Cdk9 protein kinase degraders and uses thereof

Abstract

The present application relates to compounds having CDK9 degradation activity and uses thereof. The compounds of the present application can effectively degrade CDK9 protein kinase in cells, and can be used for preparing drugs for treating and / or preventing diseases or disorders related to CDK9 mediation.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, and specifically relates to compounds, pharmaceutical compositions and their uses for degrading CDK9 protein kinase. Background Technology

[0002] Cyclin-dependent kinases (CDKs) belong to the serine / threonine protein kinase family, which has been identified to contain approximately 20 highly homologous subproteins. CDKs bind to regulatory subunits of cyclin to form functional heterodimeric complexes, thereby regulating related cellular biological functions such as cell proliferation, cell differentiation, DNA repair, and apoptosis. CDK9, a member of the CDK subfamily, is responsible for cellular transcription. Therefore, targeting CDK9 can effectively disrupt the gene expression network of diseased cells and induce apoptosis in diseased cells (such as various tumor cells), making it a potential target for the development of therapeutic drugs, particularly anti-tumor drugs.

[0003] Currently, the most advanced research and only marketed drugs targeting CDKs (CDK4 / 6) are those targeting CDK6, such as palbociclib, ribociclib, abemaciclib, and trilaciclib. No drugs targeting other CDK proteins have yet been approved for marketing, indicating significant potential and room for development in this field. To date, an increasing number of CDK9 inhibitors have shown good antitumor activity in vitro, primarily pyrazoles, pyrimidines, and flavonoids. However, these compounds currently face limitations in further clinical research and application due to poor selectivity, low activity, and poor clinical tolerability.

[0004] As a revolutionary emerging technology, protein degradation targeting chimeras (PROTACs) have significant advantages in overcoming drug resistance, improving activity, and enhancing targeting selectivity. PROTACs are bifunctional molecules comprising an E3 ligase ligand, a target protein ligand, and a linker. They achieve the degradation of disease-related target proteins by hijacking the ubiquitin-proteasome system (UPS) in the body. Since its initial proposal in 2001, PROTAC technology has developed rapidly over the past 20 years, achieving significant breakthroughs in anti-tumor drug development. Several PROTAC degraders are already in clinical trials, notably ARV-471 and ARV-766, developed by Yale University and Arvinas, which are in Phase II-III clinical trials. Internationally renowned pharmaceutical companies such as Pfizer and Merck have invested approximately $2 billion in Arvinas to develop new PROTAC protein degradation drugs. Furthermore, an increasing number of degraders have entered or are about to enter Phase I clinical trials. Kymera successfully raised $148 million and went public on Nasdaq, and KT-474 is currently in Phase I clinical trials, demonstrating the feasibility of this technology. With the increasing number of protein degraders entering clinical trials and the emergence of various PROTAC-derived technologies, the demand for E3 ligase ligands is growing rapidly. VHL and CRBN are currently the two most commonly used and effective E3 ligases for PROTAC design; however, their ligand selection is relatively limited, which significantly restricts the application scope of PROTACs and their derivative technologies. Developing novel and efficient E3 ligase ligands could not only help degrade a wider range of proteins, but also potentially induce less systemic and more selective (e.g., tissue or organ specific) target protein degradation by leveraging different biological properties and E3 ligase expression levels.

[0005] Therefore, to address the shortcomings of existing technologies, it is necessary to develop drugs with novel structures that can efficiently degrade the CDK9 protein. Summary of the Invention

[0006] The purpose of this invention is to provide a drug that can efficiently degrade CDK9 to treat or prevent diseases in which the activity of diseased cells or related pathogenic cells depends on CDK9.

[0007] In a first aspect of the invention, a compound of general formula (I) or a stereoisomer, enantiomer, transisomer, optical isomer, racemate, tautomer or a pharmaceutically acceptable salt thereof, its hydrate, or its isotopically labeled compound is provided:

[0008]

[0009] in,

[0010] Z is selected from 5-10 N-containing heterocycles;

[0011] L is -L 1 -[W 1 -L'-W 2 ] s -L 3 -;

[0012] L 1 Connected to Z, and L 1 Selected from chemical bonds, -CO-, -O-, substituted or unsubstituted -(CH2). m - Substituted or unsubstituted C3-C8 carbon rings, substituted or unsubstituted 4-10 membered heterocycles;

[0013] L 3 Selected from chemical bonds, -C(=O)-(CH2) p -、-(CH2) p NH-, -O- or

[0014] s can be 0, 1, 2, 3, 4, 5, or 6;

[0015] Each L' is independently selected from: chemical bonds, substituted or unsubstituted C3-C8 carbon rings, substituted or unsubstituted 4-10 membered heterocycles, and -NHCO- in each occurrence;

[0016] Each W 1 and W 2 Each time it appears, it is independently selected from chemical bonds, -CO-, and -(CH2). m -、-Y-(CH2) m -、-(CH2) n -Y-, -Y-(CH2) m -Y-(CH2) n -、-(CH2) m -Y-(CH2CH2O) n -;

[0017] Each p is independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 each time it appears;

[0018] Each occurrence of m is independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12;

[0019] Each n is independently selected from 0, 1, 2, 3, 4, 5, or 6 in each occurrence;

[0020] Each Y is independently selected from -O- and -S- for each occurrence;

[0021] R1 is selected from: H, halogen, C1-C6 alkyl, prodrug group, such as ester group, amide group, alcohol group, phosphate ester group, glycoside, sustained-release group, etc.

[0022] In another preferred embodiment, R1 is selected from: H, methyl, ethyl.

[0023] In another preferred embodiment, the compound has the structure shown in formula (II):

[0024]

[0025] Z and L are defined as above.

[0026] In another preferred embodiment, Z is selected from... In another preferred embodiment, the compound has the structure shown in formula (III-1) or formula (III-2):

[0027]

[0028] In another preferred embodiment, each L' is independently selected from: chemical bonds, -NHCO-.

[0029] In another preferred embodiment, s is 0, 1, 2 or 3.

[0030] In another preferred embodiment, L is selected from:

[0031]

[0032]

[0033] In another preferred embodiment, L is selected from:

[0034]

[0035]

[0036] In another preferred embodiment, the compound is selected from the group consisting of:

[0037]

[0038]

[0039]

[0040]

[0041]

[0042]

[0043]

[0044]

[0045]

[0046] A second aspect of the present invention provides a pharmaceutical composition comprising the compound described in the first aspect of the present invention or a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof, and a pharmaceutically acceptable carrier.

[0047] A third aspect of the invention provides the use of the compounds described in the first aspect of the invention, or pharmaceutically acceptable salts thereof, in the preparation of medicaments for treating CDK9-mediated diseases or conditions.

[0048] In another preferred embodiment, the CDK9-mediated related diseases or conditions include tumors.

[0049] A fourth aspect of the present invention provides a method for treating a tumor disease, comprising the steps of administering to a subject in need a compound of formula (I) as described in the first aspect of the present invention, or a pharmaceutically acceptable salt, solvate, stereoisomer, or prodrug thereof.

[0050] In another preferred embodiment, the object includes humans and non-human mammals.

[0051] In another preferred embodiment, the tumor is selected from the group consisting of: soft tissue sarcoma, primitive neuroectodermal tumor, pulmonary sarcoma, osteosarcoma, liposarcoma, Kaposi's sarcoma, midline carcinoma, breast cancer, lung cancer, colorectal cancer, liver cancer, pancreatic cancer, peritoneal cancer, thyroid tumor, parathyroid tumor, soft tissue hyperplasia, warts, Langerhans cell histiocytosis, hamartoma, gestational trophoblastic disease, skin cancer, Merkel cell carcinoma, bone tumor, melanoma, leukemia, salivary gland tumor. Neuroendocrine tumors, lymphoma, brain tumors, neuroblastoma, mesothelioma, esophageal cancer, medulloblastoma, glioblastoma, meningioma, retinoblastoma, kidney cancer, bladder cancer, stomach cancer, uterine cancer, vulvar cancer, prostate cancer, testicular cancer, penile cancer, bile duct cancer, gallbladder cancer, ureteral cancer, adrenocortical cancer, head and neck cancer, oral cancer, nasopharyngeal cancer, myeloma, ovarian cancer, cervical cancer, germ cell tumors, thymic tumors, myelodysplastic syndrome.

[0052] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Attached Figure Description

[0053] Figure 1 The study showed that CDK9 degrading agents significantly reduced CDK9 protein levels in TC-71 cells.

[0054] Figure 2 This invention demonstrates the inhibitory activity of the CDK9 degrading agent of the present invention and the control CDK9 degrading agent THAL-SNS032 on tumor cell growth.

[0055] Figure 3 The CDK9 degrading agent of the present invention exhibits good inhibitory activity against a variety of tumor cells.

[0056] Figure 4 Compound 4 of the present invention showed that it had a good inhibitory effect on tumors in mice. Detailed Implementation

[0057] Through extensive and in-depth research, the inventors have developed, for the first time, a class of CDK9 degrading compound compounds with a novel structure. One end of these compounds targets the CDK9 protein requiring degradation, while the other end, linked by a series of chain linkers, targets and binds to an E3 ubiquitin ligase, forming a complete PROTAC molecule. Compared to existing CDK9 inhibitors, the compounds of this invention possess the ability to rapidly inhibit tumor proliferation and have the potential to become an effective treatment for malignant tumors.

[0058] the term

[0059] In this invention, the term "halogen" refers to F, Cl, Br, or I.

[0060] In this invention, "C1-C6 alkyl" refers to a straight-chain or branched alkyl group comprising 1 to 6 (1, 2, 3, 4, 5 or 6) carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, neopentyl or similar groups.

[0061] In this invention, the term "C3-C8 cycloalkyl" refers to a cyclic alkyl group having 3-8 (1, 2, 3, 4, 5, 6, 7, or 8) carbon atoms on a ring, and non-limitingly includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc. The term "C..." 3-6 "Cycloalkyl" has a similar meaning.

[0062] In this invention, the term "C1-C6 alkoxy" refers to a straight-chain or branched alkoxy group having 1-6 carbon atoms, and includes, without limitation, methoxy, ethoxy, propoxy, isopropoxy, and butoxy. Preferably, it is a C1-C4 alkoxy group.

[0063] "Carbocyclic group," "carbocyclic," or "cycloalkyl" refers to a saturated or partially unsaturated (i.e., non-aromatic) group containing all carbon ring atoms. A carbocyclic group typically contains one ring of 3 to 7 carbon atoms or two fused rings, each containing 3 to 7 carbon atoms. It includes monocyclic, bicyclic, and tricyclic systems, wherein bicyclic and tricyclic systems include spirocyclic, fused, and bridged rings. The cycloalkyl substituent can be a self-substituted nitrogen or carbon atom side group, or a substituted carbon atom may have two substituents connected as a cycloalkyl group. Examples of carbocyclic groups include cyclohexenyl, cyclohexyl, cyclopentenyl, cyclopentyl, cyclobutenyl, cyclobutyl, and cyclopropyl rings. In one embodiment, the carbocyclic group is optionally substituted as described above. In one embodiment, the cycloalkyl group is a partially unsaturated (i.e., non-aromatic) group containing all carbon ring atoms.

[0064] In this invention, the term "heterocyclic group" or "heterocycle" refers to a 4-10 membered heterocyclic group containing 1, 2, or 3 heteroatoms selected from N, O, and S. It may be used alone or in combination with other terms to refer to a saturated or partially saturated cyclic group consisting of 4-10 (4, 5, 6, 7, 8, 9, or 10) ring atoms, wherein 1, 2, or 3 of the ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the carbon, nitrogen, and sulfur atoms may be optionally oxidized (i.e., C(=O), NO, SO, and SO2). It includes monocyclic, bicyclic, and tricyclic systems, wherein bicyclic and tricyclic systems include spirocyclic, fused, and bridged rings. Furthermore, heteroatoms may occupy the connection positions between the heterocyclic group and the rest of the molecule, and the heterocyclic group includes saturated and partially unsaturated heterocyclic groups. Non-limiting examples of heterocyclic groups include azirrobutyl, oxacyclobutyl, thioheterobutyl, pyrrolidinyl, pyrazolyl, imidazoalkyl, tetrahydrothiophene (including tetrahydrothiophene-2-yl and tetrahydrothiophene-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuranyl-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperidinyl and 2-piperidinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxyl, dithiaalkyl, isoxazolyl, isothiazolyl, 1,2-oxazinyl, 1,2-thiaazinyl, hexahydropyridazinyl, homopiperidinyl, and homopiperidinyl.

[0065] In this invention, the term "substitution" refers to the substitution of one or more hydrogen atoms on a specific group by a specific substituent. The specific substituent is the substituent described accordingly above, or the substituent appearing in the various embodiments. Unless otherwise specified, a substituted group may have a substituent selected from a specific group at any substituted site of that group, and the substituents may be the same or different at each position. Those skilled in the art will understand that the combinations of substituents contemplated in this invention are those that are stable or chemically feasible. Such substituents include, but are not limited to: halogens, hydroxyl groups, carboxyl groups (-COOH), cyano groups (-CN), C1-C6 alkyl groups, C2-C6 alkenyl groups, C3-C8 cycloalkyl groups, 3- to 12-membered heterocyclic groups, aryl groups, heteroaryl groups, C1-C8 aldehyde groups, C2-C10 acyl groups, C2-C10 ester groups, amino groups, C1-C6 alkoxy groups, C1-C10 sulfonyl groups, etc.

[0066] In this invention, the terms 1-6 refer to 1, 2, 3, 4, 5, or 6. Other similar terms each have a similar meaning independently. The term "multiple" refers to 2-6 or more, such as 2, 3, 4, 5, or 6.

[0067] It should be understood that when a certain group exists simultaneously at multiple different positions in a compound, its definition at each position is independent of each other; they can be the same or different.

[0068] The compounds of this invention can exist in specific geometric or stereoisomeric forms. This invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, (R)- and (S)- enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof, as well as other mixtures, such as mixtures where enantiomers are enriched in diastereomers, all of which fall within the scope of this invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this invention.

[0069] As used herein, the term "pharmaceutically acceptable salt" refers to a salt formed by the compounds of the present invention with an acid or base that is suitable for use as a medicine. Pharmaceutically acceptable salts include both inorganic and organic salts. A preferred class of salts are those formed by the compounds of the present invention with an acid. Suitable acids for forming salts include, but are not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, and naphthalenesulfonic acid; and amino acids such as proline, phenylalanine, aspartic acid, and glutamic acid.

[0070] Another preferred class of salts are salts formed by the compounds of the present invention with a base, such as alkali metal salts (e.g., sodium or potassium salts), alkaline earth metal salts (e.g., magnesium or calcium salts), ammonium salts (such as lower alkanol ammonium salts and other pharmaceutically acceptable amine salts), such as methylamine salts, ethylamine salts, propylamine salts, dimethylamine salts, trimethylamine salts, diethylamine salts, triethylamine salts, tert-butylamine salts, ethylenediamine salts, hydroxyethylamine salts, dihydroxyethylamine salts, trihydroxyethylamine salts, and amine salts formed from morpholine, piperazine, and lysine, respectively.

[0071] Compounds and their preparation methods

[0072] The present invention provides a compound of formula (I) or a tautomer, optical isomer, hydrate, solvate or pharmaceutically acceptable salt thereof.

[0073]

[0074] The groups are defined above.

[0075] In another preferred embodiment, in the compound, any one of Z, L, and R1 is independently the corresponding group in the specific compound of the present invention.

[0076] The main advantages of this invention include:

[0077] (1) The compounds of the present invention have excellent degradation activity against CDK9, and their activity is significantly better than that of existing CDK9 degrading agents.

[0078] (2) The compounds of the present invention have significant inhibitory effects on a variety of cancer cells.

[0079] (3) The compounds of the present invention have significant inhibitory effects on a variety of tumors.

[0080] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and parts by weight.

[0081] Example

[0082] The following synthetic routes describe the preparation of the general formula compounds of the present invention. All final compounds of the present invention are prepared by the methods described in these routes or by similar methods, which are well known to those skilled in the art of organic chemistry. All variable groups used in these routes are as defined in the claims.

[0083] Example 1

[0084]

[0085] Synthesis of Example 1: (a) TEA, DCM, 0℃; (b) POCl3, reflux; (c) MeOH, rt; (d) 4, NaBH4, EtOH, Acetone, rt~reflux; (e) HATU, DIPEA, DMF, rt; (f) TFA, DCM, rt; (g) HATU, DIPEA, DMF, rt; (h) DMP, DCM, rt; (i) NaBH3CN, DCM, MeOH, rt.

[0086] Specific experimental steps:

[0087] a: Compound 1 (1.51 g, 10 mmol) was dissolved in 35 mL of anhydrous dichloromethane. The system was cooled to -5 °C, and triethylamine (4.82 mL, 35 mmol) was added dropwise. The mixture was stirred at -5 °C for 30 min, and then compound 2 (0.88 mL, 11 mmol) was slowly added dropwise while maintaining the temperature at -5 °C. The reaction mixture was stirred for 1 h. The reaction was quenched with 1 N HCl (20 mL), and extracted three times with dichloromethane (3 × 50 mL). The separated organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by column chromatography (petroleum ether / ethyl acetate = 20 / 1) to give compound 3 as a white solid. Yield: 95%.

[0088] b: Compound 3 (958 mg, 5 mmol) was weighed and dissolved in 3 mL of POCl3. The reaction system was stirred at 105 °C for 1 h. After cooling the reaction solution to room temperature, it was added dropwise to ice water and extracted five times with ethyl acetate (5 × 10 mL). The organic phases were combined and the pH was adjusted to 7–8 with saturated sodium bicarbonate solution. The mixture was then washed successively with water and saturated brine, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by column chromatography (petroleum ether / ethyl acetate = 9 / 1) to give compound 4, a pale yellow solid. Yield: 76%.

[0089] c: Compound 5 (1.30 g, 5 mmol) and compound 6 (1.94 g, 20 mmol) were weighed and dissolved in 34 mL of methanol. The reaction system was stirred at room temperature for 48 h. After the reaction was completed, the solvent was removed by vacuum distillation, and then water was added to the residue. The pH was adjusted to approximately 12 with 10% sodium hydroxide solution. The residue was filtered to obtain a reddish-brown solid. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 1 / 9) to give compound 7, a brown solid. Yield: 32%.

[0090] d: Compound 7 (314 mg, 2 mmol) was dissolved in 20 mL of anhydrous ethanol. Sodium borohydride (160 mg, 4 mmol) was slowly added to the reaction system at room temperature, and the mixture was stirred for 1 h at room temperature. Acetone (10 mL) was slowly added dropwise to the system, and stirring was continued for 1 h. Then, an ethanol solution (3 mL) of compound 4 (350 mg, 2 mmol) was added dropwise to the reaction solution, and the reaction system was refluxed for 1 h. After the reaction was completed, the reaction solution was cooled to room temperature, the solvent was removed by vacuum distillation, the solvent was quenched with water, and the mixture was extracted three times with ethyl acetate (3 × 15 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by column chromatography (petroleum ether / ethyl acetate = 4 / 1) to give compound 8, a brown solid. Yield: 60%.

[0091] e,f: Compound 8 (539 mg, 2 mmol) and compound 9 (460 mg, 2 mmol) were weighed and dissolved in 10 mL of DMF. DIPEA (0.87 mL, 5 mmol) and HATU (1.14 g, 3 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was quenched with water and extracted three times with ethyl acetate (3 × 20 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by prep-HPLC to obtain a white solid. This solid was dissolved in 10 mL of anhydrous dichloromethane, and TFA (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min. The solvent was removed by vacuum distillation to obtain the trifluoroacetate of compound 10 (SNS-032), a brown oily substance. Overall yield of the two steps: 59%.

[0092] g: Compound 10 (190 mg, 0.5 mmol) and Compound 11 (59 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 10 mL), and the organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by prep-HPLC to give Compound 12 as a white solid. Yield: 78%.

[0093] h: Weigh compound 12 (240 mg, 0.5 mmol), dissolve in 5 mL of anhydrous dichloromethane, add Dys-Martin reagent (233 mg, 0.55 mmol), and stir the reaction mixture overnight at room temperature. Quench with water, filter with diatomaceous earth, extract the filtrate three times with dichloromethane (3 × 10 mL), combine the organic phases and wash with saturated brine, dry over anhydrous sodium sulfate, concentrate, and purify the crude product by prep-HPLC to give compound 13, a white solid. Yield: 70%.

[0094] i: Compound 13 (120 mg, 0.25 mmol) and compound 14 (75 mg, 0.25 mmol) were weighed and dissolved in 3 mL of a mixture of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (31 mg, 0.5 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 1, a white solid. Yield: 65%. Purity: 98%, Rt = 5.1 min. 1 H NMR(600MHz,DMSO-d6)δ12.32(s,1H),11.14(s,1H),7.86(s,2H),7.39(s, 1H),6.71(s,1H),5.18-5.12(m,1H),4.45-4.16(m,5H),4.05(s,2H),3.96- 3.90(m,1H),3.08-2.87(m,4H),2.74(s,1H),2.64-2.55(m,3H),2.39(s,2 H),2.07(s,1H),1.82(s,2H),1.59(s,5H),1.44-1.38(m,1H),1.17(s,9H). LC / MS(ESI,m / z):[M+H] + =762.57.

[0095] Example 2

[0096]

[0097] Synthesis of Example 2: (a) HATU, DIPEA, DMF, rt; (b) DMP, DCM, rt; (c) NaBH3CN, DCM, MeOH, rt.

[0098] Specific experimental steps:

[0099] a: Compound 10 (190 mg, 0.5 mmol) and compound 15 (66 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The mixture was quenched with water and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 16 as a white solid. Yield: 74%.

[0100] b: Weigh compound 16 (247 mg, 0.5 mmol), dissolve in 5 mL of anhydrous dichloromethane, add Dys-Martin reagent (233 mg, 0.55 mmol), stir the reaction mixture overnight at room temperature, quench with water, filter with diatomaceous earth, extract the filtrate three times with dichloromethane (3 × 10 mL), combine the organic phases and wash with saturated brine, dry over anhydrous sodium sulfate, concentrate to obtain crude product, and purify by prep-HPLC to obtain compound 17, a white solid. Yield: 65%.

[0101] c: Compound 17 (123 mg, 0.25 mmol) and compound 14 (75 mg, 0.25 mmol) were weighed and dissolved in 3 mL of a mixture of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (31 mg, 0.5 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 2, a white solid. Yield: 52%. Purity: 98%, Rt = 5.6 min. 1 H NMR(600MHz,DMSO-d6)δ12.34(s,1H),11.16(s,1H),7.97(s,2H),7.39(s,1H),6.72(s ,1H),5.18(d,J=11.2Hz,1H),4.76(s,4H),4.44-4.37(m,1H),4.06(s,2H),3.92(d,J= 9.4Hz,1H),3.05(s,1H),2.90(s,1H),2.75(s,1H),2.60(t,J=21.2Hz,3H),2.36(s,2H ),2.11-2.06(m,1H),1.83(s,2H),1.71(s,2H),1.56(s,4H),1.39(s,4H),1.18(s,9H). LC / MS(ESI,m / z):[M+H] + =776.60.

[0102] Example 3

[0103]

[0104] Synthesis of Example 3: (a) HATU, DIPEA, DMF, rt; (b) DMP, DCM, rt; (c) NaBH3CN, DCM, MeOH, rt.

[0105] Specific experimental steps:

[0106] a: Compound 10 (190 mg, 0.5 mmol) and compound 18 (73 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The mixture was quenched with water and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. Prep-HPLC purification yielded compound 19 as a white solid. Yield: 78%.

[0107] b: Weigh compound 19 (254 mg, 0.5 mmol), dissolve in 5 mL of anhydrous dichloromethane, add Dys-Martin reagent (233 mg, 0.55 mmol), and stir the reaction mixture overnight at room temperature. Quench with water, filter with diatomaceous earth, and extract the filtrate three times with dichloromethane (3 × 10 mL). Combine the organic phases and wash with saturated brine. Dry the mixture on anhydrous sodium sulfate and concentrate to obtain the crude product. Purify the crude product by prep-HPLC to give compound 20 as a white solid. Yield: 55%.

[0108] c: Compound 20 (127 mg, 0.25 mmol) and compound 14 (75 mg, 0.25 mmol) were weighed and dissolved in 3 mL of a mixed solution of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (31 mg, 0.5 mmol) was added, and the reaction system was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 3, a white solid. Yield: 45%. Purity: 98%, Rt = 4.4 min. 1H NMR (600MHz, DMSO-d6) δ12.33(s,1H),11.16(s,1H),7.99(s,2H),7.39(s,1H),6.71(s,1H),5.18(dd,J=12.9,5.4Hz,1 H),4.98(d,J=14.9Hz,2H),4.67(d,J=15.0Hz,2H),4.40(d,J=12.9Hz,1H),4.05(s,2H),3.92(d,J=13.5Hz,1H),3.08- 3.01(m,1H),2.90(ddd,J=19.4,13.9,5.3Hz,1H),2.78-2.68(m,2H),2.64-2.53(m,3H),2.33(dt,J=18.6,7.6Hz,2H), 2.09(t,J=5.1Hz,1H),1.82(t,J=15.7Hz,2H),1.70(s,2H),1.54(d,J=8.3Hz,3H),1.37(q,J=11.0Hz,6H),1.18(s,9H). LC / MS(ESI,m / z):[M+H] + =790.63.

[0109] Example 4

[0110]

[0111] Synthesis of Example 4: (a) HATU, DIPEA, DMF, rt; (b) DMP, DCM, rt; (c) NaBH3CN, DCM, MeOH, rt.

[0112] Specific experimental steps:

[0113] a: Compound 10 (190 mg, 0.5 mmol) and compound 21 (80 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 0.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The mixture was quenched with water and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The extract was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 22 as a white solid. Yield: 78%.

[0114] b: Weigh compound 22 (261 mg, 0.5 mmol), dissolve in 5 mL of anhydrous dichloromethane, add Dys-Martin reagent (233 mg, 0.55 mmol), and stir the reaction mixture overnight at room temperature. Quench with water, filter with diatomaceous earth, extract the filtrate three times with dichloromethane (3 × 10 mL), combine the organic phases, wash with saturated brine, dry over anhydrous sodium sulfate, concentrate to obtain crude product, and purify by prep-HPLC to give compound 23, a white solid. Yield: 62%.

[0115] c: Compound 23 (130 mg, 0.25 mmol) and compound 14 (75 mg, 0.25 mmol) were weighed and dissolved in 3 mL of a mixture of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (31 mg, 0.5 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 4, a white solid. Yield: 54%. Purity: 98%, Rt = 4.7 min. 1 H NMR (600MHz, DMSO-d6) δ12.33(s,1H),11.15(s,1H),7.96(s,2H),7.39(s,1H),6.71(s,1H),5.17(dd,J= 12.9,5.3Hz,1H),4.71(s,4H),4.39(d,J=12.5Hz,1H),4.05(s,2H),3.91(d,J=13.2Hz,1H),3.03(t,J=12 .2Hz,1H),2.89(d,J=13.6Hz,1H),2.73(t,J=11.2Hz,1H),2.63-2.56(m,4H),2.33(dt,J=16.5,8.1Hz,2 H), 2.11-2.06 (m, 1H), 1.81 (t, J = 14.5Hz, 2H), 1.67 (s, 1H), 1.58-1.48 (m, 4H), 1.34 (s, 8H), 1.17 (s, 9H). LC / MS(ESI,m / z):[M+H] + =804.60.

[0116] Example 5

[0117]

[0118] Synthesis of Example 5: (a) HATU, DIPEA, DMF, rt; (b) DMP, DCM, rt; (c) NaBH3CN, DCM, MeOH, rt.

[0119] Specific experimental steps:

[0120] a: Compound 10 (190 mg, 0.5 mmol) and compound 24 (87 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The mixture was quenched with water and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. Prep-HPLC purification yielded compound 25 as a white solid. Yield: 73%.

[0121] b: Weigh compound 25 (268 mg, 0.5 mmol), dissolve in 5 mL of anhydrous dichloromethane, add Dys-Martin reagent (233 mg, 0.55 mmol), and stir the reaction mixture overnight at room temperature. Quench with water, filter with diatomaceous earth, extract the filtrate three times with dichloromethane (3 × 10 mL), combine the organic phases, wash with saturated brine, dry over anhydrous sodium sulfate, concentrate to obtain crude product, and purify by prep-HPLC to give compound 26, a white solid. Yield: 60%.

[0122] c: Compound 26 (134 mg, 0.25 mmol) and compound 14 (75 mg, 0.25 mmol) were weighed and dissolved in 3 mL of a mixture of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (31 mg, 0.5 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 5, a white solid. Yield: 49%. Purity: 98%, Rt = 5.1 min. 1H NMR (600MHz, DMSO-d6) δ12.32(s,1H),11.15(s,1H),7.93(s,2H),7.39(s,1H),6.71(s,1H),5.32(t,J=4.7Hz,1 H),5.16(dd,J=12.9,5.4Hz,1H),4.62-4.36(m,4H),4.05(s,2H),3.91(d,J=13.2Hz,1H),3.05-3.00(m,1H),2.9 2-2.87(m,1H),2.73(td,J=7.7,3.8Hz,1H),2.65-2.54(m,4H),2.29(s,2H),2.08(dd,J=9.3,4.3Hz,1H),1.99( dd,J=13.7,6.7Hz,2H),1.82(d,J=14.4Hz,2H),1.66-1.63(m,1H),1.54-1.47(m,4H),1.31(s,8H),1.17(s,9H). LC / MS(ESI,m / z):[M+H] + =818.70.

[0123] Example 6

[0124]

[0125] Synthesis of Example 6: (a) HATU, DIPEA, DMF, rt; (b) DMP, DCM, rt; (c) NaBH3CN, DCM, MeOH, rt.

[0126] Specific experimental steps:

[0127] a: Compound 10 (190 mg, 0.5 mmol) and compound 27 (94 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 10 mL), and the organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. Prep-HPLC purification yielded compound 28 as a white solid. Yield: 70%.

[0128] b: Weigh compound 28 (275 mg, 0.5 mmol), dissolve in 5 mL of anhydrous dichloromethane, add Dys-Martin reagent (233 mg, 0.55 mmol), and stir the reaction mixture overnight at room temperature. Quench the reaction mixture with water, filter with diatomaceous earth, and extract the filtrate three times with dichloromethane (3 × 10 mL). Combine the organic phases and wash with saturated brine. Dry the mixture on anhydrous sodium sulfate and concentrate to obtain the crude product. Purify the crude product by prep-HPLC to give compound 29, a white solid. Yield: 68%.

[0129] c: Compound 29 (137 mg, 0.25 mmol) and compound 14 (75 mg, 0.25 mmol) were weighed and dissolved in 3 mL of a mixture of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (31 mg, 0.5 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 6, a white solid. Yield: 55%. Purity: 98%, Rt = 6.5 min, 1H NMR(600MHz,DMSO-d6)δ12.32(s,1H),11.15(s,1H),7.91(s,2H),7.39(s,1H),6.71(s,1H ),5.20-5.13(m,1H),4.53-4.30(m,5H),4.05(s,2H),3.93-3.88(m,1H),3.15(s,2H),3.03 (s,1H),2.90(t,J=13.8Hz,1H),2.73(s,1H),2.59(dd,J=26.4,15.1Hz,3H),2.32-2.25(m ,2H),2.11-1.97(m,2H),1.81(s,3H),1.62(s,2H),1.49(s,4H),1.29(s,8H),1.17(s,9H). LC / MS(ESI,m / z):[M+H] + =832.72.

[0130] Example 7

[0131]

[0132] Synthesis of Example 7: (a) HATU, DIPEA, DMF, rt; (b) DMP, DCM, rt; (c) NaBH3CN, DCM, MeOH, rt.

[0133] Specific experimental steps:

[0134] a: Compound 10 (190 mg, 0.5 mmol) and compound 30 (101 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 10 mL), and the organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 31 as a white solid. Yield: 70%.

[0135] b: Weigh compound 31 (282 mg, 0.5 mmol), dissolve in 5 mL of anhydrous dichloromethane, add Dys-Martin reagent (233 mg, 0.55 mmol), and stir the reaction mixture overnight at room temperature. Quench with water, filter with diatomaceous earth, and extract the filtrate three times with dichloromethane (3 × 10 mL). Combine the organic phases and wash with saturated brine. Dry the mixture on anhydrous sodium sulfate and concentrate to obtain the crude product. Purify the crude product by prep-HPLC to give compound 32, a white solid. Yield: 64%.

[0136] c: Compound 32 (141 mg, 0.25 mmol) and compound 14 (75 mg, 0.25 mmol) were weighed and dissolved in 3 mL of a mixed solution of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (31 mg, 0.5 mmol) was added, and the reaction system was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 7, a white solid. Yield: 57%. Purity: 98%, Rt = 6.4 min. 1 H NMR(600MHz,DMSO-d6)δ12.30(s,1H),11.13(s,1H),7.81(s,2H),7.39(s,1H),6.71(s,1H ),5.14(d,J=11.0Hz,1H),4.42-4.35(m,1H),4.07(d,J=23.0Hz,6H),3.92-3.87(m,1H),3. 02(s,1H),2.88(d,J=13.8Hz,1H),2.75(d,J=39.5Hz,3H),2.64-2.55(m,3H),2.33-2.26( m,2H),2.10-2.02(m,1H),1.80(s,3H),1.51(d,J=33.0Hz,6H),1.27(s,11H),1.17(s,9H). LC / MS(ESI,m / z):[M+H] +=846.78.

[0137] Example 8

[0138]

[0139] Synthesis of Example 8: (a) HATU, DIPEA, DMF, rt; (b) DMP, DCM, rt; (c) NaBH3CN, DCM, MeOH, rt.

[0140] Specific experimental steps:

[0141] a: Compound 10 (190 mg, 0.5 mmol) and compound 33 (108 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 10 mL), and the organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 34 as a white solid. Yield: 73%.

[0142] b: Weigh compound 34 (289 mg, 0.5 mmol), dissolve in 5 mL of anhydrous dichloromethane, add Dys-Martin reagent (233 mg, 0.55 mmol), and stir the reaction mixture overnight at room temperature. Quench the reaction mixture with water, filter with diatomaceous earth, and extract the filtrate three times with dichloromethane (3 × 10 mL). Combine the organic phases and wash with saturated brine. Dry the mixture on anhydrous sodium sulfate and concentrate to obtain the crude product. Purify the crude product by prep-HPLC to give compound 35 as a white solid. Yield: 56%.

[0143] c: Compound 35 (144 mg, 0.25 mmol) and compound 14 (75 mg, 0.25 mmol) were weighed and dissolved in 3 mL of a mixture of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (31 mg, 0.5 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 8, a white solid. Yield: 50%. Purity: 98%, Rt = 6.8 min. 1H NMR(600MHz,DMSO-d6)δ12.31(s,1H),11.14(s,1H),7.91(d,J=23.6Hz,2H),7.42-7.37(m, 1H),6.71(s,1H),5.18-5.13(m,1H),4.38(t,J=41.1Hz,5H),4.05(s,2H),3.92-3.88(m,1H ),3.17-3.01(m,3H),2.90(s,1H),2.75-2.71(m,1H),2.63-2.55(m,3H),2.36-2.23(m,3H) ,2.05(d,J=35.4Hz,2H),1.81(s,3H),1.61(s,2H),1.48(s,4H),1.27(s,11H),1.17(s,9H). LC / MS(ESI,m / z):[M+H] + =860.54.

[0144] Example 9

[0145]

[0146] Synthesis of Example 9: (a) Boc2O, NaHCO3, THF, rt; (b) MeI, K2CO3, ACN, rt; (c) 4M HClin 1,4-dioxane, DCM, rt; (d) 23, NaBH3CN, AcONa, DCM, MeOH, rt.

[0147] Specific experimental steps:

[0148] a: Weigh compound 14 (300 mg, 1 mmol), dissolve in 15 mL of anhydrous dichloromethane, add sodium bicarbonate (210 mg, 2.5 mmol) and Boc₂O (240 mg, 1.1 mmol), and stir the reaction mixture overnight at room temperature. Remove the solvent by vacuum distillation, dissolve the resulting solid in dichloromethane (15 mL), wash with 1N HCl (5 mL), wash the separated organic phase with saturated brine, dry with anhydrous sodium sulfate, and concentrate to obtain the crude product. The crude product is purified by column chromatography to give compound 36, a gray solid. Yield: 87%.

[0149] b: Weigh compound 36 (399 mg, 1 mmol), dissolve in 15 mL of acetonitrile, add K₂CO₃ (276 mg, 2 mmol) and iodomethane (156 mg, 1.1 mmol), and stir the reaction mixture overnight at room temperature. Quench with water, extract three times with ethyl acetate (3 × 10 mL), combine the organic phases, wash with saturated brine, dry over anhydrous sodium sulfate, concentrate to obtain crude product, and purify by column chromatography to give compound 37, a gray solid. Yield: 56%.

[0150] c: Compound 37 (207 mg, 0.5 mmol) was weighed and dissolved in 10 mL of anhydrous dichloromethane. 2 mL of 4 M dioxane hydrochloride solution was added, and the reaction mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum distillation to give compound 38, a gray solid. Yield: 92%.

[0151] d: Compound 38 (78 mg, 0.25 mmol), compound 23 (130 mg, 0.25 mmol), and sodium acetate (41 mg, 0.5 mmol) were weighed and dissolved in 3 mL of a mixture of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (31 mg, 0.5 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 9, a white solid. Yield: 60%. Purity: 98%, Rt = 5.5 min. 1 H NMR(600MHz,)δ12.31(s,1H),7.77(d,J=11.0Hz,2H),7.39(s,1H),6.71(s,1H),4.39(d,J=1 2.3Hz,1H),4.05(s,2H),3.98(s,4H),3.91(d,J=13.8Hz,2H),3.02(s,4H),2.97-2.92(m,1H ),2.71-2.68(m,2H),2.58(t,J=12.9Hz,2H),2.34-2.28(m,2H),2.10-2.06(m,1H),2.03-1. 97(m,1H),1.81(t,J=14.6Hz,2H),1.53-1.49(m,4H),1.32(s,6H),1.23(s,3H),1.17(s,9H). LC / MS(ESI,m / z):[M+H] + =818.61.

[0152] Example 10

[0153]

[0154] Synthesis of Example 10: (a) TsCl, TEA, DMAP, DCM, rt; (b) TFA, DCM, rt; (c) 10, HATU, DIPEA, DMF, rt; (d) 14, DIPEA, NMP, 100℃.

[0155] Specific experimental steps:

[0156] a: Compound 39 (441 mg, 2 mmol) was weighed and dissolved in 10 mL of anhydrous dichloromethane. Triethylamine (1.1 mL, 8 mmol), p-toluenesulfonyl chloride (419 mg, 2.2 mmol), and DMAP (5 mg, 0.04 mmol) were added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched with water and extracted three times with dichloromethane (3 × 10 mL). The organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by column chromatography to give compound 40, a white oily liquid. Yield: 88%.

[0157] b: Weigh compound 40 (749 mg, 2 mmol), dissolve in 5 mL of anhydrous dichloromethane, add TFA (5 mL), and stir the reaction mixture at room temperature for 2 h. Remove the solvent by vacuum distillation to give compound 41, a pale yellow oil. Yield: 75%.

[0158] c: Weigh out compound 41 (159 mg, 0.5 mmol) and compound 10 (190 mg, 0.5 mmol) and dissolve them in 2 mL of DMF. Add DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol). Stir the reaction mixture at room temperature for 30 min. Quench with water and extract three times with ethyl acetate (3 × 10 mL). Combine the organic phases and wash with saturated brine. Dry the mixture on anhydrous sodium sulfate and concentrate to obtain the crude product. Purify the crude product by prep-HPLC to obtain compound 42, a pale yellow solid. Yield: 68%.

[0159] d: Compound 42 (680 mg, 1.0 mmol) and compound 14 (150 mg, 0.5 mmol) were weighed and dissolved in 2 mL of NMP. DIPEA (0.26 mL, 1.5 mmol) was added. The reaction mixture was heated to 100 °C, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 10, a white solid. Yield: 35%. Purity: 98%, Rt = 5.4 min. 1H NMR (600MHz, DMSO-d6) δ12.29(s,1H),11.12(s,1H),7.77(s,2H),7.38(s,1H),6.71(s,1H),5.13(dd,J=12.9,5.4Hz,1H ),4.34(d,J=12.6Hz,1H),4.23(d,J=13.7Hz,1H),4.12(d,J=13.7Hz,1H),4.04(s,2H),4.01(s,3H),3.86(d,J=11.9Hz, 1H),3.59(d,J=6.7Hz,6H),3.01(t,J=12.3Hz,1H),2.88(t,J=5.8Hz,3H),2.75-2.70(m,1H),2.58(ddd,J=34.3,19.1,1 0.5Hz,3H),2.09-2.03(m,1H),1.81(d,J=12.6Hz,2H),1.65-1.57(m,1H),1.47-1.41(m,1H),1.23(s,1H),1.17(s,9H). LC / MS(ESI,m / z):[M+H] + =808.61.

[0160] Example 11

[0161]

[0162] Synthesis of Example 11: (a) TsCl, TEA, DMAP, DCM, rt; (b) DIPEA, NMP, 100℃; (c) TFA, DCM, rt; (d) 10, HATU, DIPEA, DMF, rt.

[0163] Specific experimental steps:

[0164] a: Compound 43 (380 mg, 2 mmol) was weighed and dissolved in 15 mL of anhydrous dichloromethane. Triethylamine (1.1 mL, 8 mmol), p-toluenesulfonyl chloride (419 mg, 2.2 mmol), and DMAP (5 mg, 0.04 mmol) were added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched with water and extracted three times with dichloromethane (3 × 10 mL). The organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by column chromatography to give compound 44, a white oily liquid. Yield: 89%.

[0165] b: Compound 44 (344 mg, 1.0 mmol) and compound 14 (150 mg, 0.5 mmol) were weighed and dissolved in 2 mL of NMP. DIPEA (0.26 mL, 1.5 mmol) was added. The reaction mixture was heated to 100 °C, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. Prep-HPLC purification yielded compound 45 as a yellow solid. Yield: 43%.

[0166] c: Weigh compound 45 (236 mg, 0.5 mmol), dissolve in 2 mL of anhydrous dichloromethane, add TFA (2 mL), and stir the reaction mixture at room temperature for 2 h. Remove the solvent by vacuum distillation to give compound 46, a yellow solid. Yield: 80%.

[0167] d: Compound 46 (208 mg, 0.5 mmol) and compound 10 (190 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 10 mL), and the organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 11, a white solid. Yield: 65%. Purity: 98%, Rt = 5.8 min. 1 H NMR (600MHz, DMSO-d6) δ12.34(s,1H),11.15(s,1H),7.99(s,2H),7.39(s,1H),6.72(s,1H),5.17(dd,J=12.8,5.1Hz ,1H),4.96(s,2H),4.74(s,2H),4.38(d,J=12.3Hz,1H),4.05(s,2H),3.94(d,J=13.0Hz,1H),3.81(s,2H),3.74(t,J= 5.7Hz,2H),3.68(s,2H),3.07(t,J=12.7Hz,1H),2.90(t,J=17.7Hz,1H),2.75(t,J=11.2Hz,1H),2.65(dd,J=38.4,9. 8Hz, 4H), 2.09 (d, J = 11.0Hz, 1H), 1.87-1.79 (m, 2H), 1.62-1.54 (m, 1H), 1.46-1.37 (m, 1H), 1.24 (s, 1H), 1.17 (s, 9H). LC / MS(ESI,m / z):[M+H] + =778.56.

[0168] Example 12

[0169]

[0170] Synthesis of Example 12: (a) NaBH3CN, DCM, MeOH, rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt.

[0171] Specific experimental steps:

[0172] a: Compound 47 (101 mg, 0.5 mmol) and compound 14 (150 mg, 0.5 mmol) were weighed and dissolved in 4 mL of a mixture of anhydrous dichloromethane and methanol (1:1, v / v). Sodium cyanoborohydride (62 mg, 1.0 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 48 as a white solid. Yield: 70%.

[0173] b: Weigh compound 48 (243 mg, 0.5 mmol), dissolve in 2 mL of anhydrous dichloromethane, add TFA (2 mL), and stir the reaction mixture at room temperature for 2 h. Remove the solvent by vacuum distillation to give compound 49, a pale yellow solid. Yield: 78%.

[0174] c: Compound 49 (215 mg, 0.5 mmol) and compound 10 (190 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 10 mL), and the organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 12, a white solid. Yield: 61%. Purity: 98%, Rt = 5.5 min. 1H NMR (600MHz, DMSO-d6) δ12.32(d,J=4.7Hz,1H),11.15(s,1H),7.98(s,2H),7.37(s,1H),6.71(s,1H),5.17(dd,J=12.8,5.4Hz ,1H),4.97-4.65(m,4H),4.24-4.17(m,1H),4.04(s,2H),3.91(d,J=13.2Hz,1H),3.64(t,J=6.0Hz,2H),3.57-3.49(m,4H),3.0 1(dd,J=15.4,9.0Hz,1H),2.89(td,J=13.6,13.1,4.4Hz,1H),2.72(d,J=11.5Hz,1H),2.66-2.54(m,4H),2.07(dd,J=10.0,4.3 Hz,1H),1.98-1.94(m,2H),1.81(d,J=12.1Hz,1H),1.71(d,J=12.2Hz,1H),1.57-1.51(m,1H),1.27-1.23(m,2H),1.17(s,9H). LC / MS(ESI,m / z):[M+H] + =792.64.

[0175] Example 13

[0176]

[0177] Synthesis of Example 13: (a) TsCl, TEA, DMAP, DCM, rt; (b) DIPEA, NMP, 100℃; (c) TFA, DCM, rt; (d) NaBH3CN, DCM, MeOH, rt; (e) 4M HCl in 1,4-dioxane, DCM, rt; (f) 53, HATU, DIPEA, DMF, rt

[0178] Specific experimental steps:

[0179] a: Compound 50 (320 mg, 2 mmol) was weighed and dissolved in 15 mL of anhydrous dichloromethane. Triethylamine (1.1 mL, 8 mmol), p-toluenesulfonyl chloride (419 mg, 2.2 mmol), and DMAP (5 mg, 0.04 mmol) were added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched with water and extracted three times with dichloromethane (3 × 10 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by column chromatography to give compound 51, a white oily liquid. Yield: 92%.

[0180] b: Compound 51 (314 mg, 1.0 mmol) and compound 14 (150 mg, 0.5 mmol) were weighed and dissolved in 2 mL of NMP. DIPEA (0.26 mL, 1.5 mmol) was added. The reaction mixture was heated to 100 °C, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by column chromatography to give compound 52 as a yellow solid. Yield: 42%.

[0181] c: Weigh compound 52 (221 mg, 0.5 mmol), dissolve in 2 mL of anhydrous dichloromethane, add TFA (2 mL), and stir the reaction mixture at room temperature for 2 h. Remove the solvent by vacuum distillation to give compound 53, a yellow solid. Yield: 70%.

[0182] d: Compound 54 (399 mg, 2.0 mmol) and compound 10 (380 mg, 1.0 mmol) were weighed and dissolved in 8 mL of a mixture of anhydrous DCM and methanol (1:1, v / v). Sodium cyanoborohydride (124 mg, 2.0 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 55 as a white solid. Yield: 50%.

[0183] e: Compound 55 (564 mg, 1.0 mmol) was weighed and dissolved in 10 mL of anhydrous dichloromethane. 5 mL of 4 M dioxane hydrochloride solution was added, and the reaction mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum distillation to give compound 56, a white solid. Yield: 93%.

[0184] f: Compound 56 (282 mg, 0.5 mmol) and compound 53 (193 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 10 mL), and the organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 13, a white solid. Yield: 54%. Purity: 98%, Rt = 4.5 min. 1H NMR(500MHz,DMSO-d6)δ12.29(s,1H),11.12(s,1H),7.79(s,2H),7.39(s,1H), 6.72(s,1H),5.14(dd,J=12.8,5.5Hz,1H),4.47(d,J=12.5Hz,1H),4.05(s,3H) ,3.98(s,5H),3.06(s,3H),2.71(t,J=7.0Hz,2H),2.55(s,2H),2.44-2.39(m,4 H), 1.86 (d, J = 11.2Hz, 5H), 1.78-1.66 (m, 6H), 1.27-1.22 (m, 3H), 1.18 (s, 9H). LC / MS(ESI,m / z):[M+H] + =832.45.

[0185] Example 14

[0186]

[0187] Synthesis of Example 14: (a) HATU, DIPEA, DMF, rt; (b) 14, DIPEA, NMP, 100℃.

[0188] Specific experimental steps:

[0189] a: Compound 56 (282 mg, 0.5 mmol) and compound 57 (100 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The mixture was quenched with water and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. Prep-HPLC purification yielded compound 58 as a pale yellow solid. Yield: 30%.

[0190] b: Compound 58 (646 mg, 1.0 mmol) and compound 14 (150 mg, 0.5 mmol) were weighed and dissolved in 2 mL of NMP. DIPEA (0.26 mL, 1.5 mmol) was added. The reaction mixture was heated to 100 °C, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by column chromatography to obtain Example 14, a white solid. Yield: 32%. Purity: 98%. Rt = 4.5 min. LC / MS (ESI, m / z): [M+H] + =817.56.

[0191] Example 15

[0192]

[0193] Synthesis of Example 15: (a) 10, NaBH3CN, DCM, MeOH, rt; (b) 4M HCl in 1,4-dioxane, DCM, rt; (c) 57, HATU, DIPEA, DMF, rt; (d) 14, DIPEA, NMP, 100℃.

[0194] Specific experimental steps:

[0195] a: Compound 59 (213 mg, 1 mmol) and compound 10 (190 mg, 0.5 mmol) were weighed and dissolved in 4 mL of a mixed solution of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (62 mg, 1.0 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 60 as a white solid. Yield: 72%.

[0196] b: Compound 60 (578 mg, 1.0 mmol) was weighed and dissolved in 10 mL of anhydrous dichloromethane. 5 mL of 4 M dioxane hydrochloride solution was added, and the reaction mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum distillation to give compound 61, a white solid. Yield: 88%.

[0197] c: Weigh out compound 61 (239 mg, 0.5 mmol) and compound 57 (100 mg, 0.5 mmol) and dissolve them in 2 mL of DMF. Add DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol). Stir the reaction mixture at room temperature for 30 min. Quench with water and extract three times with ethyl acetate (3 × 10 mL). Combine the organic phases and wash with saturated brine. Dry the mixture on anhydrous sodium sulfate and concentrate it to obtain the crude product. Purify the crude product by prep-HPLC to obtain compound 62, a pale yellow solid. Yield: 22%.

[0198] d: Compound 62 (660 mg, 1.0 mmol) and compound 14 (150 mg, 0.5 mmol) were weighed and dissolved in 2 mL of NMP. DIPEA (0.26 mL, 1.5 mmol) was added. The reaction mixture was heated to 100 °C, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 15, a white solid. Yield: 10%. Purity: 98%. Rt = 4.7 min. LC / MS (ESI, m / z): [M+H] + =832.32.

[0199] Example 16

[0200]

[0201] Synthesis of Example 16: (a) 14, K2CO3, DMF, rt; (b) TFA, DCM, rt; (c) 61, HATU, DIPEA, DMF, rt.

[0202] Specific experimental steps:

[0203] a: Compound 63 (390 mg, 2 mmol) and compound 14 (599 mg, 2 mmol) were weighed and dissolved in 8 mL of DMF. K2CO3 (553 mg, 4 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The reaction mixture was quenched with water and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by column chromatography to give compound 64 as a pale yellow solid. Yield: 65%.

[0204] b: Weigh compound 64 (413 mg, 1 mmol), dissolve in 3 mL of anhydrous dichloromethane, add 3 mL of TFA, and stir the reaction mixture at room temperature for 2 h. Remove the solvent by vacuum distillation to give compound 65, a yellow solid. Yield: 76%.

[0205] c: Weigh compound 65 (179 mg, 0.5 mmol) and compound 61 (239 mg, 0.5 mmol) and dissolve them in 2 mL of DMF. Add DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol). Stir the reaction mixture at room temperature for 30 min. Quench the reaction mixture with water and extract three times with ethyl acetate (3 × 10 mL). Combine the organic phases and wash with saturated brine. Dry the mixture on anhydrous sodium sulfate and concentrate it to obtain the crude product. Purify the crude product by prep-HPLC to obtain Example 16, a white solid. Yield: 46%. Purity: 98%, Rt = 4.4 min.1 H NMR(500MHz,DMSO-d6)δ12.26(s,1H),11.12(s,1H),7.81(s,2H),7.39(s,1H),6.72(s,1H),5.14(dd,J= 12.8,5.4Hz,1H),4.37(d,J=12.3Hz,1H),4.11(s,4H),4.05(s,3H),3.94(d,J=12.2Hz,1H),3.68(d,J=1 4.5Hz,1H),3.59(d,J=14.5Hz,1H),3.06-2.96(m,3H),2.94-2.88(m,1H),2.64-2.53(m,4H),2.33(d,J= 5.6Hz, 2H), 2.15 (s, 2H), 2.10-2.04 (m, 1H), 1.81 (d, J = 11.2Hz, 3H), 1.70 (t, J = 13.4Hz, 5H), 1.18 (s, 9H). LC / MS(ESI,m / z):[M+H] + =817.73.

[0206] Example 17

[0207]

[0208] Synthesis of Example 17: (a) 11, HATU, DIPEA, DMF, rt; (b) DMP, DCM, rt; (c) 14, NaBH3CN, DCM, MeOH, rt.

[0209] Specific experimental steps:

[0210] a: Compound 56 (232 mg, 0.5 mmol) and compound 11 (59 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.26 mL, 1.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The mixture was quenched with water and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 66 as a white solid. Yield: 57%.

[0211] b: Weigh compound 66 (282 mg, 0.5 mmol), dissolve in 5 mL of anhydrous dichloromethane, add Dys-Martin reagent (233 mg, 0.55 mmol), and stir the reaction mixture overnight at room temperature. Quench with water, filter with diatomaceous earth, and extract the filtrate three times with dichloromethane (3 × 10 mL). Combine the organic phases and wash with saturated brine. Dry the mixture on anhydrous sodium sulfate and concentrate to obtain the crude product. Purify the crude product by prep-HPLC to give compound 67, a white solid. Yield: 42%.

[0212] c: Compound 67 (281 mg, 0.5 mmol) and compound 14 (150 mg, 0.5 mmol) were weighed and dissolved in 4 mL of a mixture of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (62 mg, 1.0 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 17, a white solid. Yield: 58%. Purity: 98%, Rt = 4.5 min. 1 H NMR(500MHz,DMSO-d6)δ12.31(s,1H),11.14(s,1H),7.81(s,2H),7.41(s,1H),6.73(s,1H),5.15(dd,J=12.8,5 .4Hz,1H),4.49(d,J=12.3Hz,1H),4.07(s,2H),4.00(s,5H),3.13(d,J=10.1Hz,2H),3.01-2.87(m,3H),2.73(t ,J=6.4Hz,2H),2.65-2.55(m,4H),2.37(d,J=7.0Hz,2H),2.07(dd,J=12.1,7.0Hz,1H),1.89(d,J=11.0Hz,5H), 1.73(q,J=10.9Hz,2H),1.61-1.54(m,4H),1.45(d,J=9.6Hz,1H),1.30(dd,J=32.2,14.2Hz,2H),1.19(s,9H). LC / MS(ESI,m / z):[M+H] + =846.56.

[0213] Example 18

[0214]

[0215] Synthesis of Example 18: (a) HATU, DIPEA, DMF, rt.

[0216] Specific experimental steps:

[0217] a: Compound 53 (193 mg, 0.5 mmol) and compound 61 (239 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF solution. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The mixture was quenched with water and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 18, a white solid. Yield: 55%. Purity: 98%, Rt = 4.6 min. 1 H NMR(500MHz,DMSO-d6)δ12.28(s,1H),11.12(s,1H),7.80(s,2H),7.39(s,1H),6.72(s,1H),5.1 4(dd,J=12.8,5.5Hz,1H),4.39(d,J=12.7Hz,1H),4.05(s,3H),3.99(s,5H),3.87(d,J=13.2Hz,1 H),3.05(s,2H),2.72(t,J=6.9Hz,2H),2.63-2.61(m,1H),2.59(d,J=4.5Hz,1H),2.54(s,2H),2. 40-2.34(m,4H),2.22(s,2H),1.83(d,J=10.7Hz,4H),1.73(tt,J=20.5,9.6Hz,8H),1.18(s,9H). LC / MS(ESI,m / z):[M+H] + =846.56.

[0218] Example 19

[0219]

[0220] Synthesis of Example 19: (a) 56, NaBH3CN, DCM, MeOH, rt; (b) 4M HCl in 1,4-dioxane, DCM, rt; (c) 65, HATU, DIPEA, DMF, rt.

[0221] Specific experimental steps:

[0222] a: Compound 54 (199 mg, 1 mmol) and compound 56 (232 mg, 0.5 mmol) were weighed and dissolved in 4 mL of a mixed solution of anhydrous dichloromethane and methanol (1:1, v / v). Sodium cyanoborohydride (62 mg, 1.0 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 68 as a white solid. Yield: 47%.

[0223] b: Compound 68 (647 mg, 1.0 mmol) was weighed and dissolved in 10 mL of anhydrous dichloromethane. 5 mL of 4 M dioxane hydrochloride solution was added, and the reaction mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum distillation to give compound 69, a white solid. Yield: 82%.

[0224] c: Weigh compound 65 (179 mg, 0.5 mmol) and compound 69 (273 mg, 0.5 mmol) and dissolve them in 2 mL of DMF. Add DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol). Stir the reaction mixture at room temperature for 30 min. Quench with water and extract three times with ethyl acetate (3 × 10 mL). Combine the organic phases and wash with saturated brine. Dry the mixture on anhydrous sodium sulfate and concentrate to obtain the crude product. Purify the crude product by prep-HPLC to obtain Example 19, a white solid. Yield: 51%. Purity: 98%, Rt = 3.3 min. 1 H NMR (500MHz, DMSO-d6) δ12.34(s,1H),11.15(s,1H),7.83(s,2H),7.41(s,1H),6.74(s,1H),5.16(dd,J=12.7,5 .4Hz,1H),4.47(d,J=11.2Hz,1H),4.12(s,4H),4.06(d,J=20.1Hz,4H),3.71(d,J=14.4Hz,1H),3.64-3.59(m,1 H),2.68(d,J=32.5Hz,3H),2.62-2.55(m,4H),2.46(d,J=12.1Hz,4H),2.09(q,J=5.1Hz,1H),1.89(s,7H),1.80 -1.68(m,3H),1.66-1.58(m,2H),1.47(d,J=9.3Hz,1H),1.33(d,J=11.8Hz,1H),1.30-1.23(m,2H),1.20(s,9H). LC / MS(ESI,m / z):[M+H] + =887.45.

[0225] Example 20

[0226]

[0227] Synthesis of Example 20: (a) 56, NaBH3CN, DCM, MeOH, rt; (b) 4M HCl in 1,4-dioxane, DCM, rt; (c) 14, K2CO3, DMF, 100℃; (d) TFA, DCM, rt; (e) 72, HATU, DIPEA, DMF, rt.

[0228] Specific experimental steps:

[0229] a: Compound 70 (171 mg, 1 mmol) and compound 56 (232 mg, 0.5 mmol) were weighed and dissolved in 4 mL of a mixture of anhydrous dichloromethane and methanol (1:1, v / v). Sodium cyanoborohydride (62 mg, 1.0 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 71 as a white solid. Yield: 63%.

[0230] b: Compound 71 (619 mg, 1.0 mmol) was weighed and dissolved in 10 mL of anhydrous dichloromethane. 5 mL of 4 M dioxane hydrochloride solution was added, and the reaction mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum distillation to give compound 72, a white solid. Yield: 80%.

[0231] c: Weigh out compound 73 (418 mg, 2 mmol) and compound 14 (599 mg, 2 mmol) and dissolve them in 8 mL of DMF solution. Add K2CO3 (553 mg, 4 mmol) and heat the reaction system to 100 °C with stirring overnight. Quench the reaction system with water, extract three times with ethyl acetate (3 × 10 mL), combine the organic phases and wash with saturated brine. Dry the mixture on anhydrous sodium sulfate and concentrate it to obtain the crude product. Purify the crude product by column chromatography to give compound 74, a pale yellow solid. Yield: 46%.

[0232] d: Weigh compound 74 (427 mg, 1 mmol), dissolve in 3 mL of anhydrous dichloromethane, add 3 mL of TFA, and stir the reaction mixture at room temperature for 2 h. Remove the solvent by vacuum distillation to give compound 75, a yellow solid. Yield: 78%.

[0233] e: Weigh compound 75 (186 mg, 0.5 mmol) and compound 72 (259 mg, 0.5 mmol) and dissolve them in 2 mL of DMF solution. Add DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol). Stir the reaction mixture at room temperature for 30 min. Quench with water and extract three times with ethyl acetate (3 × 10 mL). Combine the organic phases and wash with saturated brine. Dry the mixture on anhydrous sodium sulfate and concentrate to obtain the crude product. Purify the crude product by prep-HPLC to obtain Example 20, a white solid. Yield: 43%. Purity: 98%, Rt = 3.2 min. 1 H NMR(500MHz,DMSO-d6)δ12.26(s,1H),11.12(s,1H),7.81(s,2H),7.38(s,1H),6.72(s,1H),5.14(dd,J=12.8 ,5.4Hz,1H),4.17(t,J=7.7Hz,1H),4.09(s,4H),4.05(s,3H),3.97(dd,J=9.0,5.1Hz,1H),3.91-3.87(m,2H), 3.42(d,J=3.2Hz,2H),2.80(d,J=9.6Hz,1H),2.65-2.58(m,1H),2.54(s,2H),2.34(t,J=11.0Hz,3H),2.10-2 .04(m,1H),1.85-1.74(m,8H),1.66(q,J=11.1Hz,3H),1.51-1.43(m,3H),1.24(d,J=6.4Hz,1H),1.18(s,9H). LC / MS(ESI,m / z):[M+H] + =873.52.

[0234] Example 21

[0235]

[0236] Synthesis of Example 21: (a) 10, NaBH3CN, DCM, MeOH, rt; (b) DIBAL, DCM, 0℃; (c) DMP, DCM, rt; (d) 14, NaBH3CN, DCM, MeOH, rt.

[0237] Specific experimental steps:

[0238] a: Compound 76 (170 mg, 1 mmol) and compound 10 (190 mg, 0.5 mmol) were weighed and dissolved in 4 mL of a mixed solution of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (62 mg, 1.0 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The organic phases were washed again with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 77 as a white solid. Yield: 44%.

[0239] b: Weigh compound 77 (267 mg, 0.5 mmol), dissolve in 5 mL of anhydrous dichloromethane, cool to 0 °C, add DIBAL (0.2 mL, 2 mmol) dropwise, and stir the reaction mixture overnight at room temperature. Quench with water, filter with diatomaceous earth, extract the filtrate three times with dichloromethane (3 × 10 mL), combine the organic phases, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate to obtain the crude product. The crude product is purified by prep-HPLC to give compound 78, a white solid. Yield: 12%.

[0240] c: Weigh compound 78 (253 mg, 0.5 mmol), dissolve in 5 mL of anhydrous dichloromethane, add Dys-Martin reagent (233 mg, 0.55 mmol), and stir the reaction mixture overnight at room temperature. Quench with water, filter with diatomaceous earth, and extract the filtrate three times with dichloromethane (3 × 10 mL). Combine the organic phases and wash with saturated brine. Dry the mixture on anhydrous sodium sulfate and concentrate to obtain the crude product. Purify the crude product by prep-HPLC to give compound 79, a white solid. Yield: 41%.

[0241] d: Compound 79 (252 mg, 0.5 mmol) and compound 14 (150 mg, 0.5 mmol) were weighed and dissolved in 4 mL of a mixture of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (62 mg, 1.0 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 21, a white solid. Yield: 30%. Purity: 98%, Rt = 4.2 min. LC / MS (ESI, m / z): [M+H] + =789.37.

[0242] Example 22

[0243]

[0244] Synthesis of Example 22: (a) NaBH3CN, DCM, MeOH, rt; (b) 4M HCl in 1,4-dioxane, DCM, rt; (c) 65, HATU, DIPEA, DMF, rt.

[0245] Specific experimental steps:

[0246] a: Compound 80 (199 mg, 1 mmol) and compound 10 (190 mg, 0.5 mmol) were weighed and dissolved in 4 mL of a mixed solution of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (62 mg, 1.0 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 81 as a white solid. Yield: 85%.

[0247] b: Compound 81 (563 mg, 1.0 mmol) was weighed and dissolved in 10 mL of anhydrous dichloromethane. 5 mL of 4 M dioxane hydrochloride solution was added, and the reaction mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum distillation to give compound 82, a white solid. Yield: 88%.

[0248] c: Weigh compound 65 (179 mg, 0.5 mmol) and compound 82 (232 mg, 0.5 mmol) and dissolve them in 2 mL of DMF. Add DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol). Stir the reaction mixture at room temperature for 30 min. Quench the reaction mixture with water and extract three times with ethyl acetate (3 × 10 mL). Combine the organic phases and wash with saturated brine. Dry the mixture on anhydrous sodium sulfate and concentrate it to obtain the crude product. Purify the crude product by prep-HPLC to obtain Example 22, a white solid. Yield: 56%. Purity: 98%. Rt = 4.0 min. 1 H NMR (500MHz, DMSO-d6) δ12.31(s,1H),11.12(s,1H),7.81(s,2H),7.39(s,1H),6.72(s,1H),5.14(dd,J=12.8,5.4Hz,1H),4.16(d,J=2.5Hz, 4H),4.05(s,4H),3.61-3.56(m,3H),3.51(s,1H),2.65-2.54(m,6H),2 .10-1.96(m,2H),1.87(s,1H),1.35(s,1H),1.23(s,8H),1.18(s,9H). LC / MS(ESI,m / z):[M+H]+ =804.14.

[0249] Example 23

[0250]

[0251] Synthesis of Example 23: (a) NaBH3CN, DCM, MeOH, rt; (b) 4MHCl in 1,4-dioxane, DCM, rt; (c) 65, HATU, DIPEA, DMF, rt.

[0252] Specific experimental steps:

[0253] a: Compound 83 (185 mg, 1 mmol) and compound 10 (190 mg, 0.5 mmol) were weighed and dissolved in 4 mL of a mixed solution of anhydrous dichloromethane and methanol (1:1, v / v). Sodium cyanoborohydride (62 mg, 1.0 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 84 as a white solid. Yield: 87%.

[0254] b: Compound 84 (550 mg, 1.0 mmol) was weighed and dissolved in 10 mL of anhydrous dichloromethane. 5 mL of 4 M dioxane hydrochloride solution was added, and the reaction mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum distillation to give compound 85, a white solid. Yield: 85%.

[0255] c: Weigh compound 65 (179 mg, 0.5 mmol) and compound 85 (225 mg, 0.5 mmol) and dissolve them in 2 mL of DMF. Add DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol). Stir the reaction mixture at room temperature for 30 min. Quench the reaction mixture with water and extract three times with ethyl acetate (3 × 10 mL). Combine the organic phases and wash with saturated brine. Dry the mixture on anhydrous sodium sulfate and concentrate it to obtain the crude product. Purify the crude product by prep-HPLC to obtain Example 23, a white solid. Yield: 60%. Purity: 98%, Rt = 3.8 min. 1H NMR(500MHz,DMSO-d6)δ12.28(s,1H),11.12(s,1H),7.81(s,2H),7.38(s,1H),6.72(s,1H),5.18-5.1 2(m,1H),4.25(t,J=8.4Hz,1H),4.08(d,J=24.2Hz,8H),3.97(t,J=8.9Hz,1H),3.85(dd,J=8.2,5.7Hz, 1H),3.56(dd,J=9.3,5.5Hz,1H),3.41(s,2H),3.00-2.85(m,3H),2.70(s,1H),2.64-2.54(m,2H),2.1 2(s,1H),2.09-2.04(m,1H),1.80(d,J=11.5Hz,2H),1.66(d,J=10.1Hz,2H),1.23(s,2H),1.17(s,9H). LC / MS(ESI,m / z):[M+H] + =790.45.

[0256] Example 24

[0257]

[0258] Synthesis of Example 24: (a) HATU, DIPEA, DMF, rt.

[0259] Specific experimental steps:

[0260] a: Compound 65 (179 mg, 0.5 mmol) and compound 56 (232 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 10 mL), and the organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 24, a white solid. Yield: 57%. Purity: 98%, Rt = 3.6 min. 1H NMR(500MHz,DMSO-d6)δ12.32(s,1H),11.12(s,1H),7.81(s,2H),7.39(s,1H),6.72(s,1H), 5.14(dd,J=12.8,5.4Hz,1H),4.46(d,J=11.4Hz,1H),4.08(d,J=27.4Hz,7H),3.70(d,J=14.7 Hz,1H),3.60(d,J=14.4Hz,1H),3.01-2.89(m,2H),2.63-2.53(m,6H),2.09-2.04(m,1H),1. 95-1.87(m,4H),1.78(s,2H),1.56-1.46(m,1H),1.44-1.33(m,2H),1.23(s,2H),1.8(s,9H). LC / MS(ESI,m / z):[M+H] + =804.12.

[0261] Example 25

[0262]

[0263] Synthesis of Example 25: (a) NaBH3CN, DCM, MeOH, rt; (b) 4M HCl in 1,4-dioxane, DCM, rt; (c) 65, HATU, DIPEA, DMF, rt.

[0264] Specific experimental steps:

[0265] a: Compound 70 (171 mg, 1 mmol) and compound 10 (190 mg, 0.5 mmol) were weighed and dissolved in 4 mL of a mixed solution of anhydrous dichloromethane and methanol (1:1, v / v). Sodium cyanoborohydride (62 mg, 1.0 mmol) was added, and the reaction mixture was stirred at room temperature for 6 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 86 as a white solid. Yield: 55%.

[0266] b: Compound 86 (268 mg, 1.0 mmol) was weighed and dissolved in 10 mL of anhydrous dichloromethane. 5 mL of 4 M dioxane hydrochloride solution was added, and the reaction mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum distillation to give compound 87, a white solid. Yield: 90%.

[0267] c: Weigh compound 65 (179 mg, 0.5 mmol) and compound 87 (218 mg, 0.5 mmol) and dissolve them in 2 mL of DMF. Add DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol). Stir the reaction mixture at room temperature for 30 min. Quench the reaction mixture with water and extract three times with ethyl acetate (3 × 10 mL). Combine the organic phases and wash with saturated brine. Dry the mixture on anhydrous sodium sulfate and concentrate it to obtain the crude product. Purify the crude product by prep-HPLC to obtain Example 25, a white solid. Yield: 66%. Purity: 98%. Rt = 4.0 min. 1 H NMR(500MHz,DMSO-d6)δ12.34(s,1H),11.14(s,1H),7.81(s,2H),7.38(s,1H),6.72(s,1H),5.15( dd,J=12.7,5.4Hz,1H),4.17(s,1H),4.12(s,3H),4.05(s,3H),3.93(t,J=8.4Hz,1H),3.67(d,J=9. 0Hz,1H),3.59(dt,J=13.1,6.6Hz,2H),3.11(q,J=7.3Hz,1H),2.89(d,J=6.5Hz,2H),2.74(s,1H),2 .69(s,5H),2.55(s,1H),2.08(dd,J=9.7,4.2Hz,1H),1.83(s,2H),1.68-1.62(m,2H),1.18(s,9H). LC / MS(ESI,m / z):[M+H] + =775.45.

[0268] Example 26

[0269]

[0270] Synthesis of Example 26: (a) HATU, DIPEA, DMF, rt; (b) TFA, DCM, rt; (c) 15, HATU, DIPEA, DMF, rt; (d) DMP, DCM, rt; (e) 14, NaBH3CN, DCM, MeOH, rt.

[0271] Specific experimental steps:

[0272] a, b: Compound 8 (539 mg, 2 mmol) and compound 88 (511 mg, 2 mmol) were weighed and dissolved in 10 mL of DMF. DIPEA (0.87 mL, 5 mmol) and HATU (1.14 g, 3 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was quenched with water and extracted three times with ethyl acetate (3 × 20 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by prep-HPLC to obtain a white solid. This solid was dissolved in 10 mL of anhydrous dichloromethane, and TFA (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min. The solvent was removed by vacuum distillation to obtain trifluoroacetate of compound 89, a brown oily substance. Overall yield of the two steps: 52%.

[0273] c: Compound 89 (203 mg, 0.5 mmol) and compound 15 (66 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The mixture was quenched with water and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The extracts were dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 90 as a white solid. Yield: 70%.

[0274] d: Weigh 260 mg (0.5 mmol) of compound 90, dissolve it in 5 mL of anhydrous dichloromethane, add Dys-Martin reagent (233 mg, 0.55 mmol), stir the reaction mixture overnight at room temperature, quench with water, filter with diatomaceous earth, extract the filtrate three times with dichloromethane (3 × 10 mL), combine the organic phases and wash with saturated brine, dry over anhydrous sodium sulfate, concentrate to obtain crude product, and purify by prep-HPLC to obtain compound 91, a white solid. Yield: 60%.

[0275] e: Compound 91 (130 mg, 0.25 mmol) and compound 14 (75 mg, 0.25 mmol) were weighed and dissolved in 3 mL of a mixture of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (31 mg, 0.5 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 26, a white solid. Yield: 44%. Purity: 98%, Rt = 5.8 min. 1H NMR(500MHz,DMSO-d6)δ12.50(s,1H),11.12(s,1H),7.79(s,2H),7.37(s,1H), 6.70(s,1H),5.16-5.11(m,1H),4.30(q,J=7.1,6.4Hz,1H),4.05(s,3H),3.96( d,J=7.3Hz,4H),2.93-2.86(m,2H),2.70-2.55(m,5H),2.37-2.25(m,4H),2.08 -1.99(m,2H),1.93-1.87(m,2H),1.55-1.50(m,6H),1.23(s,2H),1.17(s,9H). LC / MS(ESI,m / z):[M+H] + =802.48.

[0276] Example 27

[0277]

[0278] Synthesis of Example 27: (a) 18, HATU, DIPEA, DMF, rt; (b) DMP, DCM, rt; (c) 14, NaBH3CN, DCM, MeOH, rt.

[0279] Specific experimental steps:

[0280] a: Compound 89 (203 mg, 0.5 mmol) and compound 18 (73 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The mixture was quenched with water and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The extracts were dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 92 as a white solid. Yield: 62%.

[0281] b: Weigh compound 92 (267 mg, 0.5 mmol), dissolve in 5 mL of anhydrous dichloromethane, add Dys-Martin reagent (233 mg, 0.55 mmol), stir the reaction mixture overnight at room temperature, quench with water, filter with diatomaceous earth, extract the filtrate three times with dichloromethane (3 × 10 mL), combine the organic phases and wash with saturated brine, dry over anhydrous sodium sulfate, concentrate to obtain crude product, and purify by prep-HPLC to obtain compound 93, a white solid. Yield: 56%.

[0282] c: Compound 93 (133 mg, 0.25 mmol) and compound 14 (75 mg, 0.25 mmol) were weighed and dissolved in 3 mL of a mixture of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (31 mg, 0.5 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 27, a white solid. Yield: 40%. Purity: 98%, Rt = 6.0 min, LC / MS (ESI, m / z): [M+H] + =817.18.

[0283] Example 28

[0284]

[0285] Synthesis of Example 28: (a) 21, HATU, DIPEA, DMF, rt; (b) DMP, DCM, rt; (c) 14, NaBH3CN, DCM, MeOH, rt.

[0286] Specific experimental steps:

[0287] a: Compound 89 (203 mg, 0.5 mmol) and compound 21 (80 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The mixture was quenched with water and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The extract was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 94 as a white solid. Yield: 55%.

[0288] b: Weigh compound 94 (274 mg, 0.5 mmol), dissolve in 5 mL of anhydrous dichloromethane, add Dys-Martin reagent (233 mg, 0.55 mmol), stir the reaction mixture overnight at room temperature, quench with water, filter with diatomaceous earth, extract the filtrate three times with dichloromethane (3 × 10 mL), combine the organic phases and wash with saturated brine, dry over anhydrous sodium sulfate, concentrate to obtain crude product, and purify by prep-HPLC to obtain compound 95, a white solid. Yield: 50%.

[0289] c: Compound 95 (136 mg, 0.25 mmol) and compound 14 (75 mg, 0.25 mmol) were weighed and dissolved in 3 mL of a mixture of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (31 mg, 0.5 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 28, a white solid. Yield: 41%. Purity: 98%, Rt = 6.2 min. 1 H NMR(400MHz,DMSO-d6)δ12.54(s,1H),11.14(s,1H),7.88(s,2H),7.38(s,1H),6.71( s,1H),5.16(dd,J=12.8,5.4Hz,1H),4.38(s,3H),4.06(s,2H),3.72-3.55(m,2H),3.1 6-3.08(m,1H),3.02(s,2H),2.96-2.84(m,1H),2.66-2.55(m,2H),2.34-2.26(m,2H) ,2.12-2.03(m,1H),1.93(s,1H),1.67-1.41(m,8H),1.38-1.22(m,10H),1.17(s,9H). LC / MS(ESI,m / z):[M+H] + =831.34.

[0290] Example 29

[0291]

[0292] Synthesis of Example 29: (a) 24, HATU, DIPEA, DMF, rt; (b) DMP, DCM, rt; (c) 14, NaBH3CN, DCM, MeOH, rt.

[0293] Specific experimental steps:

[0294] a: Compound 89 (203 mg, 0.5 mmol) and compound 24 (87 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The mixture was quenched with water and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The extracts were dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 96 as a white solid. Yield: 49%.

[0295] b: Weigh compound 96 (281 mg, 0.5 mmol), dissolve in 5 mL of anhydrous dichloromethane, add Dys-Martin reagent (233 mg, 0.55 mmol), stir the reaction mixture overnight at room temperature, quench with water, filter with diatomaceous earth, extract the filtrate three times with dichloromethane (3 × 10 mL), combine the organic phases and wash with saturated brine, dry over anhydrous sodium sulfate, concentrate to obtain crude product, and purify by prep-HPLC to obtain compound 97, a white solid. Yield: 50%.

[0296] c: Compound 97 (139 mg, 0.25 mmol) and compound 14 (75 mg, 0.25 mmol) were weighed and dissolved in 3 mL of a mixture of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (31 mg, 0.5 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 29, a white solid. Yield: 37%. Purity: 98%, Rt = 6.3 min. 1 H NMR(400MHz,DMSO-d6)δ12.52(s,1H),11.14(s,1H),7.84(s,2H),7.39(s,1H),6.71(s,1H), 5.15(dd,J=12.9,5.3Hz,1H),4.21(d,J=7.9Hz,4H),4.07-4.05(m,2H),3.65(d,J=6.8Hz,2H) ,3.17-3.09(m,1H),3.08-2.98(m,1H),2.93-2.85(m,3H),2.66-2.55(m,2H),2.30(d,J=14.3 Hz,2H),2.10-2.04(m,1H),1.95-1.90(m,1H),1.62-1.45(m,8H),1.29(s,10H),1.18(s,9H). LC / MS(ESI,m / z):[M+H] + =845.02.

[0297] Example 30

[0298]

[0299] Synthesis of Example 30: (a) 27, HATU, DIPEA, DMF, rt; (b) DMP, DCM, rt; (c) 14, NaBH3CN, DCM, MeOH, rt.

[0300] Specific experimental steps:

[0301] a: Compound 89 (203 mg, 0.5 mmol) and compound 27 (94 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The mixture was quenched with water and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The extracts were dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 98 as a white solid. Yield: 42%.

[0302] b: Weigh compound 98 (288 mg, 0.5 mmol), dissolve in 5 mL of anhydrous dichloromethane, add Dys-Martin reagent (233 mg, 0.55 mmol), stir the reaction mixture overnight at room temperature, quench with water, filter with diatomaceous earth, extract the filtrate three times with dichloromethane (3 × 10 mL), combine the organic phases and wash with saturated brine, dry over anhydrous sodium sulfate, concentrate to obtain crude product, and purify by prep-HPLC to obtain compound 99, a white solid. Yield: 45%.

[0303] c: Compound 99 (142 mg, 0.25 mmol) and compound 14 (75 mg, 0.25 mmol) were weighed and dissolved in 3 mL of a mixture of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (31 mg, 0.5 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 30, a white solid. Yield: 31%. Purity: 98%, Rt = 6.4 min. 1H NMR(400MHz,DMSO-d6)δ12.52(s,1H),11.14(s,1H),7.81(s,2H),7.39(s,1H),6.71(s, 1H),5.15(dd,J=13.0,5.3Hz,1H),4.07(d,J=12.9Hz,6H),3.70-3.53(m,3H),3.17-3.09 (m,1H),2.95-2.85(m,1H),2.78(s,2H),2.65-2.56(m,2H),2.33-2.25(m,2H),2.12-2.0 3(m,1H),1.92(t,J=6.9Hz,1H),1.60-1.40(m,8H),1.27(d,J=6.3Hz,12H),1.17(s,9H). LC / MS(ESI,m / z):[M+H] + =859.25.

[0304] Example 31

[0305]

[0306] Synthesis of Example 31: (a) 30, HATU, DIPEA, DMF, rt; (b) DMP, DCM, rt; (c) 14, NaBH3CN, DCM, MeOH, rt.

[0307] Specific experimental steps:

[0308] a: Compound 89 (203 mg, 0.5 mmol) and compound 30 (101 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The mixture was quenched with water and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The extracts were dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 100 as a white solid. Yield: 42%.

[0309] b: Weigh compound 100 (295 mg, 0.5 mmol), dissolve in 5 mL of anhydrous dichloromethane, add Dys-Martin reagent (233 mg, 0.55 mmol), stir the reaction mixture overnight at room temperature, quench with water, filter with diatomaceous earth, extract the filtrate three times with dichloromethane (3 × 10 mL), combine the organic phases and wash with saturated brine, dry over anhydrous sodium sulfate, concentrate to obtain crude product, and purify by prep-HPLC to obtain compound 101, a white solid. Yield: 40%.

[0310] c: Compound 101 (145 mg, 0.25 mmol) and compound 14 (75 mg, 0.25 mmol) were weighed and dissolved in 3 mL of a mixture of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (31 mg, 0.5 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 31, a white solid. Yield: 25%. Purity: 98%, Rt = 6.6 min. 1 H NMR(400MHz,DMSO-d6)δ12.51(s,1H),11.13(s,1H),7.79(s,2H),7.38(s,1H),6.71(s, 1H),5.15(dd,J=13.0,5.6Hz,1H),4.03(d,J=22.2Hz,7H),3.71-3.53(m,3H),3.18-3.00 (m,2H),2.95-2.86(m,1H),2.75-2.67(m,2H),2.66-2.56(m,2H),2.33-2.25(m,2H),2.1 0-2.04(m,1H),1.91(t,J=6.2Hz,1H),1.49(s,8H),1.26(d,J=6.6Hz,12H),1.17(s,9H). LC / MS(ESI,m / z):[M+H] + =873.43.

[0311] Example 32

[0312]

[0313] Synthesis of Example 32: (a) 33, HATU, DIPEA, DMF, rt; (b) DMP, DCM, rt; (c) 14, NaBH3CN, DCM, MeOH, rt.

[0314] Specific experimental steps:

[0315] a: Compound 89 (203 mg, 0.5 mmol) and compound 33 (108 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The mixture was quenched with water and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The extracts were dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 102 as a white solid. Yield: 42%.

[0316] b: Weigh compound 102 (151 mg, 0.25 mmol), dissolve in 5 mL of anhydrous dichloromethane, add Dys-Martin reagent (117 mg, 0.27 mmol), stir the reaction mixture overnight at room temperature, quench with water, filter with diatomaceous earth, extract the filtrate three times with dichloromethane (3 × 10 mL), combine the organic phases and wash with saturated brine, dry over anhydrous sodium sulfate, concentrate to obtain crude product, and purify by prep-HPLC to obtain compound 103, a white solid. Yield: 39%.

[0317] c: Compound 103 (59 mg, 0.10 mmol) and compound 14 (30 mg, 0.10 mmol) were weighed and dissolved in 3 mL of a mixed solution of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (12 mg, 0.2 mmol) was added, and the reaction system was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 5 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 32, a white solid. Yield: 23%. Purity: 98%, Rt = 6.8 min. 1H NMR(400MHz,DMSO-d6)δ12.51(s,1H),11.13(s,1H),7.80(s,2H),7.38(s,1H),6.71(s,1H),5.15(d d,J=13.0,5.4Hz,1H),4.05(d,J=4.4Hz,7H),3.71-3.52(m,3H),3.13(d,J=12.8Hz,1H),3.06-2.99( m,1H),2.89(dd,J=13.4,4.9Hz,1H),2.78-2.71(m,2H),2.65-2.55(m,2H),2.28(dd,J=14.0,7.6Hz ,2H),2.11-2.02(m,1H),1.95-1.88(m,1H),1.59-1.41(m,8H),1.25(d,J=6.1Hz,14H),1.17(s,9H). LC / MS(ESI,m / z):[M+H] + =886.98.

[0318] Example 33

[0319]

[0320] Synthesis of Example 33: (a) NaBH3CN, DCM, MeOH, rt; (b) 14, NaBH3CN, DCM, MeOH, rt.

[0321] Specific experimental steps:

[0322] a: Compound 104 (140 mg, 1 mmol) and compound 10 (190 mg, 0.5 mmol) were weighed and dissolved in 4 mL of a mixed solution of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (62 mg, 1.0 mmol) was added, and the reaction system was stirred at room temperature for 24 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain compound 105, a white solid, with a yield of 80%.

[0323] b: Compound 105 (50 mg, 0.10 mmol) and compound 14 (30 mg, 0.10 mmol) were weighed and dissolved in 3 mL of a mixed solution of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (12 mg, 0.2 mmol) was added, and the reaction system was stirred at room temperature for 3 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 5 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 33, a white solid. Yield: 56%. Purity: 98%, Rt = 4.8 min. 1 H NMR(500MHz,DMSO-d6)δ12.25(s,1H),11.12(s,1H),7.78(s,2H),7.38(s,1H),6.72(s,1H),5. 14(dd,J=12.8,5.5Hz,1H),4.05(s,2H),3.94(s,4H),2.97-2.86(m,4H),2.64-2.54(m,3H),2. 21(d,J=6.2Hz,1H),2.10-1.97(m,4H),1.85(d,J=9.2Hz,2H),1.79(d,J=11.6Hz,4H),1.66(t, J=16.6Hz, 3H), 1.49 (dd, J=17.9, 8.3Hz, 3H), 1.23 (s, 1H), 1.18 (s, 9H), 0.89 (d, J= 9.4Hz, 2H). LC / MS(ESI,m / z):[M+H] + =788.86.

[0324] Example 34

[0325]

[0326] Synthesis of Example 34: (a) NaBH3CN, DCM, MeOH, rt; (b) 4M HCl in 1,4-dioxane, DCM, rt; (c) 65, HATU, DIPEA, DMF, rt

[0327] Specific experimental steps:

[0328] a, b: Compound 106 (227 mg, 1 mmol) and compound 10 (190 mg, 0.5 mmol) were weighed and dissolved in 4 mL of a mixture of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (62 mg, 1.0 mmol) was added, and the reaction mixture was stirred at room temperature for 24 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC, dissolved in 10 mL of anhydrous dichloromethane, and 4 M dioxane hydrochloride solution (5 mL) was added. The reaction mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum distillation to obtain compound 107, a white solid. Two-step yield: 68%.

[0329] c: Compound 65 (36 mg, 0.10 mmol) and compound 107 (49 mg, 0.10 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (86 μL, 0.5 mmol) and HATU (557 mg, 0.15 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 10 mL), and the organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain the product in Example 34, a white solid. Yield: 57%. Purity: 98%, Rt = 4.7 min. 1 H NMR (500MHz, DMSO-d6) δ12.27(s,1H),11.12(s,1H),7.81(s,2H),7.38(s,1H),6.72(s,1H),5.14(dd,J=12. 8,5.4Hz,1H),4.35(d,J=12.4Hz,1H),4.10(s,5H),4.05(s,3H),3.92(d,J=13.0Hz,1H),3.65(d,J=14.5Hz, 1H),3.58(d,J=14.5Hz,1H),2.96(t,J=11.6Hz,4H),2.57(d,J=20.6Hz,2H),2.42-2.38(m,2H),2.07-1.98( m,4H),1.78(d,J=10.9Hz,2H),1.70-1.64(m,4H),1.62(s,1H),1.51(s,1H),1.41-1.37(m,2H),1.18(s,9H). LC / MS(ESI,m / z):[M+H] + =832.11.

[0330] Example 35

[0331]

[0332] Synthesis of Example 35: (a) NaBH3CN, DCM, MeOH, rt; (b) 4M HCl in 1,4-dioxane, DCM, rt; (c) 65, HATU, DIPEA, DMF, rt

[0333] Specific experimental steps:

[0334] a, b: Compound 108 (213 mg, 1 mmol) and compound 10 (190 mg, 0.5 mmol) were weighed and dissolved in 4 mL of a mixture of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (62 mg, 1.0 mmol) was added, and the reaction mixture was stirred at room temperature for 24 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC, dissolved in 10 mL of anhydrous dichloromethane, and 4 M dioxane hydrochloride solution (5 mL) was added. The reaction mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum distillation to obtain compound 109, a white solid. Two-step yield: 70%.

[0335] c: Compound 65 (36 mg, 0.10 mmol) and compound 109 (48 mg, 0.10 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (86 μL, 0.5 mmol) and HATU (557 mg, 0.15 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 10 mL), and the organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 35, a white solid. Yield: 60%. Purity: 98%, Rt = 4.4 min. 1H NMR(500MHz,DMSO-d6)δ12.21(s,1H),11.12(s,1H),7.81(s,2H),7.38(s,1H),6.72(s,1H),5.13(dd, J=11.1,5.3Hz,1H),4.12-4.01(m,8H),3.84(dt,J=35.4,13.3Hz,2H),3.68-3.56(m,1H),3.53-3.43(m ,1H),2.94-2.81(m,4H),2.64-2.54(m,2H),2.45-2.39(m,1H),2.24-2.15(m,1H),2.05(dt,J=15.7,9. 5Hz, 2H), 1.97-1.91 (m, 1H), 1.79-1.69 (m, 4H), 1.66-1.60 (m, 4H), 1.24 (d, J = 6.6Hz, 1H), 1.18 (s, 9H). LC / MS(ESI,m / z):[M+H] + =818.31.

[0336] Example 36

[0337]

[0338] Synthesis of Example 36: (a) NaBH3CN, DCM, MeOH, rt; (b) 4M HCl in 1,4-dioxane, DCM, rt; (c) 65, HATU, DIPEA, DMF, rt

[0339] Specific experimental steps:

[0340] a, b: Compound 110 (227 mg, 1 mmol) and Compound 10 (190 mg, 0.5 mmol) were weighed and dissolved in 4 mL of a mixture of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (62 mg, 1.0 mmol) was added, and the reaction mixture was stirred at room temperature for 24 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC, dissolved in 10 mL of anhydrous dichloromethane, and 4 M dioxane hydrochloride solution (5 mL) was added. The reaction mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum distillation to obtain Compound 111, a white solid. Two-step yield: 32%.

[0341] c: Compound 65 (36 mg, 0.10 mmol) and compound 111 (49 mg, 0.10 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (86 μL, 0.5 mmol) and HATU (557 mg, 0.15 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 10 mL), and the organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 36, a white solid. Yield: 50%. Purity: 98%, Rt = 4.6 min. 1 H NMR(500MHz,DMSO-d6)δ12.22(s,1H),11.12(s,1H),7.81(s,2H),7.38(s,1H),6.71(s,1H),5.14(dd,J=12.8,5.4Hz,1H) ,4.39(d,J=12.0Hz,1H),4.08(d,J=26.7Hz,8H),3.96(d,J=12.8Hz,1H),3.66(d,J=14.5Hz,1H),3.60-3.57(m,1H),2.74( d,J=7.4Hz,1H),2.60(d,J=16.2Hz,2H),2.56-2.53(m,1H),2.43(t,J=10.1Hz,2H),2.27-2.23(m,1H),2.07-2.01(m,2H) ,1.96(d,J=13.3Hz,1H),1.74(d,J=9.7Hz,2H),1.66-1.52(m,4H),1.24(d,J=6.6Hz,3H),1.18(s,9H),0.88-0.86(m,3H). LC / MS(ESI,m / z):[M+H] + =832.42.

[0342] Example 37

[0343]

[0344] Synthesis of Example 37: (a) HATU, DIPEA, DMF, rt; (b) DMP, DCM, rt; (c) 14, NaBH3CN, DCM, MeOH, rt.

[0345] Specific experimental steps:

[0346] a: Compound 10 (190 mg, 0.5 mmol) and compound 112 (80 mg, 0.5 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The mixture was quenched with water and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to give compound 113 as a white solid. Yield: 35%.

[0347] b: Weigh compound 113 (130 mg, 0.25 mmol), dissolve in 5 mL of anhydrous dichloromethane, add Dys-Martin reagent (117 mg, 0.27 mmol), stir the reaction mixture overnight at room temperature, quench with water, filter with diatomaceous earth, extract the filtrate three times with dichloromethane (3 × 10 mL), combine the organic phases and wash with saturated brine, dry over anhydrous sodium sulfate, concentrate to obtain crude product, and purify by prep-HPLC to obtain compound 114, a white solid. Yield: 40%.

[0348] c: Weigh out compound 114 (52 mg, 0.10 mmol) and compound 14 (30 mg, 0.10 mmol), dissolve them in 3 mL of a mixed solution of anhydrous dichloromethane and methanol (volume ratio 1:1), add sodium cyanoborohydride (16 mg, 0.25 mmol), and stir the reaction system at room temperature for 3 h. Remove the solvent by vacuum distillation, quench with water, extract three times with ethyl acetate (3 × 10 mL), combine the organic phases and wash with saturated brine, dry to anhydrous sodium sulfate, concentrate to obtain crude product, and purify by prep-HPLC to obtain Example 37, a white solid. Yield: 22%. Purity: 98%, Rt = 3.5 min. 1H NMR(500MHz,DMSO-d6)δ12.31(s,1H),11.13(s,1H),7.82(s,2H),7.39(s,1H),6.72(s,1H),5.15(d d,J=12.7,5.4Hz,1H),4.41(d,J=12.4Hz,1H),4.22-3.93(m,8H),3.06(t,J=12.2Hz,1H),2.94-2.87 (m,1H),2.78-2.71(m,2H),2.61-2.55(m,4H),2.07(dt,J=10.2,5.2Hz,1H),1.86(t,J=14.6Hz,4H) ,1.70(d,J=11.9Hz,2H),1.58-1.51(m,2H),1.45-1.36(m,3H),1.18(s,9H),1.04(d,J=10.0Hz,2H). LC / MS(ESI,m / z):[M+H] + =803.12.

[0349] Example 38

[0350]

[0351] Synthesis of Example 38: (a) NaBH3CN, DCM, MeOH, rt; (b) 4M HCl in 1,4-dioxane, DCM, rt; (c) 65, HATU, DIPEA, DMF, rt

[0352] Specific experimental steps:

[0353] a, b: Compound 115 (254 mg, 1 mmol) and compound 10 (190 mg, 0.5 mmol) were weighed and dissolved in 4 mL of a mixture of anhydrous dichloromethane and methanol (volume ratio 1:1). Sodium cyanoborohydride (62 mg, 1.0 mmol) was added, and the reaction mixture was stirred at room temperature for 24 h. The solvent was removed by vacuum distillation, quenched with water, and extracted three times with ethyl acetate (3 × 10 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by prep-HPLC, dissolved in 10 mL of anhydrous dichloromethane, and 4 M dioxane hydrochloride solution (5 mL) was added. The reaction mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum distillation to obtain compound 116, a white solid. Two-step yield: 20%.

[0354] c: Compound 65 (36 mg, 0.10 mmol) and compound 116 (52 mg, 0.10 mmol) were weighed and dissolved in 2 mL of DMF. DIPEA (86 μL, 0.5 mmol) and HATU (557 mg, 0.15 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 10 mL), and the organic phases were combined and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain the crude product. The crude product was purified by prep-HPLC to obtain Example 38, a white solid. Yield: 52%. Purity: 98%, Rt = 3.3 min. 1 H NMR(500MHz,DMSO-d6)δ12.26(s,1H),11.12(s,1H),7.81(s,2H),7.38(s,1H),6.72(s,1H),5.14(dd,J=12.8 ,5.4Hz,1H),4.17(t,J=7.7Hz,1H),4.09(s,4H),4.05(s,3H),3.97(dd,J=9.0,5.1Hz,1H),3.91-3.87(m,2H), 3.42(d,J=3.2Hz,2H),2.80(d,J=9.6Hz,1H),2.65-2.58(m,1H),2.54(s,2H),2.34(t,J=11.0Hz,3H),2.10-2 .04(m,1H),1.85-1.74(m,8H),1.66(q,J=11.1Hz,3H),1.51-1.43(m,3H),1.24(d,J=6.4Hz,1H),1.18(s,9H). LC / MS(ESI,m / z):[M+H] + =859.55.

[0355] Example 39 Bioactivity Test

[0356] I. Cell Culture

[0357] NCI-H226 (H226), NALM6, and 94T778 (T778) cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum (Novizan) and 1% penicillin / streptomycin (Beyotime). TC-71, A-375, A549, A-673, SKUT1, SJSA-1, MES-SA, and U87 cells were cultured in DMEM high-glucose medium (containing L-glutamine and 110 mg / L sodium pyruvate) containing 10% fetal bovine serum (Novizan) and 1% penicillin / streptomycin (Beyotime). All cells were cultured at 37°C in a 5% CO2 incubator and passaged after digestion with 0.05% trypsin solution containing EDTA (Pronotin). All cell lines were commercially available.

[0358] II. Half-cell activity inhibition rate (IC50) 50 )test

[0359] Day 1: Based on the cell growth characteristics, lay 1.2 x 10⁻⁶ cells. 3 Up to 1.0x10 4 One cell per well was added to a 96-well plate with 90 μL of culture medium. A blank control well containing no cells was also included.

[0360] Day 2: Add 10 μL of culture medium containing different concentrations of the compound to each well. For each experiment, set up at least two replicates for each concentration.

[0361] Day 5: Add 10 μL of thiazolyl blue (MTT) solution to each well, incubate at 37°C for 3 hours, discard the supernatant, add 100 μL of MTT stop reagent per well, shake at room temperature for 30 minutes, and read the absorbance at OD570 nm using a microplate reader. Subtract the absorbance (background value) of the blank control well from the readings of each experimental well to calculate the relative activity. Then, use GraphpadPrism 7 to perform nonlinear fitting and calculate the IC50. 50 .

[0362] III. Western blot (WB) detection of protein degradation effects

[0363] Step 1: Lay 5x10 5 Cells were placed in a 6cm dish.

[0364] Step 2: After 24 hours, add DMSO or a compound of different concentrations to each petri dish.

[0365] Step 3: After 6 hours of drug treatment, discard the culture medium, wash twice with pre-cooled PBS, scrape off cells and centrifuge. Discard the PBS, retain the cell pellet for total protein extraction.

[0366] Step 4: Prepare High Salt cell lysis buffer (50 mmol / L Tris pH 8.0, 420 mmol / L NaCl, 5% Glycerol, 0.5% NP-40, 1 mmol / L EDTA), and add DTT (1 mmol / L), PMSF (1 mmol / L), MgCl2 (1 mmol / L), BenzoNuclease (0.5 U / mL, nearshore protein), 1X protease inhibitor mixture (Taoshu Biotechnology), and 1X phosphatase inhibitor mixture (Taoshu Biotechnology). Add an appropriate volume of lysis buffer to the cell pellet, resuspend thoroughly, incubate on ice for 10 min, vortex for 2 s, and repeat the incubation for 10 min and vortexing twice. Centrifuge at 12000 rpm for 10 min using a 4-degree high-speed centrifuge, collect the supernatant sample and store temporarily on ice.

[0367] Step 5: Use the Bradford Protein Assay Kit (Beyotime) to determine the protein concentration of each sample.

[0368] Step 6: Add 1 / 3 volume of 4X protein loading buffer (containing β-mercaptoethanol) to each protein sample, mix well, and then denature at 95°C for 5 minutes in a metal bath.

[0369] Step 7: Perform SDS-PAGE (SDS denaturing polyacrylamide gel electrophoresis) and wet transfer at a sample loading rate of 15 μg / well, following the standard procedure.

[0370] Step 8: Block the transferred PVDF membrane (Millipore) with a 5% skim milk PBST solution, and proceed with primary antibody incubation, PBST washing, secondary antibody incubation, and PBST washing. Finally, protein development is performed using a chemiluminescence immunoassay on the Qinxiang gel imaging system. Antibody information is as follows:

[0371]

[0372] Step 9: If protein quantification is required, use ImageJ for grayscale analysis to calculate the degradation efficiency of each compound at different concentrations of intracellular CDK9. Use GraphPad Prism7 to perform nonlinear fitting on the degradation curves to obtain the DC (density precipitate). 50 .

[0373] IV. Drug Efficacy Testing in Immunodeficient Mice

[0374] Step 1: Prepare 8-week-old NCG immunodeficient mice (all purchased from commercial sources), shave their back fur and set them aside.

[0375] Step 2: After washing the in vitro cultured tumor cells TC-71 twice with PBS, digest them with 0.05% trypsin, collect the cells, and count them using a hemocytometer.

[0376] Step 3: Resuspend the cells in a 50% matrix gel (Lambolid) PBS mixture at a density of 10 million cells / mL, and inoculate the tumor cells subcutaneously on both sides of the back of immunodeficient mice at a dose of 1 million cells per tumor.

[0377] Step 4: Two weeks after inoculation, the subcutaneous xenograft model was successfully constructed and grew to a suitable size.

[0378] Step 5: Prepare the stock solution of the compound and resuspend it in 5% Solutol HS-15 (Aladdin) saline solution at a dose of 10 mg / kg. Administer via tail vein injection or solvent treatment.

[0379] Step 6: The test mice were euthanized 2h, 8h and 24h after drug administration, and the transplanted tumor tissue and spleen and other organs were collected for immunoblotting analysis.

[0380] V. Drug Treatment Experiment for Subcutaneous Xenografts in Immunodeficient Mice

[0381] Step 1: Prepare 8-week-old NCG immunodeficient mice (all purchased from commercial sources), shave their back fur and set them aside.

[0382] Step 2: After washing the in vitro cultured tumor cells TC-71 twice with PBS, digest them with 0.05% trypsin, collect the cells, and count them using a hemocytometer.

[0383] Step 3: Resuspend the cells in a 50% matrix gel (Lambolid) PBS mixture at a density of 5 million cells / mL, and inoculate the tumor cells subcutaneously on both sides of the back of immunodeficient mice at a dose of 500,000 cells per tumor.

[0384] Step 4: Three days after inoculation, the subcutaneous xenograft began to expand significantly;

[0385] Step 5: Prepare the stock solution of the compound and resuspend it in 5% Solutol HS-15 (Aladdin) saline solution at a dose of 10 mg / kg. Administer via tail vein injection or solvent treatment. The dosing regimen is every other day for two weeks. Measure the tumor volume in mice.

[0386] Step 6: At the end of the experiment, the mice were euthanized and the transplanted tumor tissue was collected for immunoblotting analysis.

[0387] like Figure 1As shown, in TC-71 cells, CDK9 series degraders have a significant ability to degrade CDK9 protein;

[0388] like Figure 2 As shown, unexpectedly, these compounds can effectively inhibit tumor cell growth, IC50. 50 It is as low as picomolar or nanomolar levels, superior to the positive control CDK9 degrader THAL-SNS032.

[0389] like Figure 3 As shown, the CDK9 degrading agent of the present invention has good inhibitory activity against a variety of tumor cells.

[0390] like Figure 4 As shown, compound 4 of the present invention has a good inhibitory effect on tumors in mice.

[0391] As shown in Table 1, these compounds exhibited CDK9 degradation activity and cell viability inhibition activity in TC-71 cells.

[0392] Table 1. Degradative activity of compounds against CDK9 protein and inhibitory activity against cell viability

[0393] compound <![CDATA[DC 50 in TC-71]]> <![CDATA[IC 50 in TC-71]]> 1 C C 2 B B 3 B B 4 B B 5 B C 6 C C 7 C C 8 B C 9 C C 10 C C 11 C B 12 B C 13 B A 14 B B 15 B C 16 A A 17 B B 18 B A 19 B C 20 B C 21 C C 22 ND B 23 ND C 24 ND B 25 ND C 26 ND C 27 ND C 28 ND C 29 ND C 30 ND C 31 ND C 32 ND C 33 ND A 34 ND B 35 ND B 36 ND A 37 ND B 38 ND C THAL-SNS032 C C

[0394] Note: DC 50 “A” indicates less than 1 nM; “B” indicates between 1 nM and 10 nM; “C” indicates greater than 10 nM; “ND” indicates not detected.

[0395] IC 50 “A” indicates less than 1 nM; “B” indicates between 1 nM and 10 nM; “C” indicates greater than 10 nM.

[0396] Example 40 Animal Pharmacokinetic (PK) Study

[0397] The aim of this study was to determine the pharmacokinetic properties of the compound after intravenous injection (IV) and oral administration (PO) in male CD1 mice.

[0398] Sample collection and analysis:

[0399] Blood volume collected: 0.03 mL

[0400] Blood collection method: orbital venous plexus

[0401] Anticoagulant: EDTA-K2

[0402] Blood collection time:

[0403] IV: 5min, 15min, 30min, 1h, 2h, 4h, 7h, 24h

[0404] PO: 15min, 30min, 1h, 2h, 4h, 7h, 24h

[0405] The collected blood samples were transferred to microcentrifuge tubes containing EDTA-K2 anticoagulant, centrifuged at 4℃ and 4000g for 5 minutes, and the supernatant was collected and stored in a refrigerator at -75℃±15℃.

[0406] Sample pretreatment was performed according to the Standard PK procedure.

[0407] The obtained data were used to calculate pharmacokinetic parameters using WinNonlin software. IV pharmacokinetic parameters included Cl and T. 1 / 2 C0, AUC, MRT, V d PO pharmacokinetic parameters include T 1 / 2 T max C max AUC, MRT, F% etc.

[0408] As shown in Table 2, compounds 4 and 16 in this invention have good pharmacokinetic parameters.

[0409] Table 2. Pharmacokinetic (PK) studies of compounds in mice

[0410]

[0411] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A compound of general formula (I) or a stereoisomer, enantiomer, transisomer, optical isomer, racemate, tautomer, or pharmaceutically acceptable salt thereof, its hydrate, or its isotopically labeled compound: in, Z is selected from 5-10 N-containing heterocycles; L is -L 1 -[W 1 -L'-W 2 ] s -L 3 -; L 1 Connected to Z, and L 1 Selected from chemical bonds, -CO-, -O-, substituted or unsubstituted -(CH2). m - Substituted or unsubstituted C3-C8 carbon rings, substituted or unsubstituted 4-10 membered heterocycles; L 3 Selected from chemical bonds, -C(=O)-(CH2) p -、-(CH2) p NH-, -O- or s can be 0, 1, 2, 3, 4, 5, or 6; Each L' is independently selected from: chemical bonds, substituted or unsubstituted C3-C8 carbon rings, substituted or unsubstituted 4-10 membered heterocycles, and -NHCO- in each occurrence; Each W 1 and W 2 Each time it appears, it is independently selected from chemical bonds, -CO-, and -(CH2). m -、-Y-(CH2) m -、-(CH2) n -Y-, -Y-(CH2) m -Y-(CH2) n -、-(CH2) m -Y-(CH2CH2O) n -; Each p is independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 each time it appears; Each occurrence of m is independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12; Each n is independently selected from 0, 1, 2, 3, 4, 5, or 6 in each occurrence; Each Y is independently selected from -O- and -S- for each occurrence; R1 is selected from: H, halogen, C1-C6 alkyl, prodrug group, such as ester group, amide group, alcohol group, phosphate ester group, glycoside, sustained-release group, etc.

2. The compound of claim 1, wherein, The compound has the structure shown in formula (II): Wherein, Z and L are as defined in claim 1.

3. The compound of claim 1, wherein, Z is selected from 4. The compound of claim 1, wherein, The compound has the structure shown in formula (III-1) or formula (III-2):

5. The compound of claim 1, wherein, Each L', in each occurrence, is independently selected from: chemical bonds, -NHCO-.

6. The compound of claim 1, wherein, L is selected from:

7. The compound of claim 1, wherein, The compounds are selected from the group consisting of:

8. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof, and a pharmaceutically acceptable carrier.

9. Use of the compound of any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating CDK9-mediated diseases or conditions.

10. The use as described in claim 9, characterized in that, The activity of diseased cells or related pathogenic cells depends on CDK9. The diseases are selected from the following group: soft tissue sarcoma, primitive neuroectodermal tumors, lung sarcoma, liposarcoma, osteosarcoma, Kaposi's sarcoma, midline carcinoma, breast cancer, lung cancer, colorectal cancer, liver cancer, pancreatic cancer, peritoneal cancer, thyroid tumors, parathyroid tumors, soft tissue hyperplasia, warts, Langerhans cell histiocytosis, hamartoma, gestational trophoblastic disease, skin cancer, Merkel cell carcinoma, bone tumors, melanoma, leukemia, salivary gland tumors, neuroendocrine tumors. Lymphoma, brain tumors, neuroblastoma, mesothelioma, esophageal cancer, medulloblastoma, glioblastoma, meningioma, retinoblastoma, kidney cancer, bladder cancer, stomach cancer, uterine cancer, vulvar cancer, prostate cancer, testicular cancer, penile cancer, bile duct cancer, gallbladder cancer, ureteral cancer, adrenocortical cancer, head and neck cancer, oral cancer, nasopharyngeal cancer, myeloma, ovarian cancer, cervical cancer, germ cell tumors, thymic tumors, myelodysplastic syndrome, granulomas, atherosclerosis, arthritis, cardiomyopathy, obesity, and autoimmune diseases.

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