MDM2 ligand-based cdks 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 relevant diseases or disorders mediated by CDK9.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceuticals, and specifically relates to CDK9 protein kinase degrading agents based on MDM2 ligands and their uses. Background Technology

[0002] The tumor suppressor protein p53 plays a central role in cancer biology, regulating key processes such as cell cycle, genome stability, cell death, metabolic reprogramming, tumor metastasis, and immune regulation. Variations in the p53 pathway promote cancer development. Typical variations in this pathway include TP53 gene mutations, TP53 gene deletions, and MDM2 (Murine double minute 2) gene amplification. The MDM2 gene encodes the E3 ubiquitin ligase for the p53 protein. MDM2 gene amplification is observed in approximately 3% of human cancers and is more common in samples with a wild-type TP53 genotype. In terms of cancer type, MDM2 amplification or overexpression is relatively common in breast cancer, glioma, urinary tract tumors, and sarcomas. Actively expressed MDM2 can induce the degradation of endogenous wild-type p53 protein in cancer cells via the ubiquitin-proteasome pathway, thereby limiting the expression and activity of p53 protein. In TP53 wild-type cancer cells, knockdown or interference with MDM2 exhibits good anti-cancer activity. Therefore, targeting and disrupting the degradation effect of MDM2 on p53 can help increase the abundance of p53 protein and stimulate its anti-cancer activity.

[0003] Currently, several classes of peptide inhibitors or small molecule inhibitors targeting the MDM2-p53 axis have been reported. Representative small molecules include cis-imidazolines (e.g., Nutlin-3a), dihydroisoquinolinones (e.g., NVP-CGM097), benzodiazepines (e.g., TDP222669), isoindolinones (e.g., NU8231), indoles (e.g., WK298), and pyrrolidones. MDM2 inhibitors such as RG7112, RG7388, NVP-CGM097, Siredalin, DS-3032, SAR405838, MK-8242, and AMG232 have been approved for clinical trials. Recently published results from a Phase 1 clinical trial of DS-3032 (Milademetan) suggest that some patients with dedifferentiated liposarcoma may benefit from MDM2 inhibitors. However, most of the reported clinical trial results are not ideal, and no drugs have been officially approved yet, suggesting that the efficacy of simply targeting the MDM2-p53 axis is relatively limited.

[0004] Protein degradation-targeting chimeras (PROTACs) utilize the intracellular ubiquitin-proteasome pathway to selectively degrade target proteins, representing an emerging direction in chemical drug development. Several molecules have already demonstrated the ability to target and degrade the MDM2 protein using the PROTAC principle, including typical molecules such as KT-253, MS3227, MD-224, MG-277, AA-265, TW-32, and SPMI-HIF2-1. PROTAC molecules generally consist of three functional modules: an E3 ubiquitin ligase-binding module, a molecular chain, and a target protein-binding module. The mechanism of action of PROTACs differs from traditional inhibitors, helping to overcome resistance in cancer cells to these inhibitors. The expression level of the E3 ubiquitin ligase and its activity-regulating complex determines the intracellular biological activity of the PROTAC molecule. Currently, most PROTAC molecules primarily utilize E3 ligases such as CRBN and VHL to degrade target proteins. However, CRBN-dependent PROTAC molecules often degrade IKZF or GSPT proteins simultaneously in addition to the target protein. Furthermore, the lack of cancer-specific expression of CRBN and VHL leads to the degradation of target proteins in both cancer cells and normal cells, resulting in strong toxic side effects. Therefore, identifying and utilizing cancer-specific E3 ligases can help improve the selectivity and therapeutic potential of PROTAC. In this context, MDM2, as a selectively upregulated oncogenic E3 ubiquitin ligase in cancer cells, becomes an ideal candidate protein.

[0005] Currently, several strategies for selectively degrading proteins of interest using the E3 ubiquitin ligase activity of MDM2 have been reported in the literature. Based on the type of MDM2 binding module, PROTAC molecules dependent on MDM2 degradation activity can be divided into two categories: peptide degraders and small molecule degraders. Successful examples of peptide degraders include hybrid designs using MDM2-p53-binding inhibitory peptides and helical peptides at the N-terminus of the BCR / Abl tetramerization domain. PMI BCR / Abl-R6; this peptide can induce the degradation of BCR / Abl and upregulate p53 protein. In the development of small molecule degraders, MDM2-binding molecules successfully used in PROTAC design include Nutlin-3, RG7112, and RG7388. PROTACs designed based on the above MDM2-binding small molecules have been reported to induce MDM2 autodegradation (homo-PROTAC) or selective degradation of other target proteins. To date, substrate proteins successfully degraded by MDM2-dependent PROTAC molecules include androgen receptor AR, BRD4, PARP1, EGFR mutants, and BCL-XL. Among these, the DC of BRD4 and EGFR degraders... 50The degradation levels have reached submicromolar levels. Some literature has also designed a series of MDM2-dependent PROTAC molecules to explore the degradation activity of MDM2 on proteins such as CDK6, TrkC, BTK, and ERRα, but the degradation effects have not been ideal. Therefore, utilizing the E3 ubiquitin ligase activity of MDM2 to selectively degrade target proteins is feasible and has the dual function of simultaneously inducing target protein degradation and stabilizing p53.

[0006] Therefore, it is still necessary to develop novel PROTAC molecules based on MDM2 to achieve efficient degradation of target proteins. Summary of the Invention

[0007] The purpose of this invention is to provide a drug that can efficiently degrade CDK9 to treat or prevent tumors or immune diseases.

[0008] 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:

[0009]

[0010] in,

[0011] D is the MDM2E3 ubiquitin ligase-binding ligand.

[0012] L is -L 1 -[W 1 -L'-W 2 ] s -L 3 -;

[0013] L 1 Connected to the piperidine ring, and L 1 Selected from chemical bonds, -CO-, or -O-;

[0014] L 3 Selected from chemical bonds, -C(=O)-(CH2) a -、-(CH2) a NH-, -O- or

[0015] s can be 0, 1, 2, 3, 4, 5, or 6;

[0016] Each L' is independently selected from: chemical bond, substituted or unsubstituted C3-C8 carbon ring, substituted or unsubstituted 4-10 membered heterocyclic group, -NHCO-;

[0017] Each W 1 and W 2Each time it appears, it is independently selected from chemical bonds, -CO-, and -(CH2). a -,-Y-(CH2) b -,-(CH2) b -Y-, -Y-(CH2) a -Y-(CH2) b -,-(CH2) a -Y-(CH2CH2O) b -;

[0018] Each 'a' is independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 each time it appears;

[0019] Each b 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-.

[0021] In another preferred embodiment, D has a structure selected from the group consisting of:

[0022]

[0023] in,

[0024] R1 and R2 are each independently selected from F, Cl, Br, and I;

[0025] R3 is selected from C1-C6 alkyl groups;

[0026] R4 is selected from chemical bonds, H, substituted or unsubstituted C1-C6 alkyl groups; the substitution refers to having one or more substituents selected from the group consisting of: deuterium, halogen, amino, hydroxyl, oxo (=O);

[0027] R5 is selected from H or oxo.

[0028] In another preferred embodiment, R3 is selected from methyl, ethyl, propyl, and isopropyl.

[0029] In another preferred embodiment, D has a structure selected from the group consisting of:

[0030]

[0031] In another preferred embodiment, D has a structure selected from the group consisting of:

[0032]

[0033] It is the site connected to L.

[0034] In another preferred embodiment, D has a structure selected from the group consisting of:

[0035]

[0036] In another preferred embodiment, each L' is independently selected from: chemical bonds, -NHCO-.

[0037] In another preferred embodiment, s is 0, 1, 2 or 3.

[0038] In another preferred embodiment, L is selected from:

[0039]

[0040]

[0041] In this example, m, n, and q are each independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In another preferred example, m, n, and q are each independently selected from 2, 3, 4, 5, or 6.

[0042] In another preferred embodiment, L is selected from the following group:

[0043]

[0044]

[0045]

[0046] In another preferred embodiment, the compound is selected from the group consisting of:

[0047]

[0048]

[0049]

[0050]

[0051]

[0052]

[0053]

[0054]

[0055]

[0056]

[0057] 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.

[0058] 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.

[0059] In another preferred embodiment, the CDK9-mediated diseases or conditions include oncological diseases and immune diseases.

[0060] In another preferred embodiment, the tumor is selected from the group consisting of: various soft tissue sarcomas, primitive neuroectodermal tumors, pulmonary sarcomas, osteosarcomas, liposarcomas, Kaposi's sarcoma, midline carcinomas, breast cancers, lung cancers, intestinal cancers, liver cancers, pancreatic cancers, peritoneal cancers, thyroid tumors, parathyroid tumors, soft tissue hyperplasia, warts, Langerhans cell histiocytosis, hamartomas, gestational trophoblastic disease, skin cancers, Merkel cell carcinomas, bone tumors, melanomas, leukemias, salivary gland tumors, neuroendocrine tumors, lymphomas, multiple myelomas, brain tumors, and neurogenic tumors. The cancers include: mesothelioma, esophageal cancer, medulloblastoma, schwannoma, malignant peripheral nerve sheath tumor, pituitary adenoma, insulinoma, glioblastoma, meningioma, retinoblastoma, renal cancer, bladder cancer, gastric cancer, uterine cancer, vulvar cancer, prostate cancer, testicular cancer, penile cancer, bile duct cancer, gallbladder cancer, ureteral cancer, adrenocortical carcinoma, head and neck cancer, oral cancer, laryngeal cancer, nasopharyngeal carcinoma, ovarian cancer, cervical cancer, germ cell tumors, thymic tumors, myelodysplastic syndromes, and other histioproliferative diseases with MDM2 amplification or high expression. The immune diseases are selected from the following group: systemic lupus erythematosus, rheumatoid arthritis, inflammatory bowel disease, multiple sclerosis, and viral infections.

[0061] 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.

[0062] In another preferred embodiment, the object includes humans and non-human mammals.

[0063] 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

[0064] Figure 1The compound of the present invention shows that it has a good degradation effect on CDK9 protein in SJSA1 cells.

[0065] Figure 2 The compound of this invention shows that it has a good degradation effect on CDK9 protein in H226 cells.

[0066] Figure 3 The compound of this invention exhibits good inhibitory activity against SJSA1 cells.

[0067] Figure 4 The compound of this invention exhibits good inhibitory activity against H226 cells.

[0068] Figure 5 The compounds of this invention exhibit good inhibitory activity in a variety of tumor cells.

[0069] Figure 6 The compounds of this invention exhibit selective degradation activity for CDK9 and Cyclin T.

[0070] Figure 7 The results showed that tail vein injection of this type of compound had good inhibitory activity against mouse xenografts of SJSA1 cells. Detailed Implementation

[0071] Through extensive and in-depth research, the inventors have developed a novel class of CDK9 degraders. These compounds target the CDK9 kinase protein to be degraded at one end, and are linked to a ligand targeting MDM2 at the other end via a series of chain linkers, forming a complete PROTAC molecule. Compared to existing CDK9 inhibitors, the compounds of this invention exhibit MDM2-dependent CDK9 and Cyclin T degradation capabilities, and possess the ability to rapidly inhibit cell proliferation, showing potential as an effective treatment for tumors and immune diseases.

[0072] the term

[0073] In this invention, the term "halogen" refers to F, Cl, Br, or I.

[0074] 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.

[0075] 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.

[0076] 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.

[0077] "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.

[0078] In this invention, the term "haloalkyl" refers to a branched or straight-chain alkyl group substituted with one or more halogen atoms, up to the maximum permissible number of halogen atoms. Examples of haloalkyl groups include, but are not limited to, trifluoromethyl, monofluoromethyl, difluoromethyl, 2-fluoroethyl, and pentafluoroethyl.

[0079] 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.

[0080] 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.

[0081] 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.

[0082] 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.

[0083] 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.

[0084] 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.

[0085] 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.

[0086] Compounds and their preparation methods

[0087] The present invention provides a compound of formula (I) or a tautomer, optical isomer, hydrate, solvate or pharmaceutically acceptable salt thereof.

[0088]

[0089] L and D are defined as above.

[0090] In another preferred embodiment, in the compound, either L or D is independently a group corresponding to the specific compound of the present invention.

[0091] The main advantages of this invention include:

[0092] (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.

[0093] (2) The compounds of the present invention have significant inhibitory effects on a variety of cancer cells.

[0094] (3) The compounds of the present invention have significant inhibitory effects on a variety of tumor models.

[0095] 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.

[0096] Example

[0097] 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.

[0098] Example 1

[0099]

[0100] 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) 4M HCl in 1,4-Dioxane, DCM, rt; (g) HATU, DIPEA, DMF, rt; (h) TFA, DCM, rt; (i) HATU, DIPEA, DMF, rt.

[0101] Specific experimental steps:

[0102] 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%.

[0103] 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 the reaction solution was cooled 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%.

[0104] 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%.

[0105] 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%.

[0106] 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: 60%.

[0107] g,h: Compound 10 (190 mg, 0.5 mmol) and Compound 11 (115 mg, 0.5 mmol) were weighed and dissolved in 10 mL of DMF. DIPEA (0.44 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 × 20 mL), and the organic phase was 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain Compound 12, a white solid. Overall yield of the two steps: 55%.

[0108] i: Compound 12 (246 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 mL of DMF solution. DIPEA (0.44 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 × 20 mL), 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%. 1H NMR (600MHz, DMSO-d6) δ12.31(s,1H),10.41(s,1H),8.41(t,J=5.7Hz,1H),8.32(d,J=8.4Hz,1H),7.74(t,J=7.2Hz,1H),7.62-7.46( m,4H),7.43-7.30(m,4H),6.71(s,1H),4.59(d,J=7.4Hz,2H),4.39(dd,J=12.7,6.9Hz,2H),4.05(s,2H),3.93(s,5H),3.26(q,J=6.7 Hz,2H),3.03(t,J=11.6Hz,1H),2.77-2.69(m,1H),2.58(t,J=11.2Hz,1H),2.32(q,J=7.5Hz,2H),1.85-1.75(m,2H),1.65(dd,J=14. 3,9.9Hz,1H),1.54(p,J=7.5Hz,5H),1.35-1.30(m,2H),1.28-1.23(m,2H),1.17(s,9H),0.98(s,9H).Rt=8.0min.MS(ESI)m / z:[M+H] + :1091.36; Purity: ≥95%.

[0109] Example 2

[0110]

[0111] Synthesis of Example 2: (a) HATU, DIPEA, DMF, rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt.

[0112] Specific experimental steps:

[0113] a, b: Compound 10 (190 mg, 0.5 mmol) and compound 13 (120 mg, 0.5 mmol) were weighed and dissolved in 10 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 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 14, a white solid. Overall yield of the two steps: 55%.

[0114] c: Compound 14 (253 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), 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: 55%. 1 H NMR (600MHz, DMSO-d6) δ12.31(s,1H),10.41(s,1H),8.40(t,J=5.7Hz,1H),8.32(d,J=8.4Hz,1H),7.74(t,J=7.2Hz,1H),7.62- 7.46(m,4H),7.44-7.30(m,4H),6.71(s,1H),4.60(d,J=7.4Hz,2H),4.39(d,J=13.1Hz,2H),4.05(s,2H),3.93(s,5H),3.25(q, J=6.5Hz,2H),3.02(t,J=11.7Hz,1H),2.77-2.69(m,1H),2.63-2.54(m,1H),2.31(q,J=7.2Hz,2H),1.87-1.76(m,2H),1.65(dd ,J=14.2,9.8Hz,1H),1.52(dq,J=14.7,7.1Hz,5H),1.33-1.23(m,6H),1.17(s,9H),0.98(s,9H).Rt=8.2min.MS(ESI)m / z:[M+H] + :1105.37; Purity: ≥95%.

[0115] Example 3

[0116]

[0117] Synthesis of Example 3: (a) HATU, DIPEA, DMF, rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt.

[0118] Specific experimental steps:

[0119] a, b: Compound 10 (190 mg, 0.5 mmol) and compound 15 (130 mg, 0.5 mmol) were weighed and dissolved in 10 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 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 16, a white solid. Overall yield of the two steps: 65%.

[0120] c: Compound 16 (260 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), 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: 60%. 1 H NMR (600MHz, DMSO-d6) δ12.31(s,1H),10.41(s,1H),8.40(t,J=5.7Hz,1H),8.32(d,J=8.2Hz,1H),7.74(t,J=7.4Hz,1H),7.62-7 .45(m,4H),7.44-7.32(m,4H),6.71(s,1H),4.60(d,J=7.1Hz,2H),4.42-4.34(m,2H),4.05(s,2H),3.98-3.88(m,5H),3.25(q,J= 6.8Hz,2H),3.02(t,J=12.0Hz,1H),2.78-2.69(m,1H),2.63-2.55(m,1H),2.30(q,J=7.1Hz,2H),1.80(t,J=13.2Hz,2H),1.65(d d,J=14.2,9.9Hz,1H),1.51(dt,J=21.5,7.2Hz,5H),1.31-1.23(m,8H),1.17(s,9H),0.98(s,9H).Rt=8.4min.MS(ESI)m / z:[M+H] + :1119.39; Purity: ≥95%.

[0121] Example 4

[0122]

[0123] Synthesis of Example 4: (a) HATU, DIPEA, DMF, rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt.

[0124] Specific experimental steps:

[0125] a, b: Compound 10 (190 mg, 0.5 mmol) and Compound 17 (136 mg, 0.5 mmol) were weighed and dissolved in 10 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 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain Compound 18, a white solid. Overall yield of the two steps: 60%.

[0126] c: Compound 18 (267 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), 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: 65%. 1H NMR (500MHz, DMSO-d6) δ12.30(s,1H),10.41(s,1H),8.39(t,J=5.7Hz,1H),8.32(d,J=8.4Hz,1H),7.74(ddd,J=8.2,6.5,1.6Hz,1H),7.60-7. 47(m,4H),7.43-7.33(m,4H),6.71(s,1H),4.60(d,J=7.2Hz,2H),4.39 (d,J=12.8Hz,2H),4.05(s,2H),3.96(t,J=11.2Hz,2H),3.93(s,3H),3. 25(q,J=6.6Hz,2H),3.02(t,J=11.5Hz,1H),2.72(ddt,J=11.4,7.6,4.0Hz,1H),2.63-2.54(m,1H),2.29(q,J=7.5Hz,2H),1.85-1.76(m,2H), 1.65(dd,J=14.1,9.8Hz,1H),1.51(dt,J=20.9,7.1Hz,5H),1.28(d,J=14.6Hz,10H),1.17(s,9H),0.98(s,9H).Rt=8.6min.MS(ESI)m / z:[M+H] + :1133.40; Purity: ≥95%.

[0127] Example 5

[0128]

[0129] Synthesis of Example 5: (a) HATU, DIPEA, DMF, rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt.

[0130] Specific experimental steps:

[0131] a, b: Compound 10 (190 mg, 0.5 mmol) and compound 19 (144 mg, 0.5 mmol) were weighed and dissolved in 10 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 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 20, a white solid. Overall yield of the two steps: 70%.

[0132] c: Compound 20 (225 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), and the organic phase was 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: 60%. 1 H NMR (600MHz, DMSO-d6) δ12.31(s,1H),10.41(s,1H),8.40(t,J=5.7Hz,1H),8.32(d,J=8.2Hz,1H),7.74(t,J=7.2Hz,1H),7.60-7 .47(m,4H),7.43-7.33(m,4H),6.71(s,1H),4.59(d,J=7.1Hz,2H),4.39(dd,J=13.2,4.5Hz,2H),4.05(s,2H),3.98-3.89(m,5H), 3.25(q,J=6.7Hz,2H),3.02(t,J=11.6Hz,1H),2.77-2.69(m,1H),2.58(t,J=11.3Hz,1H),2.29(q,J=7.6Hz,2H),1.87-1.76(m,2H ),1.65(dd,J=14.3,9.7Hz,1H),1.55-1.45(m,5H),1.32-1.25(m,12H),1.17(s,9H),0.98(s,9H).Rt=8.8min.MS(ESI)m / z:[M+H] + :1147.42; Purity: ≥95%.

[0133] Example 6

[0134]

[0135] Synthesis of Example 6: (a) HATU, DIPEA, DMF, rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt.

[0136] Specific experimental steps:

[0137] a, b: Compound 10 (190 mg, 0.5 mmol) and compound 21 (150 mg, 0.5 mmol) were weighed and dissolved in 10 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 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 22, a white solid. Overall yield of the two steps: 65%.

[0138] c: Compound 22 (281 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), and the organic phase was 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: 65%. 1H NMR(600MHz,DMSO-d6)δ12.31(s,1H),10.41(s,1H),8.39(t,J=5.9Hz,1H),8 .32(d,J=8.4Hz,1H),7.74(t,J=7.4Hz,1H),7.61-7.52(m,3H),7.48(d,J=8. 4Hz,1H),7.42-7.33(m,4H),6.71(s,1H),4.60(d,J=7.6Hz,2H),4.43-4.35( m,2H),4.05(s,2H),3.96(t,J=11.0Hz,1H),3.92(s,3H),3.90(d,J=13.6Hz,1 H),3.25(q,J=6.4Hz,2H),3.02(t,J=12.8Hz,1H),2.77-2.69(m,1H),2.58(t ,J=12.5Hz,1H),2.29(q,J=7.6Hz,2H),1.81(t,J=14.0Hz,2H),1.65(dd,J=14 .4,9.8Hz,1H),1.55-1.49(m,3H),1.47(t,J=7.1Hz,2H),1.30-1.23(m,14H) ,1.17(d,J=2.1Hz,9H),0.98(d,J=1.9Hz,9H).Rt=9.0min.MS(ESI)m / z:[M+H] + :1161.44; Purity: ≥95%.

[0139] Example 7

[0140]

[0141] Synthesis of Example 7: (a) HATU, DIPEA, DMF, rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt.

[0142] Specific experimental steps:

[0143] a, b: Compound 10 (190 mg, 0.5 mmol) and compound 23 (157 mg, 0.5 mmol) were weighed and dissolved in 10 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 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 24, a white solid. Overall yield of the two steps: 70%.

[0144] c: Compound 24 (289 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), 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: 65%. 1 H NMR (600MHz, DMSO-d6) δ12.31(s,1H),10.41(s,1H),8.39(t,J=5.7Hz,1H),8.32(d,J=8.4Hz,1H),7.74(t,J=7.2Hz,1H),7.61-7.45 (m,4H),7.42-7.32(m,4H),6.71(s,1H),4.60(d,J=7.4Hz,2H),4.39(dd,J=12.8,7.6Hz,2H),4.05(s,2H),3.98-3.88(m,5H),3.25( q,J=6.7Hz,2H),3.02(t,J=11.7Hz,1H),2.77-2.69(m,1H),2.58(t,J=11.3Hz,1H),2.34-2.23(m,2H),1.86-1.76(m,2H),1.65(dd, J=14.3,9.7Hz,1H),1.50(dt,J=27.5,7.2Hz,5H),1.27(d,J=20.7Hz,16H),1.17(s,9H),0.98(s,9H).Rt=9.2min.MS(ESI)m / z:[M+H] + :1175.45; Purity: ≥95%.

[0145] Example 8

[0146]

[0147] Synthesis of Example 8: (a) HATU, DIPEA, DMF, rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt.

[0148] Specific experimental steps:

[0149] a, b: Compound 10 (190 mg, 0.5 mmol) and compound 25 (116 mg, 0.5 mmol) were weighed and dissolved in 10 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 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 26, a white solid. Overall yield of the two steps: 65%.

[0150] c: Compound 26 (247 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), 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: 60%. 1H NMR (600MHz, DMSO-d6) δ12.31(s,1H),10.42(s,1H),8.46(t,J=6.1Hz,1H),8.33(d,J=8.4Hz,1H),7.74(t,J=7.5Hz,1H),7.60-7.47(m,4H),7 .44-7.32(m,4H),6.71(s,1H),4.60(s,2H),4.38(d,J=6.8Hz,2H),4.0 5(s,2H),3.99-3.93(m,2H),3.93(s,3H),3.67(t,J=6.7Hz,2H),3.53( t,J=6.1Hz,2H),3.43(d,J=6.5Hz,2H),3.08-2.98(m,1H),2.72(t,J=11.3Hz,1H),2.61(dd,J=10.9,5.2Hz,2H),1.80(t,J=12.5Hz,2H),1.66 (d,J=9.8Hz,1H),1.57(t,J=12.4Hz,1H),1.41(t,J=12.5Hz,1H),1.31-1.22(m,2H),1.16(s,9H),0.97(s,9H).Rt=9.3min.MS(ESI)m / z:[M+H] + :1093.34; Purity: ≥95%.

[0151] Example 9

[0152]

[0153] Synthesis of Example 9: (a) HATU, DIPEA, DMF, rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt.

[0154] Specific experimental steps:

[0155] a, b: Compound 10 (190 mg, 0.5 mmol) and compound 27 (138 mg, 0.5 mmol) were weighed and dissolved in 10 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 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 28, a white solid. Overall yield of the two steps: 70%.

[0156] c: Compound 28 (270 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), 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%. 1 H NMR (600MHz, DMSO-d6) δ12.31(s,1H),10.42(s,1H),8.50(t,J=5.7Hz,1H),8.33(d,J=8.4Hz,1H),7.74(t,J=7.2Hz,1H),7.62-7.48(m,4 H),7.44-7.29(m,4H),6.71(s,1H),4.60(d,J=6.8Hz,2H),4.38(d,J=12.5Hz,2H),4.05(s,2H),3.99-3.88(m,5H),3.63(t,J=6.7Hz,2H) ,3.56-3.50(m,6H),3.42(d,J=5.7Hz,2H),3.01(t,J=12.0Hz,1H),2.75-2.69(m,1H),2.61-2.53(m,2H),1.80(t,J=11.4Hz,2H),1.65(d d,J=14.4,9.8Hz,1H),1.62-1.52(m,1H),1.45-1.36(m,1H),1.29-1.23(m,2H),1.17(s,9H),0.98(s,9H).Rt=9.6min.MS(ESI)m / z:[M+H] + :1037.36; Purity: ≥95%.

[0157] Example 10

[0158]

[0159] Synthesis of Example 10: (a) HATU, DIPEA, DMF, rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt.

[0160] Specific experimental steps:

[0161] a, b: Compound 10 (190 mg, 0.5 mmol) and compound 29 (160 mg, 0.5 mmol) were weighed and dissolved in 10 mL of DMF. DIPEA (0.4 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 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 30, a white solid. Overall yield of the two steps: 78%.

[0162] c: Compound 30 (291 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), 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 10, a white solid. Yield: 70%. 1 H NMR(600MHz,DMSO-d6)δ12.29(s,1H),10.41(s,1H),8.49(s,1H),8.32(s,1H),7.73(s,1H),7.60-7.47(m,4H),7.37(d ,J=23.1Hz,4H),6.71(s,1H),4.59(s,2H),4.38(s,2H),4.04(s,2H),3.95(s,2H),3.92(s,3H),3.60(s,2H),3.51(d,J= 16.9Hz,10H),3.41(s,2H),3.01(t,J=14.8Hz,1H),2.71(s,1H),2.58(s,2H),1.79(s,2H),1.65(d,J=16.0Hz,1H),1.55 (d,J=12.5Hz,1H),1.39(d,J=12.7Hz,1H),1.29-1.21(m,2H),1.17(s,9H),0.98(s,9H).Rt=9.9min.MS(ESI)m / z:[M+H] + :1181.39; Purity: ≥95%.

[0163] Example 11

[0164]

[0165] Synthesis of Example 11: (a) HATU, DIPEA, DMF, rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt.

[0166] Specific experimental steps:

[0167] a, b: Compound 10 (190 mg, 0.5 mmol) and compound 31 (182 mg, 0.5 mmol) were weighed and dissolved in 10 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 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 32, a white solid. Overall yield of the two steps: 65%.

[0168] c: Compound 32 (313 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), 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 11, a white solid. Yield: 55%. 1H NMR(600MHz,DMSO-d6)δ12.31(s,1H),10.42(s,1H),8.50(t,J=6.0Hz,1H) ,8.33(d,J=8.4Hz,1H),7.74(t,J=7.2Hz,1H),7.63-7.48(m,4H),7.44-7.3 1(m,4H),6.71(d,J=2.0Hz,1H),4.60(d,J=6.8Hz,2H),4.44-4.34(m,2H),4 .05(s,2H),3.99-3.93(m,2H),3.92(s,3H),3.61(t,J=6.8Hz,2H),3.53(d, J=5.7Hz, 6H), 3.49 (d, J=8.7Hz, 8H), 3.43 (t, J=6.0Hz, 2H), 3.02 (t, J=12. 4Hz,1H),2.77-2.68(m,1H),2.60(q,J=7.6Hz,2H),1.80(t,J=12.4Hz,2H), 1.65(dd,J=14.3,9.7Hz,1H),1.60-1.52(m,1H),1.40(p,J=12.8Hz,1H),1. 29-1.23(m,2H),1.17(s,9H),0.98(s,9H).Rt=10.1min.MS(ESI)m / z:[M+H] + :1225.42; Purity: ≥95%.

[0169] Example 12

[0170]

[0171] Synthesis of Example 12: (a) NaBH3CN, DCM / MeOH, rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt; (d) TFA, DCM, rt; (e) HATU, DIPEA, DMF, rt.

[0172] Specific experimental steps:

[0173] a, b: Compound 10 (380 mg, 1 mmol) and compound 33 (200 mg, 1 mmol) were weighed and dissolved in 10 mL of DCM / MeOH (volume ratio 1:1). The reaction system was stirred at room temperature for 30 min, and then NaBH3CN (60 mg, 2 mmol) was added. The reaction system was stirred at room temperature overnight. The reaction system was quenched with water, and the solvent was removed by vacuum distillation to obtain the crude product. The crude product was then purified by prep-HPLC to obtain a white solid. This crude product was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction system was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 34, a white solid. Overall yield of the two steps: 55%.

[0174] c, d: Compound 34 (231 mg, 0.5 mmol) and compound 25 (116 mg, 0.5 mmol) were weighed and dissolved in 10 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 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 35, a white solid. Overall yield of the two steps: 65%.

[0175] e: Compound 35 (289 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), and the organic phase was 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 12, a white solid. Yield: 65%. 1H NMR (500MHz, DMSO-d6) δ10.42(s,1H),8.46(t,J=5.6Hz,1H),8.36-8.24(m,2H),7.74(t,J=6.6Hz,1H),7.62-7.48(m,4H),7 .43-7.32(m,4H),6.71(s,1H),4.60(d,J=6.4Hz,2H),4.39(d,J=11.7Hz,2H),4.05(s,2H),3.93(s,5H),3.65(t,J=6.6Hz,2H ),3.52(t,J=6.0Hz,2H),3.43(t,J=5.7Hz,2H),3.31(p,J=6.7Hz,2H),2.94-2.79(m,4H),2.60-2.54(m,2H),2.48-2.42(m, 2H),2.14(t,J=11.0Hz,2H),1.77-1.55(m,6H),1.29-1.23(m,2H),1.17(s,9H),0.98(s,9H).Rt=8.8min.MS(ESI)m / z:[M+H] + :1176.41; Purity: ≥95%.

[0176] Example 13

[0177]

[0178] Synthesis of Example 13: (a) HATU, DIPEA, DMF, rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt.

[0179] Specific experimental steps:

[0180] a, b: Compound 34 (231 mg, 0.5 mmol) and compound 27 (138 mg, 0.5 mmol) were weighed and dissolved in 10 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 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 36, a white solid. Overall yield of the two steps: 55%.

[0181] c: Compound 36 (311 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), and the organic phase was 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 13, a white solid. Yield: 65%. 1 H NMR (500MHz, DMSO-d6) δ10.41(s,1H),8.51(t,J=5.7Hz,1H),8.34(s,2H),7.74(t,J=7.3Hz,1H),7.62-7.47(m,4H), 7.43-7.31(m,4H),6.71(s,1H),4.60(d,J=6.9Hz,2H),4.39(d,J=8.5Hz,2H),4.05(s,2H),3.92(s,5H),3.61(t,J=6 .7Hz,2H),3.55-3.49(m,6H),3.42(q,J=6.0Hz,2H),3.25-3.20(m,4H),2.71(t,J=7.2Hz,4H),2.48-2.43(m,2H),2. 14(t,J=11.6Hz,2H),1.77-1.55(m,6H),1.29-1.22(m,2H),1.17(s,9H),0.98(s,9H).Rt=9.2min.MS(ESI)m / z:[M+H] + :1220.44; Purity: ≥95%.

[0182] Example 14

[0183]

[0184] Synthesis of Example 14: (a) NaH, DMF, 0℃~rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt.

[0185] Specific experimental steps:

[0186] a, b: Nutlin-3 (580 mg, 1 mmol) was weighed and dissolved in 10 mL of DMF. NaH (36 mg, 1.5 mmol) was added under ice bath conditions, and the reaction mixture was stirred at 0 °C for 30 min. Then, compound 37 (193 mg, 1 mmol) was added, and the reaction mixture was stirred at room temperature for 1 h. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), 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 a white solid. This was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 38, a white solid. Overall yield of the two steps: 65%.

[0187] c: Compound 38 (320 mg, 0.5 mmol) and compound 26 (245 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), and the organic phase was 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 14, a white solid. Yield: 55%. Rt = 9.1 min. MS (ESI) m / z: [M+H] + :1116.36; Purity: ≥95%.

[0188] Example 15

[0189]

[0190] Synthesis of Example 15: (a) HATU, DIPEA, DMF, rt.

[0191] Specific experimental steps:

[0192] a: Compound 38 (319 mg, 0.5 mmol) and compound 28 (270 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), and the organic phase was 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 15, a white solid. Yield: 60%. Rt = 9.4 min. MS (ESI) m / z: [M+H] +:1160.38; Purity: ≥95%.

[0193] Example 16

[0194]

[0195] Synthesis of Example 16: (a) HATU, DIPEA, DMF, rt.

[0196] Specific experimental steps:

[0197] a: Compound 38 (319 mg, 0.5 mmol) and compound 30 (291 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), 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 16, a white solid. Yield: 55%. Rt = 9.7 min. MS (ESI) m / z: [M+H] + :1204.41; Purity: ≥95%.

[0198] Example 17

[0199]

[0200] Synthesis of Example 17: (a) HATU, DIPEA, DMF, rt; (b) TFA, DCM, rt; (c) K2CO 3, ACN,reflux;(d)TFA,DCM,rt;(e)HATU,DIPEA,DMF,rt.

[0201] Specific experimental steps:

[0202] a, b: Compound 10 (380 mg, 1 mmol) and compound 39 (115 mg, 0.5 mmol) were dissolved in 10 mL of DMF, and DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added. 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 × 20 mL), 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 a white solid. This crude product was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 40, a white solid. Overall yield of the two steps: 75%.

[0203] c, d: Compound 40 (245 mg, 0.5 mmol) and Compound 41 (267 mg, 1 mmol) were dissolved in 10 mL of acetonitrile, and potassium carbonate (138 mg, 1 mmol) was added. The reaction mixture was refluxed overnight. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by prep-HPLC to give a white solid. This crude product was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to give Compound 42, a white solid. Overall yield of the two steps: 60%.

[0204] e: Compound 42 (290 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), and the organic phase was 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: 50%. 1 H NMR(00MHz,DMSO-d6)δ12.32(s,1H),10.42(s,1H),8.52(t,J=5.7Hz,1H),8.33(d,J=8.4Hz,1H),7.74(t,J=6.5Hz,1H), 7.61-7.48(m,4H),7.43-7.32(m,4H),6.71(s,1H),4.60(d,J=5.6Hz,2H),4.38(d,J=11.7Hz,2H),4.05(s,2H),3.93(s, 5H),3.56(dt,J=20.4,5.6Hz,4H),3.43(q,J=5.6Hz,2H),3.01(t,J=10.7Hz,3H),2.77-2.54(m,5H),2.24(d,J=12.1Hz, 2H),1.88-1.77(m,2H),1.69-1.49(m,6H),1.30-1.22(m,2H),1.18(s,9H),0.98(s,9H).Rt=8.5min.MS(ESI)m / z:[M+H] + :1176.41; Purity: ≥95%.

[0205] Example 18

[0206]

[0207] Synthesis of Example 18: (a) K2CO 3, ACN,reflux;(b)TFA,DCM,rt;(c)HATU,DIPEA,DMF,rt.

[0208] Specific experimental steps:

[0209] a, b: Compound 40 (245 mg, 0.5 mmol) and Compound 43 (311 mg, 1 mmol) were dissolved in 10 mL of acetonitrile, and potassium carbonate (138 mg, 1 mmol) was added. The reaction mixture was refluxed overnight. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by prep-HPLC to give a white solid. This crude product was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to give Compound 44, a white solid. Overall yield of the two steps: 65%.

[0210] c: Compound 44 (311 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), and the organic phase was 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 18, a white solid. Yield: 45%. 1H NMR(500MHz,DMSO-d6)δ12.32(s,1H),10.41(s,1H),8.53(t,J=5.6Hz,1H),8.3 3(d,J=8.5Hz,1H),7.74(t,J=6.6Hz,1H),7.60-7.49(m,4H),7.42-7.32(m,4H) ,6.71(s,1H),4.60(d,J=6.4Hz,2H),4.39(d,J=11.4Hz,2H),4.05(s,2H),3.97 (d,J=11.4Hz,2H),3.93(s,3H),3.62(t,J=5.6Hz,2H),3.57-3.54(m,5H),3.43( q,J=6.1Hz,2H),3.18-3.11(m,2H),3.05(t,J=12.2Hz,1H),2.78(d,J=13.0Hz, 2H),2.76-2.68(m,2H),2.57(d,J=22.9Hz,2H),2.47(s,2H),1.88-1.78(m,2H), 1.65(dd,J=14.5,9.9Hz,4H),1.54(d,J=11.0Hz,1H),1.41(d,J=11.0Hz,1H),1 .25(d,J=13.3Hz,2H),1.17(s,9H),0.98(s,9H).Rt=8.8min.MS(ESI)m / z:[M+H] + :1220.44; Purity: ≥95%.

[0211] Example 19

[0212]

[0213] Synthesis of Example 19: (a) K2CO 3, DMF,rt;(b)TFA,DCM,rt;(c)HATU,DIPEA,DMF,rt;(d)TFA,DCM,rt;(e)HATU,DIPEA,DMF,rt.

[0214] Specific experimental steps:

[0215] a, b: Compound 10 (190 mg, 0.5 mmol) and compound 37 (117 mg, 0.6 mmol) were weighed and dissolved in 10 mL of DMF. Potassium carbonate (138 mg, 1 mmol) was added, and the reaction mixture was stirred at room temperature for 2 h. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 45, a white solid. Overall yield of the two steps: 80%.

[0216] c,d: Compound 45 (219 mg, 0.5 mmol) and compound 46 (115 mg, 0.5 mmol) were weighed and dissolved in 10 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 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 47, a white solid. Overall yield of the two steps: 65%.

[0217] e: Compound 47 (225 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 mL of DMF solution. DIPEA (0.43 mL, 2.5 mmol) and HATU (285 mg, 0.75 mmol) were added. 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 phase was 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 19, a white solid. Yield: 50%. 1HNMR(500MHz,DMSO-d6)δ12.33(s,1H),10.41(s,1H),8.54(d,J=5.0Hz,1H),8.33(d,J=8.4Hz,1H),7.75(t, J=6.6Hz,1H),7.63-7.47(m,4H),7.44-7.32(m,4H),6.72(s,1H),4.60(d,J=6.0Hz,2H),4.38(d,J=8.7Hz,1H ),4.06(s,2H),3.94(s,4H),3.60(p,J=6.7Hz,2H),3.55-3.41(m,6H),3.12(q,J=7.3Hz,2H),2.63(d,J=10. 8Hz,3H),1.93-1.71(m,4H),1.28(q,J=6.6Hz,8H),1.18(s,9H),0.98(s,9H).Rt=7.0min.MS(ESI)m / z:[M+H] + :1147.40; Purity: ≥95%.

[0218] Example 20

[0219]

[0220] Synthesis of Example 20: (a) K2CO 3, ACN,reflux; (b)TFA,DCM,rt; (c)K2CO 3, ACN,reflux;(d)TFA,DCM,rt;(e)HATU,DIPEA,DMF,rt.

[0221] Specific experimental steps:

[0222] a, b: Compound 10 (190 mg, 0.5 mmol) and compound 48 (277 mg, 1 mmol) were dissolved in 10 mL of acetonitrile, and potassium carbonate (138 mg, 1 mmol) was added. The reaction mixture was refluxed overnight. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by prep-HPLC to obtain a white solid. This crude product was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 49, a white solid. Overall yield of the two steps: 80%.

[0223] c,d: Compound 49 (238 mg, 0.5 mmol) and Compound 50 (291 mg, 1 mmol) were dissolved in 10 mL of acetonitrile, and potassium carbonate (138 mg, 1 mmol) was added. The reaction mixture was refluxed overnight. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by prep-HPLC to obtain a white solid. This crude product was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain Compound 51, a white solid. Overall yield of the two steps: 60%.

[0224] c: Compound 51 (294 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), and the organic phase was 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 20, a white solid. Yield: 55%. 1 H NMR (500MHz, DMSO-d6) δ12.26(s,1H),10.37(s,1H),8.31(d,J=8.2Hz,1H),8.17(s,1H),7.75(t,J=6.6Hz,1H),7.64-7.50(m,2H),7.4 3-7.32(m,4H),7.10(d,J=1.7Hz,1H),6.96(dd,J=8.2,1.7Hz,1H),6.72(s,1H),4.60(d,J=10.7Hz,2H),4.40(t,J=10.1Hz,1H),4.05( s,2H),3.94(t,J=11.3Hz,1H),3.89(s,3H),3.32(d,J=9.3Hz,2H),2.81(d,J=108.0Hz,8H),2.48(d,J=14.2Hz,1H),2.21(d,J=6.7Hz, 2H),2.03-1.95(m,2H),1.90-1.53(m,13H),1.34-1.23(m,4H),1.19(s,9H),1.11(s,2H),0.98(s,9H).Rt=8.0min.MS(ESI)m / z:[M+H] + :1186.47; Purity: ≥95%.

[0225] Example 21

[0226]

[0227] Synthesis of Example 21: (a) K2CO 3, ACN,reflux; (b)TFA,DCM,rt; (c)K2CO 3, ACN,reflux;(d)TFA,DCM,rt;(e)HATU,DIPEA,DMF,rt.

[0228] Specific experimental steps:

[0229] a, b: Compound 10 (190 mg, 0.5 mmol) and Compound 50 (291 mg, 1 mmol) were dissolved in 10 mL of acetonitrile with potassium carbonate (138 mg, 1 mmol) added. The reaction mixture was refluxed overnight. The reaction mixture was quenched with water and extracted three times with ethyl acetate (3 × 20 mL). The organic phase was 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane with trifluoroacetic acid (3 mL). The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain Compound 52, a white solid. Overall yield of the two steps: 75%.

[0230] c, d: Compound 52 (245 mg, 0.5 mmol) and Compound 53 (224 mg, 1 mmol) were dissolved in 10 mL of acetonitrile, and potassium carbonate (138 mg, 1 mmol) was added. The reaction mixture was refluxed overnight. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain Compound 54, a white solid. Overall yield of the two steps: 70%.

[0231] e: Compound 54 (267 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), and the organic phase was 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: 65%. 1H NMR (500MHz, DMSO-d6) δ12.30(s,1H),10.42(s,1H),8.55(s,1H),8.33(d,J=8.4Hz,1H),7.78-7.69(m,1H),7.62-7.44(m ,4H),7.43-7.30(m,4H),6.72(s,1H),4.60(d,J=6.1Hz,2H),4.39(s,1H),4.05(s,2H),3.93(s,4H),3.47(d,J=5.6Hz,2H) ,3.14-2.92(m,4H),2.69(s,2H),2.50-2.35(m,2H),2.25(t,J=10.6Hz,2H),2.14(t,J=10.8Hz,2H),1.82(d,J=15.1Hz,2 H),1.77-1.62(m,5H),1.43(d,J=5.8Hz,2H),1.38-1.23(m,5H),1.17(s,9H),0.98(s,9H).Rt=7.5min.MS(ESI)m / z:[M+H] + :1132.42; Purity: ≥95%.

[0232] Example 22

[0233]

[0234] Synthesis of Example 22: (a) K2CO 3, ACN,reflux;(b)TFA,DCM,rt;(c)HATU,DIPEA,DMF,rt.

[0235] Specific experimental steps:

[0236] a, b: Compound 10 (190 mg, 0.5 mmol) and Compound 41 (268 mg, 1 mmol) were dissolved in 10 mL of acetonitrile, and potassium carbonate (138 mg, 1 mmol) was added. The reaction mixture was refluxed overnight. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by prep-HPLC to obtain a white solid. This crude product was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain Compound 55, a white solid. Overall yield of the two steps: 80%.

[0237] c: Compound 55 (233 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), 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 22, a white solid. Yield: 55%. 1 H NMR (500MHz, DMSO-d6) δ12.25(s,1H),10.41(s,1H),8.51(t,J=5.8Hz,1H),8.33(d,J=8.4Hz,1H),7.74(t,J=7.4Hz ,1H),7.61-7.43(m,4H),7.45-7.31(m,4H),6.72(s,1H),4.59(d,J=6.7Hz,2H),4.42-4.30(m,1H),4.05(s,2H),3. 92(s,4H),3.56(dt,J=16.0,5.7Hz,4H),3.43(q,J=5.8Hz,2H),3.01(d,J=11.9Hz,2H),2.60(t,J=6.0Hz,2H),2.13 (t,J=11.3Hz,2H),1.83-1.54(m,5H),1.30-1.22(m,2H),1.17(s,9H),0.98(s,9H).Rt=8.5min.MS(ESI)m / z:[M+H] + :1065.34; Purity: ≥95%.

[0238] Example 23

[0239]

[0240] Synthesis of Example 23: (a) K2CO 3, ACN,reflux;(b)TFA,DCM,rt;(c)HATU,DIPEA,DMF,rt.

[0241] Specific experimental steps:

[0242] a, b: Compound 10 (190 mg, 0.5 mmol) and compound 43 (311 mg, 1 mmol) were dissolved in 10 mL of acetonitrile, and potassium carbonate (138 mg, 1 mmol) was added. The reaction mixture was refluxed overnight. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by prep-HPLC to give a white solid. This crude product was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to give compound 56, a white solid. Overall yield of the two steps: 75%.

[0243] c: Compound 56 (256 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), and the organic phase was 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 23, a white solid. Yield: 60%. 1 H NMR (600MHz, DMSO-d6) δ12.36(s,1H),10.41(s,1H),8.54(d,J=5.7Hz,1H),8.33(d,J=8.4Hz,1H),7.74(t,J=7.2H z,1H),7.62-7.47(m,4H),7.43-7.29(m,4H),6.72(s,1H),4.59(d,J=6.4Hz,2H),4.38(dd,J=12.6,7.4Hz,1H),4.0 6(s,2H),3.92(s,4H),3.68(s,2H),3.59-3.53(m,6H),3.43(q,J=6.0Hz,2H),2.98(s,2H),2.62(s,2H),1.96-1.74 (m,4H),1.65(dd,J=14.4,9.8Hz,1H),1.35-1.21(m,4H),1.17(s,9H),0.98(s,9H).Rt=8.8min.MS(ESI)m / z:[M+H] + :1109.37; Purity: ≥95%.

[0244] Example 24

[0245]

[0246] Synthesis of Example 24: (a) K2CO 3, ACN,reflux;(b)TFA,DCM,rt;(c)HATU,DIPEA,DMF,rt.

[0247] Specific experimental steps:

[0248] a, b: Compound 10 (190 mg, 0.5 mmol) and compound 57 (355 mg, 1 mmol) were dissolved in 10 mL of acetonitrile, and potassium carbonate (138 mg, 1 mmol) was added. The reaction mixture was refluxed overnight. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by prep-HPLC to give a white solid. This crude product was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to give compound 58, a white solid. Overall yield of the two steps: 55%.

[0249] c: Compound 58 (278 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), and the organic phase was 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 24, a white solid. Yield: 45%. 1H NMR (500MHz, DMSO-d6) δ12.17(s,1H),10.34(s,1H),8.42(t,J=5.7Hz,1H),8.25(d,J=8.4Hz,1H),7.70-7.64(m,1H) ,7.52-7.40(m,4H),7.35-7.26(m,4H),6.64(s,1H),4.53(s,2H),4.31(d,J=20.3Hz,1H),3.98(s,2H),3.85(s,4H),3 .48-3.40(m,12H),3.35(q,J=6.0Hz,2H),2.92(d,J=11.9Hz,2H),2.51(t,J=5.8Hz,2H),2.11-2.03(m,2H),1.75-1.6 7(m,2H),1.58(dd,J=14.0,10.4Hz,3H),1.22-1.15(m,2H),1.10(s,9H),0.91(s,9H).Rt=9.1min.MS(ESI)m / z:[M+H] + :1153.39; Purity: ≥95%.

[0250] Example 25

[0251]

[0252] Synthesis of Example 25: (a) K2CO 3, ACN,reflux;(b)TFA,DCM,rt;(c)HATU,DIPEA,DMF,rt.

[0253] Specific experimental steps:

[0254] a, b: Compound 34 (232 mg, 0.5 mmol) and compound 41 (268 mg, 1 mmol) were dissolved in 10 mL of acetonitrile, and potassium carbonate (138 mg, 1 mmol) was added. The reaction mixture was refluxed overnight. The reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by prep-HPLC to give a white solid. This crude product was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to give compound 59, a white solid. Overall yield of the two steps: 70%.

[0255] c: Compound 59 (275 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), and the organic phase was 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 25, a white solid. Yield: 50%. 1 H NMR (600MHz, DMSO-d6) δ12.45(s,1H),10.42(s,1H),8.61(s,1H),8.35(d,J=8.4Hz,1H),7.73(t,J=7.1Hz,1H), 7.62-7.50(m,4H),7.43-7.30(m,4H),6.72(s,1H),4.60(s,2H),4.39(d,J=12.5Hz,1H),4.07(s,2H),3.94(s,4 H),3.78(s,2H),3.61(t,J=5.7Hz,3H),3.52-3.44(m,4H),3.26(s,2H),2.96(s,2H),2.76(s,1H),2.28-1.83(m ,9H),1.64(dd,J=14.3,9.9Hz,1H),1.30-1.22(m,4H),1.18(s,9H),0.97(s,9H).Rt=8.8min.MS(ESI)m / z:[M+H] + :1148.42; Purity: ≥95%.

[0256] Example 26

[0257]

[0258] Synthesis of Example 26: (a) NaBH3CN, DCM / MeOH, rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt; (d) TFA, DCM, rt; (e) HATU, DIPEA, DMF, rt.

[0259] Specific experimental steps:

[0260] a, b: Compound 10 (380 mg, 1 mmol) and compound 60 (171 mg, 1 mmol) were weighed and dissolved in 10 mL of DCM / MeOH (volume ratio 1:1). The reaction system was stirred at room temperature for 30 min, and then NaBH3CN (125 mg, 2 mmol) was added. The reaction system was stirred at room temperature overnight. The reaction system was quenched with water, and the solvent was removed by vacuum distillation to obtain the crude product. The crude product was then purified by prep-HPLC to obtain a white solid. This crude product was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction system was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 61, a white solid. Overall yield of the two steps: 65%.

[0261] c, d: Compound 61 (218 mg, 0.5 mmol) and compound 25 (116 mg, 0.5 mmol) were weighed and dissolved in 10 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 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 62, a white solid. Overall yield of the two steps: 65%.

[0262] e: Weigh compound 62 (275 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) and dissolve them in 10 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 the reaction mixture with water, extract three times with ethyl acetate (3 × 20 mL), wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, and concentrate to obtain the crude product. The crude product is purified by prep-HPLC to obtain Example 26, a white solid. Yield: 55%. 1H NMR(500MHz,DMSO-d6)δ9.51(s,1H),8.74(s,1H),7.77(dd,J=8.5,2.1Hz,1H),7.74(s,1H),7.63(d,J=8.4Hz,1H),7.50(dd,J=6.8,4.5Hz,1H),7 .44-7.33(m,3H),7.32-7.25(m,3H),7.16(dd,J=12.1,2.2Hz,1H),7.01( s,1H),4.51-4.45(m,1H),4.26-4.19(m,2H),4.20(s,2H),4.06(dd,J=6. 0,5.2Hz,1H),3.89(s,2H),3.80(dd,J=9.5,4.2Hz,2H),3.68(t,J=6.6Hz ,2H),3.60-3.48(m,6H),3.24(p,J=4.1Hz,1H),2.84(ddd,J=12.3,8.7,6 .0Hz,2H),2.79-2.67(m,3H),2.58(t,J=6.6Hz,2H),2.03(ddt,J=13.2,8.8,5.9Hz,2H),1.17(s,9H),0.86(s,9H).Rt=8.5min.MS(ESI)m / z:[M+H] + :1148.38; Purity: ≥95%.

[0263] Example 27

[0264]

[0265] Synthesis of Example 27: (a) HATU, DIPEA, DMF, rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt.

[0266] Specific experimental steps:

[0267] a, b: Compound 61 (218 mg, 0.5 mmol) and compound 27 (134 mg, 0.5 mmol) were weighed and dissolved in 10 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 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 63, a white solid. Overall yield of the two steps: 60%.

[0268] c: Compound 63 (297 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), and the organic phase was 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: 50%. 1 H NMR (500MHz, DMSO-d6) δ12.32(s,1H),10.34(s,1H),8.46(t,J=5.6Hz,1H),8.25(dd,J=8.5,2. 4Hz,1H),7.67(t,J=7.4Hz,1H),7.52-7.42(m,4H),7.34-7.26(m,4H),6.65(s,1H),4.53(s,2H ),4.31(s,2H),3.99(s,2H),3.86(s,5H),3.60-3.33(m,14H),2.22(q,J=6.9Hz,2H),1.96-1.5 4(m,5H),1.23-1.16(m,5H),1.11(s,9H),0.91(d,J=1.5Hz,9H).Rt=8.9min.MS(ESI)m / z:[M+H] + :1192.41; Purity: ≥95%.

[0269] Example 28

[0270]

[0271] Synthesis of Example 28: (a) NaBH3CN, DCM / MeOH, rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt; (d) TFA, DCM, rt; (e) HATU, DIPEA, DMF, rt.

[0272] Specific experimental steps:

[0273] a, b: Compound 34 (463 mg, 1 mmol) and compound 33 (200 mg, 1 mmol) were weighed and dissolved in 10 mL of DCM / MeOH (volume ratio 1:1). The reaction system was stirred at room temperature for 30 min, and then NaBH3CN (125 mg, 2 mmol) was added. The reaction system was stirred at room temperature overnight. The reaction system was quenched with water, and the solvent was removed by vacuum distillation to obtain the crude product. The crude product was then purified by prep-HPLC to obtain a white solid. The crude product was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction system was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 64, a white solid. Overall yield of the two steps: 40%.

[0274] c,d: Compound 64 (273 mg, 0.5 mmol) and compound 25 (116 mg, 0.5 mmol) were weighed and dissolved in 10 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 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 65, a white solid. Overall yield of the two steps: 45%.

[0275] e: Compound 65 (330 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), and the organic phase was 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: 40%. 1H NMR (500MHz, DMSO-d6) δ9.51(s,1H),8.74(s,1H),7.77(dd,J=8.5,2.1Hz,1H),7.74(s,1H),7.63(d,J=8.4Hz,1H),7.50(dd,J=6.8,4. 5Hz,1H),7.44-7.33(m,2H),7.32-7.25(m,2H),7.16(dd,J=12.1,2.2Hz,1H),7.01(s,1H),4.51-4.45(m,1H),4.25-4.19(m,2H),4.20( s,2H),4.06(dd,J=6.0,5.2Hz,1H),3.89(s,2H),3.68(t,J=6.6Hz,2H),3.58(dd,J=4.5,3.8Hz,1H),3.55-3.46(m,3H),3.38(ddd,J=12 .1,8.2,5.7Hz,2H),2.95-2.82(m,3H),2.79-2.69(m,4H),2.66-2.54(m,2H),1.17(s,9H),0.86(s,9H).Rt=8.1min.MS(ESI)m / z:[M+H] + :1259.48; Purity: ≥95%.

[0276] Example 29

[0277]

[0278] Synthesis of Example 29: (a) HATU, DIPEA, DMF, rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt.

[0279] Specific experimental steps:

[0280] a, b: Compound 64 (273 mg, 0.5 mmol) and compound 66 (95 mg, 0.5 mmol) were weighed and dissolved in 10 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 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 67, a white solid. Overall yield of the two steps: 60%.

[0281] c: Compound 67 (308 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), 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: 45%. 1H NMR(500MHz,DMSO-d6)δ9.51(s,1H),8.74(s,1H),8.23(t,J=5.5Hz,1H),7.77(dd,J=8.4,2.0Hz,1H),7.74(s,1H),7.63(d,J=8.4Hz,1H),7.50 (dd,J=6.8,4.5Hz,1H),7.44-7.33(m,3H),7.32-7.25(m,2H),7.16(dd,J=12.1,2.2Hz,1H),7.01(s,1H),4.51-4.45(m,1H),4.26-4.19(m,2H) ,4.20(s,2H),4.06(dd,J=6.0,5.2Hz,1H),3.89(s,2H),3.55-3.42(m,4H),3.38(ddd,J=12.1,8.2,5.7Hz,2H),2.95-2.82(m,4H),2.79-2.69( m,5H),2.61(ddd,J=11.9,8.1,5.4Hz,2H),2.51(t,J=6.0Hz,2H),2.08-1.53(m,3H),1.17(s,9H),0.86(s,9H).Rt=8.2min.MS(ESI)m / z:[M+H] +:1215.46; Purity: ≥95%.

[0282] Example 30

[0283]

[0284] Synthesis of Example 30: (a) NaBH3CN, DCM / MeOH, rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt; (d) TFA, DCM, rt; (e) HATU, DIPEA, DMF, rt.

[0285] Specific experimental steps:

[0286] a, b: Compound 10 (380 mg, 1 mmol) and compound 68 (227 mg, 1 mmol) were weighed and dissolved in 10 mL of DCM / MeOH (volume ratio 1:1). The reaction system was stirred at room temperature for 30 min, and then NaBH3CN (125 mg, 2 mmol) was added. The reaction system was stirred at room temperature overnight. The reaction system was quenched with water, and the solvent was removed by vacuum distillation to obtain the crude product. The crude product was then purified by prep-HPLC to obtain a white solid. This crude product was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction system was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 69, a white solid. Overall yield of the two steps: 35%.

[0287] c, d: Compound 69 (245 mg, 0.5 mmol) and compound 25 (116 mg, 0.5 mmol) were weighed and dissolved in 10 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 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 70, a white solid. Overall yield of the two steps: 40%.

[0288] e: Weigh compound 70 (303 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) and dissolve them in 10 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 the reaction mixture with water and extract three times with ethyl acetate (3 × 20 mL). Wash the organic phase with saturated brine, dry to anhydrous sodium sulfate, and concentrate to obtain the crude product. Purify the crude product by prep-HPLC to obtain Example 30, a white solid. Yield: 45%. Rt = 8.5 min. MS (ESI) m / z: [M+H] + :1204.44; Purity: ≥95%.

[0289] Example 31

[0290]

[0291] Synthesis of Example 31: (a) HATU, DIPEA, DMF, rt; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF, rt.

[0292] Specific experimental steps:

[0293] a, b: Compound 69 (245 mg, 0.5 mmol) and compound 27 (134 mg, 0.5 mmol) were weighed and dissolved in 10 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 × 20 mL), 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 a white solid. It was then dissolved in 10 mL of anhydrous dichloromethane, and trifluoroacetic acid (3 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 71, a white solid. Overall yield of the two steps: 40%.

[0294] c: Compound 71 (325 mg, 0.5 mmol) and RG7388 (308 mg, 0.5 mmol) were weighed and dissolved in 10 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 reaction mixture was quenched with water, extracted three times with ethyl acetate (3 × 20 mL), and the organic phase was 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: 45%. Rt = 8.9 min. MS (ESI) m / z: [M+H] + :1248.47; Purity: ≥95%.

[0295] Example 32 Bioactivity Test

[0296] I. Cell Culture

[0297] NCI-H226 (also known as H226), MKN-45 (also known as MKN45), LNCaP clone FGC (also known as LNCAP), and HepG2 (also known as HEPG2) cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum (Novizan) and 1% penicillin / streptomycin. ACHN, HCT116, SJSA-1 (also known as SJSA1), CADO-ES1, U-2OS (also known as U2OS), U-87MG (also known as U87), MCF7, and TC-71 cells were cultured in DMEM high-glucose medium containing 10% fetal bovine serum and 1% penicillin / streptomycin. ASC52telo mesenchymal stem cells were cultured in DMEM / F12 medium containing 10% fetal bovine serum, 1 ng / mL bFGF, and 0.2 mg / mL G418.

[0298] All cells were cultured at 37°C in a 5% CO2 incubator and passaged by digestion with 0.05% trypsin solution containing EDTA. All cells were commercially available.

[0299] II. Half-cell activity inhibition rate (IC50) 50 )test

[0300] Day 1: Based on the different cell growth rates, lay 1.2 x 10⁻⁶ cells per layer. 3 Up to 4.5x10 3 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.

[0301] Day 2: Add 10 μL of culture medium containing different concentrations of the compound to each well to achieve final concentrations of 0, 1 nmol / L, 10 nmol / L, 100 nmol / L, 1 μmol / L, and 10 μmol / L. Two technical replicates were set up for each concentration in each experiment.

[0302] Day 5: Add 10 μL of thiazolyl blue (MTT) dye 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 Graphpad Prism to perform nonlinear fitting and calculate the IC50. 50 If cells maintain a survival rate of more than 50% under the highest concentration of the compound set in the experiment, it is considered an IC50 (inactivated cell death syndrome). 50 >10μmol / L.

[0303] III. Western blot (WB) detection of protein degradation effects

[0304] Step 1: Lay 5x10 5 Cells were placed in a 6cm dish.

[0305] Step 2: After 24 hours, add DMSO or a compound of varying concentrations to each culture dish; for example, at the half-maximal degradation concentration (DC). 50 In the related experiments, different concentrations of the test compound were added to each petri dish to achieve final concentrations of 0, 32 nmol / L, 125 nmol / L, 500 nmol / L and 2 μmol / L.

[0306] Step 3: After drug treatment for a certain period of time (usually 8 hours), 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.

[0307] 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 the lysis buffer at 12,000 rpm for 10 min using a 4-degree high-speed centrifuge, collect the supernatant sample and store temporarily on ice.

[0308] Step 5: Use the Bradford Protein Assay Kit (Beyotime) to determine the protein concentration of each sample.

[0309] Step 6: Add 1 / 3 volume of 4x protein loading buffer (containing β-mercaptoethanol) to each protein sample, mix well, and denature at 95°C for 5 minutes in a metal bath.

[0310] 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.

[0311] Step 8: Block the transferred PVDF membrane (0.45 μm pore size, Millipore) with a final concentration of 5% skim milk PBST (phosphate-buffered saline containing Tween-20), and then perform primary antibody incubation, PBST washing, secondary antibody incubation, and PBST washing step by step. Finally, protein development is performed using chemiluminescence on the Qinxiang gel imaging system. Antibody information is as follows:

[0312]

[0313]

[0314] Step 9: If protein quantification is required, use ImageJ for grayscale analysis, and use GraphPad Prism to perform nonlinear fitting on the degradation curves to calculate the DC of each compound on CDK9 protein in cells. 50 IV. Drug Treatment Experiment for Subcutaneous Xenografts in Immunodeficient Mice

[0315] Step 1: Prepare 8-week-old NCG immunodeficient mice (all purchased from commercial sources), shave their back fur and set them aside.

[0316] Step 2: Digest the in vitro cultured SJSA1 tumor cells with 0.05% trypsin, collect the cells and count them using a hemocytometer.

[0317] Step 3: Resuspend the cells in a PBS mixture containing 50% matrix gel (Lambolid) at a density of 7.5 million cells / mL, and inoculate the tumor cells subcutaneously on both sides of the back of immunodeficient mice at a dose of 750,000 cells per tumor.

[0318] Step 4: Three days after inoculation, the subcutaneous xenograft begins to expand. At this point, dilute the stock solution of the compound with physiological saline containing 5% Solutol HS-15 (Aladdin). Administer the compound via intraperitoneal or tail vein injection (dose 20 mg / kg) or by solvent treatment. The administration regimen is every other day for two weeks. Measure the mouse's body weight and tumor volume.

[0319] Step 5: At the end of the experiment, the mice were euthanized, and the transplanted tumor tissue was collected and weighed.

[0320] like Figure 1 As shown, this type of compound exhibits good degradation effect on CDK9 protein in SJSA1 cells.

[0321] like Figure 2 As shown, this type of compound exhibits good degradation effect on CDK9 protein in H226 cells.

[0322] like Figure 3 As shown, this type of compound exhibits good inhibitory activity against SJSA1 cells.

[0323] like Figure 4 As shown, this type of compound exhibits good inhibitory activity against H226 cells.

[0324] like Figure 5 As shown, this type of compound exhibits good inhibitory activity in various tumor cells.

[0325] like Figure 6 As shown, this type of compound selectively degrades CDK9 and Cyclin T after 8 hours of treatment with SJSA1 cells.

[0326] like Figure 7 As shown, tail vein injection of this type of compound has good inhibitory activity against mouse xenografts of SJSA1 cells.

[0327] As shown in Table 1, these compounds exhibited CDK9 degradation activity and cell viability inhibition activity in SJSA1 and H226 cells.

[0328] Table 1. Degradative activity of compounds against CDK9 protein and inhibitory activity against cell viability

[0329] compound <![CDATA[DC 50 in SJSA1]]> <![CDATA[IC 50 in SJSA1]]> <![CDATA[DC 50 in H226]]> <![CDATA[IC 50 in H226]]> dCDK9-1 B B C B dCDK9-2 C B C B dCDK9-3 C C C C dCDK9-4 C C C C dCDK9-5 C C C C dCDK9-6 C C C C dCDK9-7 C C C C dCDK9-8 A A A A dCDK9-9 A A B A dCDK9-10 A A A A dCDK9-11 C A C A dCDK9-12 A A A A dCDK9-13 A A A A dCDK9-14 C B C B dCDK9-15 C C C B dCDK9-16 C B C B dCDK9-17 A A B A dCDK9-18 A A A A dCDK9-19 C A C A dCDK9-20 A A C A dCDK9-21 A A C A dCDK9-22 A A A A dCDK9-23 A A A A dCDK9-24 A A A A dCDK9-25 A A A A dCDK9-26 A A A A dCDK9-27 A B A A dCDK9-28 A B A A dCDK9-29 A A A A

[0330] Note:

[0331] DC 50 "A" indicates less than 0.5 μmol / L; "B" indicates between 0.5 μmol / L and 2 μmol / L; "C" indicates greater than 2 μmol / L.

[0332] IC 50 “A” indicates less than 1 μmol / L; “B” indicates between 1 μmol / L and 10 μmol / L; “C” indicates greater than 10 μmol / L.

[0333] As shown in Table 2, these compounds exhibit excellent pharmacokinetic parameters in mice.

[0334] Table 2. Pharmacokinetic (PK) studies of compound 10 in mice

[0335]

[0336] Example 33 Animal Pharmacokinetic (PK) Study

[0337] 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.

[0338] Sample collection and analysis:

[0339] Blood volume collected: 0.03 mL

[0340] Blood collection method: orbital venous plexus

[0341] Anticoagulant: EDTA-K2

[0342] Blood collection time:

[0343] IV: 5min, 15min, 30min, 1h, 2h, 4h, 7h, 24h

[0344] PO: 15min, 30min, 1h, 2h, 4h, 7h, 24h

[0345] 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℃.

[0346] Sample pretreatment was performed according to the Standard PK procedure.

[0347] 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.

[0348] 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, D is the MDM2 E3 ubiquitin ligase-binding ligand. L is -L 1 -[W 1 -L'-W 2 ] s -L 3 -; L 1 Connected to the piperidine ring, and L 1 Selected from chemical bonds, -CO-, or -O-; L 3 Selected from chemical bonds, -C(=O)-(CH2) a -、-(CH2) a 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). a -,-Y-(CH2) b -,-(CH2) b -Y-, -Y-(CH2) a -Y-(CH2) b -,-(CH2) a -Y-(CH2CH2O) b -; Each 'a' is independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 each time it appears; Each b is independently selected from 0, 1, 2, 3, 4, 5, or 6 in each occurrence; Each Y is independently selected from -O- and -S-.

2. The compound of claim 1, wherein, D has a structure selected from the following group: in, R1 and R2 are each independently selected from F, Cl, Br, and I; R3 is selected from C1-C6 alkyl groups; R4 is selected from chemical bonds, H, substituted or unsubstituted C1-C6 alkyl groups; the substitution refers to having one or more substituents selected from the group consisting of: deuterium, halogen, amino, hydroxyl, oxo (=O); R5 is selected from H or oxo.

3. The compound of claim 1, wherein, D has a structure selected from the following group:

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

5. The compound of claim 1, wherein, s can be 0, 1, 2 or 3.

6. The compound of claim 1, wherein, L is selected from: Among them, m, n, and q are each independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

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. Preferably, the CDK9-mediated diseases or conditions include oncological diseases and immune diseases.

10. The use as described in claim 9, characterized in that, The tumors were selected from the following group: various soft tissue sarcomas, primitive neuroectodermal tumors, pulmonary sarcomas, osteosarcomas, 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, multiple myeloma, brain tumors, neuroblastoma, mesothelioma, esophageal cancer, medulloblastoma, schwannoma, and malignant ectodermal tumors. Peripheral nerve sheath tumor, pituitary adenoma, insulinoma, glioblastoma, meningioma, retinoblastoma, kidney cancer, bladder cancer, gastric 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, laryngeal cancer, nasopharyngeal cancer, ovarian cancer, cervical cancer, germ cell tumors, thymic tumors, myelodysplastic syndrome, and other histioproliferative diseases with MDM2 amplification or high expression; the immune diseases are selected from the following group: systemic lupus erythematosus, rheumatoid arthritis, inflammatory bowel disease, multiple sclerosis, and viral infections.

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