hz, 2H), 2.94-3.03 (m, 2H), 2.75-2.85 (m, 2H), 2.41-2.46 (m, 5H), 2.34 (s, 2H), 1.92-2.09 (m, 4H), 1.70-1.83 (m, 1H), 1.37 (d, J = 6.8 Hz, 3H), 1.23 (s, 1H), 0.94 (s, 9H).

[0990] Synthesis of APH15:

[0991] The synthesis of PROTAC molecule APH15 followed the procedure of APH14, but with the starting material changed to compound H15-1 and 4A. The desired product APH15 (20.0 mg, 16.3 pmol, 6.55% yield, 93.6% purity) was obtained as a white solid after purification.

[0992] 1 H NMR: EC16331-167-P1L (400 MHz DMSO) d ppm 8.98 (s, 1H), 8.38 (d, J = 7.6 Hz, 1H), 8.16 (d, J = 9.2 Hz, 1H), 8.03 (d, J = 8.0 Hz, 1H), 7.79 (d, J = 8.0 Hz, 1H), 7.60 (d, J = 7.2 Hz, 1H), 7.53 (d, J = 8.8 Hz, 1H), 7.46-7.50 (m, 1H), 7.44 (s, 1H), 7.42 (s, 2H), 7.37-7.40 (m, 2H), 7.35-7.37 (m, 2H), 7.34 (s, 1H), 7.26 (d, J = 12.0 Hz, 1H), 7.17 (d, J = 8.8 Hz, 1H), 7.07-7.13 (m, 1H), 6.93 (d, J = 8.8 Hz, 1H), 4.92 (s, 3H), 4.50 (d, J = 9.2 Hz, 1H), 4.42 (t, J = 8.0 Hz, 1H), 4.28 (s, 1H), 4.02 (t, J = 6.0 Hz, 2H), 3.86 (t, J = 5.6 Hz, 2H), 3.60 (s, 2H), 3.53 (s, 3H), 2.79 (t, J = 7.2 Hz, 2H), 2.62 (dd, J = 16.8, 6.8 Hz, 2H), 2.45 (s, 4H), 2.31 (s, 1H), 2.26 (s, 3H), 1.93-2.01 (m, 3H), 1.77-1.82 (m, 1H), 1.37 (d, J = 6.8 Hz, 3H), 0.94 (s, 12H).

[0993] Synthesis of APH16:

[0994] The synthesis of PROTAC molecule APH16 followed the procedure of APH14, but with the starting material changed to compound H16-1 and 4A. The desired product APH16 (20.0 mg, 16.0 pmol, 6.49% yield, 92.9% purity) was obtained as a white solid after purification.

[0995] 1H NMR: EC16331-166-P1M (400 MHz DMSO) δ ppm 8.96-8.99 (m, 1 H), 8.37 (d, J = 7.6 Hz, 1 H), 8.04 (d, J = 7.6 Hz, 1 H), 7.77-7.84 (m, 2 H), 7.60 (d, J = 7.6 Hz, 1 H), 7.54 (d, J = 8.8 Hz, 1 H), 7.46-7.50 (m, 1 H), 7.44 (s, 1 H), 7.33-7.43 (m, 6 H), 7.23-7.29 (m, 1 H), 7.15-7.19 (m, 1 H), 7.06-7.12 (m, 1 H), 6.95 (d, J = 8.8 Hz, 1 H), 5.04-5.16 (m, 1 H), 4.88-4.94 (m, 3 H), 4.51 (d, J = 9.2 Hz, 1 H), 4.42 (t, J = 8.0 Hz, 1 H), 4.27 (s, 1 H), 4.02 (t, J = 6.4 Hz, 2 H), 3.86 (t, J = 5.6 Hz, 2 H), 3.60 (s, 2 H), 3.48 (s, 3 H), 2.98 (t, J = 5.2 Hz, 2 H), 2.80 (t, J = 7.2 Hz, 2 H), 2.42-2.47 (m, 3 H), 2.36 (t, J = 7.2 Hz, 2 H), 2.22 (s, 3 H), 2.11-2.19 (m, 1 H), 1.92-2.04 (m, 3 H), 1.79 (tt, J = 8.4, 4.0 Hz, 1 H), 1.59-1.68 (m, 2 H), 1.37 (d, J = 6.8 Hz, 3 H), 1.16-1.27 (m, 1 H), 0.93 (s, 9 H).

[0996] e) Synthesis of APH17, APH18, APH19, APH20, APH21

[0997] Synthesis of APH17:

[0998] 1. Synthesis of compound H17-2

[0999] Compound 6A (300 mg, 1.26 mmol, 1.00 equiv), compound H17-1 (212 mg, 1.89 mmol, 1.50 equiv), Pd(PPh3)2Cl2 (88.5 mg, 126 µmol, 0.10 equiv) and Cul (12.0 mg, 63.0 µmol, 0.05 equiv) were mixed in TEA (5.00 mL) and stirred at 75 °C for 2 h under N2. LCMS (EC17015-233-P1A, P1: RT = 0.355 min) detected the desired product with 62.9% yield. The reaction mixture was filtered and concentrated under reduced pressure to give the product as a crude, which was further purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 7 / 3 to 1 / 1) to give H17-2 product (220 mg, 990 µmol, 78.5% yield) as a yellow oil.

[1000] LC-MS: EC17015-233-P1A, product: RT = 0.355 min, m / z = 223.0 [M+H] +

[1001] 1 H NMR: EC17015-233-P1A (400 MHz DMSO) δ ppm 10.21 (s, 1H), 7.13 (dd, J = 12.0, 1.8 Hz, 1H), 7.01 (dd, J = 8.4, 1.0 Hz, 1H), 6.84-6.93 (m, 1H), 3.62 (s, 3H), 2.60 (dd, J = 9.6, 5.2 Hz, 4H).

[1002] 2. Synthesis of compound H17-3

[1003] A mixture of compound Q4-3 (200 mg, 329 pmol, 1.00 equiv), compound H17-2 (80.5 mg, 362 pmol, 1.10 equiv) and Cs2CO3 (322 mg, 988 pmol, 3.00 equiv) in DMA (3.00 mL) was stirred at 25 °C for 12 h. LCMS (EC17015-247-P1A, P1: RT = 0.517 min) detection showed that the desired product was generated with 68.2% yield. The reaction mixture was diluted with water (20.0 mL) and extracted with ethyl acetate (15.0 mL*3). The organic layer was washed with saturated brine (20.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product, which was further purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 7 / 3 to 1 / 1) to give H17-3 product (155 mg, 207 pmol, 62.9% yield) as a yellow solid.

[1004] LC-MS: EC17015-247-P1A, product: RT = 0.517 min, m / z = 749.2 [M+H] +

[1005] 1 H NMR: EC17015-247-P1A (400 MHz DMSO) d ppm 12.82 (s, 1H), 8.03 (d, J = 7.8 Hz, 1H), 7.78 (d, J = 8.0 Hz, 1H), 7.55 (d, J = 6.8 Hz, 1H), 7.44-7.51 (m, 2H), 7.31-7.31 (m, 1H), 7.31-7.41 (m, 2H), 7.19 (dd, J = 11.8, 1.6 Hz, 1H), 7.04-7.13 (m, 2H), 6.91 (d, J = 8.8 Hz, 1H), 4.93 (s, 2H), 3.96-4.03 (m, 2H), 3.78 (t, J = 6.0 Hz, 2H), 3.62 (s, 3H), 2.98 (t, J = 5.8 Hz, 2H), 2.60-2.66 (m, 4H), 2.54 (s, 2H), 1.86-1.96 (m, 2H), 1.30 (s, 9H).

[1006] 3. Synthesis of compound H17-4

[1007] To a solution of compound H17-3 (155 mg, 207 pmol, 1.00 equiv) in methanol (1.20 mL) and water (0.40 mL) was added LiOH.H2O (26.1 mg, 621 pmol, 3.00 equiv), and the mixture was stirred at 25 °C for 4 h. LCMS (EC17015-252-P1A2, P1 : RT = 0.483 min) showed that the desired product was generated with 92.2% purity. The reaction mixture was adjusted to pH 3 with 1 M hydrochloric acid, diluted with water (10.0 mL), extracted with DCM and methanol solution (DCM:methanol = 10:1, 15.0 mL*3), and the organic layer was dried over Na2S04, filtered and concentrated under reduced pressure to give compound H17-4 product (150 mg, crude) as a yellow solid.

[1008] LC-MS: EC17015-252-P1A2, product: RT = 0.483 min, m / z = 735.1 [M+H] +

[1009] 4. Synthesis of compound H17-5

[1010] Compound H17-4 (150 mg, 204 pmol, 1.00 equiv), compound 2 (90.8 mg, 204 pmol, 1.00 equiv), PyBOP (127 mg, 245 pmol, 1.20 equiv) and TEA (62.0 mg, 612 pmol, 85.2 pL, 3.00 equiv) were mixed in DMA (2.00 mL) and stirred at 25 °C for 1 h. LCMS (EC17015-256-P1A, P1 : RT = 0.548 min) showed that the desired product was generated with 81.8% purity. The reaction mixture was diluted with water (20.0 mL), extracted with DCM (15.0 mL*3), and the organic layer was washed with saturated brine (10.0 mL), dried over Na2S04, filtered and concentrated under reduced pressure to give compound H17-5 product crude (200 mg) as a yellow oil. The crude product was used directly in the next reaction without purification.

[1011] LC-MS: EC17015-256-P1A, product: RT = 0.548 min, m / z = 1161.4 [M+H] +

[1012] 5. Synthesis of compound APH17

[1013] Compound H17-5 (190 mg, 164 pmol, 1.00 equiv) was dissolved in HC1 / dioxane (4.00 mL), and the mixture was stirred at 25 °C for 6 h. LCMS (EC17015-261-P1A1, P1: RT = 0.597 min) detected the desired product with 30.0% purity. The reaction mixture was filtered and concentrated under reduced pressure to give the product crude, which was further purified by prep-HPLC (column: CD02-Waters Xbidge BEH C18 150*25*10 pm; mobile phase: [water (NH4HCO3)-ACN]; gradient: 28%-58% B in 13 min) to give product APH17 (24.0 mg, 21.2 pmol, 12.9% yield, 97.5% purity) as a white solid.

[1014] LC-MS: EC17015-261-P1A1, product: RT = 0.597 min, m / z = 1105.3 [M+H] +

[1015] 1 H NMR: EC17015-261-P1A (400 MHz DMSO) d ppm 8.98 (s, 1H), 8.38 (d, J = 7.8 Hz, 1H), 8.01 (dd, J = 19.2, 8.2 Hz, 2H), 7.79 (d, J = 7.8 Hz, 1H), 7.60 (d, J = 7.8 Hz, 1H), 7.53 (d, J = 8.4 Hz, 1H), 7.47 (s, 1H), 7.32 - 7.45 (m, 7H), 7.17 (d, J = 12.4 Hz, 1H), 7.06 - 7.11 (m, 1H), 6.93 (d, J = 8.6 Hz, 1H), 5.13 (s, 1H), 4.92 (s, 3H), 4.56 (d, J = 9.2 Hz, 1H), 4.42 (t, J = 8.2 Hz, 1H), 4.28 (s, 1H), 4.01 (t, J = 5.6 Hz, 2H), 3.85 (s, 2H), 3.54 - 3.69 (m, 3H), 2.98 (d, J = 5.8 Hz, 2H), 2.78 (t, J = 7.0 Hz, 2H), 2.58 (s, 3H), 2.45 (s, 3H), 2.35 - 2.42 (m, 1H), 1.89 - 2.08 (m, 3H), 1.70 - 1.85 (m, 1H) 1.37 (d, J = 6.8 Hz, 3H), 0.94 (s, 9H).

[1016] Synthesis of APH18:

[1017] The synthesis of PROTAC molecule APH18 was referred to APH17, but the starting material was replaced by compound H18-1 and 6A. After synthesis, the target product APH18 (32.7 mg, 27.5 μmol, 18.6% yield, 93.9% purity) was obtained by purification, which was a white solid.

[1018] LC-MS: EC21184-23-p1aa, P1: RT = 0.602 min, m / z = 1119.5 [M+H] +

[1019] 1 H NMR: EC21184-23-P1 QC (400 MHz, DMSO) δ ppm 8.98 (s, 1H), 8.38 (d, J = 7.6 Hz, 1H), 8.03 (d, J = 7.6 Hz, 1H), 7.91 (d, J = 9.2 Hz, 1H), 7.79 (d, J = 7.6 Hz, 1H), 7.60 (d, J = 7.2 Hz, 1H), 7.31-7.54 (m, 9H), 7.25 (dd, J = 12.0, 1.6 Hz, 1H), 7.12-7.17 (m, 1H), 7.04-7.11 (m, 1H), 6.91 (d, J = 8.8 Hz, 1H), 5.00-5.23 (m, 1H), 4.92 (s, 2H), 4.53 (d, J = 9.2 Hz, 1H), 4.42 (t, J = 8.00 Hz, 1H), 4.28 (d, J = 1.6 Hz, 1H), 4.01 (t, J = 6.4 Hz, 2H), 3.85 (t, J = 6.0 Hz, 2H), 3.61 (s, 2H), 2.94-3.04 (m, 3H), 2.78 (t, J = 7.6 Hz, 2H), 2.45 (s, 3H), 2.34-2.43 (m, 3H), 2.24-2.34 (m, 1H), 1.91-2.05 (m, 3H), 1.65-1.85 (m, 4H), 1.37 (d, J = 6.8 Hz, 3H), 0.91-0.98 (m, 9H).

[1020] Synthesis of APH19:

[1021] The synthesis of PROTAC molecule APH19 was referred to APH17, but the starting material was replaced by compound H19-1 and 6A. After synthesis, the target product APH19 (18.0 mg, 15.6 μmol, 10.0% yield, 98.5% purity) was obtained by purification, which was a white solid.

[1022] LC-MS: EC21184-25-p1b, Product: RT = 0.612 min, m / z = 1133.5 [M+H] +

[1023] 1 H NMR: EC21184-25-p1qc (400 MHz DMSO) δ ppm 8.94-9.02 (m, 1 H), 8.37 (d, J = 7.6 Hz, 1 H), 8.03 (d, J = 7.6 Hz, 1 H), 7.81 (dd, J = 17.2, 8.8 Hz, 2 H), 7.60 (d, J = 7.2 Hz, 1 H), 7.29-7.56 (m, 10 H), 7.20 (d, J = 12.0 Hz, 1 H), 7.09 (dt, J = 16.4, 8.4 Hz, 2 H), 6.8 (d, J = 8.8 Hz, 1), 5.02-5.22 (m, 1 H), 4.87-4.98 (m, 3 H), 4.52 (d, J = 9.2 Hz, 1 H), 4.42 (t, J = 8.0 Hz, 1 H), 4.28 (s, 1 H), 4.00 (t, J = 6.0 Hz, 2 H), 3.85 (s, 2 H), 3.60 (s, 2 H), 2.97 (s, 2 H), 2.79 (t, J = 6.8 Hz, 2 H), 2.42-2.47 (m, 3 H), 2.39 (t, J = 6.4 Hz, 2 H), 2.25-2.34 (m, 1 H), 2.11-2.21 (m, 1 H), 1.91-2.05 (m, 3 H), 1.74-1.84 (m, 1 H), 1.57-1.69 (m, 2 H), 1.45-1.54 (m, 2 H), 1.37 (d, J = 6.8 Hz, 3 H), 0.94 (s, 9 H).

[1024] Synthesis of APH20:

[1025] The synthesis of PROTAC molecule APH20 was referred to APH17, but the starting material was replaced by compound H20-1 and 6A. After purification, the target product APH20 (18.0 mg, 14.9 μmol, 5.96% yield, 94.7% purity) was obtained as a white solid.

[1026] LC-MS: EC17015-336-P1B, Product: RT = 0.629 min, m / z = 1147.6 [M+H] +

[1027] 1H NMR: EC17015-336-P1A (400 MHz DMSO) δ ppm 8.98 (s, 1H), 8.37 (d, J = 7.6 Hz, 1H), 8.02 (d, J = 7.6 Hz, 1H), 7.80 (dd, J = 12.8, 8.8 Hz, 2H), 7.60 (d, J = 6.8 Hz, 1H), 7.45-7.51 (m, 2H), 7.31-7.45 (m, 7H), 7.20 (dd, J = 12.0, 1.8...

Claims

1. A compound of formula I, or a pharmaceutical salt, ester, prodrug, solvate, stereoisomer, or deuterated compound thereof: ALE (I) Part A is the small molecule ligand portion targeting the Bcl-xL protein, part E is the small molecule ligand portion targeting the E3 ubiquitin ligase complex, and part L is the linker group. Part A has the structure of formula 1 or 2: R0 either does not exist or is selected from: -COOR a -CONR a R b -SO3R a -SO2NR a R b -SO2NR a COR b -CONR a SO2R b -COSO2NR a R b , X is selected from hydrogen, hydroxyl, amino, or CH(OR) a )2; R a and R b Independently selected from hydrogen, cyano, substituted or unsubstituted alkyl or substituted or unsubstituted alkoxy; Y is absent or selected from substituted or unsubstituted alkylene, substituted or unsubstituted alkyleneoxy, substituted or unsubstituted alkenyl, substituted or unsubstituted alkenyloxy, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted heterocyclic. Z1 and Z2 are either absent or selected from substituted or unsubstituted alkylene-cyclic groups, substituted or unsubstituted cycloalkyl groups, substituted or unsubstituted heterocyclic groups, substituted or unsubstituted aryl groups, and substituted or unsubstituted heteroaryl groups. Part E is selected from the following structure: Where R1 is O, NH, or absent; R2 is H or methyl; R3 is... R c It consists of F, methoxy, Cl, and cyano groups; Part L has the following structure: Part L connects to part A at L1 and to part E at L2. L1 and L2 do not exist independently or are selected from: X either does not exist or has the following structure: X1 and X2 either do not exist independently or are selected from: W does not exist or is selected from: in, R4 is independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, and substituted or unsubstituted heterocyclic groups; R5 is independently selected from hydrogen, nitro, cyano, isocyano, amino, hydroxyl, thiol, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, alkylamino, dialkylamino, carboxyl, carbonyl, amide, sulfonyl, sulfonic acid, phosphoryl, and phosphonyl. q and r are independently selected from integers from 0 to 10; m1, m2, n1, n2, s1, and s2 are independently selected from integers from 0 to 10.

2. The compound of formula I according to claim 1, wherein part A is a deuterated part having the structure of formula 1' or 2': One or more hydrogen atoms at position * are replaced by deuterium. Preferably, the two hydrogen atoms at position * are replaced with deuterium.

3. The compound of formula I according to claim 1 or 2, wherein R a and R b It is independently selected from hydrogen, cyano, alkyl, alkoxy, halogen-substituted alkyl or halogen-substituted alkoxy.

4. The compound of formula I according to claim 1 or 2, wherein R0 is absent or selected from -COOR. a -CONHR b -SO3R a -SO2NHR b -SO2NHCOR b -CONHSO2R b -COSO2NHR b , where R a and R b Independently selected from hydrogen, cyano, and alkyl groups. Preferably, R0 is selected from:

5. The compound of formula I according to claim 1, wherein Y is absent or selected from alkylene, alkyleneoxy, alkenyl, alkenyloxy, aryl, heteroaryl, cycloalkylene, and heterocyclic groups optionally substituted with one or more substituents selected from halogens and alkyl groups.

6. The compound of formula I according to claim 1, wherein Y is selected from alkylene, alkyleneoxy, alkenyl, or alkenyloxy. Preferably, Y is selected from Where m is 0, 1, 2 or 3.

7. The compound of formula I according to claim 1, wherein Y is selected from arylene groups and 5- or 6-membered heteroarylene groups optionally substituted with one or more substituents selected from halogens and alkyl groups. Preferably, Y is selected from a phenylene or pyrazolene group optionally substituted with one or more substituents selected from halogens and alkyl groups. More preferably, Y is selected from It is connected to Z1 or Z2 at the nitrogen atom of the ring.

8. The compound of formula I according to claim 1, wherein Y is selected from the following structures: It is connected to Z1 or Z2 at the nitrogen atom in the ring.

9. The compound of formula I according to claim 1, wherein Z1 and Z2 are independently selected from those optionally selected by one or more R 1’ Substituted alkylene-cyclic groups, cycloalkyl-(R2') p Heterocyclic group-(R2') p aryl-(R2') p heteroaryl-(R2') p aryl-propynyl, heteroaryl-propynyl, aryl-propynyl-N(R2')2, heteroaryl-propynyl-N(R2')2, aryl-propynyl-cycloalkyl-(R2') p Heteroaryl-propynyl-cycloalkyl-(R2') p aryl-propynyl-heterocyclic-(R2') p and heteroaryl-propynyl-heterocyclic-(R2') p , Where R 1’ Selected from deuterium, halogen, amino, hydroxyl, nitro, thiol, cyano, isocyanate, substituted or unsubstituted alkyl (e.g., alkyl substituted with halogen, amino, hydroxyl, nitro and / or carboxyl), substituted or unsubstituted alkoxy (e.g., alkoxy substituted with halogen, amino, hydroxyl, nitro and / or carboxyl), alkylamino, dialkylamino, carboxyl, carbonyl, amide, sulfonyl, sulfonic acid, phosphoryl and phosphonyl. R2' is hydrogen, alkyl, aryl, heteroaryl, cycloalkyl, or heterocyclic, and p can be 0, 1, 2, 3, or 4.

10. The compound of formula I according to claim 9, wherein for the Z1 and Z2 groups, the cyclic group is The cycloalkyl group is cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl; The heterocyclic group is a 5- or 6-membered heterocyclic group containing one or two heteroatoms independently selected from nitrogen, oxygen and sulfur, preferably piperidinyl, piperazine, morpholinyl or 3,9-diazaspiro[5.5]undecyl; The aryl group is phenyl; and / or The heteroaryl group is a 5- or 6-membered heteroaryl group containing one or two heteroatoms independently selected from nitrogen, oxygen and sulfur, preferably a pyridyl group.

11. The compound of formula I according to claim 9, wherein Z1 is selected from the following groups: Preferably, Z1 is selected from the following groups:

12. The compound of formula I according to claim 9, wherein Z2 is selected from the following groups: Preferably, Z2 is selected from the following groups:

13. The compound of formula I according to claim 1 or 2, wherein the A portion is selected from the following structures:

14. The compound of formula I according to claim 1, wherein the E portion is selected from the following structures:

15. The compound of formula I according to claim 1, wherein L1 is absent or selected from...

16. The compound of formula I according to claim 1, wherein L2 is absent or selected from...

17. The compound of formula I according to claim 1, wherein the X group is absent or selected from: u1, u2, u3, and v1 are independently selected from integers from 0 to 10.

18. The compound of formula I according to claim 1, wherein the L portion is absent or selected from the following structures: q, u1, u2, u3 and v1 are independently selected from integers from 0 to 10.

19. The compound of formula I according to claim 1, wherein the L portion is absent or selected from the following structures:

20. A composition comprising an effective amount of the compound of formula (I) according to any one of claims 1-19 and a pharmaceutical carrier.

21. A method for selectively killing one or more senescent cells or treating an aging-related disease or condition in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1-19 or a composition according to claim 20.

22. Use of the compound according to any one of claims 1-19 or the composition according to claim 20 in the preparation of a medicament for selectively killing one or more senescent cells in a subject or for treating aging-related diseases or conditions.

23. The method or use according to claim 21 or 22, wherein the age-related disease or condition is a metabolic disease, inflammatory disease or condition, lung disease or condition, neurological disease or condition, proliferative disease, kidney disease or condition, eye disease or condition, or dermatological disease or condition.

24. The method or use according to claim 21 or 22, wherein the age-related disease or condition is selected from: (i) Selected from inflammatory or autoimmune diseases or conditions such as oral mucositis, inflammatory bowel disease, kyphosis and intervertebral disc herniation; (ii) Selected from neurological diseases or conditions such as Alzheimer's disease, Parkinson's disease, Huntington's disease, dementia, mild cognitive impairment, macular degeneration, and motor neuron dysfunction; (iii) Metabolic diseases selected from diabetic ulcers, metabolic syndrome, and obesity; (iv) Selected from pulmonary fibrosis, chronic obstructive pulmonary disease, asthma, cystic fibrosis, emphysema, bronchiectasis, and age-related loss of lung function. (v) Selected from eye diseases or conditions such as macular degeneration, glaucoma, cataracts, presbyopia, and vision loss; (vi) Selected age-related conditions including renal failure, weakness, hearing loss, muscle fatigue, skin condition, skin wound healing, liver fibrosis, pancreatic fibrosis, oral submucosal fibrosis, and sarcopenia; and (vii) Selected from skin diseases or conditions such as eczema, psoriasis, hyperpigmentation, moles, rashes, atopic dermatitis, urticaria, diseases and conditions related to photosensitivity or photoaging, wrinkles, pruritus, hypoesthesia, eczematous rashes, eosinophilic dermatitis, reactive neutrophilic dermatitis, pemphigus, bullous pemphigoid, immune bullous dermatitis, dermatofibrosis, cutaneous lymphoma, and cutaneous lupus.

25. The method or use according to claim 21 or 22, wherein the subject is diagnosed with cancer and optionally is undergoing cancer treatment selected from chemotherapy and radiation therapy.

26. The method or use according to claim 21 or 22, wherein the compound or composition is described. (i) Reduce cells or cancer cells that have been pushed toward aging; (ii) Reduce one or more side effects produced by senescent cells, wherein said side effects include inflammation, promotion of cancer growth and promotion of metastasis; (iii) Reduce one or more side effects of chemotherapy; or (iv) Reduce one or more side effects of radiation therapy.

27. The method or use according to claim 21 or 22, wherein the subject has a viral infection, optionally, wherein the viral infection results in an overproduction of activated macrophages in the subject.

28. The method or use according to claim 27, wherein the viral infection is a coronavirus infection selected from coronaviruses, severe acute respiratory syndrome (SARS) coronavirus (CoV), SARS-CoV-2 (which causes COVID-19 (coronavirus disease) 2019) and Middle East respiratory syndrome (MERS)-CoV.

29. The method or use according to claim 27, wherein the subject has SARS-CoV-2 infection and has been diagnosed with coronavirus disease 2019.

30. The method of claim 21, wherein the compound or composition, after in vivo administration, yields the following structure in an effective amount to kill one or more senescent cells: R0, Y, and Z1 are as described in any one of claims 1-19.

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