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

PROTAC targeting Bcl-xL protein and its application Technical Field

[0001] The present invention relates to PROTAC targeting Bcl-xL protein and its application. Background Art

[0002] BCL-2 (B-cell lymphoma 2) gene family members play a central role in regulating programmed cell death by controlling pro-apoptotic and anti-apoptotic intracellular signals (Czabotar, PE 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, especially BCL-xL protein is specifically highly expressed in a variety of senescent cells (He et al., Nat Commun 11, 1996 (2020)). At present, 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 blood toxicity, 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. The PROTAC molecule consists of three parts: a target protein (POI) ligand, a linker, and an E3 ubiquitin ligase ligand. PROTAC is a heterogeneous bifunctional chimeric molecule, one end of which targets the POI and the other end recruits the E3 ubiquitin ligase to form a dumbbell-shaped ternary complex. The PROTAC molecule mediates a transient interaction between the E3 ligase and the POI. The E3 ubiquitin ligase attaches ubiquitin to the POI, marking it as a defective or damaged protein. The proteasome then recognizes and degrades the marked POI. The main advantages of PROTAC technology include event-driven activity (rather than the occupancy-driven activity of conventional small molecule inhibitors), targeting undruggable proteins, overcoming drug resistance, and low dosage, which is expected to improve many limitations of traditional inhibitors. Since peptides were first used to verify the artificial induction of intracellular protein degradation, and the molecular weight of such peptides is too large, cell-penetrating peptides are needed to increase cell permeability. Therefore, PROTAC technology developed slowly during the 10 years from 2001 to 2011. With the continuous discovery of small molecule E3 ligase ligands, PROTAC technology has developed rapidly. For example, mouse double minute gene 2 protein (MDM2), cell inhibitor of apoptosis protein (cIAP), VHL (VHL) and CRBN (cereblon) have been successfully used in small molecule PROTACs, especially ARV-110 and ARV-471 based on CRBN have been used in clinical studies. However, because PROTAC molecules are usually large in molecular weight and do not meet the "five rules of drug-like properties", resulting in poor drugability, drug development based on this technology usually has poor pharmacokinetic parameters in vivo, and there are challenges of low cell permeability and oral bioavailability. It is necessary to try to screen different E3 ligase ligands and carefully optimize 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 flexibility of the linker group, and the connection site are important for the formation of the active ternary complex, degradation activity and target selectivity of PROTAC. The design of the linker group can further enhance the efficacy by reducing the free energy or restricting the biologically active conformation. Optimizing the structure-activity relationship (SAR) within the linker group will provide an opportunity to adjust the pharmacokinetic properties of protein degraders. However, there is no universally applicable design strategy for the design and optimization of linkers. Often, it starts with a short and simple alkane or polyethylene glycol (PEG) chain, and its related properties are adjusted through continuous iterative trial and error.In summary, PROTACs are a powerful approach to overcoming current drug development barriers, but further research is needed to design and optimize molecules, gradually identify patterns to improve physicochemical properties, and further explore the clinical efficacy and safety of PROTACs. PROTACs offer a broad range of targets and a vast market. With the continued advancement and refinement of this technology, PROTACs could become a successful therapy like small molecule inhibitors, monoclonal antibodies, and immunotherapy, benefiting a wider range of patients.

[0004] Aging is a major risk factor for physiological degeneration, increased incidence of chronic diseases and age-related mortality (Lopez-Otin et al., Cell 153, 1194-1217 (2013)). During the aging process, 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 proinflammatory 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)). Eliminating senescent cells has become an attractive potential approach to improve 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, called "senolytics," have attracted considerable interest. BCL-xL inhibitors can effectively kill senescent cells as anti-aging drugs (Zhu et al., Aging Cell 15, 428-435 (2016)). Bcl-xL inhibitors induce apoptosis in senescent cells by removing the key anti-apoptotic mechanism that maintains the survival of senescent cells, thereby achieving the clearance of senescent cells in vitro and in vivo. They have shown great potential in the treatment of various aging-related diseases such as osteoarthritis, age-related macular degeneration, and diabetic nephropathy.

[0005] By structurally modifying BCL-xL protein inhibitors and combining them with E3 ligase ligands to form PROTACs, they effectively combine the advantages of both. On the one hand, they can specifically induce apoptosis in senescent cells, and on the other hand, they can effectively reduce the hematological toxicity of BCL-xL protein inhibitors. Simultaneously, the overall molecular structure is optimized to ensure clinical efficacy and safety, providing new insights into the treatment of a range of geriatric and aging-related diseases.

[0006] Summary of the Invention

[0007] An object of the present invention is to provide a compound of formula I, or a pharmaceutically acceptable salt, ester, prodrug, solvate, stereoisomer or deuterated compound thereof: ALE (I)

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

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

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

[0011] Where R0 does not exist or is 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, hydroxy, amino or CH(OR a )2;

[0014] where 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] Z1 and Z2 are independently absent or selected from a substituted or unsubstituted alkylene-cyclic group, a substituted or unsubstituted cycloalkyl group, a substituted or unsubstituted heterocyclic group, a substituted or unsubstituted aryl group, or a substituted or unsubstituted heteroaryl group.

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

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

[0019] In some embodiments, R a and R b Independently selected from hydrogen, cyano, substituted or unsubstituted C1-C6 alkyl or substituted or unsubstituted C1-C6 alkoxy.

[0020] In some embodiments, R a and R b Independently selected from hydrogen, cyano, alkyl (eg C1-C6 alkyl), alkoxy (eg C1-C6 alkoxy), halogen-substituted alkyl (eg C1-C6 alkyl) or halogen-substituted alkoxy (eg C1-C6 alkoxy).

[0021] In some embodiments, 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 (eg, C1-C6 alkyl).

[0022] In some embodiments, R0 is selected from:

[0023] In some embodiments, Y is absent or selected from substituted or unsubstituted C1-6 alkylene, substituted or unsubstituted C1-6 alkyleneoxy, substituted or unsubstituted C2-6 alkenylene, substituted or unsubstituted C2-6 alkenyleneoxy, substituted or unsubstituted phenylene, substituted or unsubstituted 5-membered 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, alkyleneoxy, alkenylene, alkenyleneoxy, arylene, heteroarylene, cycloalkylene, and heterocyclylene, optionally substituted with one or more substituents selected from halogen and alkyl (e.g., C1-6 alkyl).

[0025] In some embodiments, Y is selected from C1-6 alkylene, C1-6 alkyleneoxy, C2-6 alkenylene, or C2-6 alkenyleneoxy.

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

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

[0028] In some embodiments, Y is selected from phenylene and 5-membered 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] The nitrogen atom of the ring is connected to the Z1 or Z2 group.

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

[0033] The nitrogen atom of the ring is connected to the Z1 or Z2 group.

[0034] In some embodiments, Z1 and Z2 are independently selected from substituted or unsubstituted -C1-6 alkylene-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, Z1 and Z2 are independently selected from 1’ Substituted-alkylene-cyclic group, cycloalkyl-(R2') p 、Heterocyclyl-(R2')p , aryl-(R2') p , heteroaryl-(R2') p , aryl-propynylene, heteroaryl-propynylene, aryl-propynylene-N(R2')2, heteroaryl-propynylene-N(R2')2, aryl-propynylene-cycloalkyl-(R2') p 、Heteroaryl-propynylene-cycloalkyl-(R2') p , aryl-propynylene-heterocyclyl-(R2') p and heteroaryl-propynylene-heterocyclyl-(R2') p .

[0036] In some embodiments, R 1’ is deuterium, halogen, amino, hydroxy, nitro, thiol, cyano, isocyano, substituted or unsubstituted alkyl (for example, alkyl substituted by halogen, amino, hydroxy, nitro, carboxyl, etc.), substituted or unsubstituted alkoxy (for example, alkoxy substituted by halogen, amino, hydroxy, nitro, carboxyl, etc.), alkylamino, dialkylamino, carboxyl, carbonyl, acylamino, sulfonyl, sulfonic acid, phosphoryl and phosphono.

[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, Z1 and Z2 are independently selected from 1’ Substituted C5-8 cycloalkyl-(R2') p 、Heterocyclyl-(R2') p , phenyl-(R2') p , heteroaryl-(R2') p , phenyl-propynyl, heteroaryl-propynyl, phenyl-propynyl-N(R2')2, heteroaryl-propynyl-N(R2')2, aryl-propynyl-cycloalkyl-(R2') p 、Heteroaryl-propynylene-cycloalkyl-(R2') p , aryl-propynylene-heterocyclyl-(R2') p and heteroaryl-propynylene-heterocyclyl-(R2') p, wherein the heterocyclyl is a 5-membered or 6-membered heterocyclyl (e.g., piperidinyl, piperazinyl, or morpholinyl) or a 7-11-membered heterospirocyclyl (e.g., 3,9-diazaspiro[5.5]undecyl) containing one or two heteroatoms, and the heteroaryl is a 5-membered or 6-membered heteroaryl (e.g., pyridinyl) containing one or two heteroatoms, 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 (eg fluorine).

[0043] In some embodiments, R 2’ is hydrogen or an alkyl group (eg, methyl).

[0044] In some embodiments, Z1 is selected from the following groups:

[0045] In some embodiments, Z1 can be selected from the following groups:

[0046] In some embodiments, Z1 can be selected from the following groups:

[0047] In some embodiments, Z1 is selected from the following groups:

[0048] In some embodiments, Z2 is selected from the following groups:

[0049] In some embodiments, Z2 is selected from:

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

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

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

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

[0054] Small molecule ligand targeting E3 ubiquitin ligase complex

[0055] In some embodiments, the small molecule ligand portion E that targets the 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 It is F, methoxy, Cl, and cyano.

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

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

[0059] Linking group L

[0060] The linking group L has the following structure:

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

[0062] in

[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, hydroxy, thiol, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, alkylamino, dialkylamino, carboxyl, carbonyl, amido, sulfonyl, sulfonic acid, phosphoryl and phosphono;

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

[0067] X does not exist or has the following structure:

[0068] X1 and X2 are independently absent or selected from:

[0069] W is absent or selected from:

[0070] wherein m1, m2, n1, n2, s1 and s2 are independently selected from integers of 0-10 (i.e., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10).

[0071] In some embodiments, R4 is independently selected from hydrogen, substituted or unsubstituted C1-6 alkyl, substituted or unsubstituted C3-8 cycloalkyl and substituted or unsubstituted 3-8 membered heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl may be substituted by halogen, amino, hydroxy, nitro and / or carboxyl.

[0072] In some embodiments, R5 is 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 phosphonyl.

[0073] In some embodiments, L1 is absent or selected from

[0074] In some embodiments, L2 is absent or selected from

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

[0076] wherein u1, u2, u3 and v1 are independently selected from integers of 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 v1 are independently selected from integers from 0 to 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 a structure of Formula 1, the linker L is selected from the following structures:

[0084] and

[0085] The small molecule ligand portion E that targets 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 a structure of Formula 2, the linker L is absent or selected from the following structures:

[0087] and

[0088] The small molecule ligand portion E that targets 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 present invention relates to a composition comprising an effective amount of a compound of formula (I) and a pharmaceutically acceptable carrier.

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

[0092] In one aspect, the present invention relates to the use of a compound of formula (I) of the present invention or a composition of the present invention in the preparation of a medicament for selectively killing one or more senescent cells or treating a senescence-related 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 renal disorder or disease, an ocular disease or disorder, or a dermatological disorder or disease.

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

[0095] (i) an inflammatory or autoimmune disease or condition selected from oral mucositis, inflammatory bowel disease, kyphosis, and herniated 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 lung 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 condition selected from macular degeneration, glaucoma, cataracts, presbyopia, and vision loss;

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

[0101] (vii) a skin disease or condition selected from eczema, psoriasis, hyperpigmentation, moles, rashes, atopic dermatitis, urticaria, diseases and conditions associated with photosensitivity or photoaging, wrinkles, pruritus, dysesthesia, eczematous rashes, eosinophilic dermatoses, reactive neutrophilic dermatoses, pemphigus, pemphigoid, immunobullous dermatoses, cutaneous fibrohistiocytosis, cutaneous lymphoma, and cutaneous lupus.

[0102] In some embodiments, the subject is diagnosed with 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 invention or the composition of the present invention

[0104] (i) reducing cells that have been pushed into senescence or cancer cells;

[0105] (ii) reducing one or more side effects of senescent cells, wherein the side effects include inflammation, promotion of cancer growth, and promotion of metastasis;

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

[0107] (iv) reducing 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 overproduction of activated macrophages in the subject.

[0109] In some embodiments, the viral infection is a coronavirus infection selected from the group consisting of coronavirus, 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, a compound or composition of the invention, upon in vivo administration, in an amount effective to kill one or more senescent cells, yields the following structure:

[0112] wherein R0, Y and Z1 are as described herein. BRIEF DESCRIPTION OF THE DRAWINGS

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

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

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

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

[0117] Figure 5 shows the platelet toxicity results of the test compounds administered in vivo. Figure 5A compares the effects of different doses of the PROTAC molecule APQ2 and the control molecule A-1331852 on peripheral blood platelet count in mice. APQ2-0.1 mpk, APQ2-0.5 mpk, and APQ-2.5 mpk represent APQ2 doses of 0.1, 0.5, and 2.5 mg / kg, respectively; A133-0.5 mpk represents the A-1331852 dose group at 0.5 mg / kg; and Veh represents the vehicle-only control group. Blood routine tests were performed at 0, 4, 24, and 72 hours after administration. Figure 5B compares the effects of PROTAC molecules APQ9 and APQ10, the control molecule A-1331852, and vehicle alone on peripheral blood platelet count in mice. Veh, APQ9-5 mpk, APQ10-5 mpk, and A133-5 mpk represented the vehicle-only group, APQ9 at 5 mg / kg, APQ10 at 5 mg / kg, and A-1331852 at 5 mg / kg, respectively. Blood routine tests were performed 4, 24, and 48 hours after administration. Figure 5C compares the effects of APQ10 and A-1331852 on platelet count at different doses. APQ10-2 mpk, APQ10-5 mpk, and APQ10-20 mpk represented APQ10 doses of 2, 5, and 20 mg / kg, respectively; A133-2 mpk, A133-5 mpk, and A133-20 mpk represented A-1331852 doses of 2, 5, and 20 mg / kg, respectively. Veh served as the vehicle-only control group. The blood routine test time points were 24, 48, and 72 hours after administration. In Figures 5A, 5B, and 5C, there were 4 mice in each group. DETAILED DESCRIPTION

[0118] definition

[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 non-aromatic substituents of organic compounds. Exemplary substituents include, but are not limited to, halogen, hydroxy, or any other organic group containing any number of carbon atoms, preferably 1-14 carbon atoms, and optionally including one or more heteroatoms, such as oxygen, sulfur, or nitrogen groups, in the form of linear, branched, or cyclic structures. Representative substituents include alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, phenyl, substituted phenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halogen, hydroxy, 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, amido, substituted amido, 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, heterocycles, substituted heterocycles, 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, hydroxy, 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, amido, substituted amido, 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, heterocycles, substituted heterocycles, amino acids, poly(lactic-co-glycolic acid), poly(lactic-co-glycolic acid), peptides, and polypeptide groups can be further substituted.

[0120] The heteroatoms, such as nitrogen, may have hydrogen substituents and / or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. It is understood that "substituted" or "substituted" includes the implicit proviso that such substitution complies with the permissible valences of the substituted atom and substituents and that the substitution results in a stable compound, i.e., a compound that does not spontaneously undergo transformations such as by rearrangement, cyclization, elimination, etc.

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

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

[0123] "Aryl" also includes polycyclic ring systems having two or more rings in which two or more carbon atoms are common to two adjacent rings (i.e., "fused rings"), wherein at least one ring is aromatic, e.g., the other cyclic ring or 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 one or more aromatic rings are replaced by one or more substituents including, but not limited to, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxy, alkoxy, carbonyl (e.g., ketone, aldehyde, carboxyl, alkoxycarbonyl, formyl, or acyl), silyl, ether, ester, thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino (or quaternized amino), amido, amidine, imine, cyano, nitro, azido, sulfhydryl, imino, alkylthio, sulfate, sulfonate, sulfamoyl, sulfoxide, sulfonamido, sulfonyl, heterocyclyl, alkaryl, haloalkyl (e.g., CF3, -CH2-CF3, -CCl3), -CN, aryl, heteroaryl, and combinations thereof.

[0125] "Heterocycle", "heterocyclic ring" and "heterocyclyl" are used interchangeably and refer to a cyclic group connected through a monocyclic or bicyclic ring carbon or nitrogen atom containing 3-10 or 3-8 ring atoms, preferably 5-6 ring atoms, consisting of carbon and one to four heteroatoms, each heteroatom selected from non-peroxide oxygen, sulfur and N(Y), wherein Y is absent or is H, O, C1-C 10Alkyl, phenyl or benzyl, and optionally containing 1-3 double bonds and optionally substituted by one or more substituents. According to the definition, heterocyclic radical 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-1,2,5-thiadiazinyl. The heterocyclic group can be optionally substituted by one or more substituents as defined above for alkyl and aryl.

[0126] The term "heteroaryl" refers to a C5-C 26The term "heteroaryl" refers to a monocyclic aromatic ring system or a combination thereof, wherein one or more carbon atoms on one or more aromatic ring structures have been replaced by heteroatoms. Suitable heteroatoms include, but are not limited to, oxygen, sulfur, and nitrogen. As used herein, the broad definition of "heteroaryl" 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, tetrazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine. Heteroaryl groups may also be referred to as "aryl heterocycles" or "heteroaromatic compounds." "Heteroaryl" also includes polycyclic ring systems with two or more rings, wherein two or more carbon atoms are shared by two adjacent rings (i.e., "fused rings"), wherein at least one ring is heteroaromatic, such as other rings or multiple rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heterocycle, or a combination thereof. Examples of heteroaromatic rings include, but are not limited to, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, benzopyranyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isatinoyl, isobenzofuranyl, isochromanyl, isoindazolyl, isodihydroindolinyl, isoindolyl, isoquinolyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, naphthyridinyl, octahydroisoquinolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrazine The heteroaryl groups may be substituted or unsubstituted heteroaryl groups, or may be substituted or unsubstituted heteroaryl groups. The heteroaryl groups may be substituted or unsubstituted heteroaryl groups, or may be substituted or unsubstituted heteroaryl groups. The heteroaryl groups may be substituted or unsubstituted heteroaryl groups, or may be substituted or unsubstituted heteroaryl groups. The heteroaryl groups may be substituted or unsubstituted heteroaryl groups, or may be substituted or unsubstituted heteroaryl groups.

[0127] The term "substituted heteroaryl" refers to a heteroaryl group in which one or more hydrogen atoms on one or more heteroaryl rings are replaced with one or more substituents including, but not limited to, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxy, alkoxy, carbonyl (e.g., ketone, aldehyde, carboxyl, alkoxycarbonyl, formyl, or acyl), silyl, ether, ester, thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino (or quaternized amino), amido, amidine, imine, cyano, nitro, azido, sulfhydryl, imino, alkylthio, sulfate, sulfonate, sulfamoyl, sulfoxide, sulfonamido, sulfonyl, heterocyclyl, alkaryl, haloalkyl (e.g., CF3, -CH2-CF3, -CCl3), -CN, aryl, heteroaryl, and combinations thereof.

[0128] As used herein, "alkyl" refers to a group of saturated aliphatic groups, including straight-chain alkyls, branched-chain alkyls, cycloalkyls (alicyclics), alkyl-substituted cycloalkyls, and cycloalkyl-substituted alkyls. In preferred embodiments, straight-chain or branched alkyls have 30 or fewer carbon atoms in their backbone (e.g., C1-C 30 For straight chain, C3-C 30 For branched chains), preferably 20 or less, more preferably 15 or less, most preferably 10 or less. Likewise, preferred cycloalkyl groups have 3-10 carbon atoms in their ring structure, and more preferably have 5, 6 or 7 carbon atoms in the ring structure. The term "alkyl" (or "lower alkyl") used throughout the specification, examples and claims is intended to include "unsubstituted alkyl" and "substituted alkyl," the latter referring to an alkyl moiety with one or more substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone. 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), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfoxide, sulfonamido, sulfonyl, heterocyclyl, aralkyl or aromatic or heteroaromatic moieties.

[0129] Unless otherwise specified, "lower alkyl" as used herein refers to an alkyl group as defined above, but having 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms, in its backbone structure. Similarly, "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 a hydrocarbon group as well as a heteroalkyl group. Suitable substituents include, but are not limited to, halogens such as fluorine, chlorine, bromine, or iodine; hydroxy; -NRR', wherein R and R' are independently hydrogen, alkyl, or aryl, and wherein the nitrogen atom is optionally quaternized; -SR, wherein R is hydrogen, alkyl, or aryl; -CN; -NO2; -COOH; carboxylates; -COR, -COOR, or CON(R)2, wherein R is hydrogen, alkyl, or aryl; azide, aralkyl, alkoxy, imino, phosphonate, phosphinate, silyl, ether, sulfonyl, sulfonamido, heterocyclic, aromatic or heteroaromatic moieties, haloalkyl (e.g., -CF3, -CH2-CF3, -CCl3); CN; -NCOCOCH2CH2; NCOCOCH; -NCS; and combinations thereof.

[0131] Those skilled in the art will appreciate that, if appropriate, the substituted portion of the hydrocarbon chain itself may be substituted. For example, the substituents of substituted alkyl groups may include halogen, hydroxyl, nitro, thiol, amino, azido, imino, amido, phosphoryl (including phosphonates and phosphinates), sulfonyl (including sulfates, sulfonamido, sulfamoyl, sulfoxides and sulfonates), and silyl, as well as ethers, alkylthio, carbonyl (including ketones, aldehydes, carboxylates and esters), haloalkyl, -CN, etc. Cycloalkyl groups may be substituted in the same manner.

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

[0133] The term "substituted alkenyl" refers to an alkenyl moiety having one or more substituents replacing one or more hydrogen atoms 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., carboxyl, alkoxycarbonyl, formyl or acyl), silyl, ether, ester, thiocarbonyl (e.g., thioester, thioacetate or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino (or quaternized amino), amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfoxide, sulfonamido, sulfonyl, heterocyclyl, alkaryl, haloalkyl, -CN, aryl, heteroaryl, and combinations thereof.

[0134] The term "substituted alkynyl" refers to an alkynyl moiety having one or more substituents replacing one or more hydrogen atoms 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., carboxyl, alkoxycarbonyl, formyl or acyl), silyl, ether, ester, thiocarbonyl (e.g., thioester, thioacetate or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino (or quaternized amino), amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfoxide, sulfonamido, sulfonyl, heterocyclyl, alkaryl, 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 following general formula:

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

[0138] wherein X is a bond, or represents oxygen or sulfur, and 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 alkaryl, substituted or unsubstituted aralkyl, substituted or unsubstituted aryl, or 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 alkaryl, substituted or unsubstituted aralkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl or (CH2) m -R"; R" represents a hydroxyl group, a substituted or unsubstituted carbonyl group, an aryl group, a cycloalkyl ring, a cycloalkenyl ring, a heterocycle or a polycycle; m is 0 or an integer from 1 to 8. Wherein X is oxygen and R is as defined above, this moiety is also referred to as a carboxyl group. When X is oxygen and R is hydrogen, the formula represents a "carboxylic acid". Wherein X is oxygen and R' is hydrogen, the formula represents a "formic acid". Wherein X is oxygen and R or R' is not hydrogen, the formula represents an "ester". Generally, where the oxygen atom of the above formula is replaced by a sulfur atom, the formula represents a 'thiocarbonyl' group. Wherein X is sulfur and R or R' is not hydrogen, the formula represents a 'thioester'. Wherein X is sulfur and R is hydrogen, the formula represents a 'thiocarboxylic acid'. Wherein X is sulfur and R' is hydrogen, the formula represents a 'thioformate'. Wherein X is a bond and R is not hydrogen, the above formula represents a 'ketone'. Wherein 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 a portion thereof

[0140] One or more hydrogen atoms in the group to which it is attached 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), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino (or quaternized amino), amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfoxide, sulfonamido, sulfonyl, heterocyclic, alkaryl, haloalkyl, -CN, aryl, heteroaryl, and combinations thereof.

[0141] The term "carboxyl" is as defined above for the following formula

[0142] And more specifically by the formula -R iv COOH definition, where R iv is an alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, alkaryl, aralkyl, aryl or heteroaryl. In preferred embodiments, the straight or branched chain alkyl, alkenyl and alkynyl groups have 30 or fewer carbon atoms in their backbone (e.g., C1-C 30 For straight chain alkyl, C3-C 30 For branched alkyl, C2-C 30For straight-chain alkenyl and alkynyl groups, C3-C 30 For branched 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 have 5, 6, or 7 carbon atoms in the ring structure.

[0143] The term "substituted carboxyl" refers to a carboxyl group as defined above, wherein R iv One or more hydrogen atoms in the group are replaced. 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), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino (or quaternized amino), amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfoxide, sulfonamido, sulfonyl, heterocyclyl, alkaryl, haloalkyl, -CN, aryl, heteroaryl, and combinations thereof.

[0144] As used herein, "heteroalkyl" refers to a linear or branched, or cyclic carbon-containing group, or a combination 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, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl, and homologues and isomers thereof, for example, n-pentyl, n-hexyl, n-heptyl, n-octyl. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, and 3-butynyl.

[0146] The term "alkoxyl" or "alkoxy", "aroxy" or "aryloxy" generally describes a radical of the formula -OR v Represented compounds, wherein R v These include, but are not limited to, substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, cycloalkenyl, heterocycloalkenyl, aryl, heteroaryl, aralkyl, heteroalkyl, alkaryl, and alkylheteroaryl.

[0147] As used herein, the term "alkoxy" or "alkoxy" refers to an alkyl group as defined above to which an oxygen group is attached. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy, and the like. An "ether" is two hydrocarbons covalently linked by an oxygen. Thus, the substituent of the alkyl group that makes the alkyl group an ether is or is similar to an alkoxy group, and can be represented, for example, by one of -O-alkyl, -O-alkenyl, and -O-alkynyl. If valence permits, the term alkoxy also includes cycloalkyl, heterocyclyl, cycloalkenyl, heterocycloalkenyl, 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 replacing one or more hydrogen atoms on one or more carbons of the alkoxy backbone. 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), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino (or quaternized amino), amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfoxide, sulfonamido, sulfonyl, heterocyclyl, alkaryl, haloalkyl, -CN, aryl, heteroaryl, and combinations thereof.

[0149] The term "alkylthio" refers to an alkyl group as defined above to which is attached a sulfenyl group. The "alkylthio" portion is represented by -S-alkyl. Representative alkylthio groups include methylthio, ethylthio, and the like. The term "alkylthio" also includes cycloalkyl groups to which is attached a sulfenyl group.

[0150] The term "substituted alkylthio" refers to an alkylthio group having one or more substituents replacing one or more hydrogen atoms on one or more carbon atoms of the alkylthio backbone. 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), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino (or quaternized amino), amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfoxide, sulfonamido, sulfonyl, heterocyclyl, alkaryl, haloalkyl, -CN, aryl, heteroaryl, and combinations thereof.

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

[0152] The term "substituted arylthio" represents an -S-aryl or -S-heteroaryl group having one or more substituents replacing a hydrogen atom on one or more ring atoms of the aryl and heteroaryl rings as defined herein. 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), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino (or quaternized amino), amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfoxide, sulfonamido, sulfonyl, heterocyclyl, alkaryl, haloalkyl, -CN, 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 group (eg, an aromatic or heteroaromatic group) substituted with a substituted or unsubstituted alkyl group.

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

[0156] 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 cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, or substituted or unsubstituted heterocyclyl, wherein R and R', independently of E, each independently represent 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 nitrogen atom to which they are attached form a heterocycle having 3 to 14 atoms in the ring structure; R"' represents a hydroxyl group, a substituted or unsubstituted carbonyl group, an aryl group, a cycloalkyl ring, a cycloalkenyl ring, a heterocycle or a polycycle; and m is 0 or an integer from 1 to 8. In a preferred embodiment, only one of R and R' can be a carbonyl group, for example, R and R' together with nitrogen do not form an imide. In a preferred embodiment, R and R' each independently represent a hydrogen atom, a substituted or unsubstituted alkyl group, a substituted or unsubstituted alkenyl group or -(CH2) m-R"'. When E is oxygen, a carbamate is formed. As will be appreciated 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 bond.

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

[0158] wherein E is absent or E is alkyl, alkenyl, alkynyl, aralkyl, alkaryl, cycloalkyl, aryl, heteroaryl, or heterocyclyl, wherein R, independently of E, 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 heterocycle having 3 to 14 atoms in the ring structure; R"' represents a hydroxyl group, a substituted or unsubstituted carbonyl group, an aryl group, a cycloalkyl ring, a cycloalkenyl ring, a heterocycle or a polycycle; and m is 0 or an integer from 1 to 8. In a preferred embodiment, only one of E and R can be a substituted or unsubstituted amine to form a "sulfonamide" or "sulfonamido". The substituted or unsubstituted amine is as defined above.

[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), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino (or quaternized amino), amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfoxide, sulfonamido, sulfonyl, heterocyclyl, alkaryl, haloalkyl, -CN, aryl, heteroaryl, and combinations thereof.

[0160] The term "sulfonic acid" refers to a sulfonyl group as defined above, wherein R is hydroxy 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 are as defined above, and R is independently hydroxy, alkoxy, aryloxy, substituted alkoxy or substituted aryloxy, as defined above. As will be appreciated by those of ordinary skill in the art, when E is oxygen, sulfate cannot be linked to another chemical species, such as to form an oxygen-oxygen bond or other unstable bond.

[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 alkaryl, substituted or unsubstituted aralkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl, -(CH2) m -R'', R'' represents a hydroxyl group, a substituted or unsubstituted carbonyl group, an aryl group, a cycloalkyl ring, a cycloalkenyl ring, a heterocycle, or a polycycle; m is 0 or an integer from 1 to 8. As will be 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 bonds.

[0163] The term "sulfamoyl" 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 alkaryl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, or substituted or unsubstituted heterocyclyl, wherein E, R, and R' each independently represent 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 nitrogen atom to which they are attached form a heterocycle having from 3 to 14 atoms in the ring structure; R"' represents a hydroxyl group, a substituted or unsubstituted carbonyl group, an aryl group, a cycloalkyl ring, a cycloalkenyl ring, a heterocycle, or a polycycle; and m is 0 or an integer from 1 to 8. In a preferred embodiment, only one of R and R' can be a carbonyl group, for example, R and R' together with nitrogen do not form an imide.

[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, or heterocyclyl, wherein R, independently of E, 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 heterocycle having 3 to 14 atoms in the ring structure; R"' represents a hydroxyl group, a substituted or unsubstituted carbonyl group, an aryl group, a cycloalkyl ring, a cycloalkenyl ring, a heterocycle or a polycycle; 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, or 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 heterocycle, 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 heterocycle having 3 to 14 atoms in the ring structure; R"' represents a hydroxyl group, a substituted or unsubstituted carbonyl group, an aryl group, a cycloalkyl ring, a cycloalkenyl ring, a heterocycle or a polycycle; and m is 0 or an integer from 1 to 8.

[0169] The term "substituted phosphono" refers to a group in which E, R vi and R viiIndependently substituted phosphono groups. 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), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino (or quaternized amino), amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfoxide, sulfonamido, sulfonyl, heterocyclyl, alkaryl, haloalkyl, -CN, aryl, heteroaryl, and combinations thereof.

[0170] The term "phosphoryl" defines a phosphoryl group wherein E is absent, is oxygen, alkoxy, aryloxy, substituted alkoxy or substituted aryloxy as defined above, and independently of E, R vi and R vii and R are independently hydroxy, alkoxy, aryloxy, substituted alkoxy or substituted aryloxy, as defined above. When E is oxygen, the phosphoryl group cannot be linked to another chemical species, such as to form an oxygen-oxygen bond or other unstable bond, as understood by those of ordinary skill in the art. vi and R vii When substituted, the 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, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino (or quaternized amino), amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfoxide, sulfonamido, sulfonyl, heterocyclyl, alkaryl, 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, or substituted or unsubstituted heterocyclyl having from 3 to 20 carbon atoms, where geometric constraints permit. The ring structure is formed by a single ring or a fused ring system. Substituted cycloalkyl, cycloalkenyl, cycloalkynyl, and heterocyclyl groups are substituted as defined above for alkyl, alkenyl, alkynyl, and heterocyclyl groups, respectively.

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

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

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

[0175] Compound C1-7-1a (5.00 g, 21.0 mmol, 1.00 equivalent), TBSCl (4.12 g, 27.3 mmol, 3.35 mL, 1.30 equivalent) and imidazole (2.86 g, 42.0 mmol, 2.00 equivalent) were mixed in DMF (30.0 mL), the air was evacuated and the mixture was filled with N2, and the mixture was stirred at 25 ° C under N2 for 2 hours. TLC (petroleum ether: ethyl acetate = 1:0, P1: R f =0.86, R1:R f =0.05) detection showed that the reaction of C1-7-1a was complete, generating a new main peak of low polarity. The reaction mixture was terminated by adding 300 mL of water and extracted with a total of 300 mL of DCM (100 mL × 3 times). The resulting organic layer was washed with 300 mL of saturated brine, dried over Na2SO4, filtered under reduced pressure, and concentrated to give a crude product. The crude product was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 1:0) to give 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 (400MHz, CDCl3) δppm 7.16 (dd, J = 10.0, 2.0Hz, 1H), 7.07 (d, J = 8.4Hz, 1H), 6.44 (t, J = 8.4Hz, 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) and stirred at 75°C under N2 for 2 hours. LCMS (EC10607-30-p1a1, P1: RT = 0.460 min) showed that about 58.1% of the desired product was produced. 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 main peak with a large polarity. The reaction mixture was terminated by adding 200 mL of water and extracted with 210 mL of DCM (70.0 mL × 3 times). The organic layer was washed with 200 mL of saturated brine, dried over Na2SO4, filtered under reduced pressure, and concentrated to obtain a crude product. The crude product was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 3:1, R f =0.22) was purified to give compound C1-7-3a (1.98 g, 4.41 mmol, 77.7% yield) as a yellow oil.

[0179] LCMS: EC EC10607-30-p1a1, RT=0.460min, 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 equiv) dissolved in THF (20.0 mL) was added pyridine hydrofluoric acid (875 mg, 8.83 mmol, 795 μL, 2.00 equiv), and the mixture was stirred at 25°C under a N2 environment for 2 hours. LCMS (EC10607-35-p1a1, P1: RT = 0.241 min) showed that approximately 89.2% of the desired product was produced. TLC (petroleum ether:ethyl acetate = 2:1, R1:R 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 a large polarity. The reaction mixture was terminated by adding 300 mL of water and extracted with 300 mL of DCM (100 mL × 3 times). The organic layer was washed with 300 mL of brine, dried over Na2SO4, filtered under reduced pressure, and concentrated to obtain a crude product. The crude product was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 2:1, R f =0.15) was purified to give compound C1-7 (1.16 g, 3.47 mmol, 78.6% yield) as a red solid.

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

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

[0184] 1H NMR (400MHz, CDCl3) δppm 7.01-7.08 (m, 2H), 6.90 (t, J = 8.8Hz, 1H), 3.49-3.57 (m, 6H), 2.62 (br t, J = 4.5Hz, 4H), 1.47 (s, 9H).

[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-phosphatricyclo[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), the air was expelled and N2 was flushed in, and the mixture was stirred at 90°C under N2 environment for 2 hours. LCMS (EC10607-26-p1a2, P1: RT = 0.649 min) showed that about 67.5% of the target product was generated. TLC (petroleum ether: ethyl acetate = 3:1, P1: R f =0.29, R1:R f =0.25) showed that the reaction of compound C1-1 was complete, and multiple new peaks were generated. The reaction mixture was quenched by adding 300 mL of water and extracted with 300 mL of DCM (100 mL × 3 times). The organic layer was washed with 300 mL of brine, dried over Na2SO4, filtered under reduced pressure, and concentrated to obtain a crude product. The crude product was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 3:1, R f =0.29) was purified to give compound C1-2 (1.25 g, 1.90 mmol, 71.7% yield) as a yellow oil.

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

[0188] 5. Synthesis of Compound C1-3

[0189] Pd / C (1.00 g, 1.83 mmol, 10% purity, 1.00 equiv) was added 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). The reaction mixture was stirred at 25° C. under 50 psi H 2 (1.23 mg, 609 μmol) for 24 hours. TLC (petroleum ether:ethyl acetate = 3:1, P1:R f =0.36, R1:R f =0.31) detection showed that the reaction of compound C1-2 was complete, generating a new peak. Pd / C was removed by filtration using diatomaceous earth, and the crude product was obtained by concentration under reduced pressure. The crude product was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 3:1, R f =0.30) was purified to give compound C1-3 (1.01 g, 1.53 mmol, 73.7% yield) as a yellow solid.

[0190] 6. Synthesis of Compound C1-4

[0191] To a mixture of compound C1-3 (1.00 g, 1.52 mmol, 1.00 equiv) dissolved in THF (15.0 mL) was added pyridine hydrofluoric acid (694 mg, 4.55 mmol, 631 μL, 65% purity, 3.00 equiv). The mixture was stirred at 25°C under a N2 atmosphere for 2 hours. LCMS (EC10607-37-p1a1, P1: RT = 0.410 min) showed approximately 46.4% of the desired product was produced. TLC (petroleum ether:ethyl acetate = 1:1, R1:R f =0.60, P1:R f =0.16) detection showed that the reaction of C1-3 was complete, generating a new major peak with a large polarity. The reaction mixture was quenched by adding 300 mL of water and extracted with 300 mL of DCM (100 mL × 3 times). The organic layer was washed with 300 mL of brine, dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a crude product. The crude product was analyzed by preparative TLC (SiO2, petroleum ether:ethyl acetate = 1:1, R f =0.16) was purified to give compound C1-4 (700 mg, 1.29 mmol, 84.7% yield) as a white solid.

[0192] LCMS:EC EC10607-37-p1a1,RT=0.410min,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), the air was purged and the mixture was filled with N2. The mixture was stirred at 25°C under N2 for 3 hours. LC-MS (EC10607-47-p1a1, P1: RT = 0.548 min) showed that approximately 46.7% of the desired product was produced. TLC (petroleum ether:ethyl acetate = 2:1, P1: R f =0.36, R1:R f =0.20) detection showed that part of C1-4 reacted and generated multiple new peaks. The reaction mixture was quenched by adding 300 mL of water and extracted with 300 mL of DCM (100 mL × 3 times). The organic layer was washed with 300 mL of brine, dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 2:1, R f =0.36) was purified to give compound C1-5 (400 mg, 572 μmol, 45.8% yield) as a yellow solid.

[0195] LCMS:EC(EC10607-47-p1a1,RT=0.548min,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), and Cs2CO3 (69.9 mg, 215 μmol, 1.50 equiv) were mixed in DMA (5.00 mL), the air was purged and the mixture was filled with N2. The mixture was stirred at 25°C under N2 for 12 hours. LC-MS (EC10607-49-p1a1, P1: RT = 0.493 min) showed that approximately 36.9% of the desired product was produced. TLC (petroleum ether:ethyl acetate = 1:1, P1: R f =0.24, R1:R f=0.42) detection showed that C1-5 was completely consumed 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 × 3 times). The organic layer was washed with 30.0 mL of brine, dried over Na2SO4, filtered and concentrated under reduced pressure to obtain the crude product. The crude product was analyzed by preparative TLC (SiO2, petroleum ether:ethyl acetate = 1:1, R f =0.24) were purified to give compound C1-6 (106 mg, 123 μmol, 86.0% yield) as a yellow solid.

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

[0199] 9. Synthesis of Compound C1

[0200] Compound C1-6 (100 mg, 116 μmol, 1.00 equiv) was dissolved in HCl / dioxane (5.00 mL), the atmosphere was purged and replaced with N2, and the mixture was stirred at 25°C under a N2 atmosphere for 12 hours. LC-MS (EC10607-51-p1a5, P1: RT = 0.412 min) indicated the formation of approximately 86.5% of the desired product. The reaction mixture was concentrated under reduced pressure to afford the crude product, which was then purified by preparative HPLC (HCl column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (HCl)-ACN]; B%: 26%-46% over 10 minutes) to afford compound C1 (25.0 mg, 35.5 μmol, 89.3% yield) as a yellow solid. LCMS (EC10607-57-p1a1, P1: RT = 0.42 minutes) analysis showed that the product had a purity of approximately 100%. HPLC (EC10607-57-p1a2, P1: RT = 2.027 minutes) analysis showed that the product C1 had a purity of approximately 96.1%.

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

[0202] 1H NMR(400MHz,DMSO+D2O)δppm 7.99(d,J=7.6Hz,1H),7.77(d,J=8.0Hz,1H),7.57(d,J=8.8Hz,1H),7.54(d,J=7.2Hz,1 H),7.47(t,J=7.2Hz,1H),7.39-7.42(m,1H),7.32-7.37(m,3H),7.27(d,J=8.9Hz,1H),7 .06-7.12(m,1H),7.01(d,J=8.8Hz,1H),4.91(s,2H),4.22(s,2H),4.08(s,2H),3.75-3 .81(m,2H),3.33-3.43(m,8H),2.94-2.99(m,2H),2.77-2.83(m,2H),1.88-1.96(m,2H).

[0203] b) Synthesis of compound C4 as a 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 equivalent) in MeCN (20.0 mL) and TFA (3.34 mL) was added NIS (4.38 g, 19.5 mmol, 1.1 equivalent), and the reaction mixture was stirred at 25°C for 12 hours. TLC (petroleum ether:ethyl acetate = 0:1, R1:R f =0.4;P1:R f =0.8) Monitoring showed that C4-1 was completely consumed and a new compound peak was generated. The reaction mixture was filtered to obtain the crude product C4-2 (3.00 g, crude product) 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 equiv) and C1-7-3 (1.41 g, 6.28 mmol, 1.5 equiv) in DMF (20.0 mL) were added CuI (39.9 mg, 209 μmol, 0.05 equiv), TEA (1.27 g, 12.6 mmol, 1.75 mL, 3.0 equiv), and Pd(dppf)Cl2 (306 mg, 418 μmol, 0.1 equiv). The reaction mixture was stirred at 90°C for 2 hours. LCMS (EC13784-5-P1A, P1: RT = 0.242 min) indicated that the reaction of C4-2 was complete, yielding the desired product as a major peak. 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×2 times), dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product 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 equiv) in DMA (3.00 mL) were added Cs2CO3 (175 mg, 537 μmol, 2.0 equiv) and C1-3 (188 mg, 268 μmol, 1.0 equiv). The reaction mixture was stirred at 25°C for 12 hours. LCMS (EC13784-6-P1A, P1: RT = 0.495 min) showed that the reaction of compound C4-3 was complete, yielding a main peak with the expected m / z. 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. The resulting organic layer was washed with saturated brine (10.0 mL × 2 times), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain the crude product. TLC (petroleum ether:ethyl acetate = 1:2, P1:R f =0.55; P2:R f =0.30) detection and purification 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 equivalent) was dissolved in HCl-dioxane (2.00 mL), and the mixture was stirred at 25°C for 1 hour. LCMS (EC13784-11-P1A2, P1: RT = 0.418 minutes) showed that the reaction of compound C4-4 was complete, yielding the desired product as a major peak with the expected m / z. The reaction mixture was filtered and concentrated under reduced pressure to obtain the crude product, which was then purified by preparative HPLC (column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (HCl)-ACN]; gradient elution: 30%-60% B over 10 minutes) to afford 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(400MHz DMSO)δppm 8.17(d,J=1.6Hz,1H),8.02(d,J=7.6Hz,1H),7.90(dd,J=10.8,1.8Hz,1H),7.78(d,J=8.0Hz,1 H),7.59(t,J=8.8Hz,2H),7.44-7.50(m,1H),7.40-7.44(m,1H),7.32-7.38(m,2H),7.02(d,J=8 .8Hz,1H),4.92(s,2H),4.37-4.42(m,2H),4.29-4.37(m,2H),3.83(t,J=6.0Hz,3H),3.57(d,J= 4.0Hz,5H),3.42-3.49(m,4H),2.98(t,J=5.6Hz,2H),2.81(t,J=7.6Hz,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), purged with N2 to remove air, and the mixture was stirred at 90°C under N2 for 12 hours. LC-MS (EC11272-85-P1A, P1: RT = 0.415 min) showed that the reaction of compound 1 was complete. 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 × 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 a crude product, which was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 3:1 to 0:1; when petroleum ether: ethyl acetate = 0:1, R f =0.24), followed by the addition of HCl / dioxane (4 M, 10.0 mL, 21.9 equiv), the introduction of N2 to expel air, and the mixture was stirred at 25 ° C. under N2 for 2 hours. The mixture was concentrated under reduced pressure to give the crude product AF3-2 (500 mg, 1.11 mmol, 61.1% yield) as a yellow solid.

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

[0223] 2. Synthesis of Compound AF3

[0224] Compound C1 (50.0 mg, 67.4 μmol, 1.00 equiv, HCl), 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), and DIEA (43.6 mg, 337 μmol, 58.7 μL, 5.00 equiv) were mixed in DMF (1.0 mL), purged with N2, and the mixture was stirred at 20°C under an N2 atmosphere for 16 hours. LC-MS (EC13958-9-P1B1, p1: RT = 0.456 min) indicated that the reaction of C1 was complete. The reaction mixture was concentrated under reduced pressure to give the crude product, which was purified by preparative HPLC (neutral conditions) to give compound AF3 (10 mg, 8.81 μmol, 13.0% yield) as a yellow solid.

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

[0226] HNMR:EC13958-9-P1Rδppm 8.42(t,J=5.6Hz,1H),7.98(d,J=7.6Hz,1H),7.74(d,J=8.0Hz,1H),7.60(d,J=6.8Hz,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,1 H),5.03(dd,J=5.2,12.8Hz,1H),4.98(s,2H),4.01(t,J=6.8Hz,2H),3.86(t,J=5.6Hz,2H),3.54-3.57(m ,2H),3.45-3.50(m,6H),3.43(d,J=4.4Hz,8H),3.39(dd,J=8.4,5.75Hz,8H),2.93-2.99(m,4H),2.78(br t,J=4.8Hz,4H),2.47(s,2H),1.88-2.09(m,4H).

[0227] Synthesis of AF4:

[0228] The synthesis steps of PROTAC molecule AF4 refer to AF3, but the starting materials are replaced with compounds 1 and AF4-1. After synthesis and purification, the target product AF4 is obtained as a yellow solid with a yield of about 13%.

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

[0230] HNMR:EC13958-10-P1R,400MHz,DMSO-d6δppm 8.39-8.46(m,1H),7.97(d,J=8.0Hz,1H),7.74(d,J=8.0Hz,1H),7.58-7.62(m,1H),7.52-7.58(m,1H),7.49(d,J=8.8Hz,1H),7.43 (t,J=7.2Hz,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=5.6Hz,1H),5.04(dd,J=1 2.8,5.6Hz,1H),4.99(s,2H),4.01(t,J=6.4Hz,2H),3.86(t,J=6.0Hz,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=4.8Hz,4H),2.46(d,J=4.4Hz,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] 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) were mixed in DMSO (40.0 mL). The mixture was purged with nitrogen to remove air. The mixture was stirred at 100°C under a nitrogen atmosphere for 12 hours. LC-MS (EC13798-1-p1a1, P1 RT = 0.312 min) indicated the formation of a product with the desired molecular weight. 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×5 times), and the organic layer was washed with 50.0 mL of saturated brine, dried over Na 2 SO 4 , filtered and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (SiO 2 , petroleum ether: ethyl acetate = 5:1 to 0:1; when petroleum ether: ethyl acetate = 0:1, R f =0.20) to give the crude product BF3-2 (5.00 g, 11.1 mmol, 78.7% yield) as a green oil.

[0235] LCMS: EC13798-1-p1a1, P1 RT=0.312min, 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, and the mixture was stirred at 25°C for 2 hours. LC-MS (EC13798-4-p1a1, P1 RT = 0.318 min) indicated the formation of the target molecular weight product. The reaction mixture was quenched by the addition of saturated NaHCO₃ (10.0 mL) and saturated Na₂S₂O₃ (10.0 mL) at 25°C, stirred for 30.0 minutes, and extracted with DCM (20.0 mL x 3). The organic layer was washed with saturated brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure to afford crude product BF3-3 (200 mg) as a yellow oil.

[0238] LCMS: EC13798-4-p1a1, P1 RT=0.318min, 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) were added compound C1 (30.0 mg, 42.5 μmol, 1.00 equiv) and NaBH(OAc)3 (27.0 mg, 127 μmol, 3.00 equiv). The mixture was stirred at 25°C for 2 hours. LC-MS (EC13798-17-p1c8, P1 RT = 0.453 min) indicated the formation of a product with the desired molecular weight. The reaction mixture was concentrated under reduced pressure to give the crude product, which was purified by preparative HPLC (TFA conditions; chromatographic column: Phenomenex luna C18 150*25.0 mm*10.0 μm; 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=0.453min, MS(ESI)m / z=1136.4[M+H] +

[0242] 1H NMR: EC13798-17-p1a2, 400MHz, DMSO+D2Oδ8.02(d,J=7.6Hz,1H),7.77(d,J=8.0Hz,1H),7.52-7.61(m,3H),7.47(t,J=8.0Hz,1H),7.38-7. 43(m,1H),7.35(m,2H),7.26(dd,J=12.0,1.6Hz,1H),7.18(d,J=8.8Hz,1H),7.05-7.12(m,2H),7.03(d,J=7.2Hz,1H),6.96(d,J=8.4Hz,1H ),5.03(dd,J=13.2,5.6Hz,1H),4.87-4.95(m,2H),4.01(t,J=6.0Hz,2H),3.84(t,J=5.6Hz,2H),3.69-3.73(m,2H),3.64(s,2H),3.60(s,4 H),3.43(t,J=5.2Hz,3H),3.26(s,2H),2.97(t,J=5.6Hz,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 steps of the PROTAC molecule BF4 refer to BF3, but the starting materials are replaced with compound 1 and BF4-1. After synthesis and purification, the target product BF4 is obtained as a yellow solid with a purity of 94.1% and a yield of about 16%.

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

[0246] 1H NMR: EC13798-11-p1a3, 400MHz, DMSO+D2Oδ8.02(d,J=8.0Hz,1H),7.78(d,J=7.6Hz,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=11.6,2.0Hz,1H),7.19(d,J=9.2Hz,1H),7.10(d,J=8.4Hz,2H),7.03(d,J =6.8Hz,1H),6.96(d,J=8.8Hz,1H),5.03(dd,J=13.2,6.0Hz,1H),4.92(s,2H),4.01(t,J=6.40Hz,2H),3.83(t,J=6.00Hz,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) Synthetic 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 C1 (0.500 g, 709 μmol, 1.00 equiv) in DMF (5.00 mL) were added TBTU (455 mg, 1.42 mmol, 2.00 equiv) and DIEA (458 mg, 3.55 mmol, 617 mL, 5.00 equiv), and the mixture was stirred at 25 °C for 2 h. After the reaction, the mixture was terminated by adding water (10.0 mL), diluted with DCM (10.0 mL), and extracted with DCM (30.0 mL, 10.0 mL × 3 times). The obtained organic layer was washed with saturated brine (20.0 mL, 10.0 mL × 2 times), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain the crude product, which was further purified by preparative HPLC (chromatographic column: Phenomenex Luna C18 150*25mm*10μm; mobile phase: [water (FA)-ACN]; gradient elution: 30%-60% B in 10 minutes) 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=0.426min, 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), and the mixture was stirred at 25°C for 1 hour. After the reaction, the mixture was concentrated under reduced pressure to give the crude product Q1-2 (25.0 mg, 31.2 μmol, 88.1% yield) as a yellow solid, which was used directly in the next reaction without purification.

[0253] LCMS: EC13798-58-p1a1, P1 RT=0.394min, 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 HCl / methanol (4.00 M, 12.5 mL, 48.5 equiv), and the mixture was stirred at 25°C for 0.3 h. LCMS (P1 RT = 0.162 min) indicated the formation of the desired molecular weight product. The reaction mixture was concentrated under reduced pressure to afford product Q1-4 (400 mg, crude) as a yellow solid, which was used directly in the next step without purification.

[0256] LCMS: EC13798-46-p1a1, P1 RT=0.162min, 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), and the mixture was stirred at 25°C for 1 hour. LCMS (P1 RT = 0.375 min) indicated the formation of the desired molecular weight product. After the reaction, the mixture was terminated by adding water (10.0 mL) at 25°C, diluted with DCM (10.0 mL), and extracted with DCM (30.0 mL, 10.0 mL × 3 times). The obtained organic layer was washed with saturated brine (30.0 mL, 10.0 mL × 3 times), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain the crude product, which was further purified by preparative HPLC (chromatographic column: Phenomenex luna C18 150*25mm*10μm; mobile phase: [water (HCl)-ACN]; gradient elution: 27%-47% B over 10.0 minutes) to obtain the product APQ1 (1.00 mg, crude) as a yellow solid.

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

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

[0261] d) Synthetic 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 CHCN (40.0 mL) were added KCO (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 hours. LC-MS (EC13784-1-p1a, P1 RT = 0.263 min) indicated that the reaction of Q2-1 was complete, yielding the product of the desired molecular weight. To the reaction mixture were added 40.0 mL of water and 40.0 mL of ethyl acetate for extraction. The resulting organic layer was washed with saturated brine (30.0 mL × 2 times), dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product, which was purified by reverse-phase HPLC (0.1% FA conditions) to give Q2-2 (2.30 g, 5.91 mmol, 47.0% yield, 86.4% purity) as a 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(400MHz DMSO)δ8.19(s,1H),7.26-7.42(m,5H),7.05-7.24(m,1H),5.01(s,3H),4.02(d,J=6.8Hz,1H),3.24(s,6H),3.05-3. 18(m,2H),2.90(d,J=10.8Hz,2H),2.33-2.45(m,2H),1.96(t,J=11.6Hz,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 equivalent) in MeOH (60.0 mL) was added Pd-C (190 mg, 10% purity). The mixture was hydrogenated under 50 psi H₂ at 20°C for 12 hours. LC-MS (EC13784-4-p1a, 0.446 min) indicated that Q2-2 had reacted completely to produce the desired molecular weight product. The reaction mixture was concentrated under reduced pressure to afford the crude product, which was not purified. The crude product, Q2-3 (1.00 g, 4.94 mmol, 87.53% yield), was obtained as a 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 equiv), compound 1 (1.23 g, 4.45 mmol, 1.0 equiv), and DIEA (1.15 g, 8.90 mmol, 1.55 mL, 2.0 equiv) were mixed in DMF (20.0 mL). The mixture was purged with N2 and the air was removed. The mixture was stirred at 90°C under N2 for 3 hours. LC-MS (EC13784-14-p1a1, P1: RT = 0.252 min) showed that the reaction of Q2-3 was complete, yielding a major peak of the target molecular weight. 30.0 mL of water and 90.0 mL of ethyl acetate were added to the reaction mixture for extraction. The resulting organic layer was washed with saturated brine (40.0 mL × 2 times), dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product, which was detected by TLC (dichloromethane:methanol = 10:1, P1:RT = 0.46). The crude product was purified by column chromatography (SiO2, DCM:MeOH = 10:1) to give Q2-4 (300 mg, crude product) as a 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 equiv) in THF (5.00 mL) was added H2SO4 (34.2 mg, 348 μmol, 18.6 μL, 2.00 equiv), and the mixture was stirred at 60°C for 0.5 h. LC-MS (EC13798-21-p1a1, P1 RT = 0.214 min) indicated the formation of the target molecular weight product. The reaction mixture was concentrated under reduced pressure to afford the crude product, which was not purified. The crude product Q2-5 (100 mg) was obtained as a yellow oil.

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

[0275] 5. Synthetic compound APQ2

[0276] To a solution of compound Q2-5 (100 mg, 121 μmol, 1.00 equiv) and C1 (85.4 mg, 121 μmol, 1.00 equiv) in DCM (5.00 mL), TEA (12.2 mg, 121 μmol, 16.8 μL, 1.00 equiv) was added dropwise at 25° C. over 5 minutes, followed by the addition of NaBH(OAc) 3 (77.0 mg, 363 μmol, 3.00 equiv) at 25° C. The mixture was stirred at 25° C. for 1 hour. LC-MS (EC13798-22-p1c4, P1 RT = 1.825 min) indicated the formation of a product of the desired molecular weight. The reaction mixture was concentrated under reduced pressure to give the crude product, which was further purified by preparative HPLC (TFA conditions; column: Phenomenex luna C18 150*25.0 mm*10.0 μm; mobile phase: [water (TFA)-ACN]; gradient: 32%-62% B over 10 min) to give APQ2 (12.2 mg, 10.2 μmol, 8.49% yield, 92.9% purity) as a yellow solid.

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

[0278] 1 H NMR: EC13798-22-p1c1, 400MHz, DMSO δ 12.83 (s, 1H), 11.10 (s, 1H), 9.40 (s, 1H), 8.04 (d, J = 8.0Hz, 1H), 7.80 (d, J = 8.0Hz, 1H), 7.5 5-7.65(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=8.8Hz, 1H),6.87(t,J=6.40Hz,1H),5.07(dd,J=12.8,5.2Hz,1H),4.93(s,2H),4.04(t,J=6.4Hz,2H),3.86(t,J=6.4Hz,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) Synthetic 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). The reaction was stirred at 20°C for 2 hours. LC-MS (EC7111-187-p1a, P1: RT = 0.464 min) showed that the reaction of compound C1-3 was complete. Multiple new compound peaks were generated, with the target m / z accounting for approximately 89.8%. The mixture was filtered and concentrated under reduced pressure to obtain the crude product, which was further purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 1:0 to 1:2) to afford 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=0.464min, 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) were 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 hours. LC-MS (EC7111-195-p1a, R1: RT = 0.425 min, P1: RT = 0.511 min) showed approximately 4.73% of compound Q4-2 remaining, with the formation of multiple new compound peaks, with the target m / z product accounting for approximately 48.7%. After the reaction, 50.0 mL of NaHCO3 was added to the mixture at 20°C to terminate the reaction. The mixture was extracted with 60.0 mL of ethyl acetate (20.0 mL × 3 times). The obtained organic layer was 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 = 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=0.511min, MS(M+H + )=609.3

[0286] 3. Synthesis of Compound Q4-4

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

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

[0289] 4. Synthesis of Compound Q4-5

[0290] Compound Q4-4 (200 mg, 281 μmol, 1.00 equiv) was mixed in HCl / EtOAc (3.00 mL) and stirred at 20°C under N2 for 24 hours. LC-MS (EC15303-1-p1b, P1: RT = 0.311 min) showed complete reaction of compound Q4-4, with the formation of multiple new peaks. The target m / z accounted for approximately 58.7%. The mixture was filtered and concentrated under reduced pressure to afford crude compound Q4-5 (250 mg) as a yellow solid, which was used directly in the next step without purification.

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

[0292] 5. Synthesis of compound APQ4

[0293] 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) were mixed in DCM (2.00 mL) and stirred at 20°C for 15 minutes. NaBH(OAc)3 (148 mg, 700 μmol, 3.00 equiv) was then added to the mixture, and the reaction was continued with stirring at 20°C under a nitrogen atmosphere for 1.75 hours. LC-MS (EC15303-3-P1A, P1: RT = 0.313 min) showed that the reaction of compound Q4-5 was complete, with the formation of multiple new peaks. The target m / z product accounted for approximately 53.7%. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain the crude product, which was further purified by preparative HPLC (FA conditions: column: Welch Ultimate C18 150*25mm*5μm; mobile phase: [water(FA)-ACN]; gradient elution: 10%-40% B over 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=0.313min, MS(M+H + )=477.7

[0295] 1H NMR: EC15303-3-p1a (400MHz, DMSO-d6) δ11.09 (s, 1H), 8.17 (s, 1H), 8.03 (d, J = 7.6Hz, 1H), 7.79 (d, J = 8.2Hz, 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=8.8Hz,1H),7.02(d,J=7.2Hz,1H),6.84(d,J=8.8Hz,1H),6.75(d,J=3.2Hz,1H),6. 40-6.64(m,1H),5.06(dd,J=12.8,5.2Hz,1H),4.89(s,2H),3.84(t,J=6.0Hz,3H),2.97(t,J=5.6Hz,3H),2.86(d,J=11.6Hz,3H),2.54-2.64(m ,8H),2.31-2.43(m,7H),2.10(d,J=6.8Hz,2H),1.88-2.06(m,4H),1.7 2-1.79(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 Molecules Recruiting VHL-E3 Enzymes or IAP-E3 Enzymes (APQ3 and APL-1 to APL-6)

[0297] a) Synthesis of PROTAC molecules containing VHL-E3 enzyme recruitment 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 methyl 3-bromopropionate (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 hours. TLC (DCM / MeOH = 10:1, R1:R f =0.10, P1: Rf = 0.75) showed that the reaction of compound 1A was complete and multiple new products were generated. The reaction mixture was filtered and concentrated under reduced pressure to obtain a crude product, which was then purified by column chromatography (SiO2, DCM / MeOH = 100 / 1 to 0 / 1, P1: Rf =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, 400MHZ δ4.01-4.04(m,1H),3.58(s,3H),3.23(s,6H),2.80(d,J=11.2Hz,2H),2.50-2.54(m,2H), 2.44(d,J=6.0Hz,2H),1.99(s,2H),1.83(td,J=11.6,2.0Hz,2H),1.56(d,J=13.2Hz,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) were 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 hours. 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 form a single main peak compound. After the reaction, the mixture was concentrated under reduced pressure to remove the solvent without purification to obtain crude compound Q3-3 (2.20 g) as a white solid.

[0303] 1 H NMR:EC12156-19-p1a1,D2O,400MHZδ4.16-4.26(m,1H),3.36(s,6H),2.89(d,J=11.6Hz,2H),2.53-2.65(m,2H ),2.33(dd,J=8.8,6.8Hz,2H),2.01(t,J=11.2Hz,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 μmol, 1.00 equiv) and compound Q3-3 (200 mg, 864.72 μmol, 1.00 equiv) in CH3CN (3.00 mL) were added HOBt (116 mg, 864 μmol, 1.00 equiv), Et3N (350 mg, 3.46 mmol, 481 μL, 4.00 equiv), and BTU (327 mg, 864 μmol, 1.00 equiv). The mixture was stirred at 25°C for 8 hours. LCMS (EC12156-24-p1a2, P1: RT = 0.299 min) showed that the reaction of compound Q3-3 was complete, yielding a major peak of the target m / z (m / z = 658.6 [M+H]). + ), accounting for about 61.5%. After the reaction, the mixture was concentrated under reduced pressure to remove the solvent, and further analyzed by preparative TLC (SiO2, DCM:MeOH=7:1, P1:R f =0.23) were purified to give compound Q3-4 (179 mg, 272 μmol, 31.4% yield) as a white solid.

[0306] LCMS: EC12156-24-p1a2, P1: RT=0.299min, 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 μmol, 1.00 equiv) was added H2SO4 (14.9 mg, 152 μmol, 8.10 μL, 2.00 equiv) and THF (2.00 mL), and the mixture was stirred at 60°C for 4 hours. LCMS (EC12156-30-p1a1, P1: RT = 0.298 min) showed that the reaction of compound Q3-4 was complete, with the formation of a new major peak accounting for approximately 89.4% of the total product, with an m / z consistent with the desired product (m / z = 612.5 [M+H]). + After the reaction, the mixture was diluted with 15.0 mL of water and extracted with 300 mL of DCM (100 mL×3 times). The obtained organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a crude product of compound Q3-5 (61.0 mg) as a yellow solid.

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

[0310] 1H NMR: EC12156-30-p1a1, CDCl3, 400MHZ δ9.68(s,1H),9.17(d,J=7.4Hz,1H),8.65-8.69(m,1H),7.64(d,J=7.8Hz,1H),7.39(q,J= 8.2Hz,4H),5.30(s,1H),5.07(m,J=7.2Hz,1H),4.77(t,J=7.6Hz,1H),4.48(s,1H),4.41(d,J=7.6Hz,1H),4.16(d,J=10.4Hz,1H ),3.56(dd,J=11.2,3.6Hz,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=6.8Hz,3H),1.06(s,7H),0.87(d,J=10.4Hz,5H)

[0311] 5. Synthesis of compound APQ3

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

[0313] LCMS: EC12156-36-p1a3, P1: RT=0.418min, 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,400MHZδ8.97(s,1H),8.57-8.67(m,1H),8.38(d,J=8.0H z,1H),8.03(d,J=7.4Hz,1H),7.78(d,J=8.4Hz,1H),7.60(d,J=7.2Hz,1H),7.31-7.51 (m,10H),7.26(dd,J=12.0,2.0Hz,1H),7.15-7.20(m,1H),7.09(t,J=8.8Hz,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=1.6Hz,1H ),4.01(t,J=6.8Hz,2H),3.8(t,J=5.6Hz,2H),3.55-3.65(m,3H),3.44(s,4H),2.97(t ,J=5.4Hz,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=7.2,2.8Hz,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=6.8Hz,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 recruitment 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] 2-Iodoethanol (137 mg, 801 μmol, 62.6 μL, 2.4 equivalents), compound 3 (200 mg, 334 μmol, 1.0 equivalents) and K2CO3 (46.1 mg, 334 μmol, 1.0 equivalents) were mixed in DMF (2.0 mL), and the mixture was stirred at 70 ° C under N2 environment for 48 hours. LC-MS (EC16364-17-P1A, P1, RT = 0.432 min) detection showed that compound 3 was completely consumed. After the reaction, the mixture was terminated by adding 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 × 3 times). The obtained organic layer was washed with saturated brine (20.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product L1-2 (140 mg, 217 μmol, 65.2% yield) as a white oil. The crude product was used directly in the next reaction without 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 equivalent) in DCM (3.0 mL) were added TsCl (83.0 mg, 435 μmol, 2.0 equivalent) and Et3N (66.1 mg, 653 μmol, 90.9 μL, 3.0 equivalent), and the mixture was stirred at 25°C for 12 hours. LC-MS (EC16364-18-P1A1, P1, RT = 0.495 min) monitoring showed that the reaction of compound L1-2 was complete. After the reaction, the mixture was quenched by adding water (20.0 mL) at 0°C, diluted with DCM (20.0 mL), and extracted with DCM (60.0 mL, 20.0 mL × 3 times). The obtained organic layer was washed with saturated brine (20.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product L1-3 (170 mg, 213 μmol, 97.9% yield) as a white oil. The crude product was used directly in the next reaction without 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 μmol, 1.0 equivalent) in DCM (2.0 mL) was added Et3N (63.5 mg, 627 μmol, 87.3 μL, 5.0 equivalent) and compound 1A (99.9 mg, 627 μmol, 5.0 equivalent), and the mixture was stirred at 85°C for 24 hours. LCMS (EC16364-42-p1a, P1, RT = 0.303 min) monitoring showed that the reaction of compound L1-3 was complete. After the reaction, the mixture was quenched by adding 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 × 3 times). The obtained organic layer was washed with saturated brine (20.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product L1-4, which was further analyzed by preparative TLC (SiO2, petroleum ether:ethyl acetate = 0:1, P1, R f =0.3) was purified to give the product L1-4 (20.0 mg, 25.5 μmol, 20.3% yield) as a white solid.

[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 μmol, 1.0 equiv) was added to formic acid (2 mL), and the mixture was stirred at 25°C under a nitrogen atmosphere for 2 hours. TLC (petroleum ether:ethyl acetate = 0:1) showed that L1-4 had reacted completely, forming a more polar main peak. The reaction mixture was concentrated under reduced pressure to afford crude product L1-5 (15 mg, 20.3 μmol, 79.6% yield) as a yellow oil, which was used directly in the next reaction without purification.

[0329] 5. Synthesis of APL-1

[0330] To a solution of compound L1-5 (15.0 mg, 20.3 μmol, 1.0 equiv) in DCM (1 mL) were added TEA (20.6 mg, 203 μmol, 28.3 μL, 10.0 equiv) and compound C1 (14.3 mg, 20.3 μmol, 1.0 equiv). The mixture was stirred at 25° C. for 0.5 h. NaBH(OAc) 3 (12.9 mg, 61.0 μmol, 3.0 equiv) was added, and the mixture was stirred at 25° C. for 3.5 h. LC-MS (EC16364-45-P1A, P1, RT = 0.381 min) indicated that the reaction of L1-5 was complete. After the reaction, the mixture was concentrated under reduced pressure to obtain a crude product, which was further purified by preparative HPLC column chromatography (FA conditions) to obtain compound APL-1 (10 mg, 7.34 μmol, 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(400MHz DMSO)δppm 8.51(s,1H)8.22(s,1H)8.09(d,J=8.8Hz,1H)8.00-8.06(m,1H)7.79(d,J=8. 0Hz,1H)7.71(s,1H)7.57-7.66(m,2H)7.42-7.50(m,3H)7.38-7.42(m,1H)7.3 4(td,J=7.2,3.38Hz,2H)7.22-7.29(m,2H)7.18(d,J=9.2Hz,1H)7.05-7.14( m,1H)6.84(d,J=9.2Hz,1H)5.39(dd,J=7.6,2.81Hz,1H)4.90(s,2H)4.47(t,J =7.6Hz,1H)4.13t,J=5.2Hz,2H)4.01(t,J=6.4Hz,2H)3.78-3.85(m,4H)3.46 (s,2H)3.17(d,J=6.8Hz,2H)2.97(t,J=5.2Hz,2H)2.91(d,J=10.0Hz,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=7.6Hz,2H)1.46(s,1H)1.15(d,J=6.8Hz,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 μmol, 1.0 eq) in DCM (10.0 mL) were added PhNTf2 (267 mg, 748 μmol, 1.6 eq), TEA (94.6 mg, 935 μmol, 130 μL, 2.0 eq), and DMAP (5.71 mg, 46.8 μmol, 0.1 eq), and the mixture was stirred at 25°C for 16 hours. After the 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% FA) to obtain compound L2-1 (300 mg, approximately 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 μmol, 1.0 equiv), compound 1A (174 mg, 1.09 mmol, 5.0 equiv), Cs2CO3 (214 mg, 657 μmol, 3.0 equiv) and XPhosPdG4 (18.8 mg, 21.9 μmol, 0.1 equiv) were mixed in dioxane (3.00 mL) and stirred at 90°C under a N2 atmosphere for 2 hours. After the reaction, the mixture was separated by adding water (10.0 mL) and DCM (60.0 mL, 10.0 mL × 6 times). The resulting organic layer was washed with saturated brine (60.0 mL, 30.0 mL × 2 times), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 5:1) to give compound L2-2 (120 mg, 129.57 μmol, 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(400MHz CDCl3)δppm 8.10(s,1H),7.74(d,J=1.6Hz,1H),7.56-7.66(m,1H),7.34(d,J=1.2Hz,1H),7.13-7.21(m,1H),5.56(dd ,J=7.6,2.4Hz,1H),4.64(dd,J=8.8,6.4Hz,2H),3.86-3.91(m,1H),3.78(d,J=11.6Hz,2H),3.39(s,6H), 2.81(s,3H),2.75(d,J=8.0Hz,2H),2.50(dt,J=6.4,3.2Hz,1H),2.27-2.32(m,1H),2.18(d,J=7.2Hz,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=7.2Hz,3H).

[0341] 3. Synthesis of Compound L2-3

[0342] Compound L2-2 (50.0 mg, 67.6 μmol, 1.0 equiv) was mixed with formic acid (0.5 mL) and stirred at 25°C for 1 hour. The mixture was concentrated under reduced pressure to remove the solvent to afford crude compound L2-3 (30.0 mg) as a yellow oil, which was used directly in the next reaction without 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 equiv), C1 (35.6 mg, 50.5 μmol, 1.0 equiv) and NaBH(OAc)3 (32.1 mg, 152 μmol, 3.0 equiv) were mixed in DMF (0.5 mL) and stirred at 25°C under N2 protection for 2 hours. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain the crude product, which was further purified by reverse phase HPLC (0.1% FA conditions) to obtain 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 (400MHz DMSO) δppm 8.44 (s, 1H), 8.11 (d, J=8.4Hz, 1H),8.02(d,J=7.6Hz,1H),7.75-7.82(m,1H),7.69(s,1H),7.60(d,J=7.13H z,1H),7.44-7.50(m,2H),7.39(d,J=5.2Hz,2H),7.35(dt,J=7.2,3.6Hz,3H), 7.14-7.30(m,4H),7.04-7.14(m,1H),6.77-6.85(m,1H),5.38(dd,J=6.6,3. 6Hz,1H),4.90(s,2H),4.43-4.50(m,2H),3.98-4.05(m,4H),3.83(d,J=5.2Hz ,6H),3.70-3.79(m,10H),3.20(s,2H),2.97(d,J=4.8Hz,2H),2.71-2.76(m, 4H),2.37-2.42(m,2H),2.23(s,4H),2.15(d,J=6.13Hz,2H),2.00(dd,J=16.8 ,6.8Hz,5H),1.78(d,J=10.4Hz,2H),1.68(d,J=3.2Hz,2H),1.61(s,2H),1.5 4(d,J=9.6Hz,2H),1.24(s,2H),1.15(d,J=6.8Hz,2H),1.06(t,J=6.8Hz,3H).

[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 equiv) in MeCN (1 mL) was added K2CO3 (69.2 mg, 501 μmol, 3.0 equiv) at 25°C, and the mixture was stirred at 25°C for 30 minutes. 1,4-dibromobutane (180 mg, 835 μmol, 100 μL, 5.0 equiv) was then added dropwise at 25°C. The mixture was stirred at 80°C for 12 hours. After the reaction, the mixture was quenched by adding 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 × 3 times). The resulting organic layer was washed with saturated brine (20.0 mL), dried over Na2SO4, filtered under reduced pressure, and concentrated to give the crude product L3-2, which was further analyzed by preparative TLC (SiO2, petroleum ether:ethyl acetate = 1:1, P1, R f =0.3) was purified to give the product L3-2 (100 mg, 136 μmol, 40.8% yield, 100% purity) as a white solid.

[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 equiv), L3-2 (80.0 mg, 109 μmol, 1.0 equiv), and DIEA (42.3 mg, 327 μmol, 56.9 μL, 3.0 equiv) were mixed in DMF (1.0 mL) and stirred at 80°C under a nitrogen atmosphere for 6 hours. LC-MS (EC16364-70-P1A1, P1, RT = 0.504 min) indicated the complete reaction of L3-2. The reaction mixture was concentrated under reduced pressure to yield the crude product, which was further purified by preparative HPLC (FA conditions) to afford 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) was mixed with HCl / dioxane (4 M, 5.0 mL, 905 equiv) and stirred at 25°C under a N2 atmosphere for 1 hour. LC-MS (EC16364-80-P1B2, P1, RT = 0.428 min) indicated complete reaction of L3-3. The reaction mixture was concentrated under reduced pressure to afford the crude product, which was then purified by preparative HPLC (HCl) to afford 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(400MHz DMSO)δppm 9.30(s,1H)9.07(s,1H)8.87(s,1H)8.75(d,J=7.6Hz,1H)8.49(s,1H)8.04(d,J= 8.0Hz,1H)7.79(d,J=8.0Hz,1H)7.68(d,J=7.6Hz,1H)7.53-7.63(m,4H)7.47(br t,J=7.6Hz,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=7.6Hz,1H)4.18(s,1H)4.01-4.10(m,6H)3.86(t,J=5.6Hz,4H)3.76-3.82(m ,4H)2.98(t,J=5.2Hz,2H)2.81(d,J=7.2Hz,2H)2.32(t,J=4.8Hz,1H)2.13-2.29(m,3H)1.94-2.06(m,5H)1.87 -1.92(m,2H)1.82(d,J=6.4Hz,2H)1.59-1.72(m,5H)1.52-1.58(m,3H)1.35(d,J=6.8Hz,3H)0.99-1.15(m,6H).

[0360] Synthesis of APL-4:

[0361] The synthesis steps of PROTAC molecule APL-4 refer to APL-3, but the starting materials are replaced with compounds 3 and L4-1. After synthesis, the target product APL-4 (5 mg, 3.59 μmol, 32.8% yield, 91.3% purity) is obtained by preparative HPLC column chromatography (HCl conditions) 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 (400MHz DMSO) δppm 9.18(s,1H)8.84(s,1H)8.73-8.73(m,1H)8.74(d,J=8.0Hz,1H)8.49(s,1H)8.04(d,J=8.0Hz,1H)7.79(d,J=8.0Hz,1H)7.68(d,J=7.6Hz, 1H)7.54-7.62(m,4H)7.47(td,J=7.6,4.13Hz,3H)7.41(d,J=7.2Hz,2H)7.36(d,J=6.0Hz,2H)7.32(d,J=9.6Hz,1H)7.23-7.29(m,2H)7.17 -7.22(m,1H)7.13(t,J=8.8Hz,1H)6.97(d,J=8.8Hz,1H)5.39(dd,J=7.6,2.69Hz,1H)4.92(s,2H)4.47(t,J=7.6Hz,1H)4.05(d,J=9.2Hz, 4H)3.86(d,J=5.2Hz,4H)3.81(dd,J=6.4,2.25Hz,2H)3.10-3.13(m,2H)2.98(t,J=5.2Hz,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=6.8Hz,3H)1.16-1.25(m,2H)1.12(d,J=7.2Hz,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 μmol, 1.0 equiv) in MeCN (3 mL) was added KCO (69.2 mg, 501 μmol, 3.0 equiv) at 25°C and stirred for 30 minutes. L5-1 (230 mg, 835 μmol, 5.0 equiv) was then added dropwise at 25°C. The mixture was stirred at 80°C for 12 hours. LC-MS (EC16364-78-P1A, P1, RT = 0.495 min) indicated the complete reaction of compound 3. 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). The obtained organic layer was washed with saturated brine (20.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product L5-2, which was further analyzed by preparative TLC (SiO2, petroleum ether:ethyl acetate = 1:2, P1, R f =0.3) was purified to give the product L5-2 (90.0 mg, 113 μmol, 67.9% yield, 100% purity) as a white solid.

[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 μmol, 1.0 equiv), C1 (35.5 mg, 50.4 μmol, 0.8 equiv), and DIEA (24.4 mg, 188 μmol, 32.9 μL, 3.0 equiv) were mixed in DMF (1.0 mL) and stirred at 80°C under a nitrogen atmosphere for 6 hours. LC-MS (EC16364-91-P1A1, P1, RT = 0.500 min) indicated the complete reaction of L5-2. The reaction mixture was concentrated under reduced pressure to obtain the crude product, which was further purified by preparative HPLC (FA conditions) to afford L5-3 (20.0 mg, 14.1 μmol, 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 μmol, 1.0 equiv) was mixed with HCl / ethyl acetate (4 M, 3.53 μL) and stirred at 25°C under a N2 atmosphere for 1 hour. LC-MS (EC16364-100-P1A1, P1, RT = 0.413 min) indicated complete reaction of L5-3. The reaction mixture was concentrated under reduced pressure to yield the crude product, which was further purified by preparative HPLC (HCl column chromatography) to yield APL-5 (13.1 mg, 9.07 μmol, 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(400MHz DMSO)δppm 9.26(s,1H)8.80-8.93(m,1H)8.74(d,J=8.0Hz,1H)8.46-8.51(m,1H)8.04(d,J=7.6 Hz,1H)7.79(d,J=7.6Hz,1H)7.68(d,J=7.6Hz,1H)7.54-7.62(m,3H)7.39-7.50(m,4 H)7.31-7.39(m,3H)7.26(dd,J=8.0,2.63Hz,2H)7.12(t,J=8.8Hz,1H)6.97(d,J=9. 2Hz, 1H) 5.38 (dd, J = 7.6, 3.31Hz, 1H) 5.32 (t, J = 4.4Hz, 1H) 4.92 (s, 2H) 4.47 (t, J = 7. 6Hz,1H)4.15-4.19(m,2H)4.03(t,J=6.4Hz,2H)3.85(dd,J=11.6,5.50Hz,6H)3.77- 3.80(m,4H)3.33(s,4H)2.98(t,J=5.6Hz,2H)2.78-2.83(m,2H)2.47(s,2H)2.46(s, 1H)2.15-2.24(m,2H)2.00(dd,J=15.2,7.63Hz,8H)1.60-1.70(m,4H)1.56(d,J=8.4 Hz,2H)1.41-1.49(m,2H)1.34(d,J=6.8Hz,3H)1.03-1.16(m,6H)0.83-0.88(m,2H).

[0376] Synthesis of APL-6:

[0377] The synthesis steps of PROTAC molecule APL-6 refer to APL-5, but the starting materials are replaced with compound 3 and L6-1. After synthesis and purification, the target product APL-6 (11.0 mg, 7.32 μmol, 53.5% yield, 90.6% purity) is obtained as a white solid.

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

[0379] 1H NMR:EC16364-101-P1C(400MHz DMSO)δppm 9.24-9.36(m,1H)8.80-8.94(m,1H)8.48(s,1H)8.04(d,J=7.6Hz,1H)7.79(d,J=7.6Hz,1H)7.6 8(d,J=7.6Hz,1H)7.54-7.62(m,3H)7.40-7.50(m,4H)7.33-7.38(m,2H)7.23-7.31(m,2H)7.13 (t,J=8.8Hz,1H)6.97(d,J=8.8Hz,1H)5.38(dd,J=7.6,3.06Hz,1H)5.32(t,J=4.8Hz,1H)4.92( s,2H)4.47(t,J=7.6Hz,1H)4.14-4.17(m,2H)4.01-4.04(m,2H)3.87(d,J=5.6Hz,4H)3.82(d,J= 4.4Hz,4H)3.76(br s,4H)3.59-3.61(m,4H)3.57(s,4H)3.34(d,J=1.2Hz,2H)2.98(t,J=5.6Hz,2H)2. 80(t,J=7.2Hz,2H)2.46(s,1H)2.33(s,3H)2.19-2.23(m,1H)2.02(d,J=7.2Hz,2H) 1.98(d,J=7.6Hz,2H)1.64(d,J=8.0Hz,2H)1.55(dd,J=8.0,1.69Hz,2H)1.43-1.4 8(m,1H)1.35(d,J=6.8Hz,3H)1.23(s,8H)1.08(d,J=6.4Hz,2H)0.83-0.86(m,1H).

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

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

[0382] Synthesis of APF1:

[0383] 1. Synthesis of compound APF1-2

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

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

[0386] 2. Synthesis of Compound APF1

[0387] Compound A1331852 (60.0 mg, 91.0 μmol, 1.00 equiv), APF1-2 (49.2 mg, 136 μmol, 1.50 equiv), HATU (51.9 mg, 136 μmol, 1.50 equiv), and DIEA (58.8 mg, 455 μmol, 79.3 μL, 5.00 equiv) were mixed in DCM (1 mL), purged with N2 to expel air, and stirred at 25°C for 2 hours. LC-MS (EC11272-23-P1B) analysis showed that compound A1331852 was fully reacted. The reaction solution was concentrated under vacuum to obtain a crude product, which was further purified by preparative HPLC (basic conditions) to obtain compound APF1 (21.6 mg, 20.0 μmol, 22.0% yield, 93.0% purity) as a yellow solid.

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

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

[0390] HNMR:EC11272-23-P1C,DMSO+D2O,400MHzδ11.08(s,1H),8.14(t,J=5.69Hz,1H), 7.99(d,J=7.6Hz,1H),7.77(d,J=8.0Hz,1H),7.58(d,J=7.6Hz,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=8.4Hz,1H),6.99(d,J =7.2Hz,1H),6.94(d,J=8.8Hz,1H),6.50-6.57(m,1H),4.92-5.05(m,3H),3.86(t,J=5.2Hz, 2H),3.64(s,2H),3.38(d,J=6.4Hz,4H),3.24(d,J=5.2Hz,2H),2.96-3.01(m,2H),2.70-2.9 2(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 PROTAC molecule APF2 refer to APF1, but the starting materials are replaced with compound 1 and APF2-1. After synthesis and purification, the target product APF2 is obtained as a yellow solid with a purity of 93.7% and a yield of approximately 23.9%.

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

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

[0395] HNMR:EC11272-24-P1A,DMSO,400MHzδ11.07-11.11(m,1H),8.18(t,J=5.6Hz,1H),7.97-8.02(m,1H),7.73-7.79(m,1H),7.58-7.64(m,1H) ,7.54(t,J=7.2Hz,1H),7.41-7.48(m,2H),7.29-7.41(m,3H),7.25(s,1H),7.08(d,J=8.4Hz,1H),7.01(d,J=7.2Hz,1H),6.94(d,J=8.8Hz,1 H),6.57(t,J=6.0Hz,1H),4.94-5.08(m,3H),3.89(t,J=6.0Hz,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 steps of PROTAC molecule APF3 refer to APF1, but the starting materials are replaced with compound 1 and APF3-1. After synthesis and purification, the target product APF3 is obtained as a yellow solid with a purity of 97.4% and a yield of about 11.4%.

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

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

[0400] HNMR:EC12165-1-P1C,DMSO,400MHzδ12.85(s,1H),11.09(s,1H),8.16(t,J=6 .0Hz,1H),8.01(d,J=7.6Hz,1H),7.78(d,J=8.0Hz,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 =8.8Hz,1H),7.02(d,J=7.2Hz,1H),6.94(d,J=8.8Hz,1H),6.58(t,J=5.6Hz,1H ),5.04(dd,J=12.8,5.4Hz,1H),4.97(s,2H),3.89(t,J=6.0Hz,2H),3.67(s,2 H),3.55-3.60(m,2H),3.47-3.53(m,4H),3.41-3.46(m,4H),3.38(dd,J=6.0, 3.6Hz,4H),3.22-3.26(m,2H),3.00(t,J=5.6Hz,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 steps of PROTAC molecule APF4 refer to APF1, but the starting materials are replaced with compound 1 and APF4-1. After synthesis and purification, the target product APF4 is obtained as a yellow solid with a purity of 93.4% and a yield of about 20.7%.

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

[0404] HNMR:EC12165-2-P1C,DMSO,400MHzδ12.84(s,1H),11.09(s,1H),8.17(t,J=5.6Hz ,1H),8.01(d,J=8.0Hz,1H),7.78(d,J=8.0Hz,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=8.4Hz,1H),7. 02(d,J=7.2Hz,1H),6.95(d,J=8.8Hz,1H),6.58(t,J=5.6Hz,1H),5.04(dd,J=12.8, 5.2Hz,1H),4.98(s,2H),3.89(t,J=6.0Hz,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,4 H),3.22-3.26(m,2H),3.00(t,J=5.6Hz,2H),2.87(ddd,J=17.2,14.0,5.2Hz,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). The mixture was stirred at 100°C for 20 hours. LCMS (EC11255-19-P1a, P1 RT = 0.283 min) showed that compound 1 had reacted sufficiently, with multiple new compound peaks appearing. The target compound accounted for approximately 55.5% of the detected peaks. The mixture was diluted with 10.0 mL of water and extracted with 30 mL of ethyl acetate (10.0 mL x 3). The resulting organic layer was washed with 10 mL of brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to yield the crude product. The crude product was purified by column chromatography (SiO2, PE:EA = 1 / 0 to 9 / 1) to afford compound APF1A-2 (140 mg, 345 μmol, 19.1% yield) as a pale yellow oil. LC-MS (EC11255-19-P1z, P1 RT = 0.272 min) revealed a purity of approximately 97.2%.

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

[0409] LCMS: EC11255-19-P1z, P1 RT=0.272min, 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 EDCl (116 mg, 607 μmol, 4.00 equiv) were mixed in DCM (1 mL), purged with N2, and stirred at 25°C for 16 hours. LC-MS (EC11272-31-P1A, P1, RT = 0.594 min) indicated that compound A1331852 had fully reacted. The reaction mixture was concentrated under vacuum to obtain a crude product, which was further purified by preparative HPLC (basic conditions) to afford compound APF1A (20.2 mg, 18.8 μmol, 12.4% yield, 97.3% purity) as a yellow solid.

[0412] LCMS:EC11272-31-P1A,RT=0.569min,M / Z(M / 2+H + )=524.2

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

[0414] HNMR:EC11272-31-P1C,DMSO,400MHzδ12.85(s,1H),11.08(s,1H),8.02(d,J=7.6Hz,1H),7.78(d,J=8.0Hz,1H),7.60(d,J=8.0Hz,1H),7.50- 7.56(m,2H),7.40-7.48(m,2H),7.35(q,J=7.6Hz,2H),7.24(s,1H),7.08(d,J=8.2Hz,1H),6.94-7.03(m,2H),6.56(t,J=5.6Hz,1H),5.00-5. 07(m,1H),4.95(s,2H),4.02-4.09(m,2H),3.84(t,J=5.6Hz,2H),3.68(s,2H),3.56(t,J=5.2Hz,2H),3.45-3.49(m,2H),3.40(d,J=5.2Hz,4H ),3.29(s,2H),3.00(t,J=5.6Hz,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) Synthetic PROTAC molecules APR1, APR2, APR3, and 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 equiv) and compound APR1-1 (538 mg, 5.43 mmol, 1.00 equiv) in DMSO (15.0 mL) was added DIPEA (1.40 g, 10.8 mmol, 1.89 mL, 2.00 equiv), and the mixture was stirred at 70°C for 16 hours. LC-MS (EC7111-89-p1a1, rt[P1] = 0.362 min) showed complete reaction of compound 1, with the appearance of multiple new compound peaks. The target compound accounted for approximately 35.3% of the detected reaction. The reaction mixture was diluted with 30.0 mL of water and extracted with 60 mL of ethyl acetate (20.0 mL×3 times). The resulting organic layer was washed with 30 mL of brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give a crude product, which was purified by column chromatography (SiO2, PE:EA=5 / 1 to 0 / 1) and further purified by preparative HPLC (neutral conditions, 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 product) as a yellow solid.

[0419] LCMS:EC7111-89-p1a1,RT=0.362min.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 sodium (2R)-2-[(1S)-1,2-dihydroxyethyl]-4-hydroxy-5-oxo-2H-furan-3-ate (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). The mixture was purged with N2 and stirred at 25°C for 3 hours. LC-MS (EC12165-6-P1A, P1: RT = 0.508 min) showed that compound 2A was fully reacted. The reaction mixture was terminated by adding 20.0 mL of water at 0°C, diluted with 20 mL of DCM, and extracted with 60 mL of ethyl acetate (20.0 mL x 3). The resulting organic layer was washed with 20 mL of saturated brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by preparative HPLC (neutral conditions) 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) determined the purity to be 96.0%.

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

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

[0424] HNMR: EC12165-6-P1C, DMSO, 400MHz δ12.85(s,1H),11.09(s,1H),8.48(t,J=6.0Hz,1H),7.99(d,J=7.8Hz,1H),7.93(s,1H),7.76(d,J=8.0Hz ,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=8.4Hz,1H),7.02(d,J=7.2Hz,1H),6.97(d,J=8.8Hz,1H ),6.57(t,J=6.0Hz,1H),4.96-5.09(m,3H),4.55(s,2H),4.41(t,J=6.4Hz,2H),3.90(t,J=6.0Hz,2H),3.67(s,2H),3.59-3.63(m,2H),3.53- 3.57(m,2H),3.43(q,J=5.6Hz,2H),3.01(t,J=5.6Hz,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 steps of PROTAC molecule APR2 refer to APR1, but the starting materials are replaced with compound 1 and APR2-1. After synthesis and purification, the target product APR2 is obtained as a yellow solid with a purity of 96.4% and a yield of about 24.4%.

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

[0428] HNMR: EC12165-5-P1C, DMSO, 400MHz δ 12.85 (s, 1H), 11.09 (s, 1H), 8.48 (t, J = 6.0Hz, 1H), 7.99 (d, J = 8.0Hz, 1H), 7.90 (s, 1H), 7.75 (d, J = 8.4Hz, 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=8.8Hz,1H),7.02(d,J=7.2Hz,1H),6.96(d,J=8.8Hz,1H),6.58(t,J=5. 6Hz,1H),5.04(dd,J=12.8,5.2Hz,1H),4.99(s,2H),4.49(s,2H),4.39(t, J=6.0Hz,2H),3.90(t,J=6.0Hz,2H),3.67(s,2H),3.54-3.59(m,4H),3.52( s,4H),3.42(q,J=5.6Hz,2H),3.00(t,J=5.6Hz,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 steps of PROTAC molecule APR3 refer to APR1, but the starting materials are replaced with compound 1 and APR3-1. After synthesis and purification, the target product APR3 is obtained as a yellow solid with a purity of 97.7% and a yield of about 18.2%.

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

[0432] 1H NMR: EC12886-2-P1C (400MHz, DMSO) δ12.01 (s, 1H), 10.25 (s, 1H), 7.58-7.67 (m, 1H), 7.14 (d, J=7.6Hz, 1H), 7.05 (s, 1H), 6.91 (d, J= 8.0Hz,1H),6.67-6.78(m,2H),6.59(dd,J=18.0,8.4Hz,3H),6.45-6.54(m,2H),6.37(s,1H),6.26(d,J=8.4Hz,1H),6.17(d,J=7.2H z,1H),6.12(d,J=8.8Hz,1H),5.73(t,J=5.6Hz,1H),4.08-4.25(m,3H),3.62(s,2H),3.50-3.58(m,2H),3.05(t,J=5.2Hz,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 steps of PROTAC molecule APR4 refer to APR1, but the starting materials are replaced with compound 1 and APR4-1. After synthesis and purification, the target product APR4 is obtained as a light yellow solid with a purity of 92.8% and a yield of about 27.4%.

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

[0436] 1 H NMR: 1HNMR(EC12886-1-P1B4)(400MHz,DMSO)δ12.00(s,1H),10.26(s,1H), 7.65(t,J=5.6Hz,1H),7.15(d,J=5.2Hz,1H),7.07(s,1H),6.86-6.98(m,1H),6.77(d,J=7.2Hz,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=8.8Hz,1H),6.19(d,J=7.00Hz,1H),6.05-6.17(m,1H),5.76(t,J=5.6Hz,1H),4.22(dd,J=12.8, 5.2Hz,1H),4.16(s,2H),3.64(s,2H),3.57(d,J=6.6Hz,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=5.6Hz,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 hours. TLC (PE:EA=5:1, P1, R f =0.4) showed that compound 2A-1 had completely reacted, generating a major peak with a large polarity. After the reaction was completed, the mixture was returned to room temperature, and 16.5 g of KOH was gradually added in small amounts. After returning to room temperature, the mixture was extracted with 200 mL of ethyl acetate (40 mL × 5 times). The resulting organic layer was dried over Na2SO4, filtered under reduced pressure, and concentrated to obtain a crude product. The crude product was purified by column chromatography (SiO2, PE / EA = 10 / 1 to 5 / 1, PE:EA = 5:1, P1, R f =0.4) to afford compound 2A-2 (7.50 g, 40.3 mmol, 90.2% yield) as a 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), and the mixture was filled with N2 to expel air. The mixture was stirred at 25°C under N2 for 1 hour. TLC (PE:EA=5:1, R1, R f =0.4) showed that compound 2A-2 was fully reacted. The reaction solution was concentrated under vacuum to obtain a crude product, which was directly used in the next step without further purification to obtain compound 2A-3 (1.20 g, 9.79 mmol, 91.16% yield, HCl) as a white solid.

[0442] 3. Synthesis of Compound 2A

[0443] Compound A1331852 (200 mg, 303.58 μmol, 1 eq), 2-ethylazide (186 mg, 1.52 mmol, 5.00 eq, HCl), HATU (173 mg, 455 μmol, 1.5.0 eq) and DIEA (196 mg, 1.52 mmol, 264 μL, 5.00 eq) were mixed in DCM (5 mL), the mixture was purged with N2 to expel air, and the mixture was stirred at 25°C under N2 for 1 hour. LC-MS (EC11272-32-P1A, P1, RT = 0.590 min) showed that compound A1331852 was fully reacted. The reaction solution was concentrated under vacuum to obtain a 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 afford compound 2A (90 mg, 112 μmol, 36.9% yield, 90.6% purity) as an off-white solid.

[0444] LCMS:EC11272-32-P1A,RT=0.590min.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 molecule A13D6-PF3)

[0446] Synthesis of A13D-1:

[0447] 1. Synthesis of Compound D1-2

[0448] 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) were added to compound D1-1 (4.00 g, 12.8 mmol, 1.00 equiv) at 25°C, and the mixture was stirred at 110°C under a CO (2.00 MPa) atmosphere for 24 hours. TLC (petroleum ether:ethyl acetate = 2:1) analysis showed that the reaction of compound D1-1 was complete, generating a new compound (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 × 2 times). The obtained organic layer was washed with saturated brine (50.0 mL × 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 give compound D1-2 (3.61 g, 12.4 mmol, 96.7% yield) as a yellow oil.

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

[0450] 2. Synthesis of Compounds D1-3

[0451] Under N2 environment, ethyl acetate (2.00 mL) and HCl / ethyl acetate (7.00 mL) were added to compound D1-2 (1.02 g, 3.50 mmol, 1.00 equivalent) at 25°C, and the mixture was stirred at 25°C for 2 hours. TLC (petroleum ether:ethyl acetate = 3:1) showed that the reaction of compound D1-2 was complete, and a new compound (R f =0.00). After the reaction, the mixture was concentrated to obtain a crude product of compound D1-3 (0.80 g).

[0452] 3. Synthesis of Compounds D1-4

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

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

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

[0456] 4. Synthesis of Compound D1-5

[0457] Methanol (2.50 mL), NaOH (42.9 mg, 1.07 mmol, 2.00 equiv) and H2O (0.50 mL) were added to compound D1-4 (320 mg, 536 μmol, 1.00 equiv) at 25°C in sequence, and the mixture was stirred at 25°C for 12 hours. LCMS (EB9483-12-IPCL2) analysis showed that the reaction of compound D1-4 was complete, and the target molecular weight product (R t =1.038 min). Water (3.00 mL) was added to the reaction mixture, and the pH was adjusted to about 5 with acetic acid. Extraction was performed with ethyl acetate (20.0 mL × 3 times). The resulting 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 =1.038min,MS(ESI)m / z=583.3[M+H] +

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

[0460] 5. Synthesis of Compound D1-6

[0461] To compound D1-5 (150 mg, 257 μmol, 1.00 equivalent) was added CH2Cl2 (2.00 mL), EDCI (148 mg, 772 μmol, 3.00 equivalent), DMAP (94.4 mg, 772 μmol, 3.00 equivalent) and compound D1-0 (59.6 mg, 386 μmol, 1.50 equivalent) at 0°C in sequence, and the mixture was stirred at 25°C for 12 hours. LCMS (EB9483-13-IPCL2) detection showed that the reaction of compound D1-5 was complete to produce the target molecular weight product (Rt =2.057 min). After the reaction, the mixture was adjusted to pH 6 with acetic acid and extracted with CH2Cl2 (20.0 mL x 3 times). The resulting organic layer was washed with saturated brine (30.0 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%: 58%-98%, 25 min) to obtain compound D1-6 (110 mg, 139 μmol, 54.04% yield) as a yellow oil.

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

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

[0464] 6. Synthesis of Compound A13D-1

[0465] To compound D1-6 (100 mg, 139 μmol, 1.00 eq) was added CH2Cl2 (1.00 mL) and TFA (5.12 g, 44.9 mmol, 3.33 mL, 323 eq) at 25°C in sequence, and the mixture was stirred at 25°C for 12 hours. LCMS (EB9484-16-IPCL1) showed that the reaction of compound D1-6 was complete, and the target molecular weight product (R t =1.833 min). After the reaction, the mixture was concentrated to obtain a crude product, which was purified by preparative HPLC (column: Welch Xtimate C18 150*30 mm*5 μm; mobile phase: [water (FA)-ACN]; B%: 56%-96%, 25 min) to obtain compound A13D-1 (30.0 mg, 43.4 μmol, 34.64% yield, 95.8% purity) as a white solid.

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

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

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

[0469] 1 H NMR:EB9483-16-P1N1(400MHz,DMSO-d6)δppm 1.50-1.60(m,10H)1.62-1.70(m,3H)1.93(br s,3H)2.10(s,3H)2.99(br t,J=5.60Hz,2H)3.71(s,2H)3.86(t,J=6.00Hz,2H)4.88(s,2H)6.91(d,J=8.8 0Hz,1H)7.20(s,1H)7.32-7.41(m,2H)7.44-7.52(m,2H)10.69(s,1H)12.80(br s,1H)

[0470] Synthesis of A13D-2:

[0471] The synthesis steps of the deuterated molecule A13D-2 refer to A13D-1, but the starting materials are replaced with intermediate compounds D2-1 and D0-0. The target product A13D-2 is purified as a yellow solid with a purity of 99.1% and a yield of approximately 18.8%.

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

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

[0474] 1HNMR: EB9776-40-P1N5 (400MHz, DMSO-d6) δ8.21 (br s, 1H), 8.03 (br d, J = 7.6Hz, 1H), 7.79 (br d, J = 8.0Hz, 1H), 7.62 (br d,J=7.6Hz,1H),7.51-7.32(m,5H),7.28(s,1H),6.89(br d,J=8.8Hz,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 steps of the deuterated molecule A13D-5 refer to A13D-1, but the starting materials are replaced with intermediate compounds D5-1 and D0-0. The target product A13D-5 is purified as a yellow solid with a purity of 98.9% and a yield of approximately 42.9%.

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

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

[0479] 1 HNMR: EB9776-26-P1N3 (400MHz, DMSO-d6) δ13.27-12.28 (m, 1H), 8.04 (d, J = 7.6Hz, 1H) ,7.79(d,J=8.0Hz,1H),7.62(d,J=7.6Hz,1H),7.52-7.32(m,5H),7.29(s,1H),6.92(br d,J=8.8Hz,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 steps of the deuterated molecule A13D-6 refer to A13D-1, but the starting materials are replaced with intermediate compounds D6-1 and D0-0. The target product A13D-6 is purified as a white solid with a purity of 98.4% and a yield of approximately 18.8%.

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

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

[0484] 1 HNMR: EB9776-48-P1N1 (400MHz, DMSO-d6) 8.15 (s, 1H), 8.04 (d, J = 7.6Hz, 1H), 7.79 (d, J = 8.0Hz, 1H), 7.61 (d, J = 7.2Hz, 1 H),7.52-7.32(m,5H),7.27(s,1H),6.93(d,J=8.8Hz,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] Under N2 environment, 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) were added to compound 3A-1 (11.0 g, 66.2 mmol, 1.00 equiv) at 25°C, and the mixture was stirred at 25°C for 36 hours. LCMS (EB8953-61-IPCL4) detection showed that the reaction of compound 3A-1 was complete to produce the target molecular weight product (R t =1.344 min). After the reaction, the mixture was concentrated to obtain a crude product, which was purified by column chromatography (ethyl acetate: petroleum ether = 5:1, Rf = 0.71) to obtain compound 3A-3 as a white solid.

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

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

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

[0491] DIPA (6.24 g, 61.7 mmol, 8.72 mL, 1.10 equivalents) was added to THF (160 mL) at 25°C under N2 environment, and n-BuLi (2.50 M, 29.2 mL, 1.30 equivalents) was added at -78°C and stirred for 30 minutes. Compound 3A-3 (19.2 g, 56.1 mmol, 1.00 equivalents) dissolved in THF (80.0 mL) was added at -78°C and stirred for 1 hour. MeI (9.08 g, 64.0 mmol, 3.98 mL, 1.14 equivalents) was added at -78°C and stirred for 4 hours. LCMS (EB8953-61-IPCL4) detection showed that the reaction of compound 3A-3 was complete to produce the target molecular weight product (R t =1.130 min). Saturated NH4Cl solution was added to the reaction mixture at 0°C to terminate the reaction. Water (100 mL) and ethyl acetate (200 mL) were added, and extraction was performed with ethyl acetate (200 mL x 2). The resulting organic layer was washed with saturated brine (100 mL), dried over Na2SO4, filtered, and concentrated to obtain the crude product. The crude product was triturated in MeOH (100 mL) at 25°C to obtain the crude product of compound 3A-4 as a white solid.

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

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

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

[0495] Toluene (100 mL), compound 3A-4 (15.6 g, 43.8 mmol, 1.00 equivalent) and triisopropyl borate (14.8 g, 78.8 mmol, 18.1 mL, 1.80 equivalent) were added to THF (100 mL) at 25°C under N2 environment. n-BuLi (2.50 M, 26.3 mL, 1.50 equivalent) was added at -78°C and stirred for 4 hours. 2,3-dimethylbutane-2,3-dione (7.76 g, 65.7 mmol, 1.50 equivalent) dissolved in toluene (80.0 mL) was added at -78°C. The mixture was slowly heated to 20°C and maintained for 1 hour. The reaction was stirred at 25°C for 12 hours. LCMS (EB8953-69-IPCL1) detection showed that the reaction of compound 3A-4 was complete to produce the target molecular weight product (R t =1.145 min). Saturated NH4Cl solution was added to the reaction mixture at 0°C to terminate the reaction. Water (100 mL) and THF (200 mL) were added, and extraction was performed with THF (300 mL). The resulting organic layer was washed with saturated brine (200 mL), dried over Na2SO4, filtered, and concentrated to obtain a crude product. The crude product was recrystallized from DMSO (180 mL) at 25°C to obtain compound 3A-6 as a white solid.

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

[0497] 1 H NMR: EB8953-69-P1N2 (400MHz, CDCl3) δppm 1.31 (s, 12H), 1.56-1.72 (m, 12H), 1.97 (br s, 3H), 2.42 (s, 3H), 3.71 (s, 2H), 7.69 (s, 1H)

[0498] 4. Synthesis of Compound 3A

[0499] Under N2 environment, 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-dimethylaniline dichloropalladium (556 mg, 786 μmol, 556 μL, 0.10 equiv) were added to compound 3A-7 (2.76 g, 9.43 mmol, 1.20 equiv) at 25°C. The mixture was stirred at 80°C for 12 hours. TLC (petroleum ether: ethyl acetate = 3:1) showed that the reaction of compound 3A-6 was complete, and a new compound (R f =0.56). Water (200.0 mL) was added to the reaction mixture, and ethyl acetate was used for extraction (200.0 mL×2 times). 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 give compound 3A (2.70 g, 6.11 mmol, 77.7% yield) as a white solid.

[0500] 1 H NMR:EB9483-4-P1N1(400MHz,DMSO-d6)δppm 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=8.40Hz,1H)7.85(d,J=8.40Hz,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 equivalent), dimethyl malonate (19.2 g, 145 mmol, 16.7 mL, 1.50 equivalent) and K2CO3 (33.5 g, 242 mmol, 2.50 equivalent) were mixed in DMF (100 mL) at 25°C and stirred at 60°C for 12 hours. TLC (petroleum ether: ethyl acetate = 3:1) showed that the reaction of compound D2-1-1 was complete, and a new compound (R f=0.39). Water (200 mL) and ethyl acetate (200 mL) were added to the reaction mixture, the organic layer was separated, and extracted with ethyl acetate (100 mL × 2 times). The resulting organic layer was washed with saturated brine (200 mL), dried over Na2SO4, filtered, and concentrated to obtain a 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 (400MHz, DMSO-d6) δ7.92 (dd, J=1.2, 8.1Hz, 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 equivalent) was added ethyl acetate (190 mL) and LiI (81.8 g, 611 mmol, 23.4 mL, 10.0 equivalent) at 25°C, and the mixture was stirred at 80°C for 4 hours. TLC (petroleum ether: ethyl acetate = 2:1) showed that the reaction of compound D2-1-2 was complete, and a new compound (R f =0.19). After the reaction, the mixture was quenched by adding citric acid (100 mL) and extracted with ethyl acetate (200 mL × 2 times). The resulting 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 = 2:1, R f =0.19) and purified by column chromatography (ethyl acetate / petroleum ether = 0-10%) to give compound D2-1-3 (5.60 g, 23.3 mmol, 38.2% yield) as a white solid.

[0507] 1 H NMR: EB9915-4-P1N1 (400MHz, DMSO-d6) δ12.82 (br s, 1H), 7.79 (dd, J=1.2, 7.9Hz, 1H), 7.64-7.50 (m, 2H), 3.90 (s, 2H)

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

[0509] Compound D2-1-3 (1.00 g, 4.17 mmol, 1.00 equiv) and K2CO3 (691 mg, 5.00 mmol, 1.20 equiv) were added to DMF at 25°C, and MeI (887 mg, 6.25 mmol, 389 μL, 1.50 equiv) was added at 0°C. The mixture was stirred at 25°C for 1 hour. TLC (petroleum ether: ethyl acetate = 1:1) showed that the reaction of compound D2-1-3 was complete, and a new compound main peak (Rf = 0.40) was generated. After the reaction, the mixture was quenched by adding water (10.0 mL) and extracted with ethyl acetate (20.0 mL × 2 times). The resulting organic layer was washed with saturated brine (20.0 mL × 3 times), dried over Na2SO4, filtered, and concentrated to obtain a crude product. The crude product (TLC: petroleum ether: ethyl acetate = 1:1, R f =0.40) was purified by column chromatography (ethyl acetate / petroleum ether = 0-20%) to give compound D2-1-4 (640 mg, 2.52 mmol, 60.5% yield) as a white solid.

[0510] 1 H NMR: EB9915-10-P1N1 (400MHz, DMSO-d6) δ7.81 (dd, J=1.2, 8.0Hz, 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) was added THF (1.00 mL) at 25°C, and KCO (17.4 mg, 126 μmol, 0.05 equiv) dissolved in D2O (1.01 g, 50.4 mmol, 20.0 equiv) was added at 0°C, and the mixture was stirred at 25°C for 12 hours. The reaction mixture was added with D2O (5.00 mL) and extracted with ethyl acetate (20.0 mL × 2 times). The resulting organic layer was dried over Na2SO4, filtered, and concentrated to give the crude product of compound D2-1-5 (590 mg, 2.30 mmol, 91.5% yield) as a white solid.

[0513] 1 H NMR: EB9915-11-P1N1 (400MHz, DMSO-d6) δ7.81 (dd, J=1.2, 7.9Hz, 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] BD3.THF (0.75M, 7.65mL, 3.00 equivalents) was added to compound D2-1-5 (490mg, 1.91mmol, 1.00 equivalents) at 0°C under N2 environment, and the mixture was stirred at 25°C for 12 hours. LCMS (EB9776-22-IPCL1) showed that compound D2-1-5 was fully reacted to produce the target molecular weight product (R t =0.335 min). CD3OD (15.0 mL) was added to the reaction mixture, and the mixture was stirred at 80°C for 16 hours. The mixture was concentrated to obtain a crude product of compound D2-1-6 (2.30 g) as a yellow oil.

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

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

[0518] Methanol (15.0 mL), DIEA (1.62 g, 12.5 mmol, 2.18 mL, 1.20 equiv) and (Boc)2O (2.27 g, 10.4 mmol, 2.39 mL, 1.00 equiv) were added to compound D2-1-6 (2.46 g, 10.4 mmol, 1.00 equiv) and stirred at 25°C for 16 hours. LCMS (EB9776-30-IPCL1) detection showed that compound D2-1-6 was fully reacted to produce the target molecular weight product (R t =0.657min). After the reaction, the mixture was concentrated to obtain a crude product. The crude product (petroleum ether:ethyl acetate=1:1, R f =0.25) and purified by column chromatography (ethyl acetate:petroleum ether=0-20%) to give compound D2-1-7 (250 mg, 743 μmol, 7.14% yield).

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

[0520] 1H NMR: EB9776-30-P1N1 (400MHz, DMSO-d6) δ7.46 (dd, J=1.2, 7.9Hz, 1H), 7.20 (dd, J=1.2, 7.6Hz, 1H), 7.14-7.09 (m, 1H), 5.20 (br s, 1H), 4.52 (br d,J=5.6Hz,1H),3.80(s,1H),3.12-3.03(m,1H),1.45(s,9H)

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

[0522] Under N2 environment, DCM (15.0 mL) and 4-methylmorpholine (749 mg, 7.41 mmol, 814 μL, 3.00 equiv) were added to compound D2-1-7 (830 mg, 2.47 mmol, 1.00 equiv) at 0°C, stirred at 0°C for 0.5 hours, and methylsulfonyl chloride (650 mg, 5.67 mmol, 439 μL, 2.30 equiv) was added, and stirred at 0°C for 2 hours. LCMS (EB9776-32-IPCL1) detection showed that compound D2-1-7 was fully reacted to produce the target molecular weight product (R t =0.613min). Water (10.0 mL) was added to the reaction mixture, and the mixture was extracted with CH2Cl2 (10.0 mL×3 times). The obtained organic layer was washed with saturated brine (10.0 mL), dried over Na2SO4, filtered, and concentrated to obtain a crude product. The crude product (petroleum ether:ethyl acetate=1:1, R f =0.34) was purified by column chromatography (ethyl acetate:petroleum ether = 0-20%) to give compound D2-1-8 (780 mg, 1.88 mmol, 76.3% yield) as a yellow oil.

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

[0524] 1 H NMR: EB9776-32-P1N1 (400MHz, DMSO-d6) δ7.51 (dd, J=1.2, 8.0Hz, 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 atmosphere, and the mixture was stirred at 25°C for 3 hours. LCMS (EB9776-34-IPCL1) showed that the target molecular weight product (R t =0.813 min). Water (10.0 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (10.0 mL×3 times). The obtained organic layer was washed with saturated brine (10.0 mL), dried over Na2SO4, filtered, and concentrated to obtain the product D2-1 (590 mg) as a yellow oil.

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

[0528] 1 H NMR: EB9776-34-P1N1 (400MHz, CHLOROFORM-d) δ7.42 (dd, J=1.2, 7.6Hz, 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 equivalent) dissolved in CCl4 (500 mL), peroxybenzoate (4.76 g, 19.6 mmol, 0.10 equivalent) and NBS (35.0 g, 196 mmol, 1.00 equivalent) were added at 25 ° C under N2 environment, and the reaction was stirred at 80 ° C for 12 hours. TLC (petroleum ether: ethyl acetate = 5:1) detection showed that the reaction of compound D5-1-1 was complete, and a new compound main peak (R f=0.48). The reaction mixture was returned to room temperature and filtered. The filtrate was washed with saturated brine (600 mL), dried over Na2SO4, filtered under reduced pressure, and concentrated to obtain a crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 100:1 to 10:1) to obtain compound D5-1-2 (39.0 g, 127 mmol, 64.5% yield) as a colorless oil.

[0532] 1 H NMR: EB9495-2-P1N1 (400MHz, DMSO-d6) δ7.69 (dd, J=0.8, 8.1Hz, 1H), 7.59 (d, J=7.6Hz, 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 equivalent) and NaCN (6.43 g, 131 mmol, 1.19 equivalent) were mixed in DMF (170 mL) at 25 ° C and stirred at 60 ° C for 6 hours. TLC (petroleum ether: ethyl acetate = 10: 1) detection showed that the reaction of compound D5-1-2 was complete, and a new compound main peak was generated. The reaction mixture was terminated by adding water (300 mL) and extracted with ethyl acetate (200 mL × 2 times). The organic layer was washed with saturated brine (200 mL), dried over Na2SO4, filtered under reduced pressure, and concentrated to give a crude product. The crude product was subjected to TLC detection (petroleum ether: ethyl acetate = 10: 1, R f =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.

[0535] 1 H NMR: EB9527-2-P1N1 (400MHz, DMSO-d6) δ7.73 (d, J=8.0Hz, 1H), 7.57-7.52 (m, 1H), 7.51-7.45 (m, 1H), 4.05 (s, 2H), 3.91 (s, 3H)

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

[0537] To a mixture of compound K2CO3 (136 mg, 984 μmol, 0.05 eq) dissolved in D2O (7.88 g, 394 mmol, 20.0 eq), compound D5-1-3 (5.00 g, 19.7 mmol, 1.00 eq) and THF (10.0 mL) were added at 25°C under a N2 atmosphere, and the reaction was stirred at 25°C for 12 hours. HNMR analysis showed that the reaction of compound D5-1-3 was complete to produce the target product. The reaction mixture was transferred to a separatory funnel to obtain the aqueous phase, which was extracted with MTBE (10.0 mL × 2 times). The resulting organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to obtain crude compound D5-1-4 (4.80 g, 10.0 mmol, 50.9% yield, 53.4% ​​purity) as a colorless oil.

[0538] 1 H NMR: EB9495-10-P1N1 (400MHz, DMSO-d6) δ7.73 (dd, J=1.2, 8.0Hz, 1H), 7.56-7.52 (m, 1H), 7.50-7.45 (m, 1H), 3.91 (s, 3H)

[0539] EB9495-10-P1N2(400MHz,DMSO-d6)δ7.73(dd,J=1.2,7.9Hz,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 equiv) was added BH3.THF (1.00 M, 77.3 mL, 6.00 equiv) at 0°C under N2 atmosphere, and the mixture was stirred at 25°C for 16 hours. LCMS (EB9776-9-IPCL2) showed that the reaction of compound D5-1-4 was complete, and a new compound (R t =1.070min). The reactant was transferred to a new bottle, methanol (155mL) was added thereto, and the reaction was stirred at 80°C for 16 hours. The mixture was concentrated under reduced pressure to obtain a crude product (dichloromethane:methanol=10:1, R f =0.20), and purified by column chromatography (petroleum ether:ethyl acetate=1:0 to 1:1) to give compound D5-1-5 (1.69 g, 7.41 mmol, 57.5% yield) as a yellow oil.

[0542] 1H NMR: EB9776-9-P1N1 (400MHz CHLOROFORM-d) δ7.61 (dd, J=1.6, 7.8Hz, 1H), 7.25-7.16 (m, 2H), 6.54 (br s, 1H), 3.48 (d, J=3.6Hz, 2H)

[0543] LCMS:EB9776-9-IPCL2, Product:R T =1.070min,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), BH3.THF (1.00 M, 51.9 mL, 7.00 equiv) was added at 0°C under N2 environment and stirred at 25°C for 16 hours, and then BH3.THF (1.00 M, 22.2 mL, 3.00 equiv) was added at 0°C and stirred at 25°C for 16 hours, and then BH3.THF (1.00 M, 14.8 mL, 2.00 equiv) was added at 0°C and stirred at 25°C for 4 hours, and then BH3.THF (1.00 M, 22.2 mL, 3.00 equiv) was added at 0°C and stirred at 25°C for 4 hours, and then BH3.THF (1.00 M, 14.8 mL, 2.00 equiv) was added at 0°C and stirred at 25°C for 4 hours. LCMS (EB9776-11-IPCL15) detection showed that the target molecular weight product (R t =0.441 min). The reactant was transferred to a new bottle, methanol (220 mL) was added, and the mixture was stirred at 80°C for 16 hours. The mixture was concentrated under reduced pressure to obtain the crude product, compound D5-1-6 (2.62 g), as a yellow oil.

[0546] LCMS:EB9776-11-IPCL15, Product:R T =0.441min,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) were added DIEA (1.90 g, 14.7 mmol, 2.56 mL, 1.20 equiv) and (Boc)2O (2.67 g, 12.2 mmol, 2.81 mL, 1.00 equiv) and stirred at 25°C for 16 hours. LCMS (EB9776-14-IPCL2) analysis showed that the reaction of compound D5-1-6 was complete to produce the target molecular weight product (R t =1.803min). The mixture was concentrated under reduced pressure to obtain a crude product. The crude product (petroleum ether:ethyl acetate=5:1, R f =0.62) was purified by column chromatography 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 =1.803min,MS(ESI)m / z=214.0[M-Boc+H] +

[0550] 1 H NMR: EB9776-14-P1N1 (400MHz CHLOROFORM-d) δ7.42 (dd, J=0.8, 7.7Hz, 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] Compound D6-1-1 (10.0 g, 50.0 mmol, 1.00 equivalent), dimethyl malonate (9.91 g, 75.0 mmol, 8.59 mL, 1.50 equivalent) and K2CO3 (17.3 g, 125 mmol, 2.50 equivalent) were mixed in DMF (50.0 mL) at 25°C and stirred at 60°C for 16 hours. TLC (petroleum ether:ethyl acetate = 3:1) showed that the reaction of compound D6-1-1 was complete, and a new compound (R f=0.53). Water (200 mL) and ethyl acetate (200 mL) were added to the reaction mixture, the organic layer was separated, and extracted with ethyl acetate (100 mL × 2 times). The resulting organic layer was washed with saturated brine (200 mL), dried over Na2SO4, filtered, and concentrated to obtain a 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 (400MHz, DMSO-d6) δ7.91 (d, J=7.6Hz, 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 equivalent) was added ethyl acetate (70 mL) and LiI (28.7 g, 215 mmol, 8.23 ​​mL, 10.0 equivalent) at 25°C, and the mixture was stirred at 80°C for 4 hours. TLC (petroleum ether: ethyl acetate = 3:1) showed that the reaction of compound D6-1-2 was complete, and a new compound (R f =0.34). After the reaction, the mixture was quenched by adding citric acid (3.0 mL) and extracted with ethyl acetate (20 mL × 2 times). The resulting organic layer was washed with saturated brine (10 mL), dried over Na2SO4, filtered, and concentrated to obtain a crude product. The crude product (TLC: petroleum ether: ethyl acetate = 3:1, R f =0.34) and purified by column chromatography (ethyl acetate / petroleum ether = 0-10%) to give compound D6-1-3 (3.90 g, 16.3 mmol, 75.7% yield) as a white solid.

[0557] 1 H NMR: EB9495-20-P1N1 (400MHz, DMSO-d6) δ7.79 (dd, J=0.8, 7.9Hz, 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 equivalent) was added THF (10.0 mL) at 0°C, and trideuterated borane (0.75 M, 13.2 mL, 1.40 equivalent) was added at 25°C, and the mixture was stirred at 0°C for 4 hours. TLC (petroleum ether: ethyl acetate = 1:1) showed that the reaction of compound D6-1-3 was complete, and a new compound (R f =0.41). After the reaction, the mixture was concentrated to obtain a crude product (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 give 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 =1.108min,MS(ESI)m / z=227.8[M+H] +

[0561] 1 H NMR: EB9495-21-P1N1 (400MHz, DMSO-d6) δ7.72 (dd, J=0.8, 7.8Hz, 1H), 7.62-7.47 (m, 2H), 4.81 (s, 1H), 2.95 (s, 2H)

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

[0563] Under N2 environment, THF (3.00 mL) was added to compound D6-1-4 (400 mg, 1.75 mmol, 1.00 equivalent) at 25°C, BH3.THF (1.00 M, 5.26 mL, 3.00 equivalent) was added at 0°C, and the mixture was stirred at 25°C for 10 hours. BH3.THF (1.00 M, 3.51 mL, 2.00 equivalent) was added at 0°C, and the mixture was stirred at 25°C for 10 hours. BH3-Me2S (10.0 M, 5.00 mL, 28.5 equivalent) was added at 0°C, and the mixture was stirred at 25°C for 10 hours. LCMS (EB9495-38-IPCL10) detection showed that the reaction of compound D6-1-4 was complete to produce the target molecular weight product (R T = 0.965 min). Methanol (10.0 mL) was added to the mixture at 80°C, and the reaction was terminated after 12 hours. The mixture was concentrated to obtain compound D6-1-5 (820 mg, crude product) as a yellow oil.

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

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

[0566] Under N2 environment, THF (5.00 mL), DIEA (548 mg, 4.24 mmol, 738 μL, 1.20 equiv) and (Boc)2O (771 mg, 3.53 mmol, 812 μL, 1.00 equiv) were added to compound D6-1-5 (820 mg, 3.53 mmol, 1.00 equiv) at 25°C, and the mixture was stirred at 25°C for 4 hours. TLC (petroleum ether: ethyl acetate = 1:1) showed that the reaction of compound D6-1-5 was complete, and a new compound (R f =0.30). After the reaction, the mixture was concentrated to obtain a crude product (TLC: petroleum ether: ethyl acetate = 1:1, R f =0.30) was purified by column chromatography (ethyl acetate:petroleum ether = 0-20%) to give compound D6-1-6 (150 mg, 452 μmol, 12.8% yield) as a yellow oil.

[0567] 1 H NMR: EB9495-42-P1N4 (400MHz, DMSO-d6) δ7.45 (d, J=7.6Hz, 1H), 7.23 (d, J=7.6Hz, 1H), 7.17-7.11 (m, 1H), 6.90 (br s, 1H), 4.74-4.67 (m, 1H), 4.33 (br d,J=4.8Hz,2H),2.85(s,2H),1.44-1.36(m,11H)

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

[0569] Under N2 environment, DCM (10.0 mL) and 4-methylmorpholine (365 mg, 3.61 mmol, 397 μL, 3.00 equiv) were added to compound D6-1-6 (400 mg, 1.20 mmol, 1.00 equiv) at 0°C, and stirred at 0°C for 0.5 hours. Methanesulfonyl chloride (340 mg, 2.97 mmol, 230 μL, 2.47 equiv) was added at 0°C, and stirred at 0°C for 2 hours. LCMS (EB9776-41-IPCL1) detection showed that the reaction of compound D6-1-6 was complete to produce the target molecular weight product (R T=0.702min). Ice water (10.0 mL) was added to the reaction mixture, and the mixture was extracted with CH2Cl2 (10.0 mL×3 times). The obtained organic layer was washed with saturated brine (10 mL), dried over Na2SO4, filtered, and concentrated to obtain a crude product. The crude product (TLC: petroleum ether: ethyl acetate = 1:1, R 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 =0.702min,MS(ESI)m / z=354.0[Mt-Bu+H] +

[0571] 1 H NMR: EB9776-41-P1N1 (400MHz, CHLOROFORM-d) δ7.51 (dd, J=0.8, 7.9Hz, 1H), 7.23 (d, J=7.2Hz, 1H), 7.16-7.10 (m, 1H), 5.10 (br s,1H),4.51(d,J=6.0Hz,2H),3.31(s,2H),2.94(s,3H),1.47-1.42(m,9H)

[0572] 7. Synthesis of Compound D6-1

[0573] Under N2 environment, THF (8.00 mL) and NaH (48.3 mg, 1.21 mmol, 60.0% purity, 1.10 eq) were added to compound D6-1-7 (450 mg, 1.10 mmol, 1.00 eq) at 0°C and stirred at 25°C for 3 hours. LCMS (EB9776-42-IPCL2) showed that the reaction of compound D6-1-7 was complete to produce the target molecular weight product (R T Water (10.0 mL) was added to the mixture, and the mixture was extracted with ethyl acetate (10.0 mL×3 times). The obtained organic layer was washed with saturated brine (10 mL), dried over Na2SO4, filtered, and concentrated to obtain crude compound D6-1 (330 mg, 1.05 mmol, 95.8%) as a yellow oil.

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

[0575] 1 H NMR: EB9776-42-P1N1 (400MHz, CHLOROFORM-d) δ7.42 (d, J=7.6Hz, 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] Under N2 environment, water (60.0 mL), compound D1-0-1 (10.0 g, 78.0 mmol, 1.00 equivalent), Fe (21.8 g, 390 mmol, 5.00 equivalent) and NH4Cl (20.9 g, 390 mmol, 5.00 equivalent) were added to ethanol (180 mL) at 25°C, and the mixture was stirred at 60°C for 12 hours. TLC (petroleum ether: ethyl acetate = 10:1) showed that the reaction of compound D1-0-1 was complete, and a new compound (R f =0.15). After the reaction, the mixture was returned to room temperature and filtered. The filter residue was washed with ethyl acetate (500 mL × 2 times), extracted with ethyl acetate (300 mL × 2 times), and the obtained organic layer was washed with saturated brine (500 mL), dried over Na2SO4, filtered, and concentrated to obtain a crude product. The crude product (TLC: petroleum ether: ethyl acetate = 10:1, R f =0.15) and 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 (400MHz, DMSO-d6) δppm 4.98 (br s, 2H).

[0580] 2. Synthesis of Compound D1-0

[0581] Under N2 environment, KSCN (4.98 g, 51.3 mmol, 4.98 mL, 3.00 equivalents) and compound D1-0-2 (2.00 g, 17.1 mmol, 1.96 mL, 1.00 equivalents) were added to acetic acid (30.0 mL) at 10 ° C. and stirred for 30 minutes. Br2 (2.73 g, 17.1 mmol, 881 μL, 1.00 equivalents) dissolved in acetic acid (10.0 mL) was then added at 10 ° C. The mixture was stirred at 25 ° C for 6 hours. LCMS (EB8953-44-IPCL2) detection showed that the reaction of compound D1-0-2 was complete to produce the target molecular weight product (R t =1.068min). After the reaction, the mixture was adjusted to pH 8 with ammonia (50%), water (100mL) was added, and extraction was performed with ethyl acetate (100mL×2 times). The obtained organic layer was washed with saturated brine (100mL), dried over Na2SO4, filtered, and concentrated to obtain a crude product. The crude product (TLC: petroleum ether:ethyl acetate = 2:1, R f =0.54) and purified by column chromatography (ethyl acetate / petroleum ether = 0-10%) to give 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 (400MHz, DMSO-d6) δppm 5.78 (s, 2H).

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

[0585] The synthesis steps of the deuterated PROTAC molecule A13D6-PF3 refer to the undeuterated PROTAC molecule APF3, but the starting material A-1331852 is replaced with the deuterated molecule A13D-6. The final product is purified by reverse phase HPLC (neutral conditions) 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=0.567min,M / Z(M+H + )=1091.4

[0587] 1HNMR:EC15866-4-P1A(400MHz,DMSO-d6)δppm 12.84(s,1H)11.08(s,1H)8.17(t,J=5.6Hz,1H)8.01(d,J=7.6Hz,1H)7.77(d,J=8.4Hz,1H)7.52-7.62(m,2H)7.43-7.48(m,2H)7.38-7.4 2(m,1H)7.31-7.37(m,2H)7.25(s,1H)7.10(d,J=8.4Hz,1H)6.99-7.05(m,1H)6.91-6.98(m,1H)6.58(t,J=5.6Hz,1H)5.04(dd,J=12.8,5. 32Hz,1H)4.93-5.01(m,2H)3.67(s,2H)3.55-3.62(m,2H)3.47-3.53(m,4H)3.41-3.46(m,4H)3.37-3.40(m,2H)3.33-3.36(m,2H)3.22-3 .26(m,2H)2.95-3.01(m,3H)2.81-2.92(m,1H)2.58(s,2H)2.05(s,3H)1.98-2.02(m,1H)1.90(s,3H)1.60-1.65(m,3H)1.49-1.56(m,9H).

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

[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 equiv) in DMF (20.0 mL) were added DMAP (22.3 mg, 182 μmol, 0.05 equiv), EDCI (769 mg, 4.01 mmol, 1.1 equiv), and acrylic acid (525 mg, 7.29 mmol, 500 μL, 2.0 equiv). The mixture was stirred at 25°C for 8 hours. LC-MS (EC16331-35-P1G, P1: RT = 0.390 min, R1: RT = 0.341 min) showed that the reaction of compound 5A was complete, yielding a major peak with the expected m / z. After the reaction, the mixture was concentrated under reduced pressure to remove the solvent to obtain a crude product, which was further purified by reverse phase HPLC (0.1% NH 3 · H 2 O) column chromatography to obtain compound Q5a-2 (200 mg, 609 μmol, 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 μmol, 1.0 equiv) in THF (2.00 mL) were added TEA (216 mg, 2.13 mmol, 296 μL, 10 equiv) and compound Q5a-2 (70.0 mg, 213 μmol, 1.0 equiv). The mixture was stirred at 25°C for 16 hours. LC-MS (EC16331-41-P1E, P1: RT = 0.413 min) showed complete reaction of compound Q5a-2, yielding a major peak with the expected m / z. After the reaction, the mixture was concentrated under reduced pressure to remove the solvent to obtain a crude product, which was further purified by preparative HPLC (column: Phenomenex luna C18 150*25 mm*10 μm; mobile phase: [water (FA)-ACN]; gradient elution: 31%-51% B over 15 minutes) to obtain compound APQ5a (20.0 mg, 17.9 μmol, 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(400MHz DMSO)δppm 10.27(s,1H),8.20(s,1H),8.03(d,J=7.8Hz,1H),7.79(d,J=7.8Hz,1H),7.59(d,J=6.6Hz,1H),7.44-7.52(m,2H),7.38-7.42( m,1H),7.32-7.37(m,2H),7.26(dd,J=12.0,2.0Hz,1H),7.14-7.19(m,3H),7.05-7.11(m,1H),6.94(d,J=9.2Hz,2H),6.89(d,J= 8.8Hz,1H),4.91(s,2H),4.00(t,J=6.4Hz,2H),3.85(t,J=5.6Hz,2H),3.69(t,J=6.6Hz,3H),3.59(d,J=3.6Hz,5H),3.45(s,3H) ,3.13(s,3H),3.07(s,3H),2.97(t,J=5.6Hz,3H),2.77(t,J=7.6Hz,3H),2.67(t,J=6.6Hz,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 equivalent) in methanol (10.0 mL) was added NaOH (1 M, 6.17 mL, 1.0 equivalent), and the reaction was stirred at 25° C. for 2 hours. After the reaction, the mixture was adjusted to a pH of less than 6 using HCl (1 M), and separated and extracted using H 2 O (20.0 mL) and DCM (30.0 mL, 3 times). The resulting organic layer was washed with saturated brine (30.0 mL, 2 times), dried over Na 2 SO 4, and concentrated under reduced pressure to give the crude product of compound Q5b-2 (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 equivalent), compound 5A (925 mg, 3.37 mmol, 1.0 equivalent), DMAP (20.6 mg, 168 μmol, 0.05 equivalent) and EDCI (711 mg, 3.71 mmol, 1.1 equivalent) were mixed in DCM (10.0 mL) and stirred at 25 ° C. under N2 environment for 8 hours. After the reaction, the mixture was separated and extracted with H2O (10.0 mL) and DCM (10.0 mL, 3 times). The resulting organic layer was washed with saturated brine (20.0 mL, 2 times), dried over Na2SO4, and concentrated under reduced pressure to give the crude product of 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 equiv) was dissolved in formic acid (5.00 mL) and stirred at 25°C for 2 hours. After the reaction, the mixture was concentrated under reduced pressure to obtain a crude product, which was further purified by reverse-phase HPLC (0.1% FA) to obtain compound Q5b-4 (30.0 mg, approximately 75% purity) as a brown solid.

[0605] 4. Synthesis of APQ5b

[0606] Compound Q5b-4 (20.0 mg, 55.8 μmol, 1.0 equiv), compound C1 (39.3 mg, 55.8 μmol, 1.0 equiv), and NaBH(OAc)3 (35.5 mg, 167 μmol, 3.0 equiv) were mixed in DMF (1.00 mL) and stirred at 25°C under a nitrogen atmosphere for 2 hours. After the reaction, the mixture was concentrated under reduced pressure to remove the solvent to obtain the crude target product, which was then purified by preparative HPLC (Phenomenex luna C18 150*25 mm*10 μm column; mobile phase: [water (FA)-ACN]; gradient: 32% to 52% B over 10 minutes) to afford 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(400MHz DMSO)δppm 10.26(s,1H),8.18(s,1H),8.03(d,J=7.6Hz,1H),7.79(d,J=8.4Hz,1H)7.60(d,J=7.2Hz,1H) ,7.51(d,J=8.8Hz,1H),7.45-7.48(m,1H),7.39-7.42(m,1H),7.32-7.38(m,2H),7.26(dd,J= 11.6,1.6Hz,1H),7.13-7.19(m,3H),7.08(d,J=8.8Hz,1H),6.89-6.94(m,3H),4.91(s,2H),4 .00(t,J=6.4Hz,2H),3.85(t,J=5.6Hz,2H),3.69(t,J=6.8Hz,3H),3.57-3.60(m,4H),3.45(br s,3H),3.12(s,3H),3.06(s,2H),2.97(t,J=5.6Hz,3H),2.78(t,J=7.38Hz,2H)2.67(t,J=6. 8Hz,3H),2.32-2.37(m,3H),2.28(t,J=7.2Hz,2H),1.91-2.00(m,3H),1.67(q,J=7.2Hz,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 the mixture was stirred at 110° C. for 3 hours. Subsequently, CO(NH 2 ) 2 (4.48 g, 74.6 mmol, 4.01 mL, 6.40 equiv) and AcOH (20 mL) were added, and the mixture was stirred at 120° C. for 12 hours. The reaction mixture was added to water (50.0 mL) at 0° C., filtered to obtain a residue, washed with water (20.0 mL), and concentrated under reduced pressure to give the crude product of compound Q6a-2 (2.20 g, 8.18 mmol, 70.3% yield) as a brown solid.

[0613] LCMS: EC16416-3-p1a, P1 RT=0.265min, 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, HCl), Q6a-2 (2.00 g, 7.43 mmol, 1.00 equiv), Cs2CO3 (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 μmol, 0.100 equiv) were mixed in dioxane (20.0 mL) and the mixture was stirred at 100° C. under N2 for 16 hours. After the reaction, the mixture was concentrated under reduced pressure to give the crude product, which was further purified by preparative HPLC (neutral conditions) to give the product Q6a-4 (300 mg, 1.15 mmol, 15.4% yield) as a white solid.

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

[0617] 3. Synthesis of Q6a-5

[0618] To a solution of Py-SO3 (122 mg, 765 μmol, 4.00 equiv) in DMSO (1.00 mL) was added TEA (96.8 mg, 957 μmol, 133 μL, 5.00 equiv) dropwise at 0°C. The mixture was stirred at 0°C for 15 minutes, followed by the dropwise addition of compound Q6a-4 (50.0 mg, 191 μmol, 1.00 equiv) dissolved in DMSO (1.00 mL) at 0°C. The mixture was stirred at 25°C for 3.75 hours. LCMS (P1, RT = 0.331 min) monitoring indicated the completion of the reaction of Q6a-4. The reaction mixture was concentrated under reduced pressure to afford the crude product, which was purified using preparative HPLC (FA conditions) to afford the product Q6a-5 (25.0 mg, 96.4 μmol, 50.4% yield) as a white solid.

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

[0620] 4. Synthesis of Q6a

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

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

[0623] 1 H NMR:EC16364-125-P1C(400MHz DMSO-d6)δ10.22(s,1H)8.03(d,J=7.6Hz,1H)7.79(d,J=8.0Hz,1H)7.59(d,J=7.2Hz,1H)7.44-7.54(m,2H)7.38-7.42(m,1H)7.32- 7.38(m,2H)7.26(dd,J=12.0,1.88Hz,1H)7.14-7.20(m,1H)7.09(d,J=8.8Hz,3H)6.90(d,J=8.8Hz,1H)6.41(d,J=8.8Hz,2H)4.91(s ,2H)4.01(t,J=6.4Hz,2H)3.85(t,J=6.0Hz,2H)3.90(t,J=7.2Hz,2H)3.66(t,J=6.8Hz,2H)3.54-3.58(m,2H)3.48(s,2H)3.21-3.2 6(m,5H)2.97(t,J=5.6Hz,2H)2.77(t,J=7.2Hz,2H)2.66(t,J=6.8Hz,2H)2.52-2.57(m,2H)2.38(d,J=2.4Hz,2H)1.92-2.00(m,2H).

[0624] Synthesis of APQ6b:

[0625] 1. Synthesis of Q6b-2

[0626] Compound Q6a-2 (2.00 g, 7.43 mmol, 1.00 equiv), Q6b-1 (1.01 g, 8.18 mmol, 1.01 equiv, HCl), Cs2CO3 (7.26 g, 22.3 mmol, 3.00 equiv) and RuPhosPdG3 (1.24 g, 1.49 mmol, 0.200 equiv) were mixed in dioxane (20.0 mL) and stirred at 100°C under N2 environment for 16 hours. After the reaction, the mixture was diluted with water (30.0 mL) and extracted with ethyl acetate (120.0 mL, 40.0 mL × 3 times). The obtained organic layer was washed with saturated brine (120.0 mL, 60.0 mL × 2 times), dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product, which was further purified by reverse phase HPLC (FA conditions) to give the product Q6b-2 (300 mg, crude product) as a brown solid.

[0627] LCMS: EC16331-20-P1B, P1 RT=0.297min, m / z=275.9[M+H] +

[0628] 2. Synthesis of Q6b-3

[0629] To a solution of compound Q6b-2 (100 mg, 363 μmol, 1.00 equiv) in DCM (1.00 mL) were added TEA (110 mg, 1.09 mmol, 152 μL, 3.00 equiv), TsCl (208 mg, 1.09 mmol, 3.00 equiv) and DMAP (178 mg, 1.45 mmol, 4.00 equiv), and the mixture was stirred at 50° C. for 4 hours. After the reaction, the mixture was terminated by adding 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 × 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, which was further purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 10:1 to 5:1) to give the product Q6b-3 (70.0 mg, 163 μmol, 44.9% yield) as a white solid.

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

[0631] 3. Synthesis of APQ6b

[0632] To a solution of compound Q6b-3 (50.0 mg, 116 μmol, 1.00 equiv) and C1 (49.2 mg, 69.8 μmol, 0.600 equiv) in DMF (0.500 mL) were added DIEA (60.2 mg, 465 μmol, 81.1 μL, 4.00 equiv) and KI (48.3 mg, 291 μmol, 2.50 equiv), and the mixture was stirred at 60°C for 2 hours. After the reaction, the mixture was filtered and concentrated under reduced pressure to give the crude product, which was further purified by preparative HPLC (FA and neutral conditions) to give the product APQ6b (2.40 mg, 2.49 μmol, 2.14% yield) as a white solid.

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

[0634] 1 H NMR:EC16364-64-P1D(400MHz DMSO-d6)δ10.23(s,1H)7.87(d,J=7.2Hz,1H)7.68(d,J=8.8Hz,1H)7.57-7.63(m,1H)7.54(d,J=8.8Hz,1H)7.2 3-7.50(m,3H)7.22-7.28(m,2H)7.18(d,J=8.8Hz,1H)7.08-7.12(m,1H)7.04(d,J=7.2Hz,2H)6.94-6.98(m,1H) 6.31(d,J=7.6Hz,1H)4.95(s,2H)4.28(d,J=6.0Hz,2H)3.98-4.03(m,2H)3.75(d,J=6.0Hz,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=5.6Hz,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 mixture of compound 4A (6.58 g, 55.3 mmol, 4.77 mL, 1.5 eq) dissolved in H2O (50.0 mL) and CHCl3 (50.0 mL) were 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 hours. After the reaction, the organic layer was separated from the mixture, washed with saturated brine (30.0 mL, twice), dried over Na2SO4, and concentrated under reduced pressure to obtain the crude product, which was further purified by column chromatography (SiO2, dichloromethane:methanol = 97:3) to obtain compound Q7a-2 as a yellow oil.

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

[0640] 2. Synthesis of Compound Q7a-3

[0641] Compound 6A (867 mg, 3.64 mmol, 1.0 equiv), Pd(PPh3)2Cl2 (51.2 mg, 72.9 μmol, 0.02 equiv), CuI (27.8 mg, 146 μmol, 0.04 equiv) and TEA (10.0 mL) were mixed, and compound Q7a-2 (500 mg, 3.64 mmol, 1.0 equiv) was added under N2 environment. The mixture was stirred at 25 ° C for 8 hours. After the reaction, the mixture was extracted with H2O (30.0 mL) and ethyl acetate (40.0 mL), filtered and concentrated under reduced pressure to give the crude product Q7a-3 (1.00 g, crude product) 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 (400MHz CDCl3) δppm 7.09-7.15 (m, 2H), 6.93 (t, J = 8.8Hz, 1H), 3.64 (s, 2H), 2.95 (t, J = 6.0Hz, 4H), 2.55 (t, J = 6.0Hz, 4H).

[0644] 3. Synthesis of Compound Q7a-4

[0645] To a mixture of compound Q7a-3 (400 mg, 1.62 mmol, 1.0 equiv), compound 5A (466 mg, 1.70 mmol, 1.05 equiv), and NaBH(OAc) (686 mg, 3.24 mmol, 2.0 equiv) in DCE (10.0 mL) was added HOAc (117 mg, 1.94 mmol, 111 μL, 1.2 equiv) dropwise at 25°C over 30 minutes. The mixture was stirred at 25°C for 1.5 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude product, which was further purified by reverse-phase HPLC (0.1% NH·H O) to afford 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] Compound Q7a-4 (80.0 mg, 158 μmol, 1.0 equiv), compound Q4-3 (106 mg, 174 μmol, 1.1 equiv), and Cs2CO3 (103 mg, 316 μmol, 2.0 equiv) were mixed in DMA (2.00 mL) and stirred at 25°C under a N2 atmosphere for 2 hours. After the reaction, the mixture was concentrated under reduced pressure to obtain the crude product, which was further purified by reverse-phase HPLC column chromatography (column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (FA)-ACN]; gradient elution: 27%-57% B over 10 minutes) to obtain 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 equivalent) was dissolved in HCl-dioxane (2.00 mL) and stirred at 25°C for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude product, which was further purified by reverse-phase HPLC (column: Daisogel SP ODS RPS 150*25 mm*5 μm; mobile phase: [water(NH4HCO3)-ACN]; gradient elution: 28%-58% B over 10 minutes) 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)δppm 10.25(s,1H),8.03(d,J=7.50Hz,1H),7.78(d,J=7.6Hz,1H),7.60(d,J=7.2Hz,1H),7.48-7.51(m, 1H),7.47(s,1H),7.38-7.42(m,1H),7.35(dd,J=7.2,2.8Hz,2H),7.27(dd,J=11.6,2.0Hz,1H),7. 16-7.19(m,1H),7.14(d,J=9.2Hz,2H),7.10(d,J=8.8Hz,1H),6.92(d,J=9.2Hz,2H),6.89(s,1H), 4.91(s,2H),4.01(t,J=6.4Hz,2H),3.85(t,J=6.0Hz,2H),3.69(t,J=6.4Hz,3H),3.45(s,3H),3.11 (s,4H),2.97(t,J=5.6Hz,3H),2.89(d,J=10.8Hz,3H),2.77(t,J=7.2Hz,3H),2.67(t,J=6.8Hz,3 H),2.62(s,5H),2.13-2.23(m,4H),1.93-2.00(m,2H),1.80(d,J=11.6Hz,2H),1.38-1.52(m,3H).

[0653] Synthesis of APQ7b:

[0654] The synthesis steps of the PROTAC molecule APQ7b refer to APQ7a, except that the starting material compound Q7a-1 is replaced with compound 1A. After synthesis, the final product is purified by preparative HPLC (column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (HCl)-ACN]; gradient elution: 18%-48% B in 8 minutes) to obtain compound APQ7b (13.7 mg, 13.4 μmol, 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 (400MHz DMSO) δppm 8.04(d,J=7.8Hz,1H),7.79(d,J=8.0Hz,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=6.4Hz,2H),3.86(t,J=5.6Hz,2H),3. 79(d,J=12.0Hz,2H),3.70(t,J=6.6Hz,2H),3.64(s,4H),3.22-3.30(m,2H ),3.04-3.18(m,6H),2.98(t,J=5.8Hz,2H),2.78-2.86(m,2H),2.69(t,J= 6.6Hz,2H),2.13(d,J=13.8Hz,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 equiv) in DMF (35.0 mL) were added compound Q8-1 (3.00 g, 14.5 mmol, 1.0 equiv), KI (2.66 g, 16.00 mmol, 1.1 equiv), and KCO (4.02 g, 29.1 mmol, 2.0 equiv). The mixture was stirred at 115° C. for 16 hours. Q8-1 (1.50 g, 8.87 mmol, 1.04 mL, 0.6 equiv) was then added, and the reaction was continued at 115° C. for 3 hours. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain the crude product, which was purified by preparative HPLC column chromatography (TFA conditions) to obtain 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 equivalent) was dissolved in formic acid (5.00 mL), and the mixture was stirred at 25° C. for 2 hours. After the reaction, the mixture was concentrated under reduced pressure to obtain a crude product of compound Q8-4 (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 equiv) in DCM (10.0 mL) were added TEA (1.17 g, 11.6 mmol, 1.61 mL, 10 equiv) and compound 1A (184 mg, 1.16 mmol, 1.0 equiv), and the mixture was stirred at 25°C for 0.5 hour, followed by the addition of NaBH(OAc)3 (737 mg, 3.48 mmol, 3.0 equiv) and the reaction was stirred at 25°C for 1 hour. The mixture was terminated by adding water (5.00 mL) at 25°C and extracted with DCM (20.0 mL, 5.00 mL × 4 times). The resulting organic layer was washed with saturated brine (10.0 mL, 5.00 mL × 2 times), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the crude product of compound Q8-5 (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 equivalent) was dissolved in formic acid (0.5 mL), and the mixture was stirred at 25° C. for 2 hours. After the reaction, the mixture was concentrated under reduced pressure to remove the solvent to obtain a crude product of compound Q8-6 (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 μmol, 1.0 equiv) and C1 (163 mg, 231 μmol, 1 equiv) in DMF (2.00 mL) were added TEA (234 mg, 2.32 mmol, 322 μL, 10.0 equiv) and NaBH(OAc)3 (147 mg, 695 μmol, 3.0 equiv), and the mixture was stirred at 25°C for 2 hours. After the reaction, the mixture was concentrated under reduced pressure to obtain a crude product, which was then purified using preparative HPLC column chromatography (Phenomenex luna C18 150*25 mm*10 μm; mobile phase: [water (FA)-ACN]; gradient elution: 20%-50% B over 10 minutes) to obtain compound APQ8 (30.0 mg, 28.6 μmol, 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(400MHz DMSO)δppm 10.29(s,1H),8.18(s,1H),8.03(d,J=7.6Hz,1H),7.79(d,J=8.0Hz,1H),7.59( d,J=7.6Hz,1H),7.51(d,J=8.8Hz,1H),7.45-7.49(m,1H),7.39-7.42(m,1H),7 .35(s,1H),7.34(s,1H),7.26(dd,J=11.6,1.6Hz,1H),7.21(d,J=9.2Hz,2H),7 .17(d,J=10.4Hz,1H),7.06-7.12(m,1H),6.94(s,1H),6.92(s,1H),6.89-6.92 (m,1H),4.91(s,2H),4.03(dt,J=16.8,6.4Hz,5H),3.85(t,J=6.0Hz,2H),3.70 (t,J=6.8Hz,3H),3.44(s,3H),2.97(t,J=5.6Hz,2H),2.90(d,J=11.6Hz,2H),2 .78(t,J=7.6Hz,3H),2.65-2.71(m,5H),2.29-2.40(m,4H),2.09(d,J=7.2Hz,2 H),1.92-2.04(m,5H),1.60-1.67(m,2H),1.41-1.49(m,1H),1.02-1.14(m,2H).

[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 equivalent) in DMA (2 mL) were added Cs2CO3 (134 mg, 411 μmol, 2.5 equivalents) and compound 6A (58.7 mg, 246 μmol, 1.5 equivalents), and the reaction was stirred at 25 ° C for 16 hours. The mixture was quenched by adding water (10.0 mL) at 0 ° C, extracted with ethyl acetate (30.0 mL, 10.0 mL × 3 times), and the resulting 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 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] 3-Bromo-1-propyne (104 mg, 874 μmol, 75.4 μL, 1.2 equiv), compound 5A (200 mg, 729 μmol, 1.0 equiv) and DIEA (565 mg, 4.37 mmol, 762 μL, 6.0 equiv) were mixed in DMF (5 mL) and stirred at 60° C. under N2 for 16 hours. After the reaction, the mixture was concentrated under reduced pressure to give the crude product, which was further purified by preparative HPLC column chromatography (basic conditions) 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] Compound Q9-1 (70.0 mg, 91.5 μmol, 1.0 equiv), Pd(PPh3)2Cl2 (1.29 mg, 1.83 μmol, 0.02 equiv), CuI (697 μg, 3.66 μmol, 0.04 equiv) and TEA (1 mL) were mixed and charged with N2 protection. Compound Q9-2 (42.9 mg, 137.3 μmol, 1.5 equiv) was then added, and the mixture was stirred at 25°C for 8 hours. The mixture was concentrated under reduced pressure and triturated with water to remove impurities to obtain a crude product of compound Q9-3 (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(400MHz DMSO)δppm 12.54-13.15(m,1H),10.26(s,1H),8.01(d,J=7.8Hz,1H),7.76(d,J=7.8Hz,1H),7.56(d,J=7.4Hz,1H),7.50(d,J=8.8H z,1H),7.42-7.48(m,1H),7.37-7.42(m,1H),7.31-7.36(m,2H),7.29(dd,J=12.0,1.8Hz,1H),7.17-7.22(m,1H),7.15( d,J=8.8Hz,2H),7.07-7.13(m,1H),6.94(d,J=9.0Hz,2H),6.91(d,J=8.8Hz,1H),4.93(s,2H),4.02(t,J=6.0Hz,2H),3. 79(t,J=6.0Hz,2H),3.69(t,J=6.6Hz,2H),3.55(s,2H)3.17(d,J=3.8Hz,4H),2.98(t,J=6.0Hz,2H),2.63-2.70(m,8H), 1.88-1.98(m,2H),1.30(s,9H).

[0686] 4. Synthesis of compound APQ9

[0687] Compound Q9-3 (50.0 mg, 52.7 μmol, 1.0 equiv) and TFA (1.54 g, 13.5 mmol, 1 mL, 255 equiv) were mixed in DCM (1 mL) and stirred at 25°C under a N2 atmosphere for 2 hours. The mixture was concentrated under reduced pressure to obtain a crude product, which was further purified by preparative HPLC column chromatography (HCl and neutral conditions) to obtain 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(400MHz DMSO)δppm 10.26(s,1H)8.03(d,J=8.0Hz,1H)7.78(d,J=8.0Hz,1H)7.60(d,J=7.2Hz,1H)7.43-7.55(m,2H)7. 38-7.42(m,1H)7.32-7.37(m,2H)7.28(dd,J=12.0,1.75Hz,1H)7.13-7.21(m,3H)7.07-7.12(m,1H) 6.88-6.96(m,3H)4.92(s,2H)4.02(t,J=6.4Hz,2H)3.85(t,J=6.0Hz,2H)3.69(t,J=6.8Hz,2H)3.55 (s,2H)3.16(s,4H)2.97(t,J=5.6Hz,2H)2.78(t,J=7.2Hz,2H)2.65-2.70(m,6H)1.91-2.02(m,2H).

[0690] Synthesis of APQ10:

[0691] 1. Synthesis of Compound Q10-2

[0692] Compound 1A (218 mg, 1.11 mmol, 3.0 equiv, HCl), 1-(4-bromophenyl)hexahydropyrimidine-2,4-dione (100 mg, 371.62 μmol, 1 equiv), Cs2CO3 (363 mg, 1.11 mmol, 3.0 equiv), and RuPhosPdG3 (31.1 mg, 37.1 μmol, 0.1 equiv) were mixed in dioxane (5 mL), purged with N2, and stirred at 90°C for 2 hours. The mixture was concentrated under reduced pressure to give the crude product, which was further purified by preparative HPLC column chromatography (FA conditions) to give compound Q10-2 (60.0 mg, 172 μmol, 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 μmol, 1 equivalent) was mixed with formic acid (1.0 mL) and stirred at 25°C under a N2 atmosphere for 1 hour. The mixture was concentrated under reduced pressure to obtain a crude product, which was used directly in the next step without purification. The resulting crude product, compound Q10-3 (35.0 mg, 116.1 μmol, 67.2% yield), was obtained as a yellow oil.

[0696] 3. Synthesis of compound APQ10

[0697] To a solution of compound Q10-3 (35 mg, 116 μmol, 1.0 equiv) in DCM (5.0 mL) were added TEA (117 mg, 1.16 mmol, 161 μL, 10.0 equiv) and compound C1 (90 mg, 127.69 μmol, 1.1 equiv). The mixture was stirred at 25°C for 0.5 h, followed by the addition of NaBH(OAc) (73.81 mg, 348.26 μmol, 3.0 equiv) and the reaction was 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 chromatography (FA conditions) to give compound APQ10 (13 mg, 12.8 μmol, 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(400MHz DMSO)δppm 12.85(s,1H),11.69(s,1H),10.44(s,1H),8.04(d,J=8.0Hz,1H),7.79(d,J=8.0Hz,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=8.0Hz,1H),7.29-7.39(m,3H),7.10 -7.19(m,1H),6.98(d,J=8.8Hz,1H),4.93(s,2H),4.30(s,2H),4.02-4.08(m, 2H),3.86(t,J=6.0Hz,2H),3.80(t,J=6.4Hz,4H),3.60-3.64(m,2H),3.33-3.5 3(m,4H),3.14-3.27(m,2H),2.98(t,J=5.6Hz,2H),2.78-2.86(m,2H),2.71(t ,J=6.4Hz,2H),2.16-2.27(m,2H),1.97(dt,J=14.4,7.04Hz,2H),1.88(s,1H).

[0700] Synthesis of APQ11:

[0701] The synthesis steps of the PROTAC molecule APQ11 refer to APQ2, but the intermediate raw material compound 1 is replaced with compound 4. After synthesis, it is purified by column chromatography (FA conditions) to obtain compound APQ11 (30.0 mg, 26.5 μmol, 18.6% yield, 97.1% purity) as a white solid.

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

[0703] 11H NMR: EC16364-14-P1C (400 MHz, DMSO) δ ppm 11.12 (s, 1H) 8.19 (s, 1H) 8.03 (d, J = 8.0 Hz, 1H) 7.89 (d, J = 9.2 Hz, 1H) 7.78 (d, J = 8.0 Hz, 1H) 7.59 (d, J = 7.2 Hz, 1H) 7.44 - 7.53 (m, 2H) 7.38 - 7.42 (m, 1H) 7.30 - 7.38 (m, 3H) 7.24 - 7.29 (m, 1H) 7.13 - 7.20 (m, 2H) 7.04 - 7.13 (m, 2H) 6.90 (d, J = 8.8 Hz, 1H) 5.84 (dd, J = 12.4, 5.25 Hz, 1H) 4.91 (s, 2H) 4.01 (t, J = 6.4 Hz, 2H) 3.85 (t, J = 6.0 Hz, 2H) 3.44 (s, 2H) 3.30 (dd, J = 12.0, 5.57 Hz, 4H) 2.86 - 3.00 (m, 6H) 2.77 (t, J = 7.6 Hz, 2H) 2.64 - 2.71 (m, 2H) 2.29 - 2.41 (m, 4H) 2.21 (d, J = 5.2 Hz, 2H) 2.10 (d, J = 6.8 Hz, 2H) 1.92 - 2.01 (m, 4H) 1.65 (d, J = 12.0 Hz, 2H) 1.40 - 1.54 (m, 2H) 1.04 - 1.18 (m, 2H).

[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 μmol, 1.00 equiv) in methanol (5.0 mL) was added SOCl (13.3 mg, 112 μmol, 8.13 μL, 5.00 equiv), and the mixture was stirred at 80°C for 3 hours. LCMS (EC17015-82-P1D, P1: RT = 0.449 min) indicated a 47.6% yield of the desired compound. After the reaction, the mixture was extracted with CH2Cl2 (10.0 mL, twice). The obtained organic layer was washed with saturated brine (10.0 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product, which was further purified by preparative HPLC (chromatographic column: Phenomenex luna C18 150*25mm*10μm; mobile phase: [water (FA)-ACN]; gradient elution: 37%-57% B in 13 minutes) to obtain 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(400MHz DMSO)δppm 10.26(s,1H),8.03(d,J=7.6Hz,1H),7.78(d,J=8.0Hz,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=12.0,1.8Hz,1H),7.20(d,J=9.0Hz,1H),7.15(d,J=8.8Hz,2H),7.05-7 .12(m,1H),6.95(dd,J=8.8,5.0Hz,3H),4.90(s,2H),4.00(t,J=6.4Hz,2H),3.77-3.86(m,2H),3.69(t,J=6. 8Hz,2H),3.62(s,3H),3.55(s,2H),3.17(s,4H),2.98(t,J=5.8Hz,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), methanesulfonamide (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. under a nitrogen atmosphere for 3 hours. LCMS (EC17015-45-P1A, P1: RT = 0.435 min) indicated a 75.6% yield of the title compound. After the reaction, the mixture was concentrated under reduced pressure to obtain a crude product, which was further purified by preparative HPLC (column: Phenomenex luna C18 150*25mm*10μm; mobile phase: [water(FA)-ACN]; gradient elution: 30%-60% B over 10 minutes) to obtain 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)δppm 10.25(s,1H),8.14(s,1H),8.02(d,J=7.2Hz,1H),7.78(d,J=8.2Hz,1H),7.59(d,J=7.6Hz,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 .0Hz,3H),4.86(s,2H),4.02(t,J=6.0Hz,2H),3.84-3.93(m,2H),3.69(t,J=6.6Hz,2H),3.54(s,2H), 3.16(s,4H),2.99(d,J=5.6Hz,2H),2.72-2.78(m,2H),2.67(dd,J=3.8,1.8Hz,9H),1.92-2.01(m,2H).

[0713] Synthesis of APQ10-1:

[0714] 1. Synthetic 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) were 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, followed by the addition of NaBH(OAc)3 (907 mg, 4.28 mmol, 3.00 equiv) and continued stirring at 25°C for 1 h. After the reaction, the mixture was diluted with water (5.0 mL) and extracted with DCM (5.0 mL*4). The resulting organic layer was washed with saturated brine (10.0 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by reverse phase HPLC (0.1% FA conditions) to give compound Q10-1-2 (160 mg, 308 μmol, 21.6% yield) as a yellow solid.

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

[0717] 2. Synthetic APQ10-1

[0718] Compound Q4-3 (117 mg, 192 μmol, 1.00 equiv), Q10-1-2 (100 mg, 192 μmol, 1.00 equiv), and CsCO (157 mg, 481 μmol, 2.50 equiv) were mixed in DMA (2.00 mL) and stirred at 25°C under a N atmosphere for 16 hours. LCMS (EC17015-91-P1A, P1: RT = 0.433 min) indicated a yield of 57.6% of the desired compound. After the reaction, the mixture was concentrated under reduced pressure to obtain a crude product, which was further purified by preparative HPLC (column: Phenomenex luna C18 150*25 mm*10 μm; mobile phase: [water (FA)-ACN]; gradient elution: 36%-66% B over 10 minutes) to obtain 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] 11H 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] Compound APQ10 (50.0 mg, 50.5 μmol, 1.00 equiv), methanesulfonamide (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) were mixed in DCM (2.00 mL) and stirred at 25°C under a nitrogen atmosphere for 1 hour. LCMS (EC17015-89-P1B, P1: RT = 0.416 min) showed approximately 76.1% yield of the title compound. The reaction mixture was diluted with water (20.0 mL) and extracted with DCM (20.0 mL x 3). The resulting organic layer was washed with saturated brine (30.0 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to yield the crude product. The crude product was purified by preparative HPLC (column: Phenomenex luna C18 150*25mm*10μm; mobile phase: [water (FA)-ACN]; gradient elution: 28%-58% B in 10 minutes) 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] δppm 10.24(s,1H),8.16(s,1H),8.03(d,J=8.4Hz,1H),7.78(d,J=8.2Hz,1H),7.58(d,J=7.8Hz,1H),7.44-7.50(m,1H),7.40(d,J =7.8Hz,2H),7.31-7.37(m,2H),7.26(d,J=12.6Hz,1H),7.09-7.19(m,4H),6.91(d,J=9.0Hz,2H),6.67-6.75(m,1H),4.83(s ,2H),4.01(t,J=6.8Hz,2H),3.86(t,J=5.8Hz,2H),3.63-3.73(m,4H),3.46(s,2H),2.97(t,J=5.2Hz,2H),2.91(s,3H),2.58 -2.73(m,8H),2.44(s,4H),2.17(d,J=6.4Hz,2H),1.91-2.03(m,2H),1.76(d,J=12.6Hz,2H),1.65(s,2H),1.08-1.29(m,3H).

[0727] Synthesis of APQ10-3:

[0728] 1. Synthetic 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 hours. Acetic acid (50.0 mL) and urea (7.11 g, 118 mmol, 6.37 mL, 4.50 equiv) were then added, and the mixture was stirred at 120°C for 12 hours. LC-MS (EC21184-306-P1B, P1: RT = 0.598 min) detected approximately 76.5% yield of the desired product. The reaction mixture was diluted with water (200 mL), filtered, and concentrated under reduced pressure to afford compound Q10-3-2 (2.50 g, 8.11 mmol, 30.8% yield, 93.1% purity) as a white solid.

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

[0731] 2. Synthetic Q10-3-3

[0732] 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) were mixed in dioxane (25.0 mL) and stirred at 90°C under a nitrogen atmosphere for 12 hours. LC-MS (EC21184-315-P1, P1: RT = 0.272 min) detected the production of approximately 12.0% of the desired product. The reaction mixture was filtered and concentrated under reduced pressure, and purified by preparative HPLC (neutral conditions) to afford 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=0.272min, m / z=366.2[M+H] +

[0734] 3. Synthetic 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 the reaction was stirred at 25°C under a nitrogen atmosphere for 0.5 hour. LC-MS (EC21184-321-P1A, P1: RT = 0.221 min) detected approximately 60.9% yield of the desired product. The reaction mixture was filtered and concentrated under reduced pressure to afford the crude product of compound Q10-3-4 (50.0 mg, crude) as a white solid.

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

[0737] 4. Synthetic APQ10-3

[0738] To a solution of compound Q10-3-4 (50.0 mg, 157 μmol, 1.00 equiv) and compound C1 (66.2 mg, 93.9 μmol, 0.60 equiv) in DCM (1.00 mL) were added TEA (158 mg, 1.57 mmol, 218 μL, 10.0 equiv) and NaBH(OAc)₃ (100 mg, 470 μmol, 3.00 equiv). The mixture was stirred at 25°C for 4 hours. LC-MS (EC21184-322-P1B, P1: RT = 1.447 min) detected approximately 44.1% yield of the desired product. The reaction mixture was diluted with water (2.0 mL), filtered, and concentrated under reduced pressure to obtain a crude product, which was further purified by preparative HPLC (column: CD02-Waters Xbidge BEH C18 150*25*10 μm; mobile phase: [water(NH4HCO3)-ACN]; gradient: 28%-58% B over 10 minutes) to obtain compound APQ10-3 (8.00 mg, 7.94 μmol, 4.57% yield, 90.3% purity) as a white solid.

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

[0740] 1H NMR:EC17015-651-P1A(400MHz DMSO) δ10.36(s,1H),8.03(d,J=7.6Hz,1H),7.78(d,J=8.0Hz,1H),7.60(d,J=6.8Hz,1H),7.52(d,J=8.8Hz,1H),7.47(t,J=7.2Hz,1H),7.38 -7.43(m,1H),7.31-7.38(m,2H),7.27(dd,J=11.6,1.6Hz,1H),7.13-7.22(m,2H),7.05-7.12(m,1H),6.92(d,J=8.4Hz,1H),6.70-6.82(m,2H ),4.92(s,2H),4.02(t,J=6.4Hz,2H),3.86(t,J=6.0Hz,2H),3.68-3.77(m,2H),3.61(t,J=6.8Hz,2H),3.43-3.48(m,4H),2.94-3.01(m,2H), 2.79(t,J=7.2Hz,2H),2.63-2.74(m,6H),2.30-2.42(m,4H),2.14(d,J =7.2Hz,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 steps of the PROTAC molecule APQ901 refer to APQ902. The raw material Q901-1 can be purchased directly. The starting materials are replaced with Q901-1 and acrylic acid. After synthesis and purification, the target product APQ901 (17.0 mg, 16.0 μmol, 19.3% yield, 85.6% purity) is obtained as a white solid.

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

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

[0746] δ12.40-13.11(m,2H),10.37(s,1H),8.04(d,J=8.0Hz,1H),7.79(d,J=8.0Hz,1H),7.59(d,J=7.2Hz,1H),7.54(d,J=8.8Hz, 1H),7.44-7.50(m,1H),7.39-7.43(m,1H),7.32-7.38(m,2H),7.30(dd,J=12.0,2.0Hz,1H),7.15-7.22(m,2H),7.11(d,J=8 .8Hz,1H),7.03-7.09(m,2H),6.95(d,J=8.8Hz,1H),4.92(s,2H),4.02(t,J=6.4Hz,2H),3.86(t,J=6.0Hz,2H),3.74(t,J=6 .8Hz,2H),3.55(s,2H),3.04(s,4H),2.98(t,J=5.6Hz,2H),2.80(t,J=7.6Hz,2H),2.68(t,J=6.4Hz,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) and the mixture was stirred at 90° C. under a nitrogen atmosphere for 12 hours. After the reaction, the mixture was added with water (600 mL) and extracted with ethyl acetate (200 mL*3). The resulting organic layer was washed with saturated brine (200 mL), dried over Na2SO4, filtered under reduced pressure, and concentrated to obtain a crude product. The crude product was further purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 9 / 1) to obtain compound Q902-2 (13.1 g, crude product) as a yellow solid.

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

[0751] 1H NMR: EC17015-352-P1A (400MHz CDCl3) δppm 8.01 (t, J=9.2Hz, 1H), 6.38-6.64 (m, 2H), 3.53-3.71 (m, 4H), 3.36-3.47 (m, 4H), 1.48 (s, 9H).

[0752] 2. Synthesis of Compound Q902-3

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

[0754] 1 H NMR:EC17015-356-P1A(400MHz CDCl3)δppm 6.69-6.77(m,1H),6.65(dd,J=13.2,2.4Hz,1H),6.58(dd,J=8.4,1.8Hz,1H) ,3.52-3.63(m,4H),3.48(d,J=10.4Hz,2H),2.90-3.04(m,4H),1.49(s,9H).

[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 and stirred at 110°C for 3 hours. Urea (6.53 g, 109 mmol, 5.84 mL, 6.42 equiv) and AcOH (50.0 mL) were then added, and the mixture was stirred at 120°C for 12 hours. The reaction mixture was concentrated under reduced pressure to afford compound Q902-4 (6.60 g, crude product) as a dark 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 hours. LCMS (EC17015-374-P1A, P1: RT = 0.509 min) indicated the production of approximately 49.1% of the desired product. The reaction mixture was concentrated under reduced pressure to afford the crude product, which was then purified using preparative HPLC (column: CD19-Daisogel SP-100-8-ODS-PK 200*50*10 μm; mobile phase: [water (NH₃H₂O)-ACN]; gradient elution: 8% to 36% B over 10 minutes) to afford compound Q902-5 (160 mg, 547 μmol, 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 μmol, 1.00 equiv), compound 4A (57.0 mg, 479 μmol, 41.3 μL, 1.00 equiv), and NaHCO (120.7 mg, 1.44 mmol, 55.9 μL, 3.00 equiv) were combined in DMF (2.00 mL), and the mixture was stirred at 25° C. for 12 hours. LCMS (EC17015-400-P1W, P1: RT = 0.566 min) indicated a yield of approximately 81.3% of the desired product. After the reaction, the mixture was filtered and concentrated under reduced pressure to give the crude product, which was purified using preparative HPLC (column: CD02-Waters Xbidge BEH C18 150*25*10 μm: mobile phase: [water (NH3H2O)-ACN]; gradient elution: 12%-42% B in 10 minutes) to give compound Q902-6 (90.0 mg, 272 μmol, 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(400MHz DMSO)

[0764] δppm 10.24-10.44(m,1H),7.20(t,J=9.0Hz,1H),6.72-6.90(m,2H),3.62(t,J=6.6Hz,2 H),3.33-3.34(m,2H),3.15-3.24(m,5H),2.68(t,J=6.6Hz,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) and stirred at 25°C under a nitrogen atmosphere for 12 hours. LCMS (EC17015-412-P1A, P1: RT = 0.449 min) indicated a yield of approximately 31.6% of the desired product. After the reaction, the mixture was added with ice water (20.0 mL), filtered and concentrated under reduced pressure to obtain the crude product, which was further purified by column chromatography (SiO2, dichloromethane / ethyl acetate = 1 / 4 to 0 / 1) to obtain compound Q902-7 (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 HCl / dioxane (5.00 mL) and stirred at 20°C for 12 hours. LCMS (EC17015-419-P1B1, P1: RT = 0.422 min) indicated the yield of approximately 71.4% of the desired product. The reaction mixture was concentrated under reduced pressure to afford the crude product, which was then purified by preparative HPLC (column: CD03-Welch Xtimate C18 150*25*5 μm; mobile phase: [water (FA)-ACN]; gradient elution: 28%-58% B over 10 minutes) to afford compound APQ902 (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(400MHz DMSO)

[0772] δppm 10.38(s,1H),8.17(s,1H),8.03(d,J=7.8Hz,1H),7.79(d,J=8.0Hz,1H),7.59(d,J=7.2Hz,1H),7.52(d,J=8.8Hz,1H),7.47 (t,J=7.6Hz,1H),7.39-7.42(m,1H),7.32-7.38(m,2H),7.28(dd,J=12.0,1.6Hz,1H),7.16-7.24(m,2H),7.07-7.14(m,1H) ,6.92(d,J=8.6Hz,1H),6.81-6.87(m,1H),6.74-6.80(m,1H),4.91(s,2H),4.02(t,J=6.4Hz,2H),3.85(t,J=5.6Hz,2H),3. 60-3.64(m,2H),3.55(s,2H),3.21(s,4H),2.97(t,J=5.4Hz,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 steps of the PROTAC molecule APQ902a refer to APQ902, but the starting materials are replaced with compounds Q902a-1 and 9A. After synthesis and purification, the target product APQ902a (8.00 mg, 8.46 μmol, 6.90% yield, 97.6% purity) is obtained as a white solid.

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

[0776] 1 H NMR:EC17015-420-P1A(400MHz DMSO)δppm 10.20(s,1H),8.03(d,J=8.0Hz,1H),7.78(d,J=7.6Hz,1H),7.60(d,J=7.6Hz,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=8.8Hz,1H),7.10(t,J=8.8Hz,1H),7.03(d,J=8.4Hz,1H),6.90(d,J=9 .2Hz,1H),6.62(d,J=1.6Hz,1H),6.49(dd,J=8.8,2.4Hz,1H),4.91(s,2H),4.02(t,J=6.4Hz,2H),3.85(t,J=6.0Hz,2H),3.77(s, 3H),3.55(s,2H),3.49(d,J=6.4Hz,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 steps of the PROTAC molecule APQ903 refer to APQ902, but the starting materials are replaced with compounds Q903-1 and 9A. After synthesis and purification, the target product APQ903 (12.0 mg, 12.0 μmol, 29.5% yield, 92.6% purity) is obtained as a white solid.

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

[0780] 1 H NMR:EC17015-421-P1A(400MHz DMSO)δppm 10.47(s,1H),8.03(d,J=8.4Hz,1H),7.78(d,J=8.0Hz,1H),7.59(d,J=7.2Hz,1H),7.52(d,J=8.8Hz,1H), 7.47(t,J=7.2Hz,1H),7.38-7.43(m,1H),7.27-7.37(m,3H),7.20(d,J=8.8Hz,1H),7.06-7.13(m,3H),6. 92(d,J=8.0Hz,1H),4.85-4.99(m,2H),4.02(t,J=6.4Hz,2H),3.86(t,J=5.6Hz,2H),3.76(t,J=6.8Hz,2H ),3.54(s,2H),3.13(s,4H),2.95-3.00(m,2H),2.76-2.81(m,2H),2.62-2.68(m,6H),1.92-2.02(m,2H).

[0781] Synthesis of APQ904:

[0782] The synthesis steps of the PROTAC molecule APQ904 were similar to those of APQ902. The intermediate APQ904-3 was synthesized using a different method. The synthesis from APQ904-3 to APQ904 was the same as that of APQ902. After synthesis and purification, the target product APQ904 (18.7 mg, 19.5 μmol, 32.0% yield, 96.9% purity) was obtained as a white solid.

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

[0784] 1H NMR:EC21184-159-p1dg1(400MHz DMSO) δ12.41-13.04(m,2H),10.47(s,1H),8.04(d,J=8.0Hz,1H),7.79(d,J=8.0Hz,1H),7.60(d,J=7.2Hz,1H) ,7.55(d,J=8.8Hz,1H),7.47(t,J=7.2Hz,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=6.4Hz,2H),3.86(t,J=6.0Hz,2H),3.66(t,J=6.8Hz,2H ),3.55(s,2H),3.08(s,4H),2.98(t,J=5.6Hz,2H),2.80(t,J=7.6Hz,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) were added BnBr (59.2 g, 346 mmol, 41.1 mL, 3.0 equiv) and KCO (47.8 g, 346 mmol, 3.00 equiv). The mixture was stirred at 85°C for 16 hours. LC-MS (EC21184-81-p1d, P1: RT = 0.577 min) indicated a yield of approximately 65.7% of the desired product. The reaction mixture was concentrated under reduced pressure to afford the crude product, which was then purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 0) to afford compound Q904-1 (40.0 g, crude) as a white solid.

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

[0789] 1H NMR: EC21184-81-p1gz (400MHz DMSO) δ7.54 (dd, J=12.4, 6.8Hz, 1H), 7.16-7.40 (m, 10H), 6.91 (dd, J=11.2, 7.6Hz, 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), CsCO (57.1 g, 175.2 mmol, 2.00 equiv), BINAP (10.9 g, 17.5 mmol, 0.20 equiv), and Pd(OAc) (1.97 g, 8.76 mmol, 0.10 equiv) were mixed in toluene (340 mL) and stirred at 100° C. under nitrogen for 16 hours. LC-MS (EC21184-91-p1a, P1: RT = 0.602 min) indicated a yield of approximately 52.7% of the desired product. After the reaction, the mixture was concentrated under reduced pressure to obtain the crude product, which was further purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 0 to 85 / 15) to obtain compound Q904-2 (36.0 g, 67.8 mmol, 77.5% yield, 93.0% purity) as a white solid.

[0792] LC-MS: EC21184-91-p1a, P1: RT=0.602min, 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 a nitrogen atmosphere. H was then introduced, and the reaction was stirred at 20°C under a hydrogen atmosphere (40 psi) for 16 hours. LC-MS (EC21184-96-p1b2, R1:RT = 0.391 min, P1:RT = 0.391 min) indicated the production of approximately 81.3% of the desired product. The reaction mixture was concentrated under reduced pressure to afford the crude product, which was then purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 0 to 88 / 12) to afford compound Q904-3 (18.0 g, 57.5 mmol, 83.4% yield, 100% purity) as a white solid.

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

[0796] 1 H NMR: EC21184-96-p1gz (400MHz CHLOROFORM) δ6.62-6.88(m,1H),6.52(dd,J=12.8,8.0Hz,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 enzymes (APH1-APH20 and APH101-APH106)

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

[0799] Synthesis of APH1:

[0800] 1. Synthesis of Compound H1-2

[0801] To a solution of compound 2 (200 mg, 450 μmol, 1.0 equiv) in DMF (2.00 mL) were added HATU (257 mg, 674 μmol, 1.5 equiv), DIEA (174 mg, 1.35 mmol, 235 μL, 3.0 equiv), and H1-1 (200 mg, 1.35 mmol, 3.0 equiv). The mixture was stirred at 25°C for 3 hours. LCMS (EC16331-51-P1A6, P1: RT = 0.359 min) showed that the reaction of compound 2 was complete, yielding a major peak with the expected m / z. After the reaction, the mixture was separated and extracted by adding water (5.00 mL) and DCM (5.00 mL*3). The obtained organic layer was washed with saturated brine (5.00 mL*2), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain the crude product of compound H1-2 (120 mg, 186.97 μmol, 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 (400MHz DMSO) δppm 8.99 (s, 1H), 8.46 (br d, J=7.6Hz, 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=12.8,8.8,4.4Hz,1H),1.37(br d,J=6.8Hz,3H),0.93(s,9H).

[0804] 2. Synthesis of Compound APH1

[0805] To a solution of compound H1-2 (100 mg, 200 μmol, 1.0 equiv) in DMF (2.00 mL) were added TEA (202 mg, 2.00 mmol, 278 μL, 10.0 equiv), NaBH(OAc)3 (127 mg, 600 μmol, 3.0 equiv), and compound C1 (70.4 mg, 99.9 μmol, 0.5 equiv). The mixture was stirred at 25°C for 2 hours. LCMS (EC16331-55-P1A, P1: RT = 0.462 min; R2: RT = 0.409 min) showed that the reaction of compound C1 was complete, yielding a major peak with the expected m / z. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain a crude compound APH1 product. The crude product was purified by preparative HPLC (FA conditions; chromatographic column: Phenomenex luna C18 150*25mm*10μm; mobile phase: [water (FA)-ACN]; gradient: 23%-53% B in 10 minutes) to obtain the product APH1 (7.00 mg, 5.31μmol, 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(400MHz DMSO)δppm 8.94-8.99(m,1H),8.39-8.46(m,1H),8.21(s,1H),8.03(br d,J=7.6Hz,1H),7.73-7.81(m,2H),7.59(br d,J=6.8Hz,1H),7.47(br dd,J=14.0,8.2Hz,2H),7.40(br t,J=7.2Hz,3H),7.32-7.36(m,3H),7.26(br d,J=12.0Hz,1H),7.19(br d,J=8.2Hz,1H),7.06-7.12(m,1H),6.88(br d,J=8.4Hz,1H),4.83-4.93(m,3H),4.37-4.53(m,3H),4.28(br s,1H),4.01(br t,J=6.0Hz,3H),3.84(br s,3H),3.58(br s,3H),3.48(br s,4H),2.90-3.08(m,6H),2.72-2.82(m,3H),2.42-2.46(m,3H),2.00-2.08(m,1H),1.97(br d,J=6.0Hz,2H),1.72-1.80(m,1H),1.38(br s,1H),1.28-1.37(m,3H),1.23(br s,1H),0.93(br s,9H).

[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 equivalents) in THF (120 mL) was added NaH (3.08 g, 76.9 mmol, 60% purity, 1.5 equivalents) at 0°C under a nitrogen atmosphere. The mixture was stirred at 0°C for 0.5 hours, and then a solution of compound H2-1 (10.0 g, 51.3 mmol, 7.57 mL, 1.0 equivalents) dissolved in THF (20.0 mL) was added dropwise at 0°C. The mixture was stirred at 15°C under a nitrogen atmosphere for 2 hours. TLC (PE:EA=15:1, R1:R f =0.92; R2:R f =0.1; P1:R f=0.44) showed that the reaction of compound H2-1 was complete, generating a new product peak. The reaction mixture was terminated by the addition of NH4Cl (200 mL) at 0°C, diluted with water (200 mL), and extracted with ethyl acetate (160 mL*3). The resulting organic layer was washed with saturated brine (200 mL*2), dried over Na2SO4, filtered under reduced pressure, and concentrated to obtain the crude product. The crude product was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 98:1 to 95:1) to obtain compound H2-3 (1.30 g, 5.24 mmol, 10.21% yield) as a colorless liquid.

[0811] 1 H NMR:EC9174-340-P1A(400MHz DMSO)δppm 4.58(t,J=5.2Hz,1H),4.00(s,2H),3.56-3.65(m,2H),3.45-3.52(m,2H),3.43(d,J=5.2Hz,2H),1.42(s,9H),1.11(t,J=7.2Hz,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 H2O (2.00 mL) was added LiOH-H2O (355 mg, 8.46 mmol, 10.0 eq.), and the mixture was stirred at 25°C for 2 h. The color was determined by TLC (PE:EA=15:1, R1:R2). f =0.38) showed that the reaction of compound H2-3 was complete, generating a new compound peak. The reaction mixture was adjusted to pH 6 with hydrochloric acid (1 M) and extracted with ethyl acetate (4.00 mL*3). The resulting organic layer was washed with saturated brine (10.0 mL*2), dried over Na2SO4, filtered, and concentrated under reduced pressure to afford the crude product, compound H2-4 (110 mg, 572 μmol, 67.67% yield), as a yellow oil, which was used directly in the next reaction without purification.

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

[0815] 3. Synthesis of Compound 4

[0816] Compound H2-4 (80.0 mg, 416 μmol, 1.0 equiv), compound 2 (185 mg, 416 μmol, 1.0 equiv), HATU (237 mg, 624 μmol, 1.5 equiv), and TEA (126 mg, 1.25 mmol, 174 μL, 3.0 equiv) were mixed in DMF (2.00 mL) and stirred at 25°C under a nitrogen atmosphere for 3 hours. LC-MS (EC16331-60-P1A1, P1: RT = 0.440 min; R2: RT = 0.313 min) showed that compound H2-4 had reacted completely, yielding a major peak with the expected m / z. The reaction mixture was filtered and concentrated under reduced pressure to obtain the crude product, which was then purified by preparative HPLC (FA conditions; column: Phenomenex luna C). 18 150*25mm*10μm; mobile phase: [water (FA)-ACN]; gradient: 30%-60% B, within 10 minutes) to obtain compound H2-5 product (80.0 mg, 114μmol, 27.33% yield, 88% purity).

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

[0818] 1 H NMR: EC16331-60-P1S (400MHz DMSO) δppm 8.99 (s, 1H), 8.45 (br d,J=7.6Hz,1H),7.41-7.45(m,2H),7.35-7.40(m,3H),4.91(quin,J=7.2Hz,1H),4.63(t,J=5.2Hz,1H),4.55(br d,J=9.6Hz,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=12.8,8.8,4.4Hz,1H),1.37(br d,J=7.2Hz,3H),1.09-1.16(m,6H),0.94(s,9H).

[0819] 4. Synthesis of Compound H2-6

[0820] To compound H2-5 (70.0 mg, 113 μmol, 1.0 equiv) was added formic acid (1.00 mL), and the mixture was stirred at 25°C for 0.5 h. LC-MS (EC16331-68-P1A2, P1: RT = 0.325 min) showed that compound H2-5 was completely reacted, yielding a major peak with the expected m / z. The reaction mixture was filtered and concentrated under reduced pressure to afford crude compound H2-6 (80.0 mg) as a yellow oil, which was used directly 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 equiv) in DMF (1.00 mL) were added TEA (149 mg, 1.47 mmol, 204.44 μL, 10.0 equiv), NaBH(OAc) (93.4 mg, 441 μmol, 3.0 equiv), and compound C1 (72.5 mg, 103 μmol, 0.7 equiv). The mixture was stirred at 25°C for 1 hour. LC-MS (EC16331-69-P1A, P1: RT = 0.448 min) showed that the reaction of compound H2-6 was complete, yielding a main peak with the expected m / z. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain a crude product, which was purified by preparative HPLC (chromatographic column: Waters Xbridge 150*25mm*5μm; mobile phase: [water (NH3H2O)-ACN]; gradient: 15%-45% B within 10 minutes) to obtain compound APH2 product (20.0 mg, 14.6μmol, 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(400MHz DMSO)δppm 8.95-9.00(m,1H),8.45(br d,J=7.6Hz,1H),8.26(s,1H),8.02(d,J=7.6Hz,1H),7.78(d,J=8.0Hz,1H),7.59(br d,J=7.6Hz,1H),7.40-7.51(m,6H),7.31-7.39(m,6H),7.26(br d,J=12.0Hz,1H),7.15-7.19(m,1H),7.09-7.09(m,1H),7.05-7.12(m,1H),6.83(br d,J=8.8Hz,1H),4.90(s,3H),4.49-4.57(m,2H),4.44(br t,J=8.0Hz,2H),4.28(br s,2H),3.97-4.02(m,3H),3.92-3.96(m,2H),3.83(br t,J=5.6Hz,3H)3.56-3.61(m,5H),3.45(s,3H),2.95-2.95(m,1H),2.96(br t,J=5.2Hz,1H),2.74(br t,J=6.8Hz,2H),2.43-2.46(m,4H),2.01-2.08(m,1H),1.91-2.01(m,2H), 1.76(ddd,J=12.8,8.8,4.4Hz,1H),1.34-1.38(m,3H),0.90-0.97(m,12H).

[0826] Synthesis of APH3:

[0827] 1. Synthesis of Compound H3-2

[0828] To a solution of compound C1-8 (150 mg, 197 μmol, 1.00 equiv) and compound H3-1 (141 mg, 789 μmol, 4.00 equiv) in DMF (2.00 mL) were added HATU (113 mg, 296 μmol, 1.50 equiv) and TEA (59.8 mg, 591 μmol, 82.3 μL, 3.00 equiv). The mixture was stirred at 20°C for 1 hour. LC-MS (EC21184-30-p1a, P1: RT = 0.439 min) confirmed complete reaction of compound C1-8, yielding the desired product in approximately 84.7% of the reaction mixture. The reaction mixture was then added with water (50.0 mL) and extracted with ethyl acetate (30.0 mL x 2). The resulting organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to afford crude compound H3-2 (180 mg, crude product) as a white solid.

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

[0830] 2. Synthesis of compound APH3-3

[0831] To a solution of compound H3-2 (180 mg, 195 μmol, 1.00 equiv) and compound 2 (86.9 mg, 195 μmol, 1.00 equiv) in DMF (2.00 mL) were added HATU (111 mg, 293 μmol, 1.50 equiv) and TEA (59.3 mg, 586 μmol, 81.6 μL, 3.00 equiv). The mixture was stirred at 20°C for 1 hour. LC-MS (EC21184-31-p1a1, P1: RT = 0.810 min) showed complete reaction of compound H3-2, yielding the desired product in approximately 8.41% of the reaction mixture. After the reaction, the mixture was added with water (100 mL) and extracted with ethyl acetate (25.0 mL*3). The obtained organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to obtain the crude product, which was further purified by preparative HPLC (neutral conditions; chromatographic column: CD07-Daisogel SP-100-8-ODS-PK 150*25*10 μm; mobile phase: [water(NH4HCO3)-ACN]; gradient: 57%-87% B in 10 minutes) to obtain compound H3-3 (50.0 g, 36.1 μmol, 18.5% yield, 97.3% purity) as a white solid.

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

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

[0834] 3. Synthesis of compound APH3

[0835] Compound H3-3 (45.0 g, 33 μmol, 1.00 equiv) was mixed in HCl / dioxane (25.0 mL) and stirred at 20°C under a nitrogen atmosphere for 4 hours. LC-MS (EC21184-35-p1a1, P1:RT=0.450 min) showed approximately 88.5% yield of the desired product. The reaction mixture was filtered and concentrated under reduced pressure to obtain the crude product, which was further purified by preparative HPLC (neutral conditions; column: CD02-Waters Xbidge BEH C18 150*25*10 μm; mobile phase: [water (NH4HCO3)-ACN]; gradient: 25%-55% B over 10 minutes) to yield 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=0.450min, m / z=1313.3[M+Na] +

[0837] 1 H NMR:EC21184-35-p1qc(400MHz DMSO) δ8.97(s,1H),8.42(d,J=7.6Hz,1H),8.03(d,J=8.0Hz,1H),7.78(d,J=8.0Hz,1H),7.60(d,J=8.0Hz,1H),7.30-7.54(m,10H),7.26(dd,J=12. 0,2.0Hz,1H),7.14-7.19(m,1H),7.05-7.12(m,1H),6.89(d,J=8.4Hz,1H ),4.85-4.95(m,3H),4.54(d,J=9.6Hz,1H),4.44(t,J=8.0Hz,1H),4.28(s ,1H),4.20(d,J=1.6Hz,2H),4.01(t,J=6.4Hz,2H),3.96(s,2H),3.85(t, J=5.6Hz,2H),3.55-3.67(m,7H),3.42-3.55(m,10H),2.97(t,J=5.6Hz,2H ),2.77(t,J=7.2Hz,2H),2.44(s,5H),2.01-2.10(m,1H),1.91-2.00(m,2H ),1.77(ddd,J=13.2,8.8,4.4Hz,1H),1.37(d,J=6.8Hz,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 μmol, 1.0 equiv) in methanol (3.00 mL) were added NaHCO (189 mg, 2.25 mmol, 87.52 μL, 5.0 equiv) and compound H4-1 (105 mg, 675 μmol, 1.5 equiv). The mixture was stirred at 60°C for 0.5 h. LC-MS (EC16331-49-P1A, P1: RT = 0.497 min) showed that the reaction of compound 2 was complete, yielding a main peak with the expected m / z. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain a crude product, which was purified by preparative HPLC (neutral conditions; column: Waters Xbridge BEH C18 150*25mm*5μm; mobile phase: [water(NH4HCO3)-ACN]; gradient: 25%-55% B in 10 minutes) to obtain compound H4-2 (160 mg, 292μmol, 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(400MHz DMSO)δppm 8.98(s,1H),8.34(d,J=7.6Hz,1H),7.34-7.45(m,4H),5.07(d,J=3.6Hz,1H),4.85-4.94( m,1H),4.44(t,J=7.6Hz,1H),4.27(brd,J=2.0Hz,1H),3.51-3.62(m,3H),3.29(s,1H),3. 17(dt,J=11.6,5.6Hz,2H),2.83(dt,J=11.6,6.0Hz,2H),2.45(s,3H),2.26-2.35(m,4H), 1.99-2.07(m,1H),1.79(ddd,J=12.8,7.6,4.8Hz,1H),1.37(d,J=7.2Hz,3H),0.99(s,9H).

[0843] 2. Synthesis of compound APH4

[0844] To a solution of compound H4-2 (100 mg, 190 μmol, 1.0 eq) in DMF (2.00 mL) were added TEA (192 mg, 1.90 mmol, 264 μL, 10.0 eq), NaBH(OAc) (121 mg, 570 μmol, 3.0 eq), and compound C1 (66.9 mg, 94.9 μmol, 0.5 eq). The mixture was stirred at 25°C for 2 hours. LC-MS (EC16331-54-P1A1, P1: RT = 0.769 min; R1: RT = 0.700 min) indicated the formation of the desired product with an m / z consistent with the expected value. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain a crude product, which was purified by preparative HPLC (neutral conditions; chromatographic column: Daisogel SP ODS RPS 150*25mm*5μm; mobile phase: [water(NH4HCO3)-ACN]; gradient: 37%-67% B, within 10 minutes) to obtain compound APH4 product (20.0 mg, 16.5μmol, 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 (400MHz DMSO) δppm 8.98 (s, 1H), 8.31 (br d, J = 7.6Hz, 1H), 8.02 (d, J = 7.6Hz, 1H), 7.78 (d, J = 8.0Hz, 1H), 7.59 (br d,J=7.2Hz,1H),7.31-7.50(m,9H),7.25(dd,J=12.0,1.2Hz,1H),7.16(br d,J=8.6Hz,1H),7.04-7.11(m,1H),6.85(br d,J=8.8Hz,1H),4.86-4.93(m,3H),4.43(brt,J=7.6Hz,1H),4.26(br s,2H),4.00(br t,J=6.4Hz,2H),3.84(br t,J=5.6Hz,3H),3.57(br s,4H),3.16-3.23(m,4H),3.07(br s,2H),2.97(br d,J=5.2Hz,3H),2.85-2.92(m,2H),2.70-2.79(m,3H),2.45(s,3H),1.94(br s,6H),1.76-1.85(m,1H),1.62-1.72(m,2H),1.37(br d,J=6.8Hz,2H),1.28(br d,J=9.2Hz,1H),1.23(br s,1H),0.92(s,9H).

[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, and the mixture was stirred at -40°C for 0.5 hours. TLC (PE:EA=7:1, R1:R f =0.80; P1:R f =0.75) indicated that compound H5-1 had reacted completely, generating two new compound peaks. The reaction mixture was quenched by the addition of 1 M HCl (30.0 mL) at -60°C, dried over NaSO, filtered, and concentrated under reduced pressure to afford crude product H5-2 (5.20 g) as a brown liquid, which was used directly in the next reaction without purification.

[0850] 1H NMR: EC16331-78-P1A (400MHz DMSO) δppm 9.59 (d, J = 1.2Hz, 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 N2 atmosphere, and the mixture was stirred at 110 ° C for 12 hours. TLC (PE: EA = 7: 1, R1: R2) was used. f =0.80; P1:R f =0.75) showed that the reaction of compound H5-2 was complete, generating a new compound peak. The reaction mixture was extracted with water (100.0 mL) and ethyl acetate (80.0 mL*3). The resulting organic layer was washed with saturated brine (100 mL*2), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product. Further purification by column chromatography (SiO2, petroleum ether:ethyl acetate = 99:1) afforded compound H5-3 (700 mg, 4.99 mmol, 9.60% yield) as a yellow oil.

[0853] 1 H NMR: EC16331-82-P1A (400MHz DMSO) δppm 5.60-5.70 (m, 2H), 4.65-4.73 (m, 1H), 3.73-3.95 (m, 4H), 2.15-2.46 (m, 6H).

[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) was bubbled with O3 (1.37 g, 2.85 mmol, 1.00 equiv) at -78°C for 1 minute, and PPh3 (898.11 mg, 3.42 mmol, 1.20 equiv) was added. The mixture was stirred at -78°C for 0.5 hours and then at 25°C for 1 hour. The reaction mixture was quenched by adding HCl (1 M, 30.0 mL) at -60°C, dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product, which was further analyzed by preparative TLC (SiO2, PE:EA=0:1; P1:R f =0.30) was purified to give compound H5-4 (100 mg, crude product) as a colorless oil.

[0856] 4. Synthesis of Compound H5-5

[0857] To a solution of compound H5-4 (50.0 mg, 290 μmol, 1.00 equiv) in DCM (1.00 mL) were added TEA (2.93 g, 2.90 mmol, 404 μL, 10.0 equiv), NaBH(OAc)3 (185 mg, 871 μmol, 3.0 equiv) and compound 2 (103 mg, 232 μmol, 0.8 equiv), and the mixture was stirred at 25°C for 2 hours. After the reaction, the mixture was extracted by adding water (5.00 mL) and ethyl acetate (5.00 mL*3). The obtained organic layer was washed with saturated brine (30.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain the crude product, which was further purified by preparative HPLC (FA conditions; column: Phenomenex luna C18 150*25mm*10μm; mobile phase: [water (FA)-ACN]; gradient: 9%-39% B in 10 minutes) to obtain compound H5-5 product (60.0 mg, 99.19 μmol, 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 (400MHz DMSO) δppm 8.70 (s, 1H), 8.36 (s, 1H), 7.66 (br d,J=7.6Hz,1H),7.40(q,J=8.2Hz,4H),7.28(s,1H),5.06(quin,J=7.2Hz,1H),4.78-4.95(m,2H),4.65(d,J=4.0Hz,1H),4.60(br s,1H),3.84-3.97(m,4H),3.77-3.83(m,1H),3.66(br dd,J=10.8,4.8Hz,1H),3.52(s,1H),3.45(br d,J=12.4Hz,1H),3.18(br d,J=9.6Hz,1H),2.96(br s,9H),2.75-2.84(m,1H),2.64-2.72(m,1H),2.57(br s,1H),2.51-2.53(m,1H),2.03(ddd,J=12.8,8.4,4.0Hz,1H),1.70-1.82(m,2H),1.60(br d,J=7.2Hz,2H),1.49(br d,J=6.8Hz,3H),1.10(s,9H).

[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) indicated the formation of the desired product. The reaction mixture was filtered and concentrated under reduced pressure to afford compound H5-6 (28.0 mg, crude product), 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 equiv) in DCM (1.00 mL) were added TEA (52.4 mg, 518 μmol, 72.1 μL, 10.0 equiv) and compound C1 (18.2 mg, 25.9 μmol, 0.50 equiv). The mixture was stirred at 25°C for 0.5 h. Subsequently, NaBH(OAc)3 (32.9 mg, 155 μmol, 3.00 equiv) was added, and the reaction was stirred at 25°C for 12 h. LCMS (EC17015-217-P1B, P1: RT = 1.388 min) indicated the formation of the desired product. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain a crude product, which was further purified by preparative HPLC (column: CD02-Waters Xbidge BEH C18 150*25*10 μm; mobile phase: [water (NH3H2O)-ACN]; gradient: 24%-54% B over 10 minutes) to obtain compound APH5 (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(400MHz DMSO) δ8.98(s,1H),8.29(d,J=7.0Hz,1H),8.01(d,J=8.2Hz,1H),7.77(d,J=7.6Hz,1H),7.60(d,J=7.6Hz,1H) ,7.41-7.47(m,3H),7.38(s,3H),7.30-7.35(m,2H),7.26(d,J=12.6Hz,1H),7.17(d,J=7.6Hz,1H),7.05-7.12 (m,1H),6.79(dd,J=7.0,2.0Hz,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=4.4Hz,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=7.0Hz,4H),1.20-1.28(m,4H),0.92(s,9H).

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

[0868] Synthesis of APH6:

[0869] 1. Synthesis of compound H6-2

[0870] To a solution of compound H6-1 (100 mg, 291 μmol, 1.0 equiv) in DMSO (1.00 mL) were added EDCI (83.7 mg, 436 μmol, 1.5 equiv), 1-hydroxy-7-azobenzotriazole (59.5 mg, 436 μmol, 61.10 μL, 1.5 equiv), NMM (88.3 mg, 873 μmol, 96.0 μL, 3.0 equiv) and compound 7A (43.1 mg, 145 μmol, 0.5 equiv), and the mixture was stirred at 25°C for 2 hours. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain the crude product, which was further purified by preparative HPLC (FA conditions; column: Phenomenex luna C18 150*25mm*10μm; mobile phase: [water (FA)-ACN]; gradient: 16%-46% B in 10 minutes) to obtain the product H6-2 (80.0 mg, 129 μmol, 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(400MHz DMSO)δppm 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=12.0,5.6Hz,1H),3.55(br d,J=10.0Hz,1H),2.89-3.02(m,2H),2.42-2.47(m,3H),2.34(br d,J=5.6Hz,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=5.6Hz,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 equiv) in DCM (1.00 mL) were added TEA (65.1 mg, 643 μmol, 89.5 μL, 10.0 equiv), NaBH(OAc)3 (40.9 mg, 193 μmol, 3.0 equiv) and compound C1 (22.67 mg, 32.17 μmol, 0.5 equiv), and the mixture was stirred at 25°C for 2 hours. After the reaction, the mixture was extracted with water (5.00 mL) and ethyl acetate (5.00 mL*3). The obtained organic layer was washed with saturated brine (10.0 mL*2), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain the crude product, which was further purified by preparative HPLC (FA conditions; column: Phenomenex luna C18 150*25mm*10μm; mobile phase: [water (FA)-ACN]; gradient: 22%-52% B in 8 minutes) to obtain compound APH6 product (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] 11H NMR: EC16331-105-P1E (400 MHz, DMSO) δ ppm 8.92 - 8.99 (m, 1H), 8.46 (br t, J = 8.4 Hz, 1H), 8.20 (s, 1H), 8.03 (d, J = 7.6 Hz, 1H), 7.79 (d, J = 8.0 Hz, 1H), 7.59 (br d, J = 7.6 Hz, 1H), 7.31 - 7.53 (m, 9H), 7.23 - 7.30 (m, 1H), 7.17 (br d, J = 8.0 Hz, 1H), 7.05 - 7.13 (m, 1H), 6.90 (br d, J = 8.8 Hz, 1H), 6.21 (s, 1H), 5.13 - 5.24 (m, 1H), 4.91 (s, 2H), 4.61 - 4.72 (m, 1H), 4.22 - 4.44 (m, 5H), 4.01 (br t, J = 6.2 Hz, 2H), 3.85 (br t, J = 5.6 Hz, 3H), 3.73 (br dd, J = 9.6, 4.8 Hz, 3H), 3.41 (br s, 1H), 3.38 - 3.38 (m, 1H), 3.38 (br s, 1H), 3.17 (s, 1H), 2.97 (br t, J = 5.6 Hz, 3H), 2.85 - 2.93 (m, 3H), 2.74 - 2.81 (m, 3H), 2.46 (br d, J = 2.8 Hz, 5H), 2.36 (br s, 3H), 2.19 (s, 2H), 2.17 (s, 1H), 1.90 - 2.06 (m, 4H), 1.73 - 1.83 (m, 1H), 1.61 - 1.72 (m, 2H), 1.23 (s, 2H), 0.96 (br d, J = 6.4 Hz, 3H), 0.78 (br s, 1H), 0.76 (d, J = 6.8 Hz, 2H).

[0877] Synthesis of APH7:

[0878] 1. Synthesis of Compound H7-2

[0879] To a solution of compound H7-1 (200 mg, 724 μmol, 1.00 equiv) and compound 7A (193 mg, 651 μmol, 0.90 equiv) in DMSO (2.00 mL) were added EDCI (208 mg, 1.09 mmol, 1.50 equiv), HOAt (148 mg, 1.09 mmol, 152 μL, 1.50 equiv) and NMM (220 mg, 2.17 mmol, 239 μL, 3.00 equiv), and the mixture was stirred at 25 ° C for 1 hour. After the reaction, the mixture was diluted with water (10.0 mL) and extracted with DCM (10.0 mL * 6). The resulting 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 analyzed by preparative TLC (petroleum ether: ethyl acetate = 0:1, P1:R f =0.50) was purified to give compound H7-2 (240 mg, 433 μmol, 59.8% yield) as a yellow oil.

[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 μmol, 1.00 equiv) in methanol (3.00 mL) and water (1.00 mL) was added LiOH.H2O (54.5 mg, 1.30 mmol, 3.00 equiv), and the mixture was stirred at 25°C for 1 hour. LCMS (EC17015-206-P1A, P1: RT = 0.328 min) showed 96.0% formation of the desired product. The reaction mixture was filtered and concentrated under reduced pressure to afford crude compound H7-3 (230 mg) as a yellow solid, which was used directly in the next step 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(400MHz DMSO)

[0885] δppm 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= 14.2,6.8Hz,3H),2.10-2.19(m,3H),1.91-2.03(m,1H),1.72-1.85(m,1H ),1.24(d,J=12.2Hz,1H),0.93(dd,J=6.2,2.8Hz,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) were added HATU (243 mg, 638 μmol, 1.50 equiv) and DIEA (330 mg, 2.55 mmol, 445 μL, 6.00 equiv). The mixture was stirred at 25°C for 1 hour. LCMS (EC17015-213-P1A, P1: RT = 0.356 min) indicated the desired product was generated in 42.4% of the reaction mixture. After the reaction, the mixture was added with water (20.0 mL) and extracted with ethyl acetate (15.0 mL*3). The obtained organic layer was washed with saturated brine (20.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a crude product, which was further analyzed by preparative TLC (dichloromethane:methanol=10 / 1, P1:R f =0.45) was purified to give compound H7-4 (100 mg, 147 μmol, 34.5% yield) as a colorless oil.

[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 compound H7-4 (45.0 mg, 77.0 μmol, 1.00 equiv) was added formic acid (1.00 mL), and the mixture was stirred at 25°C for 0.5 h. LCMS (EC17015-224-P1A, P1: RT = 0.341 min) revealed 91.5% formation of the desired product. The reaction mixture was filtered and concentrated under reduced pressure to afford compound H7-5 (42.0 mg, crude) as a yellow oil, which was used directly in the next step without 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) were 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). 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 the reaction was stirred at 25°C for 1 h. LCMS (EC17015-225-P1B, P1: RT = 0.448 min) indicated the formation of the desired product in 52.0% of the reaction mixture. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain a crude product, which was further purified by preparative HPLC (chromatographic column: CD01-Phenomenex luna C18 150*25*10 μm; mobile phase: [water (FA)-ACN]; gradient: 30%-50% B in 10 minutes) to obtain 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(400MHz DMSO)

[0896] δppm 8.90-9.05(m,1H),8.18(s,1H),8.03(d,J=7.8Hz,1H),7.78(d,J=8.0Hz,1H),7.59(d,J=7.4Hz,1H),7.48-7.54(m,1H),7.46(d,J=8.2Hz,1H ),7.30-7.44(m,7H),7.26(d,J=12.4Hz,1H),7.14-7.21(m,1H),7.05-7.13(m,1H),6.91(d,J=8.8Hz,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=6.4Hz,3H),3.85(t,J=5.6Hz,4H),3.69-3.77(m,4H),2.97(t,J=5.4Hz,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=6.4Hz,3H),0.78(dd,J=14.2,6.8Hz,5H).

[0897] Synthesis of APH8:

[0898] 1. Synthesis of compound H8-2

[0899] To a solution of compound H8-1 (200 mg, 812 μmol, 1.0 equiv) in DMSO (2.00 mL) were added EDCI (233 mg, 1.22 mmol, 1.5 equiv), HOAt (166 mg, 1.22 mmol, 170 μL, 1.5 equiv), compound 7A (120 mg, 406.00 μmol, 0.5 equiv) and NMM (246 mg, 2.44 mmol, 267.82 μL, 3.0 equiv), and the mixture was stirred at 25 ° C for 1 hour. After the reaction, the mixture was added with water (15.0 mL) and extracted with ethyl acetate (15.0 mL * 3). The resulting organic layer was washed with saturated brine (25.0 mL * 2), dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product, which was further analyzed by preparative TLC (SiO2, PE:EA = 0:1, P1:R f =0.1) was purified to give compound H8-2 (100 mg, 177 μmol, 21.74% yield, 92.6% purity) as a 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 μmol, 1.0 equiv) in DCM (2.00 mL) were added TEA (135 mg, 1.33 mmol, 185.72 μL, 10.0 equiv), NaBH(OAc)3 (84.8 mg, 400 μmol, 3.0 equiv) and compound C1 (47.0 mg, 66.7 μmol, 0.5 equiv), and the mixture was stirred at 25°C for 2 hours. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain a crude product, which was further purified by preparative HPLC (FA conditions; column: Phenomenex luna C18 150*25mm*10μm; mobile phase: [water (FA)-ACN]; gradient: 29%-59% B in 10 minutes) to obtain compound APH8 product (10.0 mg, 7.57μmol, 5.67% yield, 91.86% purity) as a white solid.

[0903] LC-MS: EC16331-94-P1B1, product: RT = 0.440 min, 1 / 2 m / z = 607.5 [M+H] + ;

[0904] 11H NMR: EC16331-94-P1H (400 MHz, DMSO) δ ppm 8.39 (br d, J = 8.4 Hz, 1H), 8.32 (s, 1H), 7.98 - 8.05 (m, 1H), 7.77 (d, J = 8.0 Hz, 1H), 7.59 (br d, J = 6.8 Hz, 1H), 7.45 (br t, J = 7.2 Hz, 1H), 7.34 - 7.42 (m, 3H), 7.33 (br dd, J = 7.2, 3.75 Hz, 2H), 7.25 - 7.30 (m, 4H), 7.24 (br s, 1H), 7.21 (br d, J = 4.4 Hz, 1H), 7.18 (br d, J = 4.0 Hz, 1H), 7.15 (br s, 1H), 7.06 - 7.12 (m, 1H), 6.71 - 6.78 (m, 1H), 6.21 (s, 1H), 5.10 - 5.21 (m, 1H), 4.88 (br s, 2H), 4.21 - 4.36 (m, 5H), 3.95 - 4.02 (m, 4H), 3.81 (br d, J = 5.2 Hz, 5H), 3.71 (br d, J = 2.4 Hz, 6H), 2.95 (br s, 3H), 2.80 - 2.88 (m, 2H), 2.65 - 2.73 (m, 3H), 2.30 - 2.37 (m, 1H), 2.15 - 2.24 (m, 4H), 1.91 - 2.03 (m, 3H), 1.52 - 1.79 (m, 3H), 1.23 (br s, 2H), 1.05 (br t, J = 7.2 Hz, 1H), 0.92 - 1.01 (m, 3H), 0.71 - 0.82 (m, 3H).

[0905] Synthesis of APH9:

[0906] 1. Synthesis of Compound H9-2

[0907] 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) were mixed in DMSO (5.00 mL). The mixture was stirred at 25°C for 1 hour. LCMS (EC17015-207-PA, P1:RT = 0.357 min) detected the formation of the desired product in 99.4% of the reaction mixture. After the reaction, the mixture was added with water (10.0 mL) and extracted with DCM (10.0 mL*6). The obtained organic layer was washed with saturated brine (10.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a crude product, which was further analyzed by preparative TLC (petroleum ether:ethyl acetate = 0 / 1, P1:R f =0.50) was purified to give compound H9-2 (700 mg, 1.53 mmol, 54.8% yield) as a colorless oil.

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

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

[0910] δppm 7.16-7.31(m,5H),5.96-6.12(m,1H),5.31(d,J=8.2Hz,1H),4.39-4.56(m,2H),3.72(dd,J=10.4,4.8Hz,1H),3.44-3.64(m,5 H),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=6.4Hz,3H),0.81(d,J=6.6Hz,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), and the mixture was stirred at 25°C for 1 hour. LCMS (EC17015-210-P1A, P1: RT = 0.327 min) indicated 100% formation of the desired product. The reaction mixture was filtered and concentrated under reduced pressure to afford crude product H9-3 (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) were 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 hour. LCMS (EC17015-216-P1B, P1: RT = 0.343 min) indicated the formation of the desired product in an 18.8% yield. The reaction mixture was filtered and concentrated under reduced pressure to obtain the crude product, which was further purified by reverse-phase HPLC (neutral conditions) to afford compound H9-4 (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 compound H9-4 (60.0 mg, 103 μmol, 1.00 equiv) was added formic acid (1.00 mL), and the mixture was stirred at 25°C for 0.5 h. LCMS (EC17015-230-P1A, P1: RT = 0.303 min) showed 87.8% formation of the desired product. The reaction mixture was filtered and concentrated under reduced pressure to afford crude product H9-5 (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) were 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. NaBH(OAc)3 (61.4 mg, 290 μmol, 3.00 equiv) was then added, and the reaction was continued at 25°C with stirring for 12 h. LCMS (EC17015-231-P1A, P1: RT = 0.447 min) indicated the formation of the desired product in 63.3% of the reaction mixture. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain a crude product, which was further purified by preparative HPLC (column: CD02-Waters Xbidge BEH C18 150*25*10 μm; mobile phase: [water(NH4HCO3)-ACN]; gradient: 38%-58% B over 10 minutes) to obtain 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(400MHz DMSO)δppm 8.03(d,J=7.8Hz,1H),7.78(d,J=8.2Hz,1H),7.60(d,J=7.2Hz,1H),7.52(d ,J=8.4Hz,1H),7.47(t,J=7.6Hz,1H),7.38-7.43(m,1H),7.35(d,J=7.4Hz,2 H),7.30-7.33(m,1H),7.29(s,4H),7.15-7.22(m,2H),7.05-7.13(m,1H),6 .91(d,J=9.0Hz,1H),6.14-6.23(m,1H),5.07-5.23(m,1H),4.92(s,2H),4.1 4-4.46(m,3H),4.01(t,J=6.4Hz,2H),3.85(t,J=5.6Hz,3H),3.72(d,J=9.8 Hz,2H),3.43-3.58(m,6H),2.97(t,J=5.6Hz,2H),2.74-2.87(m,5H),2.64-2 .73(m,2H),2.28-2.36(m,3H),2.14-2.21(m,4H),2.07(d,J=5.2Hz,2H),1.9 0-2.03(m,5H),1.50-1.80(m,5H),0.95(d,J=6.6Hz,3H),0.68-0.86(m,5H).

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

[0925] Synthesis of APH10:

[0926] 1. Synthesis of compound H10-1

[0927] Compound 8A (200 mg, 418 μmol, 1.00 equiv), compound 1A (100 mg, 628 μmol, 1.50 equiv), HATU (239 mg, 628 μmol, 1.50 equiv), and DIEA (324 mg, 2.51 mmol, 437 μL, 6.00 equiv) were combined in DMF (3.00 mL). The mixture was stirred at 25°C for 2 hours. LCMS (EC17015-293-P1B, P1: RT = 0.370 min) indicated a 20.1% yield of the desired product. After the reaction, the mixture was added with water (20.0 mL) and extracted with ethyl acetate (15.0 mL * 5). The resulting 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 HPLC (chromatographic column: CD07-Daisogel SP-100-8-ODS-PK 150 * 25 * 10 μm; mobile phase: [water (NH4HCO3) -ACN]; gradient: 24% -54% B over 12 minutes) to give compound H10-1 (165 mg, 266 μmol, 63.7% yield) as a 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(400MHz CDCl3)

[0930] δppm 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=9.6Hz,4H),2.16-2.26(m,1H),2.15-2.15(m,1H),1.92-2.13(m,2H),1.71-1.8 7(m,3H),1.70-1.71(m,1H),1.09-1.22(m,3H),0.84-0.89(m,1H),0.98(t,J=6.2Hz,3H)

[0931] 2. Synthesis of Compound H10-2

[0932] To compound H10-1 (77.0 mg, 124 μmol, 1.00 equiv) was added formic acid (1.00 mL), and the mixture was stirred at 25°C for 0.5 h. LCMS (EC17015-297-P1A) indicated that compound H10-1 had reacted completely. The reaction mixture was filtered and concentrated under reduced pressure to afford crude product H10-2 (71.0 mg) as a colorless oil.

[0933] LC-MS: EC17015-297-P1A, product: RT = 0.334 min, m / z = 573.1 [M+H] +

[0934] 3. Synthesis of Compound APH10

[0935] To a solution of compound H10-2 (71.0 mg, 124 μmol, 1.00 equiv) in DCM (3.00 mL) were added TEA (125 mg, 1.24 mmol, 172 μL, 10.0 equiv) and compound C1 (43.7 mg, 62.0 μmol, 0.50 equiv). The mixture was stirred at 25°C for 0.5 h. NaBH(OAc)3 (78.8 mg, 372 μmol, 3.00 equiv) and TEA (125 mg, 1.24 mmol, 172 μL, 10.0 equiv) were then added, and the mixture was stirred at 25°C for 1 h. LCMS (EC17015-299-P1A, P1: RT = 0.610 min) indicated the formation of the desired product in 68.0% yield. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain the crude product, which was further purified by preparative HPLC (column: CD02-Waters Xbidge BEH C18 150*25*10 μm; mobile phase: [water(NH4HCO3)-ACN]; gradient: 32%-62% B over 13 minutes) to obtain compound APH10 (26.0 mg, 19.4 μmol, 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(400MHz DMSO)δppm 8.04(d,J=7.6Hz,1H),7.79(d,J=8.0Hz,1H),7.60(d,J=7.6Hz,1H),7.53(d,J=8 .8Hz,1H),7.48(t,J=7.6Hz,1H),7.39-7.43(m,1H),7.27-7.39(m,6H),7.21-7. 26(m,1H),7.06-7.13(m,1H),7.18(d,J=8.8Hz,1H),6.93(d,J=8.4Hz,1H),6.14 -6.25(m,1H),5.06-5.22(m,1H),4.92(s,2H),4.20-4.45(m,3H),4.02(t,J=6.2H z,2H),3.86(t,J=5.6Hz,2H),3.66-3.77(m,2H),3.43-3.61(m,7H),3.10-3.17( m,2H),2.98(t,J=5.4Hz,2H),2.86-2.92(m,1H),2.79(t,J=7.4Hz,2H),2.61-2. 75(m,2H),2.27-2.43(m,5H),2.16-2.21(m,3H),2.08(s,2H),1.91-2.03(m,4H) ,1.51-1.79(m,4H),1.19-1.29(m,1H),0.95(d,J=6.4Hz,3H),0.68-0.83(m,4H).

[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 MeI (19.0 g, 133 mmol, 8.31 mL, 5.00 equiv) dropwise at 25°C under N2 atmosphere over 30 minutes, and the mixture was stirred at 80°C for 2.5 hours. After the reaction, the mixture was added with water (200 mL) and extracted with ethyl acetate (120 mL*3). The resulting organic layer was washed with saturated brine (120 mL*2), dried over Na2SO4, filtered under reduced pressure, and concentrated to give a crude product of compound H11-2 (9.00 g) as a yellow solid, which was used directly in the next reaction without purification.

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

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

[0943] 2. Synthesis of Compound H11-3

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

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

[0946] 3. Synthesis of Compound H11-4

[0947] Compound H11-3 (1.80 g, 7.20 mmol, 1.00 equiv, HCl), compound 7A (1.07 g, 3.60 mmol, 0.50 equiv), EDCI (2.07 g, 10.8 mmol, 1.50 equiv), NMM (2.18 g, 21.6 mmol, 2.37 mL, 3.00 equiv), and HOAt (1.47 g, 10.8 mmol, 1.51 mL, 1.50 equiv) were mixed in DMSO (20.0 mL) and stirred at 25°C for 1 hour. LCMS (EC17015-237-P1A1, P1: RT = 0.353 min) showed that the desired product was produced in 84.0% of the reaction mixture. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain a crude product, which was further purified by reverse phase HPLC (0.1% FA) to obtain 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 equiv) in methanol (12.0 mL) and water (4.00 mL) was added LiOH.H2O (409 mg, 9.76 mmol, 3.00 equiv). The mixture was stirred at 25°C for 1 hour. LCMS (EC17015-249-P1A, P1: RT = 0.326 min) showed 100% formation of the desired product. The reaction mixture was filtered and concentrated under reduced pressure to afford compound 8A (1.80 g, crude product) as a yellow solid, which was used directly in the next reaction 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 μmol, 1.00 equiv), compound C1 (10.3 mg, 14.6 μmol, 0.70 equiv), PyBOP (13.1 mg, 25.1 μmol, 1.20 equiv), and TEA (6.35 mg, 62.8 μmol, 8.74 μL, 3.00 equiv) were mixed in DMA (0.50 mL) and stirred at 25°C for 1 hour. LCMS (EC17015-294-P1E1, P1: RT = 0.423 min) detected 18.8% formation of the desired product. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain the crude product, which was further purified by preparative HPLC (column: CD02-Waters Xbidge BEH C18 150*25*10 μm; mobile phase: [water(NH4HCO3)-ACN]; gradient: 22%-52% B over 10 minutes) to obtain compound APH11 (8.30 mg, 5.61 μmol, 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(400MHz DMSO)δppm 8.02(d,J=8.2Hz,1H),7.78(d,J=8.4Hz,1H),7.60(d,J=7.2Hz,1H),7.43-7.51(m,2H),7.38-7.42(m,1H),7.23-7.37(m,7 H),7.17(d,J=8.4Hz,1H),7.05-7.13(m,1H),6.85(d,J=7.8Hz,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=6.0Hz,3H),0.68-0.86(m,4H).

[0956] Synthesis of APH12:

[0957] 1. Synthesis of compound H12-2

[0958] Compound 8A (200 mg, 418 μmol, 1.00 equiv), compound H12-1 (66.0 mg, 628 μmol, 68.4 μL, 1.50 equiv), HATU (239 mg, 628 μmol, 1.50 equiv), and DIEA (324 mg, 2.51 mmol, 437 μL, 6.00 equiv) were mixed in DMF (3.00 mL) and stirred at 25°C for 1 hour. LCMS (EC17015-279-P1B, P1: RT = 0.337 min) indicated the formation of the desired product in 99.7% of the reaction mixture. After the reaction, the mixture was concentrated under reduced pressure to obtain a crude product, which was further purified by preparative HPLC (neutral conditions; column: CD07-Daisogel SP-100-8-ODS-PK 150*25*10 μm; mobile phase: [water(NH4HCO3)-ACN]; gradient: 22%-52% B over 10 minutes) to obtain compound H12-2 (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 μmol, 1.00 equiv) was added formic acid (1.00 mL), and the mixture was stirred at 25°C for 0.5 h. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain compound H12-3 (60.0 mg, crude product) as a yellow oil, which was used directly in the next reaction without 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 μmol, 1.00 equiv) and compound C1 (40.7 mg, 57.8 μmol, 0.50 equiv) in DCM (2.00 mL) were added TEA (117 mg, 1.16 mmol, 161 μL, 10.0 equiv) and NaBH(OAc)3 (73.5 mg, 347 μmol, 3.00 equiv), and the mixture was stirred at 25°C for 2 h. After the reaction, the mixture was filtered and concentrated under reduced pressure to give a crude product (60.0 mg), which was further purified by preparative HPLC (column: Phenomenex luna C18 150*25mm*10μm; mobile phase: [water(HCl)-ACN]; gradient: 37%-67% B in 14 minutes) to give compound APH12 (25.0 mg, 19.3μmol, 16.7% yield, 96.2% purity, HCl) as a white solid.

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

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

[0967] d) Synthesis of APH14, APH15, and 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, HCl) in ACN (15.0 mL) were 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), and the mixture was stirred at 25° C. for 12 hours. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain the crude product, which was further purified by column chromatography (SiO 2 , petroleum ether / ethyl acetate = 4 / 1 to 1 / 1) to obtain the product H14-2 (2.25 g, 15.9 mmol, 74.2% yield) as a yellow oil.

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

[0972] 2. Synthesis of Compound H14-3

[0973] Compound 6A (200 mg, 840 μmol, 1.00 equiv), compound H14-2 (178 mg, 1.26 mmol, 1.50 equiv), Pd(PPh3)2Cl2 (59.0 mg, 84.0 μmol, 0.10 equiv), and CuI (8.00 mg, 42.0 μmol, 0.05 equiv) were stirred in TEA (2.00 mL) at 75°C under a nitrogen atmosphere for 2 hours. LCMS (EC17015-226-P1A, P1: RT = 0.341 min) indicated the formation of the desired product in 49.3% yield. The reaction mixture was concentrated under reduced pressure to afford the crude product, which was then purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 7 / 3 to 1 / 1) to afford compound H14-3 (170 mg, 677 μmol, 80.5% yield) as a 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 (400MHz CDCl3) δppm 7.08-7.19(m,2H),6.89-6.97(m,1H),3.75(s,3H),3.67(s,2H),3.40(s,2H),2.48(s,3H).

[0976] 3. Synthesis of Compound H14-4

[0977] A solution of compound Q4-3 (200 mg, 329 μmol, 1.00 equiv), compound H14-3 (91.0 mg, 362 μmol, 1.10 equiv), and CsCO (322 mg, 988 μmol, 3.00 equiv) in DMA (4.00 mL) was stirred at 25°C under an N2 atmosphere for 4 hours. LCMS (EC17015-229-P1A, P1: RT = 0.469 min) showed 48.0% formation of the desired product. After the reaction, the mixture was added with water (20.0 mL) and extracted with ethyl acetate (15.0 mL*3). The obtained 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 column chromatography (SiO2, petroleum ether / ethyl acetate = 3 / 2 to 1 / 1) to give compound H14-4 (228 mg, 293 μmol, 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(400MHz DMSO)δppm 12.84(s,1H),8.03(d,J=7.8Hz,1H),7.78(d,J=8.0Hz,1H),7.55(d,J=7.2Hz,1H),7.4 3-7.52(m,2H),7.27-7.42(m,4H),7.18(d,J=8.2Hz,1H),7.05-7.13(m,1H),6.91(d,J =8.8Hz,1H),4.93(s,2H),4.02(t,J=6.07Hz,2H),3.78(t,J=6.0Hz,2H),3.56-3.64(m ,5H),3.33(s,4H),2.96-3.00(m,2H),2.66(t,J=7.6Hz,2H),2.33(s,3H),1.29(s,9H).

[0980] 4. Synthesis of Compound H14-5

[0981] To a solution of compound H14-4 (228 mg, 293 μmol, 1.00 equiv) in methanol (3.00 mL) and H2O (1.00 mL) was added LiOH.H2O (36.9 mg, 880 μmol, 3.00 equiv). The mixture was stirred at 25°C for 0.5 h. LCMS (EC17015-232-P1W, P1:RT = 0.464 min) detected the formation of the desired product at a rate of 68.5%. The reaction mixture was adjusted to pH 3 with 1 M hydrochloric acid, diluted with water (10.0 mL), and extracted with a mixture of DCM and methanol (DCM:methanol = 10:1, 15.0 mL x 3). The resulting organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to afford crude product H14-5 (200 mg, 262 μmol, 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 μmol, 1.00 equiv), compound 2 (69.8 mg, 157 μmol, 1.00 equiv), PyBOP (98.1 mg, 188 μmol, 1.20 equiv), and TEA (47.7 mg, 471 μmol, 65.6 μL, 3.00 equiv) were combined in DMA (2.00 mL) and stirred at 25°C for 1 hour. LCMS (EC17015-242-P1C, P1: RT = 0.859 min) detected the formation of the desired product in 92.3% of the reaction mixture. After the reaction, the mixture was diluted with water (20.0 mL) and extracted with DCM (15.0 mL*3). The obtained organic layer was washed with saturated brine (10.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give the crude product H14-6 (160 mg, crude product) as a yellow oil, which was used directly in the next reaction without purification.

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

[0986] 6. Synthesis of compound APH14

[0987] Compound H14-6 (150 mg, 126 μmol, 1.00 equiv) was added with HCl / dioxane (5.00 mL), and the mixture was stirred at 25°C for 6 hours. LCMS (EC17015-250-P1B2, P1:RT = 0.607 min) detected the production of the desired product at a rate of 35.2%. The reaction mixture was concentrated under reduced pressure to afford the crude product, which was then purified by preparative HPLC (column: CD02-Waters Xbidge BEH C18 150*25*10 μm; mobile phase: [water (NH4HCO3)-ACN]; gradient: 36%-66% B over 10 minutes) to afford compound APH14 (20.0 mg, 17.6 μmol, 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(400MHz DMSO)δppm 8.93-9.02(m,1H),8.44(d,J=7.6Hz,1H),8.03(d,J=8.0Hz,1H),7.76-7.81(m,1H),7.69(d,J=9.6Hz,1H) ,7.60(d,J=7.6Hz,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=9.6Hz,1H),4.39-4.48(m,1H),4.29(d,J =2.8Hz,1H),4.02(t,J=6.4Hz,2H),3.85(t,J=5.8Hz,2H),3.54-3.65(m,4H),3.11(d, J=2.8Hz,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=6.8Hz,3H),1.23(s,1H),0.94(s,9H).

[0990] Synthesis of APH15:

[0991] The synthesis steps of PROTAC molecule APH15 refer to APH14, but the starting materials are replaced with compounds H15-1 and 4A. After synthesis and purification, the target product APH15 (20.0 mg, 16.3 μmol, 6.55% yield, 93.6% purity) is obtained as a white solid.

[0992] 1 H NMR:EC16331-167-P1L(400MHz DMSO)δppm 8.98(s,1H),8.38(d,J=7.6Hz,1H),8.16(d,J=9.2Hz,1H),8.03(d,J=8.0Hz,1H) ,7.79(d,J=8.0Hz,1H),7.60(d,J=7.2Hz,1H),7.53(d,J=8.8Hz,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=12.0Hz,1H),7.17(d,J=8.8Hz,1H),7.07-7.13(m,1H),6.93(d,J= 8.8Hz,1H),4.92(s,3H),4.50(d,J=9.2Hz,1H),4.42(t,J=8.0Hz,1H),4.28(s,1 H),4.02(t,J=6.0Hz,2H),3.86(t,J=5.6Hz,2H),3.60(s,2H),3.53(s,3H),2.79( t,J=7.2Hz,2H),2.62(dd,J=16.8,6.8Hz,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=6.8Hz,3H),0.94(s,12H).

[0993] Synthesis of APH16:

[0994] The synthesis steps of PROTAC molecule APH16 refer to APH14, but the starting materials are replaced with compounds H16-1 and 4A. After synthesis and purification, the target product APH16 (20.0 mg, 16.0 μmol, 6.49% yield, 92.9% purity) is obtained as a white solid.

[0995] 1H NMR:EC16331-166-P1M(400MHz DMSO)δppm 8.96-8.99(m,1H),8.37(d,J=7.6Hz,1H),8.04(d,J=7.6Hz,1H),7.77-7.84(m,2H), 7.60(d,J=7.6Hz,1H),7.54(d,J=8.8Hz,1H),7.46-7.50(m,1H),7.44(s,1H),7.33- 7.43(m,6H),7.23-7.29(m,1H),7.15-7.19(m,1H),7.06-7.12(m,1H),6.95(d,J=8. 8Hz,1H),5.04-5.16(m,1H),4.88-4.94(m,3H),4.51(d,J=9.2Hz,1H),4.42(t,J=8. 0Hz,1H),4.27(s,1H),4.02(t,J=6.4Hz,2H),3.86(t,J=5.6Hz,2H),3.60(s,2H),3. 48(s,3H),2.98(t,J=5.2Hz,2H),2.80(t,J=7.2Hz,2H),2.42-2.47(m,3H),2.36(t, J=7.2Hz,2H),2.22(s,3H),2.11-2.19(m,1H),1.92-2.04(m,3H),1.79(tt,J=8.4,4 .0Hz,1H),1.59-1.68(m,2H),1.37(d,J=6.8Hz,3H),1.16-1.27(m,1H),0.93(s,9H).

[0996] e) Synthesis of APH17, APH18, APH19, APH20, and 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 CuI (12.0 mg, 63.0 μmol, 0.05 equiv) were mixed in TEA (5.00 mL) and stirred at 75°C under a nitrogen atmosphere for 2 hours. LCMS (EC17015-233-P1A, P1:RT=0.355 min) revealed the production of the desired product at a yield of 62.9%. The reaction mixture was filtered and concentrated under reduced pressure to obtain the crude product, which was further purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 7 / 3 to 1 / 1) to afford H17-2 (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(400MHz DMSO)δppm 10.21(s,1H),7.13(dd,J=12.0,1.8Hz,1H),7.01(dd,J=8.4,1.0Hz,1H),6.84-6.93(m,1H),3.62(s,3H),2.60(dd,J=9.6,5.2Hz,4H).

[1002] 2. Synthesis of Compound H17-3

[1003] Compound Q4-3 (200 mg, 329 μmol, 1.00 equiv), compound H17-2 (80.5 mg, 362 μmol, 1.10 equiv), and CsCO (322 mg, 988 μmol, 3.00 equiv) were combined in DMA (3.00 mL) and stirred at 25°C for 12 hours. LCMS (EC17015-247-P1A, P1: RT = 0.517 min) indicated a 68.2% yield of the desired product. After the reaction, the mixture was diluted with water (20.0 mL) and extracted with ethyl acetate (15.0 mL*3). The obtained 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 the H17-3 product (155 mg, 207 μmol, 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(400MHz DMSO)δppm 12.82(s,1H),8.03(d,J=7.8Hz,1H),7.78(d,J=8.0Hz,1H),7.55(d,J=6.8Hz,1H),7.44-7. 51(m,2H),7.31-7.31(m,1H),7.31-7.41(m,2H),7.19(dd,J=11.8,1.6Hz,1H),7.04-7.13(m ,2H),6.91(d,J=8.8Hz,1H),4.93(s,2H),3.96-4.03(m,2H),3.78(t,J=6.0Hz,2H),3.62(s, 3H), 2.98 (t, J = 5.8Hz, 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 μmol, 1.00 equiv) in methanol (1.20 mL) and water (0.40 mL) was added LiOH.H2O (26.1 mg, 621 μmol, 3.00 equiv), and the mixture was stirred at 25°C for 4 hours. LCMS (EC17015-252-P1A2, P1:RT = 0.483 min) detected the formation of the desired product at a rate of 92.2%. The reaction mixture was adjusted to pH 3 with 1M hydrochloric acid, diluted with water (10.0 mL), and extracted with DCM and methanol (DCM:methanol = 10:1, 15.0 mL x 3). The resulting organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to afford compound H17-4 (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 μmol, 1.00 equiv), compound 2 (90.8 mg, 204 μmol, 1.00 equiv), PyBOP (127 mg, 245 μmol, 1.20 equiv), and TEA (62.0 mg, 612 μmol, 85.2 μL, 3.00 equiv) were mixed in DMA (2.00 mL) and stirred at 25°C for 1 hour. LCMS (EC17015-256-P1A, P1: RT = 0.548 min) showed 81.8% yield of the desired product. After the reaction, the mixture was diluted with water (20.0 mL) and extracted with DCM (15.0 mL*3). The obtained organic layer was washed with saturated brine (10.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain the crude product of compound H17-5 (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 μmol, 1.00 equiv) was dissolved in HCl / dioxane (4.00 mL) and the mixture was stirred at 25°C for 6 hours. LCMS (EC17015-261-P1A1, P1:RT = 0.597 min) detected the formation of the desired product at a rate of 30.0%. The reaction mixture was filtered and concentrated under reduced pressure to obtain the crude product, which was further purified by preparative HPLC (column: CD02-Waters Xbidge BEH C18 150*25*10 μm; mobile phase: [water (NH4HCO3)-ACN]; gradient: 28%-58% B over 13 minutes) to obtain the product APH17 (24.0 mg, 21.2 μmol, 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(400MHz DMSO)δppm 8.98(s,1H),8.38(d,J=7.8Hz,1H),8.01(dd,J=19.2,8.2Hz,2H),7.79(d,J=7.8Hz,1H),7.60(d,J=7.8Hz,1H),7.53(d,J=8.4Hz,1H), 7.47(s,1H),7.32-7.45(m,7H),7.17(d,J=12.4Hz,1H),7.06-7.11(m,1H),6.93(d,J=8.6Hz,1H),5.13(s,1H),4.92(s,3H),4.56(d,J =9.2Hz,1H),4.42(t,J=8.2Hz,1H),4.28(s,1H),4.01(t,J=5.6Hz,2H),3.85(s,2H),3.54-3.69(m,3H),2.98(d,J=5.8Hz,2H),2.78(t ,J=7.0Hz,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=6.8Hz,3H),0.94(s,9H).

[1016] Synthesis of APH18:

[1017] The synthesis steps of PROTAC molecule APH18 refer to APH17, but the starting materials are replaced with compounds H18-1 and 6A. After synthesis and purification, the target product APH18 (32.7 mg, 27.5 μmol, 18.6% yield, 93.9% purity) is obtained as a white solid.

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

[1019] 1 H NMR:EC21184-23-P1QC(400MHz,DMSO)δppm 8.98(s,1H),8.38(d,J=7.6Hz,1H),8.03(d,J=7.6Hz,1H),7.91(d,J=9.2Hz,1H),7.79(d,J=7.6Hz,1H),7.60(d,J=7.2Hz,1H),7.31-7.54(m,9 H),7.25(dd,J=12.0,1.6Hz,1H),7.12-7.17(m,1H),7.04-7.11(m,1H),6.91(d,J=8.8Hz,1H),5.00-5.23(m,1H),4.92(s,2H),4.53(d,J=9.2H z,1H),4.42(t,J=8.00Hz,1H),4.28(d,J=1.6Hz,1H),4.01(t,J=6.4Hz,2H),3.85(t,J=6.0Hz,2H),3.61(s,2H),2.94-3.04(m,3H),2.78(t,J= 7.6Hz,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=6.8Hz,3H),0.91-0.98(m,9H).

[1020] Synthesis of APH19:

[1021] The synthesis steps of PROTAC molecule APH19 refer to APH17, but the starting materials are replaced with compounds H19-1 and 6A. After synthesis and purification, the target product APH19 (18.0 mg, 15.6 μmol, 10.0% yield, 98.5% purity) is obtained as 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(400MHz DMSO)δppm 8.94-9.02(m,1H),8.37(d,J=7.6Hz,1H),8.03(d,J=7.6Hz,1H),7.81(dd ,J=17.2,8.8Hz,2H),7.60(d,J=7.2Hz,1H),7.29-7.56(m,10H),7.20(d, J=12.0Hz,1H),7.09(dt,J=16.4,8.4Hz,2H),6.8(d,J=8.8Hz,1),5.02-5 .22(m,1H),4.87-4.98(m,3H),4.52(d,J=9.2Hz,1H),4.42(t,J=8.0Hz,1H ),4.28(s,1H),4.00(t,J=6.0Hz,2H),3.85(s,2H),3.60(s,2H),2.97(s, 2H),2.79(t,J=6.8Hz,2H),2.42-2.47(m,3H),2.39(t,J=6.4Hz,2H),2.2 5-2.34(m,1H),2.11-2.21(m,1H),1.91-2.05(m,3H),1.74-1.84(m,1H), 1.57-1.69(m,2H),1.45-1.54(m,2H),1.37(d,J=6.8Hz,3H),0.94(s,9H).

[1024] Synthesis of APH20:

[1025] The synthesis steps of the PROTAC molecule APH20 refer to APH17, but the starting materials are replaced with compounds H20-1 and 6A. After synthesis and purification, the target product APH20 (18.0 mg, 14.9 μmol, 5.96% yield, 94.7% purity) is 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 (400MHz DMSO) δppm 8.98 (s, 1H), 8.37 (d, J=7.6Hz, 1H),8.02(d,J=7.6Hz,1H),7.80(dd,J=12.8,8.8Hz,2H),7.60(d,J=6.8Hz,1H),7.45-7.51(m,2H),7.31-7.45(m,7H),7.20(dd,J=12.0,1.8 Hz,1H),7.03-7.13(m,2H),6.87(d,J=8.8Hz,1H),4.82-5.03(m,3H),4.51(d,J=9.6Hz,1H),4.42(t,J=8.0Hz,1H),4.27(s,1H),4.00(t,J=6. 4Hz,2H),3.85(t,J=5.6Hz,2H),3.60(s,2H),2.97(t,J=5.6Hz,2H),2.76(t,J=7.6Hz,2H),2.43-2.47(m,3H),2.36(t,J=6.8Hz,2H),2.21-2 .31(m,1H),2.08-2.20(m,1H),1.88-2.08(m,4H),1.78(ddd,J=12.8,8.4,4.8Hz,1H),1.44-1.57(m,4H),1.37(d,J=7.2Hz,5H),0.92(s,9H).

[1028] Synthesis of APH21:

[1029] The synthesis steps of the PROTAC molecule APH21 refer to APH17, but the starting materials are replaced with compounds H21-1 and 6A. After synthesis and purification, the target product APH21 (22.0 mg, 18.3 μmol, 7.43% yield, 96.6% purity) is obtained as a white solid.

[1030] LC-MS: EC17015-342-P1A1, product: RT = 0.642 min, m / z = 1161.3 [M+H] +

[1031] 1 H NMR:EC17015-342-P1A(400MHz DMSO)

[1032] δppm 8.98(s,1H),8.37(d,J=7.6Hz,1H),8.03(d,J=8.0Hz,1H),7.79(dd,J=8.4,5 .6Hz,2H),7.59(d,J=7.6Hz,1H),7.52(d,J=8.8Hz,1H),7.46-7.50(m,1H),7. 32-7.45(m,7H),7.20(d,J=12.0Hz,1H),7.03-7.15(m,2H),6.92(d,J=8.4Hz ,1H),4.92(s,3H),4.51(d,J=9.2Hz,1H),4.41(t,J=8.0Hz,1H),4.27(s,1H), 4.00(t,J=6.4Hz,2H),3.85(t,J=5.6Hz,2H),3.60(s,2H),2.95-3.00(m,2H) ,2.74-2.82(m,2H),2.45(s,3H),2.36(t,J=6.8Hz,2H),2.17-2.30(m,2H),2. 12(dt,J=13.6,7.2Hz,1H),1.91-2.03(m,3H),1.78(ddd,J=12.8,8.4,4.4Hz, 1H),1.44-1.54(m,4H),1.37(d,J=7.2Hz,5H),1.23-1.31(m,2H),0.93(s,9H)

[1033] f) Synthesis of APH101, APH102, APH103, APH104, APH105, and APH106

[1034] Synthesis of APH101:

[1035] 1. Synthesis of compound H101-1

[1036] Compound H101-1 (1.00 g, 5.40 mmol, 1.00 equiv) was mixed in HCl / dioxane (1.00 mL) and stirred at 20°C under a nitrogen atmosphere for 2 hours. After the reaction, the mixture was filtered and concentrated under reduced pressure to afford compound H101-2 (930 mg, crude product) as a white solid.

[1037] 1 H NMR: EC21184-134-p1a (400MHz DMSO) δ9.37 (d, J = 1.2 Hz, 1H), 3.88 (d, J = 3.2 Hz, 4H), 2.99-3.20 (m, 1H).

[1038] 2. Synthesis of Compound H101-3

[1039] To a solution of compound H101-2 (100 mg, 823 μmol, 1.00 equiv, HCl) and compound 8A (275 mg, 576 μmol, 0.70 equiv) in DMF (3.00 mL) were added HATU (469 mg, 1.23 mmol, 1.50 equiv) and DIEA (319 mg, 2.47 mmol, 430 μL, 3.00 equiv). The mixture was stirred at 20°C for 2 hours. LC-MS (EC21184-136-p1a, P1: RT = 0.344 min) indicated a yield of approximately 20.9% of the desired product. After the reaction, the mixture was concentrated under reduced pressure to obtain a crude product, which was further purified by preparative HPLC (FA conditions; column: CD01-Phenomenex luna C18 150*25*10 μm; mobile phase: [water (FA)-ACN]; gradient elution: 10%-40% B in 10 minutes) to obtain compound H101-3 (25.0 mg, 35.2 μmol, 4.28% yield, 76.7% purity) as a light yellow solid.

[1040] LC-MS: EC21184-136-p1a, P1: RT=0.344min, m / z=545.2[M+H] +

[1041] 3. Synthesis of compound APH101

[1042] To a solution of compound H101-3 (25.0 mg, 45.9 μmol, 1.00 equiv) and compound C1 (22.6 mg, 32.1 μmol, 0.70 equiv) in DCM (1.00 mL) were added TEA (46.4 mg, 459 μmol, 63.8 μL, 10.0 equiv) and NaBH(OAc) (29.2 mg, 138 μmol, 3.00 equiv). The mixture was stirred at 20°C for 4 hours. LC-MS (EC21184-144-p1b, P1: RT = 2.036 min) indicated a yield of approximately 58.8% of the desired product. After the reaction, the mixture was concentrated under reduced pressure to obtain the crude product, which was purified using preparative HPLC (neutral conditions; chromatographic column: CD02-Waters Xbidge BEH C18 150*25*10 μm; mobile phase: [water (NH4HCO3)-ACN]; gradient elution: 26%-56% B in 10 minutes) and further purified using preparative HPLC (basic conditions; chromatographic column: CD02-Waters Xbidge BEH C18 150*25*10 μm; mobile phase: [water (NH3H2O)-ACN]; gradient elution: 17%-47% B in 10 minutes) to obtain compound APH101 (6.11 mg, 4.72 μmol, 10.3% yield, 95.3% purity) as a white solid.

[1043] LC-MS: EC21184-144-p1b, P1: RT=2.036min, m / z=617.7[M / 2+H] +

[1044] 1 H NMR:EC21184-144-p1b(400MHz DMSO)

[1045] δ11.88-13.34(m,1H),8.35-8.51(m,1H),8.03(d,J=8.0Hz,1H),7.79(d,J=8 .0Hz,1H),7.60(d,J=7.6Hz,1H),7.44-7.55(m,2H),7.32-7.43(m,5H),7.23 -7.31(m,3H),7.15-7.19(m,1H),7.06-7.12(m,1H),6.92(d,J=8.8Hz,1H),6 .05-6.25(m,1H),5.01-5.15(m,1H),4.92(s,2H),4.19-4.34(m,2H),3.92-4 .11(m,3H),3.85(t,J=6.0Hz,2H),3.76-3.83(m,1H),3.68-3.75(m,2H),3.4 4(s,3H),2.97(t,J=5.6Hz,2H),2.79(t,J=7.2Hz,2H),2.60-2.68(m,1H),2. 39-2.48(m,5H),2.21-2.39(m,6H),2.11-2.21(m,4H),1.92-2.01(m,3H),1. 66-1.76(m,1H),1.18-1.34(m,2H),0.95(d,J=6.4Hz,3H),0.72-0.80(m,3H).

[1046] Synthesis of APH102:

[1047] 1. Synthesis of compound H102-2

[1048] To a solution of compound H102-1 (1.00 g, 3.53 mmol, 1.00 equiv) and compound 4A (630 mg, 4.23 mmol, 456 μL, 1.20 equiv) in ACN (10.0 mL) was added KCO (1.46 g, 10.6 mmol, 3.00 equiv), and the mixture was stirred at 60° C. for 2 hours. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain a crude product, which was further purified by column chromatography (SiO, DCM / methanol = 100 / 0 to 85 / 15) to obtain compound H102-2 (530 mg, crude) as a brown solid.

[1049] 1H NMR:EC21184-58-P1A(400MHz CDCl3)δ3.37-3.46(m,4H),3.32(d,J=2.4Hz,2H),2.90(d,J=11.2Hz,2H),2.29-2.39(m,4H), 2.14-2.28(m,5H),1.79(d,J=13.2Hz,2H),1.49-1.56(m,1H),1.46(s,9H),1.21-1.33(m,2H)

[1050] 2. Synthesis of compound H102-3

[1051] Compound H102-2 (430 mg, 1.34 mmol, 1.00 equiv) was mixed in HCl / dioxane (20.0 mL) and stirred at 20°C for 1 hour. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain compound H102-3 (300 mg, crude product) as a white solid.

[1052] 3. Synthesis of compound H102-4

[1053] Compound H102-3 (300 mg, 1.36 mmol, 1.00 equiv), compound Q9-1 (518 mg, 678 μmol, 0.50 equiv), CuI (12.9 mg, 67.8 μmol, 0.05 equiv) and Pd(PPh3)2Cl2 (95.1 mg, 135 μmol, 0.10 equiv) were mixed in a solution of TEA (5.00 mL) and DMF (1.00 mL), and the reaction was stirred at 20°C under a N2 atmosphere for 4 hours. After the reaction, the mixture was added with water (50 mL) and extracted with ethyl acetate (20.0 mL*3). The obtained organic layer was washed with saturated brine (30.0 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 = 100 / 0 to 0 / 100) to obtain compound H102-4 (300 mg, crude product) as a brown solid.

[1054] LC-MS: EC21184-69-p1a, P1: RT=0.398min, m / z=429.9[M / 2+H] +

[1055] 1 H NMR:EC21184-69-p1aa(400MHz DMSO)

[1056] δ8.03(d,J=7.6Hz,1H),7.79(d,J=8.0Hz,1H),7.56(d,J=7.2Hz,1H),7.44-7.53(m,2H),7.39-7.43(m,1H),7.35(td,J=7.2,4.0Hz ,2H),7.27(dd,J=12.0,1.6Hz,1H),7.14-7.20(m,1H),7.06-7.13(m,1H),6.92(d,J=8.8Hz,1H),4.93(s,2H),4.08-4.19(m,1H),3. 97-4.05(m,2H),3.79(t,J=6.0Hz,2H),3.47(s,2H),3.17(s,2H),2.97-2.99(m,1H),2.78-2.86(m,2H),2.62-2.72(m,3H),2.17(d, J=6.4Hz,4H),1.92-1.98(m,2H),1.69(d,J=10.8Hz,2H),1.40-1.54(m,2H),1.31(s,9H),1.16-1.23(m,6H),1.11(d,J=10.0Hz,1H)

[1057] 4. Synthesis of Compound H102-5

[1058] To a solution of compound H102-4 (200 mg, 233 μmol, 1.00 equiv) in DMF (2.00 mL) were added compound 8A (111 mg, 233 μmol, 1.00 equiv), HATU (133 mg, 350 μmol, 1.50 equiv), and DIEA (90.3 mg, 699 μmol, 122 μL, 3.00 equiv). The mixture was stirred at 20° C. for 16 hours. LC-MS (EC21184-132-P1A, P1: RT = 0.425 min) indicated a yield of approximately 55.6% of the desired product. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain the crude product, which was further purified by preparative HPLC (HCl conditions; column: CD02-Waters Xbidge BEH C18 150*25*10 μm; mobile phase: [water (HCl)-ACN]; gradient elution: 30%-60% B in 10 minutes) to obtain compound H102-5 (25.0 mg, 18.2 μmol, 7.83% yield, 96.1% purity) as a white solid.

[1059] LC-MS: EC21184-132-P1A, P1: RT=0.425min, m / z=659.9[M / 2+H] +

[1060] 5. Synthesis of compound APH102

[1061] Compound H102-5 (25.0 mg, 19.0 μmol, 1.00 equiv) was mixed in HCl / dioxane (10.0 mL) and stirred at 20°C under a nitrogen atmosphere for 4 hours. LC-MS (EC21184-142-p1c, P1: RT = 2.008 min) indicated the formation of approximately 79.5% of the desired product. The reaction mixture was filtered and concentrated under reduced pressure to obtain the crude product, which was then purified by preparative HPLC (neutral conditions; column: CD02-Waters Xbidge BEH C18 150*25*10 μm; mobile phase: [water (NH4HCO3)-ACN]; gradient elution: 32%-62% B over 10 minutes) to afford compound APH102 (7.36 mg, 5.73 μmol, 30.2% yield, 98.1% purity) as a white solid.

[1062] LC-MS: EC21184-142-p1c, P1: RT=2.008min, m / z=631.8[M / 2+H] +

[1063] 1 H NMR:EC21184-142-P1ZB(400MHz DMSO)

[1064] δ12.32-13.04(m,1H),8.37-8.76(m,1H),8.04(d,J=7.6Hz,1H),7.79(d,J=8.0Hz, 1H),7.60(d,J=7.2Hz,1H),7.53(d,J=8.8Hz,1H),7.44-7.50(m,1H),7.39-7.43(m ,1H),7.23-7.38(m,8H),7.13-7.20(m,1H),7.06-7.12(m,1H),6.94(d,J=8.8Hz,1 H),6.09-6.24(m,1H),5.08-5.17(m,1H),4.92(s,2H),4.20-4.32(m,2H),4.02(br t,J=6.4Hz,2H),3.86(t,J=6.0Hz,2H),3.66-3.76(m,2H),3.48(s,1H), 3.44(s,3H),2.98(t,J=5.2Hz,2H),2.75-2.83(m,6H),2.09-2.30(m,12H ),2.06(d,J=6.4Hz,3H),1.92-1.99(m,3H),1.64-1.76(m,3H),1.38-1. 48(m,1H),1.04-1.13(m,2H),0.94(d,J=6.63Hz,3H),0.73-0.80(m,3H).

[1065] Synthesis of APH103:

[1066] 1. Synthesis of compound H103-2

[1067] To a solution of compound H103-1 (100 mg, 540 μmol, 1.00 equiv) in DMF (3.00 mL) were added compound 8A (258 mg, 540 μmol, 1.00 equiv), HATU (246 mg, 648 μmol, 1.20 equiv) and DIEA (209 mg, 1.62 mmol, 282 μL, 3.00 equiv), and the mixture was stirred at 20°C for 2 hours. LC-MS (EC21184-116-P1A, P1: RT = 0.437 min) detection showed that approximately 73.1% of the desired product was produced. After the reaction, the mixture was filtered and concentrated under reduced pressure to obtain a crude product, which was further purified by preparative HPLC (FA conditions; column: CD19-Daisogel SP-100-8-ODS-PK 200*50*10 μm; mobile phase: [water (NH4HCO3)-ACN]; gradient elution: 47%-77% B in 10 minutes) to obtain compound H103-2 (150 mg, 232 μmol, 43.0% yield, 100% purity) as a white solid.

[1068] LC-MS: EC21184-116-P1A, P1: RT=0.437min, m / z=645.2[M+H] +

[1069] 1 H NMR:EC21184-116-p1cc(400MHz DMSO)

[1070] δ8.31-8.74(m,1H),7.25-7.40(m,4H),6.10-6.25(m,1H),5.09-5.18(m,1H),4.96-5.08(m,1H),4.20-4.33(m, 2H),4.05-4.19(m,1H),3.72(d,J=9.6Hz,2H),3.38(d,J=11.2Hz,1H),2.94-3.10(m,1H),2.81-2.93(m,1H),2. 78(d,J=6.4Hz,1H),2.61-2.70(m,1H),2.34-2.46(m,1H),2.21-2.30(m,1H),2.15-2.21(m,3H),1.95(t,J=9.2 Hz,1H),1.65-1.79(m,3H),1.38(d,J=1.2Hz,9H),1.04-1.37(m,3H),0.95(d,J=6.4Hz,3H),0.73-0.80(m,3H).

[1071] 2. Synthesis of compound H103-3

[1072] Compound H103-2 (150 mg, 232 μmol, 1.00 equiv) was mixe...

Claims

1. A compound of formula I, or a pharmaceutically acceptable salt, ester, prodrug, solvate, stereoisomer or deuterated compound thereof: ALE (I) The A part is a small molecule ligand part targeting Bcl-xL protein, the E part is a small molecule ligand part targeting E3 ubiquitin ligase complex, and the L part is a connecting group. Part A has a structure of Formula 1 or 2: Where R0 is absent or selected from: -COOR a , -CONR a R b , -SO3R a , -SO2NR a R b , -SO2NR a COR b , -CONR a S02R b , -COSO2NR a R b , X is selected from hydrogen, hydroxy, amino or CH(OR a )2; R a and R b are independently selected from hydrogen, cyano, substituted or unsubstituted alkyl or substituted or unsubstituted alkoxy; Y is absent or is 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; Z1 and Z2 are independently 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, substituted or unsubstituted heteroaryl groups; The E part is selected from the following structures: Wherein R1 is O, NH or absent; R2 is H or methyl; R3 is R c is F, methoxy, Cl, cyano; The L section has the following structure: Wherein the L part is connected to the A part at L1 and to the E part at L2, L1 and L2 are independently absent or selected from: X does not exist or has the following structure: X1 and X2 are independently absent or selected from: W is absent or selected from: in, R4 is independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl and substituted or unsubstituted heterocyclyl; 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, acylamino, 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 of 0-10.

2. The compound of formula I according to claim 1, wherein the A moiety is a deuterated moiety having a structure of formula 1' or 2': wherein one or more hydrogens at position * are deuterated, Preferably, the two hydrogens at position * are deuterated.

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

4. A 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 are independently selected from hydrogen, cyano and alkyl, Preferably, R0 is selected from:

5. A compound of formula I according to claim 1, wherein Y is absent or selected from alkylene, alkyleneoxy, alkenylene, alkenyleneoxy, arylene, heteroarylene, cycloalkylene and heterocyclylene optionally substituted by one or more substituents selected from halogen and alkyl.

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

7. A compound of formula I according to claim 1, wherein Y is selected from arylene and 5- or 6-membered heteroarylene optionally substituted by one or more substituents selected from halogen and alkyl, Preferably, Y is selected from phenylene or pyrazolylene optionally substituted by one or more substituents selected from halogen and alkyl, More preferably, Y is selected from wherein the bond is to Z1 or Z2 at the ring nitrogen atom.

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

9. The compound of formula I according to claim 1, wherein Z1 and Z2 are independently selected from 1’ Substituted-alkylene-cyclic group, cycloalkyl-(R2') p 、Heterocyclyl-(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-propynylene-cycloalkyl-(R2') p , aryl-propynylene-heterocyclyl-(R2') p and heteroaryl-propynylene-heterocyclyl-(R2') p , Where R 1’ is selected from the group consisting of deuterium, halogen, amino, hydroxy, nitro, thiol, cyano, isocyano, substituted or unsubstituted alkyl (e.g. alkyl substituted by halogen, amino, hydroxy, nitro and / or carboxyl), substituted or unsubstituted alkoxy (e.g. alkoxy substituted by halogen, amino, hydroxy, nitro and / or carboxyl), alkylamino, dialkylamino, carboxyl, carbonyl, amido, sulfonyl, sulfonic acid, phosphoryl and phosphonyl, R2' is hydrogen, alkyl, aryl, heteroaryl, cycloalkyl or heterocyclyl, and p is 0, 1, 2, 3 or 4.

10. A 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-membered or 6-membered heterocyclic group containing one or two heteroatoms independently selected from nitrogen, oxygen and sulfur, preferably a piperidinyl group, a piperazinyl group, a morpholinyl group or a 3,9-diazaspiro[5.5]undecyl group; The aryl group is a phenyl group; and / or The heteroaryl group is a 5-membered 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. A 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. A compound of formula I according to claim 1 or 2, wherein the A moiety is selected from the following structures:

14. The compound of formula I according to claim 1, wherein the E moiety 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: wherein u1, u2, u3 and v1 are independently selected from integers of 0-10.

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

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

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

21. A method of selectively killing one or more senescent cells or treating a senescence-related disease or disorder 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 a compound according to any one of claims 1 to 19 or a composition according to claim 20 in the preparation of a medicament for selectively killing one or more senescent cells or treating a senescence-related disease or disorder in a subject.

23. The method or use of claim 21 or 22, wherein 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 renal disorder or disease, an ocular disease or disorder, or a dermatological disorder or disease.

24. The method or use according to claim 21 or 22, wherein the aging-related disease or disorder is selected from: (i) an inflammatory or autoimmune disease or condition selected from oral mucositis, inflammatory bowel disease, kyphosis and intervertebral disc herniation; (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; (iii) a metabolic disease selected from diabetic ulcer, metabolic syndrome and obesity; (iv) a lung disease selected from pulmonary fibrosis, chronic obstructive pulmonary disease, asthma, cystic fibrosis, emphysema, bronchiectasis and age-related loss of lung function; (v) an ocular disease or condition selected from macular degeneration, glaucoma, cataracts, presbyopia and vision loss; (vi) an age-related disorder selected from renal failure, frailty, hearing loss, muscle fatigue, skin conditions, skin wound healing, liver fibrosis, pancreatic fibrosis, oral submucous fibrosis and sarcopenia; and (vii) a skin disease or condition selected from eczema, psoriasis, hyperpigmentation, nevus, rash, atopic dermatitis, urticaria, diseases and conditions associated with photosensitivity or photoaging, wrinkles, pruritus, dysesthesia, eczematous rash, eosinophilic dermatosis, reactive neutrophilic dermatosis, pemphigus, pemphigoid, immunobullous dermatosis, cutaneous fibrohistiocytosis, cutaneous lymphoma and cutaneous lupus.

25. The method or use according to claim 21 or 22, wherein the subject is diagnosed with cancer and is optionally undergoing a cancer treatment selected from chemotherapy and radiotherapy.

26. The method or use according to claim 21 or 22, wherein the compound or composition (i) reducing cells that have been pushed into senescence or cancer cells; (ii) reducing one or more side effects produced by senescent cells, wherein the side effects include inflammation, promotion of cancer growth, and promotion of metastasis; (iii) reducing one or more side effects of chemotherapy; or (iv) reducing one or more side effects of radiation therapy.

27. The method or use of claim 21 or 22, wherein the subject has a viral infection, optionally wherein the viral infection results in overproduction of activated macrophages in the subject.

28. The method or use of claim 27, wherein the viral infection is a coronavirus infection selected from coronavirus, 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 of claim 27, wherein the subject has a SARS-CoV-2 infection and has been diagnosed with coronavirus disease 2019.

30. The method of claim 21, wherein the compound or composition, upon in vivo administration, is in an amount effective to kill one or more senescent cells to obtain the following structure: Wherein R0, Y and Z1 are as described in any one of claims 1-19.

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