Multifunctional protein degrader and application thereof

Abstract

Disclosed are a compound having histone demethylase, deubiquitinating enzyme and related signaling pathway (such as cMyC and RNF6) degradation activity, and a use thereof. The compound is capable of efficiently downregulating levels of histone demethylases (such as KDM5, LSD), deubiquitinating enzymes (such as USP25, USP28), cMyC, TNF6, and the like, in cells, and can be used in the preparation of a drug for treating and / or preventing related diseases or conditions mediated by histone demethylases and deubiquitinating enzymes.

Description

Multifunctional protein degraders and uses thereof TECHNICAL FIELD

[0001] The present application belongs to the field of medicine, and specifically relates to compounds for degrading histone demethylase, deubiquitinase, c-MYC and / or RNF6, pharmaceutical compositions and uses thereof. BACKGROUND

[0002] Histone demethylases play an important role in various types of tumors, and abnormal expression of histone demethylases is associated with the occurrence of various tumors. KDM5A, also known as JARID1A, is a histone demethylase that mainly regulates gene expression through demethylation. For example, studies have found that the expression level of histone demethylase (such as KDM5A) in various cancer types (such as breast cancer, prostate cancer, melanoma, etc.) is significantly up-regulated, and is related to the invasiveness and metastasis of tumors. Due to the importance of histone demethylases in tumors, researchers are exploring therapeutic strategies targeting histone demethylases. Inhibiting the activity of histone demethylases can provide a new treatment approach for certain cancers. Histone demethylases also interact with various cell signaling pathways, such as cMYC, Wnt, Notch, etc., which play a driving role in the process of tumor occurrence and development. c-Myc is a key transcription factor that can promote the proliferation and metabolism of tumor cells. Studies have found that in some cases, histone demethylases can achieve transcriptional regulation of their target genes by affecting the level of c-Myc. In many types of cancer, the co-activation of KDM5A and c-Myc is positively correlated with the malignancy, invasiveness and poor prognosis of tumors. Overexpression of histone demethylases can enhance the transcriptional activity of c-Myc, thereby promoting the proliferation and survival of tumor cells. By regulating the activity of these signaling pathways, histone demethylases can promote the proliferation and metastasis of tumor cells. Since the interaction between histone demethylases and c-MYC plays an important role in tumors, co-intervention targeting both proteins can be a potential therapeutic strategy. For example, developing small molecules that can simultaneously inhibit histone demethylases and c-MYC will be more effective in inhibiting the growth of tumor cells.

[0003] Currently, there is no anti-tumor drug targeting KDM5A_c-MYC, although histone demethylase inhibitors have shown potential application value in anti-tumor research, the development of selective inhibition of histone demethylase without affecting other related enzymes is still a challenge. c-MYC is considered an "undruggable" target because of its highly complex structural and functional properties, making it a significant challenge to directly target its activity. However, despite this, researchers continue to explore effective strategies and methods to inhibit the function of c-MYC. While direct inhibition of c-MYC strategies face difficulties, some small molecules can indirectly affect its activity, for example: small molecules that inhibit the binding of c-MYC to its target DNA: Some researchers have found small molecule compounds that can interfere with the binding of c-MYC to DNA, thereby reducing its transcriptional activity. Small molecules that interfere with MYC transcription and translation: Some new small molecules can affect the transcription or translation process of c-MYC, inhibiting its function through these pathways.

[0004] PROTAC (PROTeolysis TArgeting Chimeras) is a targeted protein degradation technology that uses small molecule compounds to regulate protein levels. It works by recruiting a specific E3 ligase to a specific target protein, causing the target protein to be polyubiquitinated, so that it is recognized by the proteasome and degraded into short peptides and amino acids. Since deubiquitinating enzymes hydrolyze ubiquitin on the target protein, the pathway of degrading the target protein by ubiquitinating the target protein is inhibited, so it is necessary to reduce the influence of deubiquitinating enzymes in the process of protein degradation.

[0005] Therefore, there is an urgent need in the art to develop a highly active and highly selective drug for down-regulating diseases or disorders mediated by histone demethylase and deubiquitinating enzyme. SUMMARY

[0006] The purpose of the present application is to provide a drug capable of selectively degrading histone demethylase, deubiquitinating enzyme, c-MYC and / or RNF6 to treat or prevent tumors.

[0007] In a first aspect of the present application, a compound represented by general formula (I) or a stereoisomer, enantiomer, atropisomer, optical isomer, racemate, tautomer thereof, or a pharmaceutically acceptable salt thereof, a hydrate thereof, an isotopically labeled compound thereof is provided:

[0008] S-L-D (I)

[0009] wherein,

[0010] S is a target protein binding ligand having the structure shown below:

[0011] Ring A is selected from: substituted or unsubstituted C3-C8cycloalkyl, substituted or unsubstituted 4-10 membered heterocyclyl, substituted or unsubstituted C6-C10aryl, substituted or unsubstituted 5-10 membered heteroaryl;

[0012] R is selected from: a bond, null, halogen, Ci-C6alkyl, Ci-C6haloalkyl, Ci-C6alkoxy, Ci-C6alkylamino, C3-C8cycloalkyl, 4-10 membered heterocyclyl, C6-C10aryl, 5-10 membered heteroaryl;

[0013] Z is selected from: a bond, CO;

[0014] R m is selected from: a bond, H, halogen, Ci-C6alkyl, Ci-C6haloalkyl, Ci-C6alkoxy, Ci-C6alkylamino, substituted or unsubstituted C3-C8cycloalkyl, substituted or unsubstituted 4-10 membered heterocyclyl, substituted or unsubstituted C6-C10aryl, substituted or unsubstituted 5-10 membered heteroaryl;

[0015] L is -L1-(L2) p -L3- which connects S and D by a covalent bond;

[0016] L1, L3are each independently selected from the group consisting of a bond, null, CO, -O-, -CONH-, substituted or unsubstituted CH2, substituted or unsubstituted NH, S(O), S(O)2, ethynyl, -CH2CH2O-, substituted or unsubstituted C3-C8cycloalkyl, substituted or unsubstituted 4-10 membered heterocyclyl, substituted or unsubstituted C6-C10aryl, substituted or unsubstituted 5-10 membered heteroaryl;

[0017] each L2is independently selected from the group consisting of a bond, null, CO, -O-, -CONH-, substituted or unsubstituted CH2, substituted or unsubstituted NH, S(O), S(O)2, ethynyl, -CH2CH2O-, substituted or unsubstituted C3-C8cycloalkyl, substituted or unsubstituted 4-10 membered heterocyclyl, substituted or unsubstituted C6-C10aryl, substituted or unsubstituted 5-10 membered heteroaryl;

[0018] said substitution means having one or more substituents selected from the group consisting of halogen, amino, hydroxyl, cyano, Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, Ci-C6haloalkyl, Ci-C6alkoxy, Ci-C6alkylamino, C3-C8cycloalkyl, C6-C10aryl, 4-10 membered heterocyclyl, 5-10 membered heteroaryl;

[0019] p is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20;

[0020] D is an E3 ubiquitin ligase binding ligand.

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

[0022] Z 1 is CH or N; R1is selected from halogen, amino, hydroxyl, cyano, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C1-C6alkylamino, C3-C8cycloalkyl, C6-C10aryl, 4-10 membered heterocyclyl, 5-10 membered heteroaryl; wherein, denotes the site of covalent attachment to L.

[0023] In another preferred embodiment, the heterocyclyl group has 1, 2, or 3 heteroatoms selected from N, O, S.

[0024] In another preferred embodiment, ring A is selected from 5-10 membered nitrogen-containing heterocyclyl.

[0025] In another preferred embodiment, the compound has a structure according to Formula (II):

[0026] wherein L, D, Z, R m , A are as defined above.

[0027] In another preferred embodiment, D has a structure selected from the group consisting of:

[0028] In another preferred embodiment, the compound has a structure according to Formula (III):

[0029] wherein wherein L, Z, R m , A are as defined above.

[0030] In another preferred embodiment, the compound has a structure according to Formula (IV):

[0031] wherein wherein L, Z, R m are as defined above.

[0032] In another preferred embodiment, the compound has a structure according to Formula (V):

[0033] In another preferred embodiment, each L1, L3is independently selected from the group consisting of a covalent bond, CO, -0-, CH2, NH, -CH2CH2O-,

[0034] In another preferred embodiment, each L2is independently selected from the group consisting of CH2, NH, -CH2CH2O-,

[0035] p is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 or 12.

[0036] In another preferred embodiment, each L1, L3is independently selected from the group consisting of a covalent bond, absent, CH2, CO, NH, -CONH-, ethynyl.

[0037] In another preferred embodiment, L1is CO, -CONH-.

[0038] In another preferred embodiment, L3is selected from the group consisting of a covalent bond, absent, CO, NH, CH2, ethynyl.

[0039] In another preferred embodiment, L3is selected from the group consisting of CO, NH, CH2.

[0040] In another preferred embodiment, L1is connected to the S moiety by a covalent bond.

[0041] In another preferred embodiment, L is selected from the group consisting of:

[0042] m, n, q are each independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0043] In another preferred embodiment, n is selected from 2, 3, 4, 5 or 6.

[0044] In another preferred embodiment, L is selected from:

[0045] In another preferred embodiment, R m is selected from the group consisting of H, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 alkylamino, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-10 membered heterocyclyl.

[0046] In another preferred embodiment, R m is selected from the group consisting of H, methyl, ethyl, n-propyl, i-propyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, t-butoxy, pyridyl, tetrahydropyrrole, piperidinyl, piperazinyl, tetrahydropyranyl, phenyl.

[0047] In another preferred embodiment, the compound has a structure selected from the group consisting of:

[0048] A second aspect of the present application provides a pharmaceutical composition comprising a compound of the first aspect of the present application, or a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof, and a pharmaceutically acceptable carrier.

[0049] A third aspect of the present application provides use of a compound of the first aspect of the present application, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating a histone demethylase, deubiquitinase, c-MYC and / or RNF6 mediated related disease or disorder.

[0050] In another preferred embodiment, the histone demethylase comprises KDM5A, KDM5B, KDM5C, KDM5D, LSD1, LSD2, KDM4A, KDM4B.

[0051] In another preferred embodiment, the deubiquitinase comprises USP25, USP28.

[0052] In another preferred embodiment, the histone demethylase, deubiquitinase, c-MYC and / or RNF6 mediated related disease or disorder comprises a tumor disease.

[0053] A fourth aspect of the present application provides a method of treating a tumor disease, comprising the step of administering to a subject in need thereof a compound of formula (I) of the first aspect of the present application, or a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof.

[0054] In another preferred embodiment, the subject comprises a human and a non-human mammal.

[0055] In another preferred embodiment, the tumor is selected from the group consisting of breast cancer, leukemia, lymphoma, lung cancer, colon cancer, liver cancer, ovarian cancer, pancreatic cancer, large intestine cancer, thyroid cancer, skin cancer, bone cancer, melanoma, salivary gland tumor, neuroendocrine tumor, brain tumor, neuroblastoma, mesothelioma, esophageal cancer, lung sarcoma, medulloblastoma, glioblastoma, retinoblastoma, kidney cancer, bladder cancer, osteosarcoma, stomach cancer, uterine cancer, vulvar cancer, small intestine cancer, prostate cancer, bile duct cancer, ureter cancer, adrenal cortex cancer, head and neck cancer, colorectal cancer, nasopharyngeal cancer, myeloma, ovarian cancer, cervical cancer.

[0056] It should be understood that, within the scope of the present application, all combinations of the aforementioned technical features of the present application and the technical features specifically described below (e.g., in the examples) can be interchanged among the various embodiments of the present application, thus forming new or preferred technical solutions. Due to the limited space, they will not be listed one by one here. BRIEF DESCRIPTION OF DRAWINGS

[0057] Figure 1 shows the degradation of the compounds LD-324-LLH, LD-110-ZJZ, LD-325-LLH, LD-327-LLH, LD-326-LLH, LD-328-LLH of the present application on LSD1 and c-MYC protein in solid tumor breast cancer cells at different doses.

[0058] Figure 2 shows the degradation of the compounds LD-110-ZJZ, LD-324-LLH, LD-325-LLH, LD-326-LLH, LD-327-LLH, LD-328-LLH, LD-329-LLH, LD-330-LLH, LD-331-LLH, LD-332-LLH, LD-333-LLH of the present application and the positive compound JQKD82 on histone demethylase and c-MYC protein in hematological tumor cells at different doses.

[0059] Figure 3 shows the degradation of the compounds LD-110-ZJZ, LD-328-LLH, LD-331-LLH of the present application on histone demethylase and c-MYC protein in hematological tumor cells at different time points.

[0060] Figure 4 shows the growth inhibition activity of the compounds LD-110-ZJZ, LD-324-LLH, LD-325-LLH, LD-327-LLH, LD-328-LLH, LD-331-LLH of the present application and the positive compound JQKD82 on tumor cells.

[0061] Figure 5 shows the degradation activity of the compound LD-331-LLH of the present application on histone demethylase and deubiquitinase in hematological tumor cells at different time points and different doses.

[0062] Figure 6 shows the degradation activity of the compounds LD-331-LLH of the present application on different doses of pathogenic proteins in solid tumor lung cancer, pancreatic cancer, breast cancer, colorectal cancer, etc. cells and the killing effect on cells.

[0063] Figure 7 shows the degradation of LSD1 protein in lung cancer cell lines by the compounds LD-110-ZJZ, LD-137-ZJZ, LD-138-ZJZ, LD-139-ZJZ, LD-321-LLH, and LD-322-LLH of the present application.

[0064] Figure 8 shows the degradation of LSD1 protein in lung cancer cell lines by the compounds LD-110-ZJZ, LD-151-ZJZ, LD-152-ZJZ, LD-153-ZJZ, LD-154-ZJZ, and LD-155-ZJZ of the present application.

[0065] Figure 9 shows the degradation of LSD1 protein in lung cancer cell lines by different doses of the compounds of the present application.

[0066] Figure 10 shows the inhibitory activity of the compounds of the present application on lung cancer cell line H520.

[0067] Figure 11 shows the inhibitory activity of the compounds of the present application on lung cancer cell line A549.

[0068] Figure 12 shows the degradation of LSD1 protein in breast cancer cell lines by the compounds of the present application.

[0069] Figure 13 shows the inhibitory activity of the compounds of the present application on breast cancer cells.

[0070] Figure 14 shows the degradation of LSD1 protein in esophageal cancer cells and the inhibitory activity of the compounds of the present application on esophageal cancer cells.

[0071] Figure 15 shows the mechanism of the compounds of the present application degrading LSD1.

[0072] Figures 16 and 17 show the tumor growth inhibition curves of the compounds of the present application in mice.

[0073] Figure 18 shows that the compounds LD331, LD331-14, and LD331-18 of the present application degrade KDM5A and down-regulate c-MYC and RNF6 in various hematological malignancy cell lines.

[0074] Figure 19 shows that the compounds LD331, LD331-14, and LD331-18 of the present application significantly reduce the expression of KDM5A and c-MYC and RNF6 in acute myeloid leukemia (AML) cell line MV4;11.

[0075] Figure 20 shows that the compounds LD331, LD331-14 and LD331-23 of the present application significantly reduce the expression of KDM5A as well as c-MYC and RNF6 in various solid tumor cell lines. DETAILED DESCRIPTION

[0076] The inventors have, through extensive and in-depth research, for the first time developed a class of compounds with a completely new structure, which can efficiently and selectively inhibit the expression of histone demethylases (such as KDM5, LSD) and deubiquitinases (such as USP25, USP28) as well as c-MYC and RNF6, thereby significantly inhibiting the growth of tumor cells, and having the potential to treat or prevent tumors. On this basis, the present application is completed.

[0077] TERMS

[0078] In the present application, the term "halogen" refers to F, Cl, Br or I.

[0079] In the present application, "C1-C6 alkyl" refers to a straight-chain or branched alkyl group including 1-6 (1, 2, 3, 4, 5 or 6) carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, neopentyl or the like.

[0080] In the present application, the term "C2-C6 alkenyl" refers to a straight-chain or branched alkenyl group having 2-6 carbon atoms containing one double bond, including, without limitation, ethenyl, propenyl, butenyl, isobutenyl, pentenyl and hexenyl, etc.

[0081] In the present application, the term "C2-C6 alkynyl" refers to a straight-chain or branched alkynyl group having 2-6 carbon atoms containing one triple bond, including, without limitation, ethynyl, propynyl, butynyl, isobutynyl, pentynyl and hexynyl, etc.

[0082] In the present application, 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 in the ring, including, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc. The term "C 3-6 Cycloalkyl" has a similar meaning.

[0083] In the present application, the term "C1-C6 alkoxy" refers to a straight-chain or branched alkoxy group having 1-6 carbon atoms, including, without limitation, methoxy, ethoxy, propoxy, isopropoxy and butoxy, etc. Preferably, C1-C4 alkoxy.

[0084] 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.

[0085] In this invention, the term "alkylamino" refers to a secondary or tertiary alkylamine group, wherein the alkyl group is independently selected from alkyl groups as defined herein. The linkage site of the alkylamino group is on a nitrogen atom. Examples of alkylamino groups include ethylamine, dimethylamine, and methylpropylamino, etc.

[0086] In this invention, the term "heterocyclic group" 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 optionally be 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; heterocyclic groups include 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.

[0087] In this invention, the terms "aromatic ring" or "aryl" have the same meaning, and are preferably "C". 6- C 10 Aryl. The term "C" 6- C 10 "Aryl" refers to an aromatic cyclic group with 6-10 carbon atoms that does not contain heteroatoms on the ring, such as phenyl and naphthyl.

[0088] In this invention, the terms "aromatic heterocycle" or "heteroaryl" have the same meaning, referring to a heteroaromatic group containing one or more heteroatoms. For example, "5-10-membered heteroaryl" refers to an aromatic heterocycle containing 1-3 heteroatoms selected from oxygen, sulfur, and nitrogen, and 2-7 carbon atoms. Non-limiting examples include: furanyl, thiophene, pyridinyl, pyrazolyl, pyrroleyl, N-alkylpyrroleyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, etc. The heteroaryl ring may be fused to an aryl, heterocyclic, or cycloalkyl ring, wherein the ring connected to the parent structure is the heteroaryl ring. The heteroaryl group may be optionally substituted or unsubstituted.

[0089] 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.

[0090] 0 sulfonyl group, etc.

[0091] 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.

[0092] 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.

[0093] 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.

[0094] 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.

[0095] 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.

[0096] Pharmaceutical Compositions and Administration

[0097] The pharmaceutical compositions of the present invention comprise, within a safe and effective range, the compound of the present invention or a pharmacologically acceptable salt thereof, and a pharmacologically acceptable excipient or carrier. "Safe and effective range" refers to an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, more preferably, 5-1000 mg of the compound of the present invention per dose. Preferably, "one dose" is one capsule or tablet.

[0098] "Pharmaceutically acceptable carriers" refers to one or more compatible solid or liquid fillers or gelling substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with and with the compounds of the present invention without significantly reducing the efficacy of the compounds. Examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as... Wetting agents (such as sodium dodecyl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0099] The pharmaceutical composition is an injection, capsule, tablet, pill, powder, or granule.

[0100] There are no particular limitations on the administration of the compounds or pharmaceutical compositions of the present invention. Representative administration methods include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and local administration.

[0101] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following components: (a) fillers or compatibilizers, such as starch, lactose, sucrose, glucose, mannitol, and silica; (b) binders, such as hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; (c) humectants, such as glycerin; (d) disintegrants, such as agar, calcium carbonate, potato starch or cassava starch, alginate, certain complex silicates, and sodium carbonate; (e) slowing agents, such as paraffin wax; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or mixtures thereof. Buffers may also be included in capsules, tablets, and pills.

[0102] Solid dosage forms such as tablets, sugar pills, capsules, pellets, and granules can be prepared using coatings and shells, such as casings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compound from such compositions can be delayed in a portion of the digestive tract. Examples of encapsulating components that can be used are polymeric substances and waxes. If necessary, the active compound may also be formed into microcapsules with one or more of the excipients described above.

[0103] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

[0104] In addition to these inert diluents, the composition may also contain auxiliaries such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and fragrances.

[0105] In addition to the active compound, the suspension may contain suspending agents such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0106] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0107] Dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays, and inhalers. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be necessary.

[0108] The compounds of this invention can be administered alone or in combination with other pharmaceutically acceptable compounds.

[0109] The treatment method of the present invention can be used alone or in combination with other treatment methods or drugs.

[0110] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to the mammal (such as a human) requiring treatment. The dosage administered is the pharmaceutically considered effective dose. For a person weighing 60 kg, the daily dose is typically 1–2000 mg, preferably 5–1000 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the patient's health condition, which are all within the scope of the skill of a skilled physician.

[0111] The main advantages of this invention include:

[0112] (1) The compounds of the present invention have excellent degradation activity against a variety of histone demethylases, deubiquitinases, c-MYC and / or RNF6.

[0113] (2) The compounds of the present invention can selectively degrade histone demethylases (e.g., KDM5A, KDM5C), deubiquitinases (e.g., USP25, USP28), c-MYC and / or RNF6.

[0114] (3) The compounds of the present invention have significant inhibitory effects on a variety of cancer cells.

[0115] (4) The compounds of the present invention have significant inhibitory effects on a variety of tumor models.

[0116] 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.

[0117] Example

[0118] 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.

[0119] Synthesis of Equations 1 to 5

[0120] Where n = 1, 2 or 3

[0121] Synthesis of Equation 1:

[0122] Step 1: 2-Chloro-3-bromo-5-hydroxypyridine (1.03 g, 5.0 mmol) was dissolved in 25 mL of a mixed solution of 1,4-dioxane and water (v:v = 4:1). 4-Cyanobenzoic acid (0.735 g, 5.0 mmol), tetrakis(triphenylphosphine)palladium (0.58 g, 0.5 mmol), and potassium carbonate (1.38 g, 10 mmol) were added sequentially. The reaction mixture was stirred at 90 °C for 5 h. The reaction solution was cooled to room temperature and then quenched with water (100 mL). The mixture was extracted three times with ethyl acetate (3 × 50 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by prep-HPLC to obtain a white solid, intermediate 1 (1.04 g, yield: 90.0%).

[0123] Step 2: Intermediate 1 (1.00 g, 4.3 mmol) was dissolved in 25 mL of DMF solution, and tert-butyl 4-(bromomethyl)piperidine-1-carboxylate (1.20 g, 4.3 mmol) and potassium carbonate (1.20 g, 8.7 mmol) were added sequentially. The reaction system was stirred overnight at 100 °C. After the reaction was completed, the reaction solution was cooled to room temperature, water was added to precipitate the solid, and the solid was filtered. Then, it was purified by prep-HPLC to obtain white intermediate 2 (1.58 g, yield: 85.0%).

[0124] Step 3: Dissolve intermediate 2 (1.50 g, 3.5 mmol) and 4-carboxyphenylboronic acid (0.58 g, 3.5 mmol) in a mixed solution of 25 mL of 1,4-dioxane and water (v:v = 4:1), [1,1-bis(diphenylphosphino)ferrocene] dichloropalladium (0.51 g, 0.4 mmol), and potassium carbonate (1.46 g, 10.6 mmol). Stir the reaction system overnight at 80 °C. End the reaction, cool the reaction solution to room temperature and quench it with water, extract it three times with ethyl acetate (3 × 50 mL), wash it with saturated brine, dry it over anhydrous sodium sulfate, concentrate it, and purify the residue by prep-HPLC to obtain a gray solid, namely formula 1 (1.17 g, yield: 65.0%).

[0125] Synthesis of formula 2:

[0126] Step 1: Dissolve N-Boc-1,4-butanediamine (1.88 g, 10.0 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (2.76 g, 10.0 mmol) in 5 mL of DMF, add DIPEA (5.06 g, 50.0 mmol), and stir the reaction system at 100 °C for 5 h. Purify the reaction solution by prep-HPLC to obtain a green intermediate 3 (2.61 g, yield: 58.8%).

[0127] Step 2: Dissolve intermediate 3 (2.60 g, 5.8 mmol) in 10 mL of anhydrous dichloromethane, add trifluoroacetic acid (3 mL), stir the reaction system at room temperature for 30 min, distill off the solvent under reduced pressure to obtain a green solid, namely formula 2 (2.00 g, yield: 98.9%).

[0128] Synthesis of formula 3:

[0129] Replace the starting materials and prepare formula 3 according to steps 1 and 2 of formula 2.

[0130] Synthesis of formula 4:

[0131] Replace the starting materials and prepare formula 4 according to steps 1 and 2 of formula 2.

[0132] Synthesis of formula 5:

[0133] Step 1: 2-(2,6-Dioxopiperidin-3-yl)-6,7-dihydropyrrolo[3,4-f]isoindole-1,3(2H,5H)-dione (0.29 g, 1.0 mmol) and 3-(Boc-amino) propionaldehyde (0.24 g, 1.3 mmol) were dissolved in 10 mL of DCM / MeOH (v:v = 1:1). The reaction system was stirred at room temperature for 30 min, then NaBH3CN (0.06 g, 2.0 mmol) was added, and the reaction system was stirred overnight at room temperature. Water was added to the reaction system to quench the reaction, and the solvent was removed by distillation under reduced pressure to obtain a crude product, which was then purified by prep-HPLC to obtain a white intermediate 5 (0.10 g, yield: 28.1%).

[0134] Replace the starting materials and prepare Compound 5 according to Step 2 of Formula 2.

[0135] Example 1

[0136] Procedure: Dissolve Formula 1 (0.51 g, 1.0 mmol) and Formula 2 (0.34 g, 1.0 mmol) in 5 mL of DMF. After adding 2 - 3 drops of DIPEA, add HATU (0.46 g, 1.2 mmol). After the reaction is completed, separate and purify by preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column model: Prep C18 OBD TM 5 μm, 50×100 mm column, to obtain a yellow solid LD-150-ZJZ (0.67 g, yield: 79.8%).

[0137] Examples 2 - 3 and Example 5

[0138] Procedure: Refer to the synthesis procedure of LD-150-ZJZ. Dissolve Formula 1 and Formula 3 in DMF. After adding 2 - 3 drops of DIPEA, add HATU. After the reaction is completed, separate and purify by preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column model: Prep C18 OBD TM 5 μm, 50×100 mm column, to obtain yellow solids LD-151-ZJZ, LD-152-ZJZ, LD-154-ZJZ.

[0139] Example 4

[0140] Procedure: Dissolve Formula 1 (0.51 g, 1.0 mmol) and Formula 4 (0.36 g, 1.0 mmol) in 5 mL of DMF. After adding 2 - 3 drops of DIPEA, add HATU (0.46 g, 1.2 mmol). After the reaction is completed, separate and purify by preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column model: Prep C18 OBD TM 5 μm, 50×100 mm column, to obtain the yellow solid LD-153-ZJZ (0.68 g, yield: 79.5%).

[0141] Example 6

[0142] Procedure: Dissolve Formula 1 (0.51 g, 1.0 mmol) and Formula 5 (0.36 g, 1.0 mmol) in 5 mL of DMF. After adding 2 - 3 drops of DIPEA, add HATU (0.46 g, 1.2 mmol). After the reaction is completed, separate and purify by preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column model: Prep C18 OBD TM 5 μm, 50×100 mm column, to obtain the yellow solid LD-156-ZJZ (0.49 g, yield: 57.6%).

[0143] Synthesis of Intermediate 6:

[0144] Procedure: Similar to Formula 2. Dissolve LD-150-ZJZ in anhydrous dichloromethane, add trifluoroacetic acid, stir the reaction system at room temperature for 30 min, and remove the solvent by reduced pressure distillation to obtain the yellow Intermediate 6.

[0145] Example 7

[0146] Procedure: Dissolve Intermediate 6 (0.74 g, 1.0 mmol) and formic acid (0.05 g, 1.0 mmol) in 5 mL of DMF. After adding 2 - 3 drops of DIPEA, add HATU (0.46 g, 1.2 mmol). After the reaction is completed, separate and purify by preparative liquid chromatography. Mobile phase: methanol / water = 60 / 40. Column model: Prep C18 OBD TM 5 μm, 50×100 mm column, to obtain the yellow solid LD-324-LLH (0.56 g, yield: 73.0%), 11H NMR (500 MHz, DMSO-d6) δ 11.05 (s, 1H), 8.46 (t, J = 4.4 Hz, 2H), 8.00 (s, 1H), 7.83 - 7.77 (m, 2H), 7.71 (d, J = 8.5 Hz, 2H), 7.55 (d, J = 8.4 Hz, 1H), 7.48 (d, J = 2.8 Hz, 1H), 7.44 - 7.38 (m, 2H), 7.27 (d, J = 8.5 Hz, 2H), 7.12 (t, J = 5.3 Hz, 1H), 6.95 (d, J = 1.8 Hz, 1H), 6.85 (dd, J = 8.4, 2.0 Hz, 1H), 5.02 (dd, J = 12.7, 5.4 Hz, 1H), 4.21 (d, J = 13.0 Hz, 1H), 4.07 (t, J = 8.8 Hz, 2H), 3.72 (d, J = 13.2 Hz, 1H), 3.29 (d, J = 5.5 Hz, 2H), 3.18 (dd, J = 12.0, 5.3 Hz, 2H), 3.08 (td, J = 12.8, 2.9 Hz, 1H), 2.94 - 2.80 (m, 1H), 2.66 (td, J = 12.8, 3.1 Hz, 1H), 2.61 - 2.50 (m, 2H), 2.16 - 2.05 (m, 1H), 2.03 - 1.93 (m, 1H), 1.92 - 1.75 (m, 2H), 1.61 (s, 4H), 1.26 - 1.21 (m, 2H). LC-MS calculated for C 43 H 41 N7O7 [M + H] + : 767.31, found 768.48. UPLC retention time: 6.3 min, purity > 95%.

[0147] Example 8

[0148] Procedure: The same as the synthesis of LD-324-LLH. Dissolve intermediate 6 and acetic acid in 5 mL of DMF, add 2 - 3 drops of DIPEA and then add HATU. After the reaction is completed, purify by preparative liquid chromatography. Mobile phase: methanol / water = 60 / 40. Column model: Prep C18 OBD TM 5 μm, 50 × 100 mm column, to obtain the yellow solid LD-325-LLH, 11H NMR (500 MHz, DMSO-d6) δ 11.05 (s, 1H), 8.53 - 8.41 (m, 2H), 7.85 - 7.77 (m, 2H), 7.71 (d, J = 8.4 Hz, 2H), 7.55 (d, J = 8.4 Hz, 1H), 7.48 (t, J = 2.8 Hz, 2H), 7.45 - 7.39 (m, 2H), 7.27 (d, J = 8.4 Hz, 1H), 7.13 (t, J = 5.3 Hz, 1H), 6.95 (d, J = 1.7 Hz, 1H), 6.85 (dd, J = 8.4, 2.0 Hz, 1H), 5.02 (dd, J = 12.7, 5.5 Hz, 1H), 4.40 (d, J = 13.1 Hz, 1H), 4.05 (d, J = 6.4 Hz, 2H), 3.85 (t, J = 6.7 Hz, 1H), 3.29 (d, J = 5.6 Hz, 1H), 3.19 (d, J = 5.3 Hz, 2H), 3.12 - 2.99 (m, 1H), 2.87 (ddd, J = 16.6, 13.7, 5.3 Hz, 1H), 2.63 - 2.53 (m, 2H), 2.14 - 2.01 (m, 1H), 2.00 (s, 4H), 1.89 - 1.72 (m, 2H), 1.61 (s, 4H), 1.36 - 1.27 (m, 1H), 1.27 - 1.17 (m, 2H), 1.14 (dd, J = 12.1, 4.0 Hz, 1H). LC-MS calculated for C 44 H 43 N7O7 [M + H]+: 781.32, found 782.60. UPLC retention time: 6.5 min, purity > 95%.

[0149] Example 9

[0150] Procedure: The same as the synthesis of LD-324-LLH. Dissolve intermediate 6 and propionic acid in 5 mL of DMF, add 2 - 3 drops of DIPEA and then add HATU. After the reaction is completed, purify by preparative liquid chromatography. Mobile phase: methanol / water = 60 / 40. Column model: Prep C18 OBD TM 5 μm, 50 × 100 mm column, to obtain the yellow solid LD-326-LLH, 11H NMR (500 MHz, DMSO-d6) δ 11.06 (s, 1H), 8.46 (t, J = 4.5 Hz, 2H), 7.80 (d, J = 8.3 Hz, 2H), 7.71 (d, J = 8.3 Hz, 2H), 7.55 (d, J = 8.4 Hz, 1H), 7.47 (d, J = 2.7 Hz, 1H), 7.42 (d, J = 8.3 Hz, 2H), 7.28 (d, J = 8.3 Hz, 2H), 7.12 (t, J = 5.0 Hz, 1H), 6.95 (s, 1H), 6.85 (dd, J = 8.4, 1.8 Hz, 1H), 5.03 (dd, J = 12.8, 5.4 Hz, 1H), 4.43 (d, J = 12.9 Hz, 1H), 4.04 (d, J = 6.3 Hz, 2H), 3.89 (d, J = 13.4 Hz, 1H), 3.72 - 3.56 (m, 1H), 3.41 - 3.25 (m, 3H), 3.24 - 3.12 (m, 3H), 3.03 (t, J = 11.9 Hz, 1H), 2.87 (ddd, J = 16.6, 13.7, 5.1 Hz, 1H), 2.66 - 2.49 (m, 2H), 2.31 (q, J = 7.4 Hz, 2H), 2.10 - 2.03 (m, 1H), 2.01 - 1.91 (m, 1H), 1.81 (dd, J = 18.1, 15.0 Hz, 2H), 1.61 (s, 4H), 0.98 (t, J = 7.4 Hz, 3H). LC-MS calculated for C 45 H 45 N7O7[M + H]+: 795.34, found 796.60. UPLC retention time: 6.8 min, purity > 95%.

[0151] Example 10

[0152] Procedure: The same as the synthesis of LD-324-LLH. Dissolve intermediate 6 and cyclopropylformic acid in 5 mL of DMF, add 2 - 3 drops of DIPEA and then add HATU. After the reaction, separate and purify by preparative liquid chromatography. Mobile phase: methanol / water = 60 / 40. Column type: Prep C18 OBD TM 5 μm, 50 × 100 mm column, to obtain the yellow solid LD-327-LLH, 11H NMR (500 MHz, DMSO-d6) δ 11.06 (s, 1H), 8.56 - 8.40 (m, 2H), 7.80 (d, J = 8.3 Hz, 2H), 7.71 (d, J = 8.4 Hz, 2H), 7.55 (d, J = 8.4 Hz, 1H), 7.48 (d, J = 2.8 Hz, 1H), 7.45 - 7.38 (m, 2H), 7.28 (d, J = 8.4 Hz, 2H), 7.12 (t, J = 5.3 Hz, 1H), 6.95 (d, J = 1.7 Hz, 1H), 6.84 (dd, J = 8.4, 1.9 Hz, 1H), 5.03 (dd, J = 12.8, 5.4 Hz, 1H), 4.34 (dd, J = 58.5, 11.6 Hz, 2H), 4.15 - 3.95 (m, 2H), 3.29 (d, J = 5.4 Hz, 2H), 3.22 - 3.09 (m, 3H), 2.87 (ddd, J = 16.6, 13.7, 5.1 Hz, 1H), 2.58 (ddd, J = 19.6, 15.3, 8.1 Hz, 2H), 2.16 - 2.05 (m, 1H), 2.04 - 1.92 (m, 2H), 1.92 - 1.70 (m, 2H), 1.61 (s, 4H), 1.31 - 1.17 (m, 3H), 0.69 (dd, J = 14.2, 6.3 Hz, 4H). LC-MS calculated for C 46 H 45 N7O7 [M + H]+: 807.34, found 808.57. UPLC retention time: 7.0 min, purity > 95%.

[0153] Example 11

[0154] Procedure: The same as the synthesis of LD-324-LLH. Dissolve intermediate 6 and cyclobutylformic acid in DMF, add 2 - 3 drops of DIPEA, then add HATU. After the reaction, purify by preparative liquid chromatography. Mobile phase: methanol / water = 60 / 40. Column model: Prep C18 OBD TM 5 μm, 50×100 mm column, to obtain the yellow solid LD-328-LLH, 11H NMR (500 MHz, DMSO-d6) δ 11.06 (s, 1H), 8.46 (dd, J = 7.5, 4.2 Hz, 2H), 7.80 (d, J = 8.4 Hz, 2H), 7.71 (d, J = 8.4 Hz, 2H), 7.55 (d, J = 8.4 Hz, 1H), 7.47 (d, J = 2.8 Hz, 1H), 7.41 (d, J = 8.3 Hz, 2H), 7.27 (d, J = 8.4 Hz, 2H), 7.12 (t, J = 5.2 Hz, 1H), 6.95 (d, J = 1.6 Hz, 1H), 6.85 (dd, J = 8.4, 1.9 Hz, 1H), 5.03 (dd, J = 12.8, 5.4 Hz, 1H), 4.39 (d, J = 13.0 Hz, 1H), 4.03 (d, J = 6.4 Hz, 2H), 3.71 (d, J = 13.4 Hz, 1H), 3.66 - 3.56 (m, 1H), 3.36 - 3.24 (m, 4H), 3.23 - 3.11 (m, 3H), 2.96 (t, J = 11.9 Hz, 1H), 2.87 (ddd, J = 16.7, 13.8, 5.2 Hz, 1H), 2.60 - 2.52 (m, 2H), 2.15 (dd, J = 19.6, 10.5 Hz, 2H), 2.09 - 2.02 (m, 3H), 1.98 (ddd, J = 15.5, 6.3, 3.7 Hz, 1H), 1.94 - 1.83 (m, 1H), 1.80 (dd, J = 19.6, 9.5 Hz, 2H), 1.72 (ddd, J = 14.0, 9.7, 6.0 Hz, 1H), 1.61 (s, 4H). LC-MS calculated for C 47 H 47 N7O7[M + H]+: 821.35, found 822.63. UPLC retention time: 7.2 min, purity > 95%.

[0155] Example 12

[0156] Procedure: The same as the synthesis of LD-324-LLH. Dissolve intermediate 6 and cyclopentanecarboxylic acid in DMF, add 2 - 3 drops of DIPEA and then add HATU. After the reaction is completed, purify by preparative liquid chromatography. Mobile phase: methanol / water = 60 / 40. Column model: Prep C18 OBD TM 5 μm, 50×100 mm column, to obtain the yellow solid LD-329-LLH, 1H NMR (500MHz, DMSO-d6) δ11.06 (s, 1H), 8.46 (t, J = 4.4Hz, 2H), 7.80 (d, J = 8.2Hz ,2H),7.71(d,J=8.3Hz,2H),7.55(d,J=8.4Hz,1H),7.47(d,J=2.7Hz,1H),7.4 2(d,J=8.2Hz,2H),7.28(d,J=8.3Hz,2H),7.12(s,1H),6.95(s,1H),6.85(dd, J=8.4,1.5Hz,1H),5.03(dd,J=12.8,5.4Hz,1H),4.43(d,J=12.8Hz,1H),4.03( dd, J = 17.9, 10.1 Hz, 3H), 3.68-3.57 (m, 1H), 3.29 (d, J = 5.3 Hz, 2H), 3.24-3.16 (m, 2H), 3.13 (tt, J = 7.3, 3.7 Hz, 1H), 3.04 (t, J = 12.1 Hz, 1H), 2.96 (dd, J = 15.7, 7.8 Hz, 1H), 2.87 (ddd, J = 16.6, 13.8, 5.1 Hz, 1H), 2.59-2.53 (m, 2H), 2.12-2.02 (m, 1H), 2.03-1.93 (m, 1H), 1.89-1.70 (m, 4H), 1.70-1.45 (m, 11H). LC-MS calculation yields C 48 H 49 N7O7[M+H]+: 835.57, found 836.60. UPLC retention time: 7.4 min, purity >95%.

[0157] Example 13

[0158] Procedure: Same as the synthesis of LD-324-LLH. Intermediate 6 and cyclohexanecarboxylic acid were dissolved in DMF, followed by the addition of 2-3 drops of DIPEA and then HATU. After the reaction was complete, the mixture was purified by preparative liquid chromatography. Mobile phase: methanol / water = 60 / 40. Column type: Prep C18OBD TM A 5μm, 50×100mm column yielded a yellow solid LD-330-LLH. 1H NMR(500MHz,DMSO-d6)δ11.06(s,1H),8.46(t,J=4.5Hz,2H),7.80(d,J=8.2Hz,2H) ,7.71(d,J=8.3Hz,2H),7.55(d,J=8.4Hz,1H),7.47(d,J=2.7Hz,1H),7.42(d,J=8. 3Hz,2H),7.27(d,J=8.3Hz,2H),7.12(s,1H),6.95(s,1H),6.84(dd,J=8.4,1.8Hz, 1H),5.03(dd,J=12.8,5.4Hz,1H),4.43(d,J=12.2Hz,1H),4.05(d,J=6.3Hz,2H),3 .97(d,J=12.7Hz,1H),3.70-3.54(m,1H),3.29(d,J=5.3Hz,2H),3.23-3.10(m,3H) ,3.05(t,J=12.2Hz,1H),2.87(ddd,J=16.5,13.7,5.1Hz,1H),2.67-2.51(m,3H),2 0.05 (dt, J = 25.9, 11.1 Hz, 1H), 2.02-1.93 (m, 1H), 1.82 (dd, J = 32.6, 11.7 Hz, 2H), 1.68 (d, J = 7.9 Hz, 2H), 1.61 (d, J = 2.5 Hz, 7H), 1.27 (dd, J = 16.5, 9.5 Hz, 6H). LC-MS calculation yields C. 49 H 51 N7O7[M+H]+: 849.38, found 850.80. UPLC retention time: 7.6 min, purity >95%.

[0159] Example 14

[0160] Procedure: Same as the synthesis of LD-324-LLH. Intermediate 6 and isobutyric acid were dissolved in DMF, followed by the addition of 2-3 drops of DIPEA and then HATU. After the reaction was complete, the mixture was purified by preparative liquid chromatography. Mobile phase: methanol / water = 60 / 40. Column type: Prep C18 OBD TM A 5μm, 50×100mm column yielded a yellow solid LD-331-LLH. 1H NMR(500MHz,DMSO-d6)δ11.06(s,1H),8.51-8.44(m,2H),7.82-7.76(m,2H),7.7 1(d,J=8.4Hz,2H),7.55(d,J=8.4Hz,1H),7.47(d,J=2.8Hz,1H),7.44-7.39(m,2H ),7.28(d,J=8.4Hz,2H),7.12(t,J=5.2Hz,1H),6.95(d,J=1.7Hz,1H),6.85(dd, J=8.4,2.0Hz,1H),5.03(dd,J=12.8,5.4Hz,1H),4.44(d,J=12.4Hz,1H),4.05(d, J = 6.4 Hz, 2H), 3.99 (t, J = 12.1 Hz, 1H), 3.68-3.55 (m, 1H), 3.29 (d, J = 5.5 Hz, 2H), 3.23-3.11 (m, 3H), 3.06 (t, J = 12.2 Hz, 1H), 2.96-2.82 (m, 2H), 2.66-2.49 (m, 2H), 2.13-2.03 (m, 1H), 1.99 (ddd, J = 9.6, 7.3, 3.2 Hz, 1H), 1.83 (dd, J = 30.5, 12.0 Hz, 2H), 1.61 (s, 4H), 1.15 (dd, J = 17.6, 10.5 Hz, 1H), 0.99 (t, J = 6.5 Hz, 6H). LC-MS calculation yields C 46 H 47 N7O7[M+H]+: 809.35, found 810.62. UPLC retention time: 7.8 min, purity >95%.

[0161] Example 15

[0162] Procedure: Same as the synthesis of LD-324-LLH. Intermediate 6 and spiro[2,2]pentane-1-carboxylic acid were dissolved in DMF, followed by the addition of 2-3 drops of DIPEA and then HATU. After the reaction was complete, the mixture was purified by preparative liquid chromatography. Mobile phase: methanol / water = 60 / 40. Column type: Prep C18 OBD TM A 5μm, 50×100mm column yielded a yellow solid LD-332-LLH. 11H NMR (500 MHz, DMSO-d6) δ 11.06 (s, 1H), 8.46 (t, J = 5.4 Hz, 2H), 7.80 (d, J = 8.2 Hz, 2H), 7.71 (d, J = 8.3 Hz, 2H), 7.55 (d, J = 8.4 Hz, 1H), 7.47 (s, 1H), 7.42 (d, J = 8.2 Hz, 2H), 7.28 (d, J = 8.3 Hz, 2H), 7.12 (t, J = 5.2 Hz, 1H), 6.95 (s, 1H), 6.85 (dd, J = 8.4, 1.8 Hz, 1H), 5.03 (dd, J = 12.8, 5.4 Hz, 1H), 4.39 (s, 1H), 4.04 (s, 3H), 3.68 - 3.56 (m, 1H), 3.29 (d, J = 5.4 Hz, 2H), 3.22 - 2.94 (m, 4H), 2.87 (ddd, J = 16.6, 13.8, 5.2 Hz, 1H), 2.67 - 2.57 (m, 1H), 2.53 (dd, J = 10.9, 4.9 Hz, 1H), 2.29 (s, 1H), 2.06 (s, 1H), 2.04 - 1.91 (m, 1H), 1.84 (dd, J = 23.3, 16.0 Hz, 2H), 1.62 (s, 4H), 1.38 (d, J = 13.4 Hz, 1H), 1.13 (ddd, J = 24.5, 12.4, 7.0 Hz, 2H), 0.89 (dd, J = 8.8, 4.1 Hz, 1H), 0.86 - 0.73 (m, 2H), 0.65 (s, 1H). LC-MS calculated for C 48 H 47 N7O7[M + H]+: 833.35, found 834.70. UPLC retention time: 8.0 min, purity > 95%.

[0163] Example 16

[0164] Procedure: The same as the synthesis of LD-324-LLH. Dissolve intermediate 6 and spiro[3.3]heptane-2-carboxylic acid in DMF, add 2 - 3 drops of DIPEA and then add HATU. After the reaction is completed, purify by preparative liquid chromatography. Mobile phase: methanol / water = 60 / 40. Column model: Prep C18 OBD TM 5 μm, 50×l00 mm column, to obtain the yellow solid LD-333-LLH, 11H NMR (500 MHz, DMSO-d6) δ 11.06 (s, 1H), 8.46 (dd, J = 7.6, 4.2 Hz, 2H), 7.86 - 7.78 (m, 2H), 7.71 (d, J = 8.4 Hz, 2H), 7.55 (d, J = 8.4 Hz, 1H), 7.47 (d, J = 2.8 Hz, 1H), 7.43 - 7.37 (m, 2H), 7.27 (d, J = 8.4 Hz, 2H), 7.12 (d, J = 4.8 Hz, 1H), 6.95 (d, J = 1.6 Hz, 1H), 6.85 (dd, J = 8.4, 1.9 Hz, 1H), 5.03 (dd, J = 12.8, 5.4 Hz, 1H), 4.37 (d, J = 12.9 Hz, 1H), 4.03 (d, J = 6.3 Hz, 2H), 3.73 (d, J = 13.4 Hz, 1H), 3.68 - 3.55 (m, 1H), 3.40 - 3.31 (m, 1H), 3.24 - 3.10 (m, 4H), 3.01 - 2.81 (m, 2H), 2.58 (dd, J = 6.7, 3.1 Hz, 2H), 2.16 - 2.05 (m, 4H), 2.06 - 1.93 (m, 4H), 1.86 - 1.68 (m, 6H), 1.61 (s, 4H), 1.21 - 0.99 (m, 2H). LC-MS calculated for C 50 H 51 N7O7[M + H]+: 861.38, found 862.74. UPLC retention time: 8.2 min, purity > 95%.

[0165] Synthesis of Intermediate 7:

[0166] Procedure: Similar to Formula 2. Dissolve Formula 1 in anhydrous dichloromethane, add trifluoroacetic acid, stir the reaction system at room temperature for 30 min, and remove the solvent by distillation under reduced pressure to obtain Intermediate 7.

[0167] Example 17

[0168] Step 1: Dissolve Intermediate 7 (0.41 g, 1.0 mmol) in 2 mL of DMF, successively add potassium carbonate (0.28 g, 2.0 mmol) and methyl iodide (0.14 g, 1.0 mmol), and react at room temperature for 12 hours. After the reaction is completed, the mixed solution is separated and purified by preparative liquid chromatography. Mobile phase: methanol / water = 10 / 90. Column model: Prep C18 OBD TM 5 μm, 50 × 100 mm column to obtain gray Intermediate 8 (0.33 g, yield: 77.2%).

[0169] Step 2: Intermediate 8 (0.33 g, 0.77 mmol) and Formula 2 (0.27 g, 0.77 mmol) were dissolved in 2 mL of DMF. After adding 2 - 3 drops of DIPEA, HATU (0.35 g, 0.93 mmol) was added. After the reaction, it was separated and purified by preparative liquid chromatography. Mobile phase: methanol / water = 40 / 60. Column model: Prep C18 OBD TM A 5 μm, 50×100 mm column was used to obtain the yellow solid LD - 334 - LLH (0.44 g, yield: 75.7%).

[0170] Example 18

[0171] Step 1: Intermediate 7 (0.41 g, 1.0 mmol) was dissolved in 2 mL of DMF. Potassium carbonate (0.28 g, 2.0 mmol) and iodoethane (0.16 g, 1.0 mmol) were added successively, and the reaction was carried out at room temperature for 12 hours. After the reaction, the mixed solution was separated and purified by preparative liquid chromatography. Mobile phase: methanol / water = 10 / 90. Column model: Prep C18 OBD TM A 5 μm, 50×100 mm column was used to obtain the gray Intermediate 9 (0.31 g, yield: 70.3%).

[0172] Step 2: Intermediate 9 (0.31 g, 0.70 mmol) and Formula 2 (0.24 g, 0.70 mmol) were dissolved in 2 mL of DMF. After adding 2 - 3 drops of DIPEA, HATU (0.32 g, 0.84 mmol) was added. After the reaction, it was separated and purified by preparative liquid chromatography. Mobile phase: methanol / water = 40 / 60. Column model: Prep C18 OBD TM A 5 μm, 50×100 mm column was used to obtain the yellow solid LD - 335 - LLH (0.42 g, yield: 75.4%).

[0173] Example 19

[0174] Step 1: Intermediate 7 (0.41 g, 1.0 mmol) was dissolved in 2 mL of DMF. Potassium carbonate (0.28 g, 2.0 mmol) and 2 - iodopropane (0.17 g, 1.0 mmol) were added successively, and the reaction was carried out at room temperature for 12 hours. After the reaction, the mixed solution was separated and purified by preparative liquid chromatography. Mobile phase: methanol / water = 10 / 90. Column model: Prep C18 OBD TM A 5 μm, 50×100 mm column was used to obtain the gray Intermediate 10 (0.27 g, yield: 59.3%).

[0175] Step 2: Intermediate 10 (0.27 g, 0.59 mmol) and Formula 2 (0.20 g, 0.59 mmol) were dissolved in 2 mL of DMF. After adding 2 - 3 drops of DIPEA, HATU (0.27 g, 0.71 mmol) was added. After the reaction, it was separated and purified by preparative liquid chromatography. Mobile phase: methanol / water = 40 / 60. Column model: Prep C18 OBD TM 5 μm, 50×100 mm column, to obtain yellow solid LD - 336 - LLH (0.39 g, yield: 50.0%).

[0176] Example 20

[0177] Step 1: Intermediate 7 (0.41 g, 1.0 mmol) was dissolved in 2 mL of DMF. Potassium carbonate (0.28 g, 2.0 mmol) and iodocyclopropane (0.17 g, 1.0 mmol) were added successively, and the reaction was carried out at room temperature for 12 hours. After the reaction, the mixed solution was separated and purified by preparative liquid chromatography. Mobile phase: methanol / water = 10 / 90. Column model: Prep C18 OBD TM 5 μm, 50×100 mm column, to obtain gray Intermediate 11 (0.29 g, yield: 64.0%).

[0178] Step 2: Intermediate 11 (0.29 g, 0.64 mmol) and Formula 2 (0.22 g, 0.64 mmol) were dissolved in 2 mL of DMF. After adding 2 - 3 drops of DIPEA, HATU (0.29 g, 0.71 mmol) was added. After the reaction, it was separated and purified by preparative liquid chromatography. Mobile phase: methanol / water = 40 / 60. Column model: Prep C18 OBD TM 5 μm, 50×100 mm column, to obtain yellow solid LD - 337 - LLH (0.37 g, yield: 74.2%).

[0179] Example 21

[0180] Step 1: Intermediate 7 (0.41 g, 1.0 mmol) was dissolved in 2 mL of DMF. Potassium carbonate (0.28 g, 2.0 mmol) and iodocyclobutane (0.18 g, 1.0 mmol) were added successively, and the reaction was carried out at room temperature for 12 hours. After the reaction, the mixed solution was separated and purified by preparative liquid chromatography. Mobile phase: methanol / water = 10 / 90. Column model: Prep C18 OBD TM5 μm, 50×100 mm column, to obtain gray intermediate 12 (0.30 g, yield: 64.2%).

[0181] Step 2: Intermediate 12 (0.29 g, 0.64 mmol) and Formula 2 (0.22 g, 0.64 mmol) were dissolved in 2 mL of DMF. After adding 2 - 3 drops of DIPEA, HATU (0.29 g, 0.71 mmol) was added. After the reaction, it was separated and purified by preparative liquid chromatography. Mobile phase: methanol / water = 40 / 60. Column model: Prep C18 OBD TM 5 μm, 50×100 mm column, to obtain yellow solid LD - 338 - LLH (0.38 g, yield: 74.6%).

[0182] Example 22

[0183] Step 1: Intermediate 7 (0.41 g, 1.0 mmol) was dissolved in 2 mL of DMF. Potassium carbonate (0.28 g, 2.0 mmol) and iodocyclopentane (0.20 g, 1.0 mmol) were added successively, and the reaction was carried out at room temperature for 12 hours. After the reaction, the mixed solution was separated and purified by preparative liquid chromatography. Mobile phase: methanol / water = 10 / 90. Column model: Prep C18 OBD TM 5 μm, 50×100 mm column, to obtain gray intermediate 13 (0.29 g, yield: 60.3%).

[0184] Step 2: Intermediate 13 (0.29 g, 0.60 mmol) and Formula 2 (0.21 g, 0.60 mmol) were dissolved in 2 mL of DMF. After adding 2 - 3 drops of DIPEA, HATU (0.27 g, 0.72 mmol) was added. After the reaction, it was separated and purified by preparative liquid chromatography. Mobile phase: methanol / water = 40 / 60. Column model: Prep C18 OBD TM 5 μm, 50×100 mm column, to obtain yellow solid LD - 339 - LLH (0.34 g, yield: 69.9%).

[0185] Example 23

[0186] Step 1: Intermediate 7 (0.41 g, 1.0 mmol) was dissolved in 2 mL of DMF. Potassium carbonate (0.28 g, 2.0 mmol) and iodocyclohexane (0.22 g, 1.0 mmol) were added successively, and the reaction was carried out at room temperature for 12 hours. After the reaction, the mixed solution was separated and purified by preparative liquid chromatography. Mobile phase: methanol / water = 10 / 90. Column model: Prep C18 OBD TM 5 μm, 50 × 100 mm column, yielded gray intermediate 14 (0.27 g, yield: 54.5%).

[0187] Step 2: Intermediate 14 (0.27 g, 0.55 mmol) and Formula 2 (0.19 g, 0.55 mmol) were dissolved in 2 mL of DMF. 2–3 drops of DIPEA were added, followed by HATU (0.25 g, 0.65 mmol). After the reaction was complete, the mixture was purified by preparative HPLC. Mobile phase: methanol / water = 40 / 60. Column type: Prep C18 OBD TM A 5 μm, 50 × 100 mm column was used to obtain a yellow solid LD-340-LLH (0.30 g, yield: 67.0%).

[0188] Example 24

[0189] Compound 2 (34 mg, 0.1 mmol) and compound 1 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This crude product was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a bright yellow chlorine solid compound LD-110-ZJZ. Overall yield of the two steps: 67%.

[0190] Example 25

[0191] 1.03 g (5.0 mmol) of 2-chloro-3-bromo-5-hydroxypyridine was dissolved in 25 mL of a mixed solution of 1,4-dioxane and water (volume ratio = 4:1). Then, 0.735 g (5.0 mmol) of 4-cyanobenzoic acid, 0.58 g (0.5 mmol) of tetrakis(triphenylphosphine)palladium, and 1.38 g (10 mmol) of K₂CO₃ were added sequentially. The reaction mixture was stirred at 90 °C for 5 h. The reaction solution was cooled to room temperature and then quenched with water (10 mL). The mixture was extracted three times with ethyl acetate (3 x 50 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by prep-HPLC to give compound 1 as a white solid. Yield: 90%.

[0192] Compound 1 (690 mg, 3 mmol) was weighed and dissolved in 25 mL of DMF solution. Then, tert-butyl 4-(bromomethyl)piperidine-1-carboxylate (833 mg, 3 mmol) and potassium carbonate (828 mg, 6 mmol) were added sequentially. The reaction mixture was stirred overnight at 100 °C. After the reaction was complete, the reaction solution was cooled to room temperature, filtered, and then purified by prep-HPLC to give compound 2 as a white solid with a yield of 85%.

[0193] Compound 2 (850 mg, 2 mmol), 4-carboxyphenylboronic acid (331 mg, 2 mmol), and potassium carbonate (828 mg, 6 mmol) were dissolved in 25 mL of a mixed solution of 1,4-dioxane and water (volume ratio = 4:1), [1,1-bis(diphenylphosphine)ferrocene]palladium dichloride (290 mg, 0.2 mmol), and potassium carbonate (828 mg, 6 mmol). The reaction mixture was stirred overnight at 80 °C. After the reaction was completed, the reaction solution was cooled to room temperature and quenched with water. The mixture was extracted three times with ethyl acetate (3 x 50 mL), washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by prep-HPLC to give compound 3 as a gray solid. Yield: 65%.

[0194] a, b: Compound 4 (50 mg, 0.11 mmol) and compound 5 (26 mg, 0.11 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.09 mL, 0.5 mmol) and HATU (50 mg, 0.13 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain a white solid, compound 6. Overall yield of the two steps: 70%.

[0195] c,d: Compound 6 (40 mg, 0.07 mmol) and compound 3 (37 mg, 0.07 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.06 mL, 0.35 mmol) and HATU (32 mg, 0.084 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 × 5 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. It was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a white solid compound LD-114-ZJZ. Overall yield of the two steps: 65%. LC-MS calculated C0... 54 H 64 N8O6S[M+H] + 952.47, found 953.49. UPLC retention time: 3.5 min, purity >95%.

[0196] Example 26

[0197] a, b: Compound 4 (50 mg, 0.11 mmol) and compound 7 (27 mg, 0.11 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.09 mL, 0.5 mmol) and HATU (50 mg, 0.13 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain a white solid, compound 8. Overall yield of the two steps: 78%.

[0198] c,d: Compound 8 (49 mg, 0.08 mmol) and compound 3 (41 mg, 0.08 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.07 mL, 0.4 mmol) and HATU (38 mg, 0.1 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a white solid compound LD-115-ZJZ. Overall yield of the two steps: 65%. 1 H NMR(500MHz,DMSO-d6)δ8.98(s,1H),8.48-8.40(m,2H),8.38(d,J=7.5Hz,1H),7.80(dd,J=12.6,8.8Hz,3H),7.72(d,J=8.4Hz,2H),7.49(d,J=2.7Hz,1 H),7.46-7.40(m,4H),7.38(d,J=8.4Hz,2H),7.27(d,J=8.4Hz,2H),4.91(p ,J=7.2Hz,1H),4.52(d,J=9.3Hz,1H),4.43(t,J=8.0Hz,1H),4.28(s,1H),4 .07(d,J=6.4Hz,2H),3.61(s,2H),3.30-3.20(m,4H),2.87(t,J=12.9Hz,2H ),2.45(s,3H),2.25(dt,J=14.6,7.6Hz,1H),2.12(dt,J=14.3,7.2Hz,2H), 2.05-1.98(m,1H),1.92(d,J=14.6Hz,2H),1.83-1.73(m,1H),1.48(q,J=8. 0Hz, 6H), 1.37 (d, J = 7.0Hz, 3H), 1.32-1.18 (m, 6H), 0.94 (s, 9H). C calculated by LC-MS 55 H 66 N8O6S[M+H] + 966.48, found 967.45. UPLC retention time: 3.8 min, purity >95%.

[0199] Example 27

[0200] a, b: Compound 4 (50 mg, 0.11 mmol) and compound 9 (30 mg, 0.11 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.09 mL, 0.5 mmol) and HATU (50 mg, 0.13 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain a white solid, compound 10. Overall yield of the two steps: 70%.

[0201] c,d: Compound 10 (47 mg, 0.08 mmol) and compound 3 (41 mg, 0.08 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.07 mL, 0.4 mmol) and HATU (38 mg, 0.1 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a white solid compound LD-116-ZJZ. Overall yield of the two steps: 65%. 1H NMR(500MHz,DMSO-d6)δ8.99(s,1H),8.48-8.36(m,3H),7.80(dd,J=19.7,8.8Hz ,3H),7.73-7.69(m,2H),7.50(d,J=2.7Hz,1H),7.45-7.40(m,4H),7.38(d,J=8.3 Hz,2H),7.29-7.24(m,2H),4.91(p,J=7.0Hz,1H),4.52(d,J=9.3Hz,1H),4.43(t ,J=8.0Hz,1H),4.28(q,J=3.5Hz,1H),4.08(d,J=6.4Hz,2H),3.60(t,J=3.7Hz,2H ), 3.31(d,J=12.5Hz,2H), 3.22(q,J=6.7Hz,2H), 2.93(q,J=11.4Hz,2H), 2.45(s,3H), 2.25(dt,J=14.9,7.7Hz,1H), 2.18-2.08(m,2H), 2.01(ddd,J=11.2,7.7,2.9Hz,1H), 1.98-1.92(m,2H), 1.79(ddd,J=12.9,8.5,4.7Hz,1H), 1.49(tt,J=13.7,5.5Hz,6H), 1.37(d,J=7.0Hz,3H), 1.31-1.19(m,10H), 0.93(s,9H). LC-MS calculation yields C 57 H 70 N8O6S[M+H] + 994.51, found 995.45. UPLC retention time: 3.9 min, purity >95%.

[0202] Example 28

[0203] a, b: Compound 4 (50 mg, 0.11 mmol) and compound 11 (32 mg, 0.11 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.09 mL, 0.5 mmol) and HATU (50 mg, 0.13 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain a white solid, compound 12. Overall yield of the two steps: 68%.

[0204] c,d: Compound 12 (46 mg, 0.07 mmol) and compound 3 (36 mg, 0.07 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.06 mL, 0.35 mmol) and HATU (32 mg, 0.08 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a white solid compound LD-124-ZJZ. Overall yield of the two steps: 65%. 1 H NMR(500MHz,DMSO-d6)δ8.99(s,1H),8.48-8.31(m,3H),7.80(dd,J=16.2,8 .9Hz,3H),7.72(d,J=8.4Hz,2H),7.50(d,J=2.9Hz,1H),7.47-7.40(m,4H), 7.38(d,J=8.4Hz,2H),7.28(d,J=8.4Hz,2H),4.91(p,J=6.9Hz,1H),4.52(d ,J=9.3Hz,1H),4.42(q,J=8.0Hz,1H),4.28(d,J=3.2Hz,1H),4.07(d,J=6.4 Hz, 2H), 3.64-3.57(m, 2H), 3.29(d, J = 12.2Hz, 2H), 3.22(q, J = 6.3Hz, 2H), 2.89(t, J = 11.7Hz, 2H), 2.45(s, 3H), 2.25(dt, J = 14.6, 7.7Hz, 1H), 2.10(dt, J = 14.2, 7.1Hz, 2H), 1.97(dd, J = 34.1, 11.8Hz, 3H), 1.79(ddd, J = 13.0, 8.5, 4.7Hz, 1H), 1.59-1.34(m, 9H), 1.32-1.19(m, 12H), 0.93(s, 9H). LC-MS calculation yields C 58 H 72 N8O6S[M+H] + :1008.53, found 1009.55. UPLC retention time: 4.2 min, purity >95%.

[0205] Example 29

[0206] a, b: Compound 4 (50 mg, 0.11 mmol) and compound 13 (33 mg, 0.11 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.09 mL, 0.5 mmol) and HATU (50 mg, 0.13 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain a white solid, compound 14. Overall yield of the two steps: 75%.

[0207] c,d: Compound 14 (50 mg, 0.08 mmol) and compound 3 (42 mg, 0.08 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.07 mL, 0.4 mmol) and HATU (38 mg, 0.1 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a white solid compound LD-125-ZJZ. Overall yield of the two steps: 60%. 1H NMR(500MHz,DMSO-d6)δ8.99(s,1H),8.72(s,1H),8.50-8.32(m,3H),7.80(d d,J=20.5,8.9Hz,3H),7.72(d,J=8.5Hz,2H),7.50(d,J=2.7Hz,1H),7.47-7. 40(m,4H),7.38(d,J=8.4Hz,2H),7.28(d,J=8.5Hz,2H),4.91(p,J=6.8Hz,1H ),4.51(d,J=9.3Hz,1H),4.42(t,J=8.0Hz,1H),4.27(d,J=7.2Hz,1H),4.08( d,J=6.4Hz,2H),3.60(d,J=3.8Hz,2H),3.30(d,J=12.5Hz,2H),3.22(d,J=6. 9Hz,2H),2.91(q,J=11.8Hz,2H),2.45(s,3H),2.28-2.19(m,1H),2.09(dd,J =14.2,6.1Hz,2H),2.03-1.92(m,3H),1.79(td,J=8.4,4.3Hz,1H),1.55-1.4 3(m,6H),1.37(d,J=7.0Hz,3H),1.29-1.21(m,14H),0.93(s,9H).C calculated by LC-MS 59 H 74 N8O6S[M+H] + :1022.55, found 1023.56. UPLC retention time: 4.4 min, purity >95%.

[0208] Example 30

[0209] a, b: Compound 4 (50 mg, 0.11 mmol) and compound 15 (29 mg, 0.11 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.09 mL, 0.5 mmol) and HATU (50 mg, 0.13 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain a white solid, compound 16. Overall yield of the two steps: 70%.

[0210] c,d: Compound 16 (47 mg, 0.08 mmol) and compound 3 (42 mg, 0.08 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.07 mL, 0.4 mmol) and HATU (38 mg, 0.1 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the white solid compound LD-127-ZJZ. Overall yield of the two steps: 60%. 1 H NMR (500MHz, DMSO-d6) δ8.98(s,1H),8.50-8.32(m,3H),7.80(dd,J=17.3,8.8Hz,3H),7.71(d,J=8.4Hz,2H),7.49(d,J=2.9Hz,1H),7.46-7.40( m,4H),7.38(d,J=8.4Hz,2H),7.27(d,J=8.4Hz,2H),4.91(p,J=7.0Hz,1H),4.52(d,J=9.3Hz,1H),4.43(t,J=8.0Hz,1H),4.28(s,1H),4.07(d,J =6.6Hz,2H),3.65-3.58(m,2H),3.29(d,J=11.9Hz,2H),3.22(d,J=6.9H z,2H),2.91(t,J=11.1Hz,2H),2.45(s,3H),2.29-2.19(m,1H),2.18-2. C 56 H 68 N8O6S[M+H] + 980.50, found 981.56. UPLC retention time: 4.0 min, purity >95%.

[0211] Example 31

[0212] a, b: Compound 2 (850 mg, 2 mmol), 4-methylphenylboronic acid (272 mg, 2 mmol), and potassium carbonate (828 mg, 6 mmol) were dissolved in 25 mL of a mixed solution of 1,4-dioxane and water (volume ratio = 4:1), [1,1-bis(diphenylphosphine)ferrocene]palladium dichloride (290 mg, 0.2 mmol), and the reaction mixture was stirred overnight at 80 °C. After the reaction was completed, the reaction solution was cooled to room temperature and quenched with water. The solution was extracted three times with ethyl acetate (3 x 50 mL), washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by prep-HPLC to obtain a grayish-white solid. This solid was then dissolved in 15 mL of anhydrous dichloromethane, and trifluoroacetic acid (5 mL) was added. The reaction mixture was stirred at room temperature for 30 min. The solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a white solid, compound 17. Two-step yield: 75%.

[0213] c, d: Compound 17 (42 mg, 0.11 mmol) and compound 18 (27 mg, 0.11 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.09 mL, 0.5 mmol) and HATU (50 mg, 0.13 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1.5 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the white solid compound 19. Overall yield of the two steps: 70%.

[0214] e: Weigh compound 19 (43 mg, 0.08 mmol) and compound 4 (36 mg, 0.08 mmol) and dissolve them in 3 mL of DMF. Add DIPEA (0.07 mL, 0.4 mmol) and HATU (38 mg, 0.1 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 × 5 mL). Wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, and concentrate to obtain the crude product. Purify the crude product by prep-HPLC to obtain a white solid LD-100-ZJZ. Yield: 70%, 1H NMR (500MHz, DMSO-d6) δ8.98(s,1H),8.42(d,J=2.8Hz,1H),8.37(d,J=7.8Hz,1H),7.82-7.75(m,3H),7.46-7.35(m,7H),7.09(d,J= 8.2Hz,2H),7.05(d,J=8.1Hz,2H),4.92(p,J=7.1Hz,1H),4.52(d,J=9.3Hz,1H),4.45-4.39(m,2H),4.28(t,J=3.8Hz,1H),4.03(d,J =6.4Hz, 2H), 3.90(d, J = 13.4Hz, 1H), 3.64-3.57(m, 2H), 3.03(t, J = 12.6Hz, 1H), 2.62-2.54(m, 1H), 2.45(s, 3H), 2.28(d, J = 16.4Hz, 6H), 2.15-1.98(m, 3H), 1.86-1.75(m, 3H), 1.47(h, J = 8.5Hz, 4H), 1.37(d, J = 7.0Hz, 3H), 1.31-1.07(m, 9H), 0.93(s, 9H). LC-MS calculation yields C 57 H 69 N7O6S[M+H] + 979.50, found 980.65. UPLC retention time: 6.7 min, purity >95%.

[0215] Example 32

[0216] a, b: Compound 17 (42 mg, 0.11 mmol) and compound 20 (29 mg, 0.11 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.09 mL, 0.5 mmol) and HATU (50 mg, 0.13 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1.5 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the white solid compound 20. Overall yield of the two steps: 76%.

[0217] c: Compound 21 (47 mg, 0.08 mmol) and compound 4 (36 mg, 0.08 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.07 mL, 0.4 mmol) and HATU (38 mg, 0.1 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 × 5 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 give a white solid LD-101-ZJZ. Yield: 71%. 1 H NMR(500MHz,DMSO-d6)δ8.98(s,1H),8.42(d,J=2.7Hz,1H),8.37(d,J=7.9Hz,1H),7.82-7.75(m,3H),7.46-7.36(m,7H),7.1 2-7.03(m,4H),4.92(p,J=7.0Hz,1H),4.52(d,J=9.3Hz,1H),4.42(t,J=8.1Hz,2H),4.28(s,1H),4.03(d,J=6.6Hz,2H),3.90( d, J = 13.4 Hz, 1H), 3.63-3.56 (m, 2H), 3.03 (t, J = 11.7 Hz, 1H), 2.57 (t, J = 13.4 Hz, 1H), 2.45 (s, 3H), 2.27 (d, J = 14.5 Hz, 6H), 2.15-1.97 (m, 3H), 1.87-1.74 (m, 3H), 1.47 (q, J = 7.6 Hz, 4H), 1.37 (d, J = 7.0 Hz, 3H), 1.31-1.11 (m, 11H), 0.93 (s, 9H). LC-MS calculation yields C 58 H 71 N7O6S[M+H] +993.50, found 994.45. UPLC retention time: 6.8 min, purity >95%.

[0218] Example 33

[0219] a, b: Compound 17 (42 mg, 0.11 mmol) and compound 22 (30 mg, 0.11 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.09 mL, 0.5 mmol) and HATU (50 mg, 0.13 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1.5 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain a white solid, compound 23. Overall yield of the two steps: 70%.

[0220] c: Compound 23 (45 mg, 0.08 mmol) and compound 4 (36 mg, 0.08 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.07 mL, 0.4 mmol) and HATU (38 mg, 0.1 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 × 5 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 give a white solid LD-102-ZJZ. Yield: 69%. 1H NMR (500MHz, DMSO-d6) δ8.98(s,1H),8.42(d,J=2.7Hz,1H),8.37(d,J=7.9Hz,1H),7.88-7.69(m,3H),7.46-7.35(m,7H),7.09(d ,J=8.4Hz,2H),7.05(d,J=8.4Hz,2H),5.10(s,1H),4.92(p,J=6.9Hz,1H),4.52(d,J=9.5Hz,1H),4.42(t,J=8.1Hz,2H),4.28(s, 1H), 4.03(d,J=6.3Hz,2H), 3.90(d,J=13.4Hz,1H), 3.66-3.53(m,2H), 3.03(t,J=11.5Hz,1H), 2.45(s,3H), 2.27(d,J=15.1Hz,6H), 2.15-1.97(m,3H), 1.87-1.75(m,3H), 1.52-1.43(m,4H), 1.37(d,J=7.0Hz,3H), 1.30-1.09(m,13H), 0.93(s,9H). LC-MS calculation yielded C 59 H 73 N7O6S[M+H] + :1007.50, found 1008.35. UPLC retention time: 7.0 min, purity >95%.

[0221] Example 34

[0222] a, b: Compound 17 (42 mg, 0.11 mmol) and compound 24 (25 mg, 0.11 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.09 mL, 0.5 mmol) and HATU (50 mg, 0.13 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1.5 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the white solid compound 25. Overall yield of the two steps: 77%.

[0223] c: Compound 25 (46 mg, 0.08 mmol) and compound 4 (36 mg, 0.08 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.07 mL, 0.4 mmol) and HATU (38 mg, 0.1 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 × 5 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 give a white solid LD-103-ZJZ. Yield: 69%. 1 H NMR(500MHz,DMSO-d6)δ8.98(s,1H),8.42(d,J=2.9Hz,1H),8.37(d,J=7.8Hz,1H),7.79(d,J=10.2Hz,3H),7.46-7.36(m,7H),7 .09(d,J=8.1Hz,2H),7.05(d,J=8.2Hz,2H),5.10(s,1H),4.92(p,J=7.2Hz,1H),4.52(d,J=9.3Hz,1H),4.43(t,J=8.0Hz,2H),4. 28 (s, 1H), 4.03 (d, J = 6.4 Hz, 2H), 3.90 (d, J = 12.5 Hz, 1H), 3.60 (s, 2H), 3.04 (t, J = 11.8 Hz, 1H), 2.45 (s, 3H), 2.27 (d, J = 14.2 Hz, 6H), 2.16–1.97 (m, 3H), 1.88–1.74 (m, 3H), 1.53–1.43 (m, 4H), 1.37 (d, J = 7.0 Hz, 3H), 1.33–1.02 (m, 7H), 0.93 (s, 9H). LC-MS calculation yields C 56 H 67 N7O6S[M+H] + 965.49, found 966.55. UPLC retention time: 6.3 min, purity >95%.

[0224] Example 35

[0225] a, b: Compound 17 (42 mg, 0.11 mmol) and compound 26 (24 mg, 0.11 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.09 mL, 0.5 mmol) and HATU (50 mg, 0.13 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1.5 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the white solid compound 27. Overall yield of the two steps: 72%.

[0226] c: Compound 27 (42 mg, 0.08 mmol) and compound 4 (36 mg, 0.08 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.07 mL, 0.4 mmol) and HATU (38 mg, 0.1 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 × 5 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 give a white solid LD-104-ZJZ. Yield: 65%. 1 H NMR (500MHz, DMSO-d6) δ8.99(s,1H),8.43(d,J=2.9Hz,1H),8.37(d,J=7.9Hz,1H),7.79(d,J=8.5Hz,3H),7.46-7.36(m,7H),7 .10(d,J=8.2Hz,2H),7.05(d,J=8.4Hz,2H),4.92(p,J=6.9Hz,1H),4.52(d,J=9.3Hz,1H),4.43(t,J=8.0Hz,2H),4.29(s,1H),4 0.04 (d, J = 6.4 Hz, 2H), 3.90 (d, J = 13.3 Hz, 1H), 3.61 (d, J = 3.8 Hz, 3H), 3.04 (t, J = 11.6 Hz, 1H), 2.45 (s, 3H), 2.27 (d, J = 12.5 Hz, 6H), 2.17-1.97 (m, 3H), 1.86-1.75 (m, 3H), 1.54-1.45 (m, 4H), 1.37 (d, J = 7.0 Hz, 3H), 1.31-0.98 (m, 5H), 0.93 (s, 9H). LC-MS calculation yields C 55 H 65 N7O6S[M+H] +951.49, found 952.45. UPLC retention time: 6.0 min, purity >95%.

[0227] Example 36

[0228] a, b: Compound 17 (42 mg, 0.11 mmol) and compound 28 (24 mg, 0.11 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.09 mL, 0.5 mmol) and HATU (50 mg, 0.13 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1.5 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the white solid compound 27. Overall yield of the two steps: 72%.

[0229] c: Compound 29 (40 mg, 0.08 mmol) and compound 4 (36 mg, 0.08 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.07 mL, 0.4 mmol) and HATU (38 mg, 0.1 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 × 5 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 give a white solid LD-105-ZJZ. Yield: 68%. 1H NMR (500MHz, DMSO-d6) δ8.98(s,1H),8.42(d,J=2.9Hz,1H),8.37(d,J=7.8Hz,1H),7.83(d,J=9.3Hz,1H),7.79(d,J=8.4Hz,2H),7. 45-7.34(m,7H),7.12-7.02(m,4H),4.96-4.86(m,1H),4.52(d,J=9.3Hz,1H),4.43(t,J=8.1Hz,2H),4.29(s,1H),4.03(d,J=6.4Hz ,2H),3.87(d,J=13.3Hz,1H),3.61(s,2H),3.04(t,J=12.9Hz,1H),2.45(s,3H),2.28(d,J=19.1Hz,6H),2.10-1.97(m,2H),1.79(d C 53 H 61 N7O6S[M+H] + 923.44, found 924.45. UPLC retention time: 5.5 min, purity >95%.

[0230] Example 37

[0231] a, b: Compound 17 (42 mg, 0.11 mmol) and compound 30 (31 mg, 0.11 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.09 mL, 0.5 mmol) and HATU (50 mg, 0.13 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1.5 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the white solid compound 31. Overall yield of the two steps: 69%.

[0232] c: Compound 31 (46 mg, 0.08 mmol) and compound 4 (36 mg, 0.08 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.07 mL, 0.4 mmol) and HATU (38 mg, 0.1 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 × 5 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 give a white solid LD-106-ZJZ. Yield: 72%. 1 H NMR (500MHz, DMSO-d6) δ8.98(s,1H),8.42(d,J=2.7Hz,1H),8.37(d,J=7.8Hz,1H),7.78(t,J=8.9Hz,3H),7.45-7.34(m,7H),7.09(d ,J=8.2Hz,2H),7.05(d,J=8.2Hz,2H),5.09(d,J=3.5Hz,1H),4.91(t,J=7.4Hz,1H),4.51(d,J=9.5Hz,1H),4.42(t,J=8.0Hz,2H),4.2 7(s, 1H), 4.03(d, J = 6.4Hz, 2H), 3.90(d, J = 13.4Hz, 1H), 3.65-3.56(m, 2H), 3.03(t, J = 11.8Hz, 1H), 2.45(s, 3H), 2.27(d, J = 6.9Hz, 6H), 2.11-1.98(m, 3H), 1.81(dd, J = 22.1, 13.4Hz, 3H), 1.47(s, 4H), 1.37(d, J = 7.0Hz, 3H), 1.30-1.11(m, 15H), 0.93(s, 9H). LC-MS calculation yields C 60 H 75 N7O6 S[M+H] + :1022.55, found 1022.57. UPLC retention time: 7.9 min, purity >95%.

[0233] Example 38

[0234] a, b: Compound 4 (50 mg, 0.11 mmol) and compound 32 (23 mg, 0.11 mmol) were weighed and dissolved in 3 mL of LDMF. DIPEA (0.09 mL, 0.5 mmol) and HATU (50 mg, 0.13 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain a white solid, compound 33. Overall yield of the two steps: 70%.

[0235] c,d: Compound 33 (41 mg, 0.08 mmol) and compound 3 (42 mg, 0.08 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.07 mL, 0.4 mmol) and HATU (38 mg, 0.1 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the white solid compound LD-308-LLH. Overall yield of the two steps: 60%. LC-MS calculated CL... 52 H 60 N8O6S[M+H] + 924.44, found 924.60. UPLC retention time: 7.8 min, purity >95%.

[0236] Example 39

[0237] a, b: Compound 4 (50 mg, 0.11 mmol) and compound 34 (24 mg, 0.11 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.09 mL, 0.5 mmol) and HATU (50 mg, 0.13 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain a white solid, compound 35. Overall yield of the two steps: 75%.

[0238] c,d: Compound 35 (45 mg, 0.08 mmol) and compound 3 (42 mg, 0.08 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.07 mL, 0.4 mmol) and HATU (38 mg, 0.1 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the white solid compound LD-309-LLH. Overall yield of the two steps: 60%. 1H NMR(500MHz,DMSO-d6)δ8.98(s,1H),8.49-8.43(m,2H),8.38(d,J=7.9Hz,1H) ,7.81(t,J=8.0Hz,3H),7.72(d,J=8.4Hz,2H),7.50(d,J=2.7Hz,1H),7.46-7.4 0(m,4H),7.38(d,J=8.4Hz,2H),7.28(d,J=8.5Hz,2H),4.92(p,J=7.2Hz,1H),4 .51(d,J=9.3Hz,1H),4.43(t,J=8.0Hz,1H),4.28(s,1H),4.07(d,J=6.4Hz,2H) ,3.65-3.57(m,2H),3.29(d,J=11.7Hz,3H),3.23(d,J=6.1Hz,2H),2.90(t,J=11.6Hz,2H),2.45(s,3H),2.33-2.24(m,1H),2.14(dd,J=14.2,7.9Hz,2H),2.01(t,J=8.3Hz,1H),1.94(d,J=11.1Hz,2H),1.79(ddd,J=13.0,8.5,4.7Hz,1H),1.56-1.46(m,6H),1.37(d,J=7.0Hz,3H),1.23(s,1H),0.93(s,9H). LC-MS calculation yields C 53 H 62 N8O6S[M+H] + 938.45, found 939.47. UPLC retention time: 3.9 min, purity >95%.

[0239] Example 40

[0240] a, b: Compound 4 (50 mg, 0.11 mmol) and compound 36 (29 mg, 0.11 mmol) were weighed and dissolved in 3 mL of DMF. DIPEA (0.09 mL, 0.5 mmol) and HATU (50 mg, 0.13 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain a white solid, compound 37. Overall yield of the two steps: 70%.

[0241] c,d: Compound 70 (45 mg, 0.08 mmol) and compound 3 (42 mg, 0.08 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.07 mL, 0.4 mmol) and HATU (38 mg, 0.1 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 × 5 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 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the white solid compound LD-310-LLH. Overall yield of the two steps: 67%. LC-MS calculated CL... 56 H 68 N8O6S[M+H] + 980.50, found 981.68. UPLC retention time: 8.3 min, purity >95%.

[0242] Example 41

[0243] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 39 (68 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 40. Overall yield of the two steps: 40%.

[0244] c,d: Compound 40 (34 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a bright yellow chlorosol compound LD-110-ZJZ. Overall yield of the two steps: 67%. 1H NMR(500MHz,DMSO-d6)δ11.06(s,1H),8.54-8.43(m,2H),7.82(d,J=6.4Hz,2H),7.72(d,J=7.9Hz,2H), 7.59-7.37(m,4H),7.28(d,J=8.0Hz,2H),7.15(s,1H),6.95(s,1H),6.85(d,J=6.3Hz,1H),5.03(dd,J=1 2.7, 5.4 Hz, 1H), 4.08 (d, J = 6.3 Hz, 2H), 3.44 (d, J = 21.1 Hz, 2H), 3.20 (q, J = 5.9 Hz, 6H), 3.01-2.79 (m, 3H), 2.57 (dt, J = 19.4, 3.7 Hz, 1H), 2.14 (d, J = 13.8 Hz, 1H), 2.04-1.89 (m, 3H), 1.70-1.41 (m, 6H). LC-MS calculations yield C. 42 H 41 N7O6[M+H] + 739.31, found 740.45. UPLC retention time: 3.6 min, purity >95%.

[0245] Example 42

[0246] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 41 (84 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 40. Overall yield of the two steps: 42%.

[0247] c,d: Compound 42 (43 mg, 0.1 mmol) and compound 17 (38 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 give the yellow solid compound LD-107-ZJZ. Yield: 67%. 1H NMR (500MHz, DMSO-d6) δ11.06(s,1H),8.41(d,J=2.9Hz,1H),7.87-7.72(m,2H),7.55(d,J=8.4Hz,1H),7.46-7.37(m,3H),7.15(t,J=5. 6Hz,1H),7.11-7.03(m,4H),7.01(d,J=2.3Hz,1H),6.89(dd,J=8.4,2.1Hz,1H),5.03(dd,J=12.8,5.5Hz,1H),4.42(d,J=13.1Hz,1H),4. 01(d,J=6.4Hz,2H), 3.91(d,J=15.0Hz,1H), 3.66-3.58(m,4H), 3.57-3.50(m,4H), 3.37-3.34(m,2H), 3.02(t,J=12.9Hz,1H), 2.87(ddd,J=16.9,13.8,5.4Hz,1H), 2.56(dd,J=8.6,5.9Hz,4H), 2.26(s,3H), 2.09-1.95(m,2H), 1.86-1.74(m,2H), 1.32-1.03(m,3H). LC-MS calculation yields C 45 H 46 N6O8S[M+H] + :798.34, found 799.29. UPLC retention time: 7.5 min, purity >95%.

[0248] Example 43

[0249] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 43 (68 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 44. Overall yield of the two steps: 45%.

[0250] c,d: Compound 44 (39 mg, 0.1 mmol) and compound 17 (38 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 give the pale yellow solid compound LD-108-ZJZ. Yield: 75%. 1 H NMR (500MHz, DMSO-d6) δ11.06(s,1H),8.41(d,J=2.9Hz,1H),7.79(d,J=8.1Hz,2H),7.55(d,J=8.4Hz,1H),7.44-7.37(m,3H),7.14(t,J=5 .5Hz,1H),7.09(d,J=8.2Hz,2H),7.05(d,J=8.2Hz,2H),7.01(d,J=2.3Hz,1H),6.89(d,J=10.7Hz,1H),5.02(dd,J=12.9,5.4Hz,1H),4.43( d,J=13.1Hz,1H), 4.01(d,J=6.4Hz,2H), 3.91(d,J=14.8Hz,1H), 3.68(t,J=6.6Hz,2H), 3.60(t,J=5.5Hz,2H), 3.34(d,J=5.5Hz,4H), 3.08-2.98(m,1H), 2.93-2.80(m,1H), 2.63-2.53(m,4H), 2.26(s,3H), 2.10-1.94(m,2H), 1.80(t,J=11.1Hz,2H), 1.29-1.08(m,2H). LC-MS calculation yields C 43 H 42 N6O7[M+H] + 754.31, found 755.39. UPLC retention time: 6.8 min, purity >95%.

[0251] Example 44

[0252] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 45 (88 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 46. Overall yield of the two steps: 45%.

[0253] c,d: Compound 46 (40 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. It was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-109-ZJZ. Overall yield of the two steps: 67%. LC-MS calculated C0... 46 H 49 N7O6[M+H] + 795.37, found 796.41. UPLC retention time: 4.3 min, purity >95%.

[0254] Example 45

[0255] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 47 (73 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 48. Overall yield of the two steps: 42%.

[0256] c,d: Compound 48 (36 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. It was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-111-ZJZ. Overall yield of the two steps: 77%. LC-MS calculated C0... 43 H 43 N7O6[M+H] + 753.33, found 754.37. UPLC retention time: 3.6 min, purity >95%.

[0257] Example 46

[0258] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 49 (78 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 50. Overall yield of the two steps: 45%.

[0259] c,d: Compound 50 (37 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. It was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-112-ZJZ. Overall yield of the two steps: 70%. LC-MS calculated C0... 44 H45 N7O6[M+H] + 767.37, found 768.44. UPLC retention time: 3.8 min, purity >95%.

[0260] Example 47

[0261] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 51 (105 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 52. Overall yield of the two steps: 40%.

[0262] c,d: Compound 52 (45 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This crude product was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a yellow solid compound LD-113-ZJZ. Overall yield of the two steps: 78%. 1H NMR (500MHz, DMSO-d6) δ8.50(t,J=4.7Hz,1H),8.47-8.44(m,1H),7.81(d,J=8.5Hz,2H),7.72(d,J=8.4Hz,2H),7.56-7.46(m ,2H),7.42(dd,J=8.5,2.0Hz,2H),7.28(d,J=1.7Hz,2H),7.20-7.08(m,1H),6.98(d,J=17.1Hz,1H),6.90-6.79(m,1H),4.06( d,J=6.3Hz,2H),3.57(d,J=17.9Hz,2H),3.52(d,J=5.3Hz,8H),3.42-3.30(m,7H),3.25(d,J=12.5Hz,3H),2.90-2.81(m,2H) ,2.42-2.30(m,1H),2.27-2.19(m,1H),2.14-1.95(m,2H),1.91(d,J=10.4Hz,2H),1.54-1.42(m,2H),1.23(s,2H).LC-MS calculated C 46 H 49 N7O9[M+H] + 843.36, found 844.33. UPLC retention time: 5.8 min, purity >95%.

[0263] Example 48

[0264] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 53 (97 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 54. Overall yield of the two steps: 65%.

[0265] c,d: Compound 54 (43 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This crude product was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a yellow solid compound LD-117-ZJZ. Overall yield of the two steps: 70%. 1 H NMR (500MHz, DMSO-d6) δ11.09(s,1H),9.10(d,J=139.0Hz,2H),8.46(d,J=2.7Hz,1H),7.82(d,J= 8.2Hz,2H),7.67(d,J=8.5Hz,1H),7.51(d,J=2.7Hz,1H),7.44(d,J=8.2Hz,2H),7.39-7.20(m,6H ), 5.08 (dd, J = 12.7, 5.4 Hz, 1H), 4.12 (dd, J = 42.2, 9.2 Hz, 4H), 3.47 (s, 3H), 3.32-3.15 (m, 4H), 3.02-2.75 (m, 7H), 2.68-2.53 (m, 2H), 2.18-1.90 (m, 6H), 1.73-1.45 (m, 4H), 1.23 (s, 1H). LC-MS calculation yielded C 47 H 48 N8O6[M+H] + :820.37, found 821.41.UPLC retention time: 3.0 min, purity >95%.

[0266] Example 49

[0267] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 55 (97 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 56. Overall yield of the two steps: 68%.

[0268] c, d: Compound 56 (83 mg, 0.24 mmol) and 1-Boc-piperidine-4-carboxaldehyde (64 mg, 0.3 mmol) were weighed and dissolved in 10 mL of DCM / MeOH (volume ratio 1:1). The reaction mixture was stirred at room temperature for 30 min, and then NaBH3CN (25 mg, 0.4 mmol) was added. The reaction mixture was stirred overnight at room temperature. The reaction mixture 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 yellow 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 57, an orange-yellow solid. Overall yield of the two steps: 55%.

[0269] e,f: Compound 57 (44 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This crude product was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a yellow solid compound LD-118-ZJZ. Overall yield of the two steps: 70%. 1 H NMR (500MHz, DMSO-d6) δ11.09(s,1H),9.09(s,1H),8.79(s,1H),8.46(d,J=2.7Hz,1H),7.82(d,J=8.5 Hz,2H),7.71(d,J=7.9Hz,1H),7.50(d,J=2.7Hz,1H),7.44(d,J=8.4Hz,3H),7.26(s,5H),5.08(dd,J= 12.7 (5.4 Hz, 1H), 4.44 (s, 1H), 4.08 (d, J = 6.4 Hz, 2H), 3.82-3.41 (m, 5H), 3.33-3.17 (m, 4H), 3.10-2.67 (m, 7H), 2.63-2.54 (m, 2H), 2.25-1.64 (m, 8H), 1.61-1.47 (m, 2H), 1.16 (d, J = 29.3 Hz, 2H). LC-MS calculation yields C. 48 H 50 N8O6[M+H] +834.39, found 835.41. UPLC retention time: 3.2 min, purity >95%.

[0270] Example 50

[0271] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 58 (98 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 59. Overall yield of the two steps: 40%.

[0272] c,d: Compound 59 (42 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This crude product was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-128-ZJZ. Overall yield of the two steps: 70%. 1H NMR (500MHz, DMSO-d6) δ8.46 (s, 1H), 8.43 (t, J = 5.7Hz, 1H), 7.81 (d, J = 8.4Hz, 2H), 7.71 (d, J = 8.4Hz, 2H), 7.55 (d, J = 8. 4Hz,1H),7.49(d,J=2.9Hz,1H),7.42(d,J=8.2Hz,2H),7.27(d,J=8.4Hz,2H),7.12(t,J=5.3Hz,1H),6.94(s,1H),6.86 -6.79(m, 1H), 5.03(dd, J = 12.7, 5.5Hz, 1H), 4.07(d, J = 6.4Hz, 2H), 3.27(s, 2H), 3.22(d, J = 7.0Hz, 2H), 3.14(q, J = 6.9Hz, 2H), 2.97-2.82(m, 3H), 2.61-2.53(m, 2H), 2.15-1.89(m, 4H), 1.59-1.47(m, 6H), 1.28(d, J = 3.8Hz, 14H). LC-MS calculation yields C 48 H 53 N7O6[M+H] + 823.41, found 824.46. UPLC retention time: 4.7 min, purity >95%.

[0273] Example 51

[0274] a, b: Compound 60 (100 mg, 0.36 mmol) and compound 61 (68 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 62. Overall yield of the two steps: 45%.

[0275] c,d: Compound 62 (35 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This crude product was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a yellow solid compound LD-129-ZJZ. Overall yield of the two steps: 75%. 1 H NMR (500MHz, DMSO-d6) δ8.93(s,1H),8.49(dd,J=20.0,4.3Hz,2H),7.82(d,J=8.4Hz,2H),7.72(d,J=8.4Hz,2H),7. 59-7.47(m,2H),7.43(d,J=8.4Hz,2H),7.28(d,J=8.5Hz,2H),7.09(dd,J=22.8,8.6Hz,1H),7.04-6.94(m,1H),6.5 6 (dt, J = 17.7, 6.0 Hz, 1H), 5.05 (d, J = 18.3 Hz, 1H), 4.07 (d, J = 6.4 Hz, 2H), 3.36-3.23 (m, 7H), 2.95-2.74 (m, 3H), 2.64-2.54 (m, 1H), 2.43-2.21 (m, 1H), 2.18-2.07 (m, 1H), 2.06-1.88 (m, 3H), 1.66-1.47 (m, 6H), 1.23 (s, 1H). LC-MS calculation yields C. 42 H 41 N7O6[M+H] + 739.31, found 740.36. UPLC retention time: 3.9 min, purity >95%.

[0276] Example 52

[0277] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 63 (62 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 64. Overall yield of the two steps: 47%.

[0278] c,d: Compound 64 (33 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This crude product was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a yellow solid compound LD-130-ZJZ. Overall yield of the two steps: 70%. LC-MS calculated C0... 41 H 39 N7O6[M+H] + 725.30, found 726.44. UPLC retention time: 3.4 min, purity >95%.

[0279] Example 53

[0280] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 65 (81 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 66. Overall yield of the two steps: 67%.

[0281] c,d: Compound 66 (38 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. It was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-131-ZJZ. Overall yield of the two steps: 73%. LC-MS calculated C0... 45 H 43 N7O6[M+H] + 777.33, found 778.34. UPLC retention time: 3.3 min, purity >95%.

[0282] Example 54

[0283] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 67 (51 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 68. Overall yield of the two steps: 70%.

[0284] c,d: Compound 68 (40 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This crude product was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a yellow solid compound LD-132-ZJZ. Overall yield of the two steps: 77%. 1H NMR(500MHz,DMSO-d6)δ11.08(s,1H),8.72(s,1H),8.47(s,1H),8.41(s,1H),7.82(d,J=6.6Hz,2H),7.68 (d,J=8.2Hz,1H),7.50(d,J=2.9Hz,1H),7.35-7.23(m,4H),6.84(d,J=2.3Hz,1H),6.70(d,J=8.4Hz,1H), 5.07 (dd, J = 12.7, 5.5 Hz, 1H), 4.21 (s, 2H), 4.08 (d, J = 6.4 Hz, 4H), 3.49 (s, 5H), 3.34 (d, J = 13.0 Hz, 3H), 3.01-2.52 (m, 8H), 2.13 (s, 1H), 2.05-1.91 (m, 3H), 1.50 (t, J = 13.0 Hz, 2H), 1.24 (d, J = 7.9 Hz, 1H). LC-MS calculation yields C. 45 H 44 N8O6[M+H] + 792.36, found 793.39. UPLC retention time: 3.5 min, purity >95%.

[0285] Example 55

[0286] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 69 (77 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 70. Overall yield of the two steps: 40%.

[0287] c,d: Compound 70 (37 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-133-ZJZ. Overall yield of the two steps: 72%. LC-MS calculated C0... 44 H 43 N7O6[M+H] + 765.33, found 766.37. UPLC retention time: 3.5 min, purity >95%.

[0288] Example 56

[0289] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 71 (82 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 72. Overall yield of the two steps: 60%.

[0290] c,d: Compound 72 (39 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-134-ZJZ. Overall yield of the two steps: 75%. LC-MS calculated C0... 45 H45 N7O6[M+H] + 779.34, found 780.37. UPLC retention time: 3.5 min, purity >95%.

[0291] Example 57

[0292] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 73 (83 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 74. Overall yield of the two steps: 62%.

[0293] c,d: Compound 74 (39 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-135-ZJZ. Overall yield of the two steps: 70%. LC-MS calculated C0... 44 H 44 N8O6[M+H] + 780.34, found 781.37. UPLC retention time: 3.6 min, purity >95%.

[0294] Example 58

[0295] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 75 (82 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 76. Overall yield of the two steps: 43%.

[0296] c,d: Compound 76 (39 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. It was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-136-ZJZ. Overall yield of the two steps: 73%. LC-MS calculated C0... 46 H 46 N7O6[M+H] + 779.34, found 780.39. UPLC retention time: 3.5 min, purity >95%.

[0297] Example 59

[0298] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 77 (72 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 77. Overall yield of the two steps: 46%.

[0299] c,d: Compound 78 (36 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. It was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-137-ZJZ. Overall yield of the two steps: 60%. LC-MS calculated C0... 43 H 41 N7O6[M+H] + 751.31, found 752.39. UPLC retention time: 3.3 min, purity >95%.

[0300] Example 60

[0301] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 79 (77 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 80. Overall yield of the two steps: 43%.

[0302] c,d: Compound 80 (37 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-138-ZJZ. Overall yield of the two steps: 66%. LC-MS calculated C0... 44 H43 N7O6[M+H] + 765.33, found 766.41. UPLC retention time: 3.5 min, purity >95%.

[0303] Example 61

[0304] a, b: Compound 81 (106 mg, 0.36 mmol) and compound 82 (68 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 83. Overall yield of the two steps: 50%.

[0305] c,d: Compound 83 (36 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-139-ZJZ. Overall yield of the two steps: 69%. LC-MS calculated C0... 42 H 40 FN7O6[M+H] + 757.30, found 758.42. UPLC retention time: 3.7 min, purity >95%.

[0306] Example 62

[0307] a, b: Compound 84 (290 mg, 1 mmol) and compound 85 (260 mg, 1.3 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 overnight at room temperature. 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 86, a white solid. Overall yield of the two steps: 20%.

[0308] c,d: Compound 86 (38 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-140-ZJZ. Overall yield of the two steps: 69%. LC-MS calculated C0... 45 H 43 N7O6[M+H] + 777.33, found 778.40. UPLC retention time: 2.7 min, purity >95%.

[0309] Example 63

[0310] a, b: Compound 84 (290 mg, 1 mmol) and compound 87 (277 mg, 1.3 mmol) were weighed and dissolved in 10 mL of DCM / MeOH (1:1 v / v). The reaction mixture was stirred at room temperature for 30 min, and then NaBH3CN (60 mg, 2 mmol) was added. The reaction mixture was stirred overnight at room temperature. The reaction mixture 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 mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 88, a white solid. Overall yield of the two steps: 22%.

[0311] c,d: Compound 88 (40 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-141-ZJZ. Overall yield of the two steps: 66%. LC-MS calculated C0... 46 H 45 N7O6[M+H] + 791.34, found 792.40. UPLC retention time: 2.8 min, purity >95%.

[0312] Example 64

[0313] a, b: Compound 84 (290 mg, 1 mmol) and compound 89 (243 mg, 1.3 mmol) were weighed and dissolved in 10 mL of DCM / MeOH (1:1 v / v). The reaction mixture was stirred at room temperature for 30 min, and then NaBH3CN (60 mg, 2 mmol) was added. The reaction mixture was stirred overnight at room temperature. The reaction mixture 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 mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 90, a white solid. Overall yield of the two steps: 15%.

[0314] c,d: Compound 90 (40 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-142-ZJZ. Overall yield of the two steps: 72%. LC-MS calculated C0... 44 H 43 N7O6[M+H] + 765.33, found 766.40. UPLC retention time: 2.9 min, purity >95%.

[0315] Example 65

[0316] a, b: Compound 91 (104 mg, 0.36 mmol) and compound 92 (68 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 93. Overall yield of the two steps: 45%.

[0317] c,d: Compound 93 (36 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This crude product was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-146-ZJZ. Overall yield of the two steps: 77%. 1H NMR(500MHz,DMSO-d6)δ8.53-8.40(m,2H),7.81(d,J=8.2Hz,2H),7.71(d,J=8.4Hz,2H),7.55(d,J=8.2Hz,1H),7 .49(d,J=2.7Hz,1H),7.42(d,J=8.4Hz,2H),7.28(d,J=10.5Hz,2H),7.11(d,J=46.1Hz,1H),6.94(d,J=18.8Hz,1 6.89-6.77 (m, 1H), 5.10 (dd, J = 13.1, 5.5 Hz, 1H), 4.06 (d, J = 6.6 Hz, 2H), 3.32-3.15 (m, 6H), 3.00 (s, 3H), 2.88-2.62 (m, 3H), 2.55 (d, J = 13.0 Hz, 1H), 2.21-1.71 (m, 5H), 1.67-1.53 ​​(m, 4H), 1.47 (s, 2H), 1.23 (s, 1H). LC-MS calculation yielded C 43 H 43 N7O6[M+H] + 753.33, found 753.37. UPLC retention time: 4.0 min, purity >95%.

[0318] Example 66

[0319] a, b: Compound 84 (290 mg, 1 mmol) and compound 94 (225 mg, 1.3 mmol) were weighed and dissolved in 10 mL of DCM / MeOH (1:1 v / v). The reaction mixture was stirred at room temperature for 30 min, and then NaBH3CN (60 mg, 2 mmol) was added. The reaction mixture was stirred overnight at room temperature. The reaction mixture 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 mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain compound 95, a white solid. Overall yield of the two steps: 16%.

[0320] c,d: Compound 95 (36 mg, 0.1 mmol) and compound 17 (38 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. It was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-155-ZJZ. Overall yield of the two steps: 78%. LC-MS calculated C0... 44 H 42 N6O6[M+H] + 750.32, found 751.36. UPLC retention time: 3.9 min, purity >95%.

[0321] Example 67

[0322] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 96 (57 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 97. Overall yield of the two steps: 43%.

[0323] c,d: Compound 97 (32 mg, 0.1 mmol) and Compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This crude product was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a yellow solid compound LD-157-ZJZ. Overall yield of the two steps: 66%. 1H NMR (500MHz, DMSO-d6) δ11.06(s,1H),9.17(s,1H),8.88(s,1H),8.75(t,J=5.6Hz,1H),8.50(d,J=2.9Hz,1H),7.80( dd,J=24.4,8.5Hz,4H),7.62-7.54(m,2H),7.43(d,J=8.5Hz,2H),7.30(d,J=8.4Hz,2H),7.04(d,J=2.1Hz,1H),6.92 (dd,J=8.4,2.1Hz,1H), 5.03(dd,J=12.7,5.4Hz,1H), 4.10(d,J=6.6Hz,2H), 3.49-3.25(m,6H), 2.96-2.83(m,3H), 2.57(d,J=18.5Hz,1H), 2.14(s,1H), 1.97(dd,J=24.0,11.7Hz,3H), 1.61-1.49(m,2H), 1.34-1.21(m,1H). LC-MS calculation yields C 40 H 37 N7O6[M+H] + 711.28, found 7121.32. UPLC retention time: 3.2 min, purity >95%.

[0324] Example 68

[0325] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 98 (83 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 99. Overall yield of the two steps: 40%.

[0326] c,d: Compound 99 (39 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. It was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-158-ZJZ. Overall yield of the two steps: 71%. LC-MS calculated C0... 45 H 47 N7O6[M+H] + 781.36, found 782.39. UPLC retention time: 3.9 min, purity >95%.

[0327] Example 69

[0328] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 100 (82 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 101. Overall yield of the two steps: 42%.

[0329] c,d: Compound 101 (39 mg, 0.1 mmol) and compound 3 (51 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This crude product was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a yellow solid compound LD-159-ZJZ. Overall yield of the two steps: 73%. 1H NMR (500MHz, DMSO-d6) δ8.47(d,J=2.7Hz,1H),8.22(d,J=7.9Hz,1H),7.82(d,J=8.5Hz,2H),7.73(d,J=8.5Hz,2H),7.58-7 .47(m,2H),7.43(d,J=8.5Hz,2H),7.30-7.11(m,3H),6.96(d,J=22.3Hz,1H),6.86(dd,J=15.3,9.5Hz,1H),5.03(dd,J=12. 7, 5.4Hz, 1H), 4.07(d, J = 6.4Hz, 2H), 3.75(d, J = 4.0Hz, 1H), 3.50-3.23(m, 5H), 3.05(d, J = 6.3Hz, 2H), 2.93-2.75(m, 3H), 2.58(d, J = 15.3Hz, 1H), 2.13-1.84(m, 8H), 1.49(q, J = 11.8Hz, 3H), 1.34(q, J = 11.8Hz, 2H), 1.10(q, J = 11.8Hz, 2H). LC-MS calculation yields C 45 H 45 N7O6[M+H] + 779.36, found 779.37. UPLC retention time: 3.6 min, purity >95%.

[0330] Example 70

[0331] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 102 (52 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1.5 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 103. Overall yield of the two steps: 45%.

[0332] c, d: Compound 103 (36 mg, 0.1 mmol) and compound 17 (38 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This crude product was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a yellow solid compound LD-301-LLH. Overall yield of the two steps: 75%. 1 H NMR (500MHz, DMSO-d6) δ11.06(s,1H),8.41(d,J=2.7Hz,1H),7.79(d,J=8.3Hz,21H),7.56(d,J=8.4Hz,1H),7.41(dd,J=8.4,5.6Hz,3H),7.14 (t,J=5.6Hz,1H),7.07(dd,J=21.1,8.1Hz,4H),6.97(d,J=1.4Hz,1H),6.87(dt,J=13.2,6.6Hz,1H),5.03(dd,J=12.8,5.4Hz,1H),4.45(d,J=1 2.8Hz, 1H), 4.01(d, J = 6.3Hz, 2H), 3.88(d, J = 13.1Hz, 1H), 3.52-3.39(m, 2H), 3.04(t, J = 12.0Hz, 1H), 2.87(ddd, J = 16.9, 14.0, 5.2Hz, 1H), 2.63(dd, J = 13.4, 7.2Hz, 3H), 2.25(s, 3H), 2.05(dd, J = 12.4, 8.9Hz, 1H), 2.03-1.95(m, 1H), 1.80(d, J = 12.0Hz, 2H), 1.33-1.08(m, 4H). LC-MS calculation yields C 41 H 38 N6O6[M+H] + 710.29, found 711.34. UPLC retention time: 6.0 min, purity >95%.

[0333] Example 71

[0334] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 104 (62 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1.5 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 105. Overall yield of the two steps: 48%.

[0335] c,d: Compound 105 (37 mg, 0.1 mmol) and compound 17 (38 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This crude product was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a yellow solid compound LD-302-LLH. Overall yield of the two steps: 75%. 1H NMR (500MHz, DMSO-d6) δ11.05(s,1H),8.41(d,J=2.7Hz,1H),7.79(d,J=8.3Hz,2H),7.55(d,J=8.4Hz,1H),7.41(dd,J=9.0,5.5Hz,3H),7.13(t ,J=5.2Hz,1H),7.07(dd,J=21.1,8.1Hz,4H),6.95(s,1H),6.85(dd,J=8.4,1.6Hz,1H),5.02(dd,J=12.8,5.4Hz,1H),4.43(d,J=12.9Hz,1H),4 0.02 (d, J = 6.3 Hz, 2H), 3.92 (d, J = 13.1 Hz, 1H), 3.18 (d, J = 4.9 Hz, 2H), 3.03 (t, J = 12.0 Hz, 1H), 2.87 (ddd, J = 16.7, 13.8, 5.1 Hz, 1H), 2.66-2.54 (m, 2H), 2.37 (s, 2H), 2.26 (s, 3H), 2.13-1.93 (m, 2H), 1.91-1.73 (m, 2H), 1.60 (s, 4H), 1.33-1.16 (m, 2H), 1.13 (qd, J = 11.9, 8.0 Hz, 1H). LC-MS calculation yields C 43 H 42 N6O6[M+H] + 738.32, found 739.46. UPLC retention time: 6.2 min, purity >95%.

[0336] Example 72

[0337] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 106 (72 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1.5 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 107. Overall yield of the two steps: 43%.

[0338] c,d: Compound 107 (40 mg, 0.1 mmol) and compound 17 (38 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This crude product was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a yellow solid compound LD-303-LLH. Overall yield of the two steps: 75%. 1 H NMR (500MHz, DMSO-d6) δ11.03(s,1H),8.41(d,J=2.7Hz,1H),7.78(d,J=8.3Hz,2H),7.55(d,J=8.4Hz,1H),7.40(dd,J=8.0,5.6Hz,3H),7.15-7 .01(m,5H),6.94(s,1H),6.87-6.80(m,1H),5.03(dd,J=12.8,5.4Hz,1H),4.42(d,J=12.8Hz,1H),4.01(d,J=6.3Hz,2H),3.89(d,J=13.1Hz,1H ), 3.18-3.10(m,2H), 3.02(t,J=12.1Hz,1H), 2.87(ddd,J=17.2,14.1,5.3Hz,1H), 2.62-2.52(m,2H), 2.30(t,J=7.4Hz,2H), 2.25(s,3H), 2.09-1.93(m,2H), 1.80(dd,J=20.9,13.5Hz,2H), 1.62-1.45(m,4H), 1.44-1.29(m,4H), 1.27-1.17(m,2H), 1.12(dt,J=11.9,9.3Hz,1H). LC-MS calculation yields C 45 H 46 N6O6[M+H] + 766.35, found 767.30. UPLC retention time: 6.6 min, purity >95%.

[0339] Example 73

[0340] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 108 (78 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1.5 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 109. Overall yield of the two steps: 40%.

[0341] c, d: Compound 109 (42 mg, 0.1 mmol) and compound 17 (38 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This crude product was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-304-LLH. Overall yield of the two steps: 70%. 1H NMR (500MHz, DMSO-d6) δ11.05(s,1H),8.41(d,J=2.2Hz,1H),7.79(d,J=8.0Hz,2H),7.55(d,J=8.3Hz,1H),7.48-7.37(m,3H),7.07(dd,J=20 .8,8.0Hz,5H),6.94(s,1H),6.83(d,J=8.0Hz,1H),5.02(dd,J=12.7,5.3Hz,1H),4.42(d,J=12.4Hz,1H),4.02(d,J=6.0Hz,2H),3.89(d,J=12 .8Hz,1H),3.14(d,J=5.6Hz,2H),3.02(t,J=12.2Hz,1H),2.88(dd,J=22.1,8.9Hz,1H),2.55(dd,J=17.0,12.4Hz,2H),2.29(t,J=7.3Hz,2H) C 46 H 48 N6O6[M+H] + 780.36, found 781.57. UPLC retention time: 6.8 min, purity >95%.

[0342] Example 74

[0343] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 110 (83 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1.5 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 111. Overall yield of the two steps: 44%.

[0344] c,d: Compound 111 (43 mg, 0.1 mmol) and compound 17 (38 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This crude product was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a yellow solid compound LD-305-LLH. Overall yield of the two steps: 76%. 1 H NMR (500MHz, DMSO-d6) δ11.06(s,1H),8.40(d,J=2.8Hz,1H),7.78(d,J=8.4Hz,2H),7.54(d,J=8.4Hz,1H),7.40(dd,J=7.7,5.6Hz,3H),7.13-7.01(m ,5H),6.93(s,1H),6.83(dd,J=8.4,1.7Hz,1H),5.03(dd,J=12.8,5.4Hz,1 H),4.42(d,J=12.8Hz,1H),4.01(d,J=6.3Hz,2H),3.88(d,J=13.2Hz,1H),3 .13(dd, J = 12.4, 6.6 Hz, 2H), 3.02(t, J = 12.0 Hz, 1H), 2.87(ddd, J = 17.3, 14.1, 5.4 Hz, 1H), 2.64-2.51(m, 2H), 2.28(t, J = 7.5 Hz, 2H), 2.25(s, 3H), 2.12-1.93(m, 2H), 1.80(dd, J = 21.9, 12.6 Hz, 2H), 1.63-1.51(m, 2H), 1.47(d, J = 6.2 Hz, 2H), 1.37-1.18(m, 10H), 1.11(tt, J = 23.5, 11.9 Hz, 1H). LC-MS calculation yields C 47 H 50 N6O6[M+H] + 794.36, found 795.52. UPLC retention time: 7.8 min, purity >95%.

[0345] Example 75

[0346] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 112 (87 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1.5 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 113. Overall yield of the two steps: 40%.

[0347] c,d: Compound 113 (44 mg, 0.1 mmol) and compound 17 (38 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-306-LLH. Overall yield of the two steps: 65%. LC-MS calculated CL... 48 H 52 N6O6[M+H] + 808.39, found 809.55. UPLC retention time: 8.5 min, purity >95%.

[0348] Example 76

[0349] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 114 (93 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1.5 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 115. Overall yield of the two steps: 40%.

[0350] c,d: Compound 115 (46 mg, 0.1 mmol) and compound 17 (38 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. It was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-307-LLH. Overall yield of the two steps: 65%. LC-MS calculated CL... 49 H 54 N6O6[M+H] + :822.41, found 823.57.UPLC retention time: 8.7 min, purity >95%.

[0351] Synthesis of compounds 118-122:

[0352] Synthesis of compound 116: same as compound 1 and compound 3.

[0353] Synthesis of Compound 117: Compound 116 (1.2 mmol) and DPPBE (1.3 nmol, 0.5 M / THF) were added sequentially to a solution of tert-butyl 6-(hydroxymethyl)-2-azaspiro[3.3]heptane-2-carboxylic acid (1 mmol) in 3 mL of THF. A solution of DIAD (1.3 mmol) in 0.5 mL of THF was added dropwise at room temperature, and the reaction was allowed to proceed for 15 minutes. After the reaction was complete, the mixture was purified by preparative liquid chromatography. Mobile phase: methanol / water = 60 / 40. Column type: Prep C18 OBDTM 5μm, 50×100mm Column, to obtain white solid 117.

[0354] Synthesis of Compound 118: Compound 117 was dissolved in DCM, and TFA was added. After the reaction was complete, preparative liquid chromatography was performed for purification. The mobile phase was methanol / water = 10 / 90. Column type: Prep C18 OBDTM 5μm, 50×100mm Column, to obtain white solid 118.

[0355] Synthesis of compounds 119 and 120: same as that of compound 117.

[0356] Synthesis of compounds 122 and 124:

[0357] Synthesis of compound 122: Compound 38 and compound 121 were weighed and dissolved in 5 mL of DMF. DIPEA was added, and the reaction system was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. Then, it was dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1.5 mL) was added. The reaction system was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 122.

[0358] Synthesis of compound 124: same as compound 122.

[0359] Example 77

[0360] Compounds 119 and 122 were weighed and dissolved in 3 mL of DMF solution. DIPEA and HATU 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 × 5 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 LD-311-LLH, and C43H was calculated by LC-MS. 40 N6O6[M+H] + 736.30, found 737.48. UPLC retention time: 8.4 min, purity >95%.

[0361] Example 78

[0362] Compounds 118 and 122 were weighed and dissolved in 3 mL of DMF solution. DIPEA and HATU 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 × 5 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 LD-312-LLH. The CL was calculated by LC-MS. 41 H 36 N6O6[M+H] + 708.27, found 709.43. UPLC retention time: 8.5 min, purity >95%.

[0363] Example 79

[0364] Compound 120 and compound 122 were weighed and dissolved in 3 mL of DMF solution. DIPEA and HATU 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 × 5 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 LD-313-LLH. The C64 concentration was calculated by LC-MS. 42 H 38 N6O6[M+H] + 722.29, found 723.47. UPLC retention time: 8.6 min, purity >95%.

[0365] Example 80

[0366] Compounds 118 and 124 were weighed and dissolved in 3 mL of DMF solution. DIPEA and HATU 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 × 5 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 LD-314-LLH. The CL content was calculated by LC-MS. 42 H 38 N6O6[M+H] + 722.29, found 723.49. UPLC retention time: 8.6 min, purity >95%.

[0367] Example 81

[0368] Compounds 120 and 124 were weighed and dissolved in 3 mL of DMF solution. DIPEA and HATU 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 × 5 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 LD-315-LLH.

[0369] Synthesis of compound 126:

[0370] Synthesis of compound 125: same as compound 1 and compound 3.

[0371] Synthesis of Compound 126: Compound 125 (1.2 mmol) and DPPBE (1.3 nmol, 0.5 M / THF) were added sequentially to a solution of tert-butyl 2-(hydroxymethyl)-7-azaspiro[3.5]nonane-7-carboxylic acid (1 mmol) in 3 mL of THF. A solution of DIAD (1.3 mmol) in 0.5 mL of THF was added dropwise at room temperature, and the reaction was allowed to proceed for 15 minutes. After the reaction, the mixture was purified by preparative liquid chromatography using a mobile phase of methanol / water = 60 / 40. Column type: Prep C18 OBDTM 5μm, 50×100mm Column, yields 126 white solids.

[0372] Synthesis of compound 127: same as compound 126.

[0373] Example 82

[0374] Compound 126 and compound 40 were weighed and dissolved in 3 mL of DMF solution. DIPEA and HATU 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 × 5 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 yellow solid, 128. This was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a yellow solid, compound LD-316-LLH. LC-MS calculated the CL... 45 H 45 N7O6[M+H] + 779.34, found 780.50. UPLC retention time: 5.6 min, purity >95%.

[0375] Example 83

[0376] Compound 127 and compound 40 were weighed and dissolved in 3 mL of DMF solution. DIPEA and HATU 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 × 5 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 yellow solid, 129. This was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a yellow solid, compound LD-317-LLH. LC-MS calculated the CL... 44 H43 N7O6[M+H] + 765.33, found 766.49. UPLC retention time: 5.8 min, purity >95%.

[0377] Synthesis of compound 131:

[0378] Step 1: Under nitrogen protection at T = 25°C, 5-bromo-2-(2,6-dioxadiazine-3-yl)isoindol-1,3-dione (0.50 g, 1.48 mmol), tert-butyl penta-4-yn-1-ylcarbamate (0.27 g, 1.48 mmol), cuprous oxide (56 mg, 0.30 mmol), bis(triphenylphosphine) dichloride (104 mg, 0.15 mmol), and triethylamine (0.45 g, 4.45 mmol) were added to a round-bottom flask, followed by the addition of N,N-dimethylformamide (5 mL) and purging with nitrogen. The reaction was heated to T = 80°C and stirred for 8.0 h under nitrogen protection; the reaction was confirmed by LC-MS; the reaction was quenched by the addition of ammonium chloride aqueous solution (10 mL). The organic phase was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The concentrated crude product was purified by column chromatography (eluent: dichloromethane / methanol = 30 / 1) to give compound 130 (yellow solid, 0.47 g, yield: 66%).

[0379] Step 2: Dissolve compound 130 in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and purify the crude product by prep-HPLC to obtain yellow solid compound 131.

[0380] Synthesis of compound 133:

[0381] Same as compound 131.

[0382] Example 84

[0383] Compound 3 and compound 131a were weighed and dissolved in 3 mL of DMF solution. DIPEA and HATU 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 × 5 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 yellow solid 134. This was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a yellow solid compound LD-319-LLH. LC-MS calculated the CL... 43 H 38 N6O6[M+H] + 734.29, found 735.47. UPLC retention time: 5.6 min, purity >95%.

[0384] Example 85

[0385] Compound 3 and compound 131b were weighed and dissolved in 3 mL of DMF solution. DIPEA and HATU 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 × 5 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 yellow solid 135. This was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a yellow solid compound LD-320-LLH. The CL ratio was calculated by LC-MS. 44 H 40 N6O6[M+H] + 748.30, found 749.48. UPLC retention time: 5.9 min, purity >95%.

[0386] Example 86

[0387] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 136 (57 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1.5 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 137. Overall yield of the two steps: 40%.

[0388] c,d: Compound 137 (36 mg, 0.1 mmol) and compound 17 (38 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This crude product was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-318-LLH. Overall yield of the two steps: 65%. 1 H NMR (500MHz, DMSO-d6) δ11.06(s,1H),8.41(d,J=2.8Hz,1H),7.86-7.75(m,2H),7.56(d,J=8.4Hz,1H),7.46-7.35(m,3H),7.17(t,J=5.4H z,1H),7.07(dd,J=22.2,8.1Hz,4H),6.96(d,J=1.8Hz,1H),6.86(dd,J=8.4,2.0Hz,1H),5.02(dd,J=12.8,5.4Hz,1H),4.45(d,J=12.9Hz, 1H), 4.01(d,J=6.4Hz,2H), 3.91(d,J=13.2Hz,1H), 3.19(dd,J=13.1,6.4Hz,2H), 3.04(t,J=11.9Hz,1H), 2.86(ddd,J=17.2,14.1,5.4Hz,1H), 2.66-2.53(m,2H), 2.44(t,J=7.2Hz,2H), 2.25(s,3H), 2.13-1.93(m,2H), 1.81(dd,J=14.1,7.4Hz,4H), 1.32-1.03(m,3H). LC-MS calculation yields C 42 H 40 N6O6[M+H] + 724.30, found 725.37. UPLC retention time: 7.2 min, purity >95%.

[0389] Example 87

[0390] a, b: Compound 38 (100 mg, 0.36 mmol) and compound 138 (62 mg, 0.36 mmol) were weighed and dissolved in 5 mL of DMF. DIPEA (0.31 mL, 1.8 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to obtain a green solid. This solid was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1.5 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain the green solid compound 139. Overall yield of the two steps: 40%.

[0391] c,d: Compound 139 (39 mg, 0.1 mmol) and compound 17 (38 mg, 0.1 mmol) were weighed and dissolved in 3 mL of DMF solution. DIPEA (0.09 mL, 0.5 mmol) and HATU (46 mg, 0.12 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 × 5 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 yellow solid. This crude product was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain the yellow solid compound LD-323-LLH. Overall yield of the two steps: 65%. 1H NMR (500MHz, DMSO-d6) δ11.05(s,1H),8.41(d,J=2.8Hz,1H),7.79(d,J=8.3Hz,2H),7.55(d,J=8.4Hz,1H),7.41(dd,J=9.0,5.5Hz,3H),7.14-7.02(m ,5H),6.94(s,1H),6.84(dd,J=8.4,1.6Hz,1H),5.02(dd,J=12.8,5.4Hz,1 H),4.43(d,J=12.9Hz,1H),4.02(d,J=6.3Hz,2H),3.90(d,J=13.0Hz,1H),3 .15(dd, J = 12.4, 6.6 Hz, 2H), 3.02(t, J = 12.0 Hz, 1H), 2.87(ddd, J = 17.0, 14.0, 5.3 Hz, 1H), 2.59-2.51(m, 2H), 2.32(t, J = 7.4 Hz, 2H), 2.25(s, 3H), 2.01(ddd, J = 12.6, 9.9, 4.5 Hz, 2H), 1.90-1.71(m, 2H), 1.66-1.49(m, 4H), 1.44-1.33(m, 2H), 1.29-1.17(m, 2H), 1.12(dt, J = 11.6, 8.6 Hz, 1H). LC-MS calculation yields C 44 H 44 N6O6[M+H] + 752.33, found 753.25. UPLC retention time: 7.6 min, purity >95%.

[0392] Example 88

[0393] Compound 3 and compound 133a were weighed and dissolved in 3 mL of DMF solution. DIPEA and HATU 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 × 5 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 yellow solid 140. This was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a yellow solid compound LD-321-LLH. The CL ratio was calculated by LC-MS. 43 H 40 N6O5[M+H] + 720.31, found 721.47. UPLC retention time: 5.5 min, purity >95%.

[0394] Example 89

[0395] Compound 3 and compound 133b were weighed and dissolved in 3 mL of DMF solution. DIPEA and HATU 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 × 5 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 yellow solid 141. This was then dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation. The crude product was purified by prep-HPLC to obtain a yellow solid compound LD-322-LLH. LC-MS calculated the CL... 44 H 40 N6O6[M+H] + 748.30, found 749.46. UPLC retention time: 5.8 min, purity >95%.

[0396] Synthesis of Equation 1:

[0397] Step 1: 2-Chloro-3-bromo-5-hydroxypyridine (5 g, 24.0 mmol) was dissolved in 50 mL of a mixed solution of 1,4-dioxane and water (v:v = 4:1). 4-Cyanobenzoic acid (0.735 g, 24.0 mmol), tetrakis(triphenylphosphine)palladium (2.77 g, 2.4 mmol), and potassium carbonate (6.63 g, 48.0 mmol) were added sequentially. The reaction mixture was stirred at 90 °C for 5 h. The reaction solution was cooled to room temperature and then quenched with water (100 mL). Extraction was performed three times with ethyl acetate (3 × 50 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The residue was separated by silica gel column chromatography to obtain a white solid, intermediate 1 (4.2 g, yield: 75.9%).

[0398] Step 2: Intermediate 1 (4.00 g, 17.3 mmol) was dissolved in 40 mL of DMF solution, and tert-butyl 4-(bromomethyl)piperidine-1-carboxylate (4.82 g, 17.3 mmol) and potassium carbonate (4.79 g, 34.7 mmol) were added sequentially. The reaction system was stirred overnight at 100 °C. After the reaction was completed, the reaction solution was cooled to room temperature, water was added to precipitate the solid, and the filter cake was separated by silica gel column chromatography to obtain white intermediate 2 (6.02 g, yield: 81.1%).

[0399] Step 3: Intermediate 2 (6 g, 14.0 mmol), 4-methoxycarbonylphenylboronic acid (5.05 g, 28.0 mmol) were dissolved in 80 mL of a mixed solution of 1,4-dioxane and water (v:v = 4:1), [1,1-bis(diphenylphosphine)ferrocene]palladium dichloride (2.05 g, 2.8 mmol), and potassium phosphate (8.93 g, 42.1 mmol). The reaction system was stirred overnight at 80 °C. After the reaction was completed, the reaction solution was cooled to room temperature and quenched with water. It was extracted three times with ethyl acetate (3 × 50 mL), washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the residue was separated by silica gel column chromatography to obtain a gray solid, namely intermediate 3 (3.6 g, yield: 48.7%).

[0400] Step 4: Dissolve intermediate 3 (3.6 g, 6.8 mmol) in 20 mL of THF solution and add NaOH (2.5 M, 5 mL). Stir the reaction mixture overnight at 60 °C. After the reaction is complete, cool the reaction solution to room temperature, adjust the pH to 2 with dilute hydrochloric acid, extract three times with acetic acid (3 × 50 mL), wash with saturated brine, dry and concentrate with anhydrous sodium sulfate, and separate the residue by silica gel column chromatography to obtain white intermediate 4 (2.31 g, yield: 62.9%).

[0401] Step 5: Dissolve intermediate 4 (2.31 g, 4.5 mmol) in 10 mL of ethyl acetate, add 5 mL of 37% hydrochloric acid, and stir the reaction mixture at room temperature for 3 h. After the reaction is complete, adjust the pH of the reaction solution to 5 with saturated sodium carbonate solution; a white solid will precipitate. Filter, dry, and the filter cake is intermediate 5 (1.20 g, yield: 64.5%).

[0402] Step 6: Intermediate 6 (1.20 g, 2.90 mmol) and isobutyric acid (0.27 g, 3.05 mmol) were dissolved in 5 mL of DMF. DIPEA (2.25 g, 17.41 mmol) and HATU (1.32 g, 3.48 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. After the reaction was completed, the mixture was purified by preparative liquid chromatography with a mobile phase of methanol / water = 60 / 40 to obtain a white solid, namely Formula 1 (1.09 g, yield: 77.67%).

[0403] Synthesis of Equation 2:

[0404] Step 1: N-Boc-1,4-Butanediamine (1.88 g, 10.0 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (2.76 g, 10.0 mmol) were dissolved in 5 mL of DMF, and DIPEA (5.06 g, 50.0 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give green intermediate 3 (2.61 g, yield: 58.8%).

[0405] Step 2: Dissolve intermediate 3 (2.60 g, 5.8 mmol) in 10 mL of anhydrous dichloromethane, add trifluoroacetic acid (3 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a green solid, namely Formula 2 (2.00 g, yield: 98.9%).

[0406] Synthesis of Equation 3:

[0407] Step 1: 4-(piperazin-1-yl)piperidin-1-carboxylic acid tert-butyl ester (0.05 g, 0.18 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.05 mg, 0.18 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.14 g, 1.09 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (0.07 g, yield: 71.7%).

[0408] Step 2: Dissolve intermediate 3 (0.07 g, 0.13 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 3 (0.04 g, yield: 75.4%).

[0409] Synthesis of Equation 4:

[0410] Step 1: Formula 1 (0.05 g, 0.10 mmol) and 4-(piperazin-1-yl)piperidine-1-carboxylic acid tert-butyl ester (0.04 g, 0.11 mmol) were dissolved in 1 mL of DMF. DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. After the reaction was completed, the mixture was purified by preparative liquid chromatography with a mobile phase of methanol / water = 70 / 30. White intermediate 3 (0.05 g, yield: 58.1%) was obtained.

[0411] Step 2: Dissolve intermediate 3 (0.05 g, 0.06 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 4 (0.03 g, yield: 77.0%).

[0412] Synthesis of Equation 5:

[0413] Step 1: 3,9-diazaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (0.05 g, 0.18 mmol) and 2-(2,6-dioxadiazin-3-yl)-5-fluoroisoindoline-1,3-dione (0.05 g, 0.18 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.14 g, 1.09 mmol) was added. The reaction system was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (0.09 g, yield: 97.4%).

[0414] Step 2: Dissolve intermediate 3 (0.09 g, 0.18 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 5 (0.04 g, yield: 59.4%).

[0415] Synthesis of Formula 6:

[0416] Step 1: Dissolve Formula 1 (0.05 g, 0.10 mmol) and 1-Boc-4-piperidinylpiperazine (0.03 g, 0.01 mmol) in 1 mL of DMF, add DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol), and stir the reaction system at room temperature for 30 min. After the reaction is complete, separate and purify by preparative liquid chromatography with a mobile phase of methanol / water = 70 / 30. White intermediate 3 (0.05 g, yield: 68.4%) is obtained.

[0417] Step 2: Dissolve intermediate 3 (0.05 g, 0.07 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a white solid, namely formula 6 (0.04 g, yield: 95.7%).

[0418] Synthesis of Equation 7:

[0419] Step 1: 1-Boc-4-piperidinylpiperazine (0.05 g, 0.18 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.05 g, 0.18 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.14 g, 1.09 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (0.07 g, yield: 76.73%).

[0420] Step 2: Dissolve intermediate 3 (0.07 g, 0.14 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 7 (0.04 g, yield: 66.0%).

[0421] Synthesis of Formula 8:

[0422] Step 1: Formula 1 (0.05 g, 0.10 mmol) and tert-butyl 2,8-diazaspiro[4.5]dec-2-carboxylic acid (0.03 g, 0.11 mmol) were dissolved in 1 mL of DMF. DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol) were added, and the reaction system was stirred at room temperature for 30 min. After the reaction was completed, the mixture was purified by preparative liquid chromatography with a mobile phase of methanol / water = 70 / 30. A white intermediate 3 (0.05 g, yield: 71.2%) was obtained. Step 2: Intermediate 3 (0.05 g, 0.07 mmol) was dissolved in 3 mL of anhydrous dichloromethane. Trifluoroacetic acid (1 mL) was added, and the reaction system was stirred at room temperature for 30 min. The solvent was removed by vacuum distillation to obtain a yellow solid, namely Formula 8 (0.03 g, yield: 71.7%).

[0423] Synthesis of Equation 9:

[0424] Step 1: 3-(piperazin-1-ylmethyl)azacyclobutane-1-carboxylic acid tert-butyl ester (0.05 g, 0.18 mmol) and 2-(26-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.05 g, 0.18 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.14 g, 1.09 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (0.07 g, yield: 76.7%).

[0425] Step 2: Dissolve intermediate 3 (0.07 g, 0.14 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely Formula 9 (0.04 g, yield: 68.3%).

[0426] Synthesis of Equation 10:

[0427] Step 1: Formula 1 (0.05 g, 0.10 mmol) and tert-butyl 3-(piperazin-1-ylmethyl)azacyclobutane-1-carboxylic acid (0.03 g, 0.11 mmol) were dissolved in 1 mL of DMF. DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. After the reaction was complete, the mixture was purified by preparative liquid chromatography (MS / LC) with a mobile phase of methanol / water = 70 / 30. Yellow intermediate 3 (0.04 g, yield: 57.7%) was obtained.

[0428] Step 2: Dissolve intermediate 3 (0.04 g, 0.06 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 10 (0.03 g, yield: 81.0%).

[0429] Synthesis of Equation 11:

[0430] Step 1: 2,8-diazaspiro[4.5]dec-2-carboxylic acid tert-butyl ester (0.04 g, 0.18 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.05 g, 0.18 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.14 g, 1.09 mmol) was added. The reaction system was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (0.09 g, yield: 97.9%).

[0431] Step 2: Dissolve intermediate 3 (0.09 g, 0.18 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely Formula 11 (0.06 g, yield: 79.7%).

[0432] Synthesis of Equation 12:

[0433] Step 1: Formula 1 (0.05 g, 0.10 mmol) and trans-(4-aminocyclohexyl)carbamate tert-butyl ester (0.02 g, 0.11 mmol) were dissolved in 1 mL of DMF. DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. After the reaction was complete, the mixture was purified by preparative liquid chromatography (MS / LC) with a mobile phase of methanol / water = 70 / 30. Yellow intermediate 3 (0.04 g, yield: 51.2%) was obtained.

[0434] Step 2: Dissolve intermediate 3 (0.04 g, 0.05 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely Formula 12 (0.03 g, yield: 97.7%).

[0435] Synthesis of Equation 13:

[0436] Step 1: Formula 1 (0.05 g, 0.10 mmol) and tert-butyl 2,7-diazaspiro[3.5]nonane-7-carboxylate (0.02 g, 0.11 mmol) were dissolved in 5 mL of DMF. DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. After the reaction was complete, the mixture was purified by preparative liquid chromatography (MS / LC) with a mobile phase of methanol / water = 70 / 30. Yellow intermediate 3 (0.04 g, yield: 58.7%) was obtained.

[0437] Step 2: Dissolve intermediate 3 (0.04 g, 0.06 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely Formula 13 (0.03 g, yield: 89.1%).

[0438] Synthesis of Equation 14:

[0439] Step 1: Dissolve Formula 1 (0.05 g, 0.10 mmol) and 4-tert-butoxycarbonylaminopiperidine (0.02 g, 0.11 mmol) in 1 mL of DMF, add DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol), and stir the reaction mixture at room temperature for 30 min. After the reaction is complete, purify the mixture using preparative liquid chromatography with a mobile phase of methanol / water = 70 / 30. Yellow intermediate 3 (0.04 g, yield: 59.6%) was obtained.

[0440] Step 2: Dissolve intermediate 3 (0.04 g, 0.06 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely Formula 14 (0.03 g, yield: 86.1%).

[0441] Synthesis of Equation 15:

[0442] Step 1: 1-tert-Butoxycarbonyl-4-aminomethylpiperidine (0.04 g, 0.19 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.05 g, 0.18 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.14 g, 1.09 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (0.04 g, yield: 48.1%).

[0443] Step 2: Dissolve intermediate 3 (0.04 g, 0.09 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely Formula 15 (0.03 g, yield: 96.05%).

[0444] Synthesis of Equation 16:

[0445] Step 1: 2,7-diazaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (0.04 g, 0.19 mmol) and 2-(2,6-dioxadiazin-3-yl)-5-fluoroisoindoline-1,3-dione (0.05 g, 0.18 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.14 g, 1.09 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (0.04 g, yield: 45.8%).

[0446] Step 2: Dissolve intermediate 3 (0.04 g, 0.08 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 3 (0.03 g, yield: 95.3%).

[0447] Synthesis of Equation 17:

[0448] Step 1: 4-(azacyclobutane-3-yl)piperazine-1-carboxylic acid tert-butyl ester (0.05 g, 0.19 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.05 g, 0.18 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.14 g, 1.09 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (0.04 g, yield: 41.1%).

[0449] Step 2: Dissolve intermediate 3 (0.04 g, 0.07 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely Formula 17 (0.03 g, yield: 91.4%).

[0450] Synthesis of Equation 18:

[0451] Step 1: Formula 1 (0.05 g, 0.10 mmol) and 4-(azacyclobutane-3-yl)piperazine-1-carboxylic acid tert-butyl ester (0.03 g, 0.11 mmol) were dissolved in 1 mL of DMF. DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. After the reaction was complete, the mixture was purified by preparative liquid chromatography (MS / LC) with a mobile phase of methanol / water = 70 / 30. A white intermediate 3 (0.04 g, yield: 56.1%) was obtained.

[0452] Step 2: Dissolve intermediate 3 (0.04 g, 0.06 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 18 (0.03 g, yield: 88.1%).

[0453] Synthesis of Equation 19:

[0454] Step 1: Dissolve Formula 1 (0.05 g, 0.10 mmol) and 1-tert-butoxycarbonyl-4-aminomethylpiperidine (g, mmol) in 1 mL of DMF, add DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol), and stir the reaction system at room temperature for 30 min. After the reaction is complete, separate and purify by preparative liquid chromatography with a mobile phase of methanol / water = 70 / 30. White intermediate 3 (0.04 g, yield: 51.2%) is obtained.

[0455] Step 2: Dissolve intermediate 3 (0.04 g, 0.05 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 19 (0.03 g, yield: 97.7%).

[0456] Synthesis of Equation 20:

[0457] Step 1: 3-Aminoazacyclobutane-1-carboxylic acid tert-butyl ester (0.03 g, 0.19 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.05 g, 0.18 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.14 g, 1.09 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (0.06 g, yield: 72.3%).

[0458] Step 2: Dissolve intermediate 3 (0.06 g, 0.12 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 20 (0.03 g, yield: 80.9%).

[0459] Synthesis of Equation 21:

[0460] Step 1: 1-Boc-3-aminomethylazinecyclobutane (0.04 g, 0.19 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.05 g, 0.18 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.14 g, 1.09 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (0.05 g, yield: 64%).

[0461] Step 2: Dissolve intermediate 3 (0.05, 0.11 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 21 (0.04 g, yield: 98.2%).

[0462] Synthesis of Equation 22:

[0463] Step 1: 6-Amino-2-azaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester (0.04 g, 0.19 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.05 g, 0.18 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.14 g, 1.09 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (0.06 g, yield: 63.6%).

[0464] Step 2: Dissolve intermediate 3 (0.05 g, 0.11 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 22 (0.04 g, yield: 89.0%).

[0465] Synthesis of Equation 23:

[0466] Step 1: 4-(aminomethyl)-4-methylpiperidin-1-carboxylic acid tert-butyl ester (0.04 g, 0.19 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.05 g, 0.18 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.14 g, 1.09 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (0.05 g, yield: 55.4%).

[0467] Step 2: Dissolve intermediate 3 (0.05 g, 0.10 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 23 (0.03 g, yield: 78.2%).

[0468] Synthesis of Equation 24:

[0469] Step 1: 1-tert-Butoxycarbonyl-4-aminomethylpiperidine (0.04 g, 0.18 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5,6-difluoroisoindoline-1,3-dione (0.05 g, 0.17 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.11 g, 0.08 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (0.06 g, yield: 64.6%).

[0470] Step 2: Dissolve intermediate 3 (0.05 g, 0.10 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 24 (0.04 g, yield: 95.6%).

[0471] Synthesis of Formula 25:

[0472] Step 1: Formula 1 (0.05 g, 0.10 mmol) and ((4-fluoropiperidin-4-yl)methyl)tert-butyl carbamate (0.03 g, 0.11 mmol) were dissolved in 1 mL of DMF. DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. After the reaction was complete, the mixture was purified by preparative liquid chromatography (MS / LC) with a mobile phase of methanol / water = 70 / 30. Yellow intermediate 3 (0.03 g, yield: 55.3%) was obtained.

[0473] Step 2: Dissolve intermediate 3 (0.03 g, 0.04 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 25 (0.02 g, yield: 89.5%).

[0474] Synthesis of Equation 26:

[0475] Step 1: Dissolve 1-Boc-4-piperidinecarboxaldehyde (0.1 g, 0.47 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain yellow intermediate 3 (0.05 g, yield: 94.2%).

[0476] Step 2: Dissolve Formula 1 (0.05 g, 0.10 mmol) and Intermediate 3 (0.01 g, 0.11 mmol) in 1 mL of DMF, add DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol), and stir the reaction system at room temperature for 30 min. After the reaction is complete, separate and purify using preparative liquid chromatography with a mobile phase of methanol / water = 70 / 30. A yellow solid, Formula 26, is obtained.

[0477] Synthesis of Equation 27:

[0478] Step 1: N-ethoxycarbonyl-4-piperidinone (1.00 g, 5.8 mmol) was dissolved in 10 mL of water, and 2 mL of 37% hydrochloric acid was added. The reaction system was refluxed and stirred at 100 °C for 5 h. The solvent was removed by vacuum distillation to obtain yellow intermediate 3 (0.3 g, yield: 57.3%).

[0479] Step 2: Dissolve Formula 1 (0.05 g, 0.10 mmol) and Intermediate 3 (0.01 g, 0.11 mmol) in 1 mL of DMF, add DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol), and stir the reaction system at room temperature for 30 min. After the reaction is complete, separate and purify using preparative liquid chromatography with a mobile phase of methanol / water = 70 / 30. A yellow solid, namely Formula 27, is obtained.

[0480] Synthesis of Equation 28:

[0481] Step 1: 4-(2-aminoethyl)piperidin-1-carboxylic acid tert-butyl ester (0.04 g, 0.19 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.05 g, 0.18 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.14 g, 0.19 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (0.04 g, yield: 47.9%).

[0482] Step 2: Dissolve intermediate 3 (0.04 g, 0.08 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 28 (0.03 g, yield: 94.5%).

[0483] Synthesis of Equation 29:

[0484] Step 1: Formula 1 (0.05 g, 0.10 mmol) and tert-butyl 4-(aminomethyl)-3,3-difluoropiperidine-1-carboxylate (0.03 g, 0.11 mmol) were dissolved in 1 mL of DMF. DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. After the reaction was complete, the mixture was purified by preparative liquid chromatography (MS / LC) with a mobile phase of methanol / water = 70 / 30. A white intermediate 3 (0.04 g, yield: 56.7%) was obtained.

[0485] Step 2: Dissolve intermediate 3 (0.04 g, mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 29 (0.03 g, yield: 85.8%).

[0486] Synthesis of Formula 30:

[0487] Step 1: 4-(aminomethyl)-3,3-difluoropiperidin-1-carboxylic acid tert-butyl ester (0.05 g, 0.18 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.05 g, 0.18 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.14 g, 1.09 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (0.06 g, yield: 66.5%).

[0488] Step 2: Dissolve intermediate 3 (0.06 g, 0.12 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 30 (0.04 g, yield: 81.7%).

[0489] Synthesis of Equation 31:

[0490] Step 1: Formula 1 (0.05 g, 0.10 mmol) and tert-butyl 4-(aminomethyl)-3,3-difluoropiperidine-1-carboxylate (0.04 g, 0.11 mmol) were dissolved in 1 mL of DMF. DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. After the reaction was complete, the mixture was purified by preparative liquid chromatography (MS / LC) with a mobile phase of methanol / water = 70 / 30. A white intermediate 3 (0.04 g, yield: 56.7%) was obtained.

[0491] Step 2: Dissolve intermediate 3 (0.04 g, 0.05 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 31 (0.03 g, yield: 85.8%).

[0492] Synthesis of Equation 32:

[0493] Step 1: ((4-aminocyclohexyl)methyl)tert-butyl carbamate (0.04 g, 0.19 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.05 g, 0.18 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.14 g, 1.09 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (0.05 g, yield: 54.7%).

[0494] Step 2: Dissolve intermediate 3 (0.05 g, 0.10 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 32 (0.03 g, yield: 73.5%).

[0495] Synthesis of Equation 33:

[0496] Step 1: Dissolve Formula 1 (0.05 g, 0.10 mmol) and ((4-aminocyclohexyl)methyl)tert-butyl carbamate (g, mmol) in 1 mL of DMF. Add DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol) to the reaction mixture and stir at room temperature for 30 min. After the reaction is complete, separate and purify using preparative liquid chromatography with a mobile phase of methanol / water = 70 / 30. White intermediate 3 (0.056 g, yield: 80%) is obtained.

[0497] Step 2: Dissolve intermediate 3 (0.056 g, 0.08 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 33 (yield: 95%).

[0498] Synthesis of Equation 34:

[0499] Step 1: 5,5-Difluoro-2,7-diazaspiro[3.5]nonane-2-carboxylic acid tert-butyl ester (0.05 g, 0.19 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.05 g, 0.18 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.14 g, 1.09 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (yield: 65%).

[0500] Step 2: Dissolve intermediate 3 (0.1 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 34 (yield: 95%).

[0501] Synthesis of Formula 35:

[0502] Step 1: Dissolve Formula 1 (0.05 g, 0.10 mmol) and tert-butyl 5,5-difluoro-2,7-diazaspiro[3.5]nonane-2-carboxylic acid (0.03 g, 0.11 mmol) in 1 mL of DMF, add DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol), and stir the reaction system at room temperature for 30 min. After the reaction is complete, separate and purify by preparative liquid chromatography with a mobile phase of methanol / water = 70 / 30. White intermediate 3 (yield: 75%) is obtained. Step 2: Dissolve intermediate 3 (0.075 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), and stir the reaction system at room temperature for 30 min. Remove the solvent by vacuum distillation to obtain a yellow solid, Formula 35 (yield: 90%).

[0503] Synthesis of Equation 36:

[0504] Step 1: Formula 1 (0.05 g, 0.10 mmol) and 2-amino-7-Boc-7-azaspiro[3.5]nonane (0.03 g, 0.11 mmol) were dissolved in 1 mL of DMF. DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. After the reaction was completed, the mixture was purified by preparative liquid chromatography with a mobile phase of methanol / water = 70 / 30. White intermediate 3 was obtained (yield: 80%).

[0505] Step 2: Dissolve intermediate 3 (0.05 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely Formula 3 (yield: 90%).

[0506] Synthesis of Equation 37:

[0507] Step 1: 2-Amino-7-Boc-7-azaspiro[3.5]nonane (0.05 g, 0.19 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.05 g, 0.18 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.14 g, 1.09 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (0.05 g, yield: 56.7%).

[0508] Step 2: Dissolve intermediate 3 (0.05 g, 0.10 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (3 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 37 (0.03 g, yield: 81.1%).

[0509] Synthesis of Equation 38:

[0510] Step 1: Formula 1 (0.05 g, 0.10 mmol) and tert-butyl 4-amino-3,3-difluoropiperidine-1-carboxylate (g, mmol) were dissolved in 1 mL of DMF. DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. After the reaction was complete, the mixture was purified by preparative liquid chromatography (PCLC) with a mobile phase of methanol / water = 70 / 30. A white intermediate 3 was obtained (yield: 65%).

[0511] Step 2: Dissolve intermediate 3 (0.05 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 38 (yield: 88%).

[0512] Synthesis of Equation 39:

[0513] Step 1: 4-Amino-3,3-difluoropiperidin-1-carboxylic acid tert-butyl ester (0.04 g, 0.19 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.05 g, 0.18 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.14 g, 1.09 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (yield: 55%).

[0514] Step 2: Dissolve intermediate 3 (0.1 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 39 (yield: 90%).

[0515] Synthesis of Formula 40:

[0516] Step 1: Dissolve 0.1 g (0.04 mmol) of 2-oxo-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester in 3 mL of anhydrous dichloromethane, add 1 mL of trifluoroacetic acid, stir the reaction system at room temperature for 30 min, and remove the solvent by vacuum distillation to obtain yellow intermediate 3 (g, yield: %). Step 2: Dissolve Formula 1 (0.05 g, 0.10 mmol) and intermediate 3 (0.02 g, 0.11 mmol) in 1 mL of DMF, add DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol), and stir the reaction system at room temperature for 30 min. After the reaction, purify by preparative liquid chromatography, mobile phase: methanol / water = 70 / 30. A yellow solid, Formula 40, is obtained.

[0517] Synthesis of Equation 41:

[0518] Step 1: 2-Formyl-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (0.10 g, 0.39 mmol) was dissolved in 3 mL of anhydrous dichloromethane, and trifluoroacetic acid (1 mL) was added. The reaction system was stirred at room temperature for 30 min, and the solvent was removed by vacuum distillation to obtain yellow intermediate 3 (g, yield: %). Step 2: Formula 1 (0.05 g, 0.10 mmol) and intermediate 3 (0.02 g, 0.11 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol) were added. The reaction system was stirred at room temperature for 30 min. After the reaction was completed, the mixture was purified by preparative liquid chromatography with a mobile phase of methanol / water = 70 / 30. A yellow solid, namely Formula 41, was obtained.

[0519] Synthesis of Equation 42:

[0520] Step 1: Dissolve 0.10 g (0.47 mmol) of 6-oxo-2-azaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain 3 g (yield: %) of yellow intermediate.

[0521] Step 2: Dissolve Formula 1 (0.05 g, 0.10 mmol) and Intermediate 3 (0.01 g, 0.11 mmol) in 1 mL of DMF, add DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol), and stir the reaction system at room temperature for 30 min. After the reaction is complete, separate and purify using preparative liquid chromatography with a mobile phase of methanol / water = 70 / 30. A yellow solid, namely Formula 42, is obtained.

[0522] Synthesis of Equation 43:

[0523] Step 1: ((4-aminocyclohexyl)methyl)tert-butyl carbamate (0.04 g, 0.18 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5,6-difluoroisoindoline-1,3-dione (0.05 g, 0.17 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.13 g, 1.02 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (yield: 66%).

[0524] Step 2: Dissolve intermediate 3 (0.1 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 43 (yield: 84%).

[0525] Synthesis of Equation 44:

[0526] Step 1: 5,5-Difluoro-2,7-diazaspiro[3.5]nonane-2-carboxylic acid tert-butyl ester (0.05 g, 0.18 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5,6-difluoroisoindoline-1,3-dione (0.05 g, 0.17 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.13 g, 1.02 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (yield: 58%).

[0527] Step 2: Dissolve intermediate 3 (0.1 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 44 (yield: 83%).

[0528] Synthesis of Equation 45:

[0529] Step 1: Formula 1 (0.05 g, 0.10 mmol) and tert-butyl 5,5-difluoro-2,7-diazaspiro[3.5]nonane-2-carboxylic acid (0.03 g, 0.11 mmol) were dissolved in 1 mL of DMF. DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol) were added, and the reaction mixture was stirred at room temperature for 30 min. After the reaction was complete, the mixture was purified by preparative liquid chromatography (PCLC) with a mobile phase of methanol / water = 70 / 30. A white intermediate of 3 g (yield: %) was obtained.

[0530] Step 2: Dissolve intermediate 3 (g, mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 45 (g, yield: %).

[0531] Synthesis of Equation 46:

[0532] Step 1: Dissolve Formula 1 (0.05 g, 0.10 mmol) and 2-amino-7-Boc-7-azaspiro[3.5]nonane (0.03 g, 0.03 mmol) in 1 mL of DMF, add DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol), and stir the reaction mixture at room temperature for 30 min. After the reaction is complete, purify the mixture using preparative liquid chromatography with a mobile phase of methanol / water = 70 / 30. A white intermediate of 46 g (yield: %) was obtained.

[0533] Step 2: Dissolve intermediate 3 (g, mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 46 (g, yield: %).

[0534] Synthesis of Equation 47:

[0535] Step 1: 2-Amino-7-Boc-7-azaspiro[3.5]nonane (0.04 g, 0.18 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5,6-difluoroisoindoline-1,3-dione (0.05 g, 0.17 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.13 g, 1.02 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (yield: 59%).

[0536] Step 2: Dissolve intermediate 3 (0.1 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 47 (yield: 86%).

[0537] Synthesis of Equation 48:

[0538] Step 1: 4-(aminomethyl)-3,3-difluoropiperidin-1-carboxylic acid tert-butyl ester (0.05 g, 0.19 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5,6-difluoroisoindoline-1,3-dione (0.05 g, 0.17 mmol) were dissolved in 5 mL of DMF, and DIPEA (0.13 g, 1.02 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (yield: 62%).

[0539] Step 2: Dissolve intermediate 3 (0.1 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 48 (yield: 90%).

[0540] Synthesis of Equation 49:

[0541] Step 1: Dissolve Formula 1 (0.05 g, 0.10 mmol) and tert-butyl 4-amino-3,3-difluoropiperidine-1-carboxylate (0.03 g, 0.11 mmol) in 1 mL of DMF. Add DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction is complete, purify the mixture using preparative liquid chromatography with a mobile phase of methanol / water = 70 / 30. White intermediate 46 (yield: 54%) is obtained.

[0542] Step 2: Dissolve intermediate 3 (0.05 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 49 (yield: 85%).

[0543] Synthesis of Formula 50:

[0544] Step 1: 4-Amino-3,3-difluoropiperidin-1-carboxylic acid tert-butyl ester (0.04 g, 0.17 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5,6-difluoroisoindoline-1,3-dione (0.05 g, 0.17 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.13 g, 1.02 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (yield: 58%).

[0545] Step 2: Dissolve intermediate 3 (0.1 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 50 (yield: 88%).

[0546] Synthesis of Equation 51:

[0547] Step 1: 6-Amino-2-azaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester (0.04 g, 0.18 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5,6-difluoroisoindoline-1,3-dione (0.05 g, 0.17 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.13 g, 1.02 mmol) was added. The reaction system was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (yield: 56%).

[0548] Step 2: Dissolve intermediate 3 (0.1 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely Formula 51 (yield: 80%).

[0549] Synthesis of Equation 52:

[0550] Step 1: 4-(2-aminoethyl)piperidin-1-carboxylic acid tert-butyl ester (0.04 g, 0.19 mmol) and 2-(2,6-dioxopiperidin-3-yl)-5,6-difluoroisoindoline-1,3-dione (0.05 g, 0.17 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.13 g, 1.02 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. The reaction solution was purified by prep-HPLC to give yellow intermediate 3 (yield: 56%).

[0551] Step 2: Dissolve intermediate 3 (0.1 mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely formula 52 (yield: 85%).

[0552] Synthesis of Equation 53:

[0553] Step 1: Dissolve Formula 1 (0.05 g, 0.10 mmol) and tert-butyl 6-amino-2-azaspiro[3.3]heptane-2-carboxylate (0.23 g, 0.11 mmol) in 1 mL of DMF, add DIPEA (0.08 g, 0.62 mmol) and HATU (0.05 g, 0.12 mmol), and stir the reaction system at room temperature for 30 min. After the reaction is complete, separate and purify by preparative liquid chromatography, mobile phase: methanol / water = 70 / 30. White intermediate 3 (g, yield: %) is obtained. Step 2: Dissolve intermediate 3 (g, mmol) in 3 mL of anhydrous dichloromethane, add trifluoroacetic acid (1 mL), stir the reaction system at room temperature for 30 min, remove the solvent by vacuum distillation, and obtain a yellow solid, namely Formula 53 (g, yield: %).

[0554] Example 90

[0555] Procedure: Dissolve Formula 1 (0.20 g, 0.41 mmol) and Formula 2 (0.15 g, 0.43 mmol) in 3 mL LDM. Add 2–3 drops of DIPEA, then add HATU (0.19 g, 0.5 mmol). After the reaction is complete, purify by preparative HPLC. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5μm, 19×150mm column chromatography yielded a yellow solid LD-331 (0.07 g, yield: 19.25%). LC-MS calculations showed C0.07... 46 H 47 N7O7[M+H]+: 809.35, found 810.52, UPLC retention time: 2.81 min, purity >90%.

[0556] Example 91

[0557] Procedure: Dissolve Formula 1 (10.00 mg, 0.02 mmol) and Formula 3 (9.54 mg, 0.02 mmol) in 1 mL of DMF. Add 2–3 drops of DIPEA, then add HATU (9.44 mg, 0.02 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction is complete, purify the mixture using preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-01 (1.90 mg, yield: 10.2%). LC-MS calculated C0... 52 H 56 N8O7[M+H]+: 904.43, found 905.52, UPLC retention time: 2.27 min, purity >95%.

[0558] Example 92

[0559] Procedure: Formula 4 (30.00 mg, 0.05 mmol), 2-(2,6-dioxadiazin-3-yl)-5-fluoroisoindoline-1,3-dione (50 mg, 0.05 mmol) was dissolved in 1 mL of DMF, and DIPEA (35.85 mg, 0.28 mmol) was added. The reaction system was stirred at 100 °C for 5 h. After the reaction was completed, the mixture was purified by preparative liquid chromatography. The mobile phase was methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-02 (3.00 mg, yield: 7.2%). LC-MS calculated C0... 52 H 56 N8O7[M+H]+: 904.43, found 905.52, UPLC retention time: 2.34 min, purity >95%.

[0560] Example 93

[0561] Procedure: Dissolve Formula 1 (10.00 mg, 0.02 mmol) and Formula 5 (8.91 mg, 0.02 mmol) in 1 mL of DMF. Add 2–3 drops of DIPEA, then add HATU (9.44 mg, 0.02 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction is complete, purify the mixture using preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TMRP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-03 (2.60 mg, yield: 14.4%). LC-MS calculated C0... 51 H 53 N7O7[M+H]+: 875.40, found 876.41, UPLC retention time: 2.86 min, purity >95%.

[0562] Example 94

[0563] Procedure: Formula 6 (30.00 mg, 0.05 mmol), 2-(2,6-dioxadiazin-3-yl)-5-fluoroisoindoline-1,3-dione (13.71 mg, 0.05 mmol) was dissolved in 1 mL of DMF, and DIPEA (36.65 mg, 0.28 mmol) was added. The reaction system was stirred at 100 °C for 5 h. After the reaction was completed, the mixture was purified by preparative liquid chromatography. The mobile phase was methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-04 (12.60 mg, yield: 29.9%). LC-MS calculated C0... 51 H 54 N8O7[M+H]+: 890.41, found 892.49, UPLC retention time: 2.24 min, purity >95%.

[0564] Example 95

[0565] Procedure: Dissolve Formula 1 (10.00 mg, 0.02 mmol) and Formula 7 (9.24 mg, 0.02 mmol) in 1 mL of DMF. Add 2–3 drops of DIPEA, then add HATU (9.44 mg, 0.02 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction is complete, purify the mixture using preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-05 (1.70 mg, yield: 9.2%). LC-MS calculated C0... 51 H 54 N8O7[M+H]+: 890.41, found 891.50, UPLC retention time: 2.35 min, purity >95%.

[0566] Example 96

[0567] Procedure: Formula 8 (30.00 mg, 0.05 mmol), 2-(2,6-dioxadiazin-3-yl)-5-fluoroisoindoline-1,3-dione (14.36 mg, 0.05 mmol) were dissolved in 1 mL of DMF, and DIPEA (38.40 mg, 0.30 mmol) was added. The reaction system was stirred at 100 °C for 5 h. After the reaction was completed, the mixture was purified by preparative liquid chromatography. The mobile phase was methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-06 (6.50 mg, yield: 15.2%). LC-MS calculated C0... 50 H 51 N7O7[M+H]+: 861.38, found 862.46, UPLC retention time: 2.83 min, purity >95%.

[0568] Example 97

[0569] Procedure: Dissolve Formula 1 (10.00 mg, 0.02 mmol) and Formula 9 (8.93 mg, 0.02 mmol) in 1 mL of DMF. Add 2–3 drops of DIPEA, then add HATU (9.44 mg, 0.02 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction, purify the mixture using preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-07 (2.30 mg, yield: 12.7%). LC-MS calculated C0... 50 H 52 N8O7[M+H]+: 876.40, found 877.45, UPLC retention time: 2.27 min, purity >95%.

[0570] Example 98

[0571] Procedure: Formula 10 (30.00 mg, 0.05 mmol), 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (14.02 mg, 0.05 mmol) were dissolved in 1 mL of DMF, and DIPEA (37.48 mg, 0.30 mmol) was added. The reaction system was stirred at 100 °C for 5 h. After the reaction was completed, the mixture was purified by preparative liquid chromatography. The mobile phase was methanol / water = 70 / 30. Column type: SymmetryShield. TMRP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-08 (2.6 mg, yield: 6.1%). LC-MS calculated C0... 50 H 52 N8O7[M+H]+: 876.40, found 877.48, UPLC retention time: 2.35 min, purity >95%.

[0572] Example 99

[0573] Procedure: Dissolve Formula 1 (10.00 mg, 0.02 mmol) and Formula 11 (8.61 mg, 0.02 mmol) in 1 mL of DMF. Add 2–3 drops of DIPEA, then add HATU (9.44 mg, 0.02 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction, purify the mixture using preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-09 (3.50 mg, yield: 19.6%). LC-MS calculated C0... 50 H 51 N7O7[M+H]+: 861.38, found 862.42, UPLC retention time: 2.79 min, purity >95%.

[0574] Example 100

[0575] Procedure: Formula 12 (30.00 mg, 0.05 mmol), 2-(2,6-dioxadiazin-3-yl)-5-fluoroisoindoline-1,3-dione (15.01 mg, 0.05 mmol) were dissolved in 1 mL of DMF, and DIPEA (40.13 mg, 0.30 mmol) was added. The reaction system was stirred at 100 °C for 5 h. After the reaction was completed, the mixture was purified by preparative liquid chromatography. The mobile phase was methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-10 (3.7 mg, yield: 8.6%). LC-MS calculated C0... 48 H 49 N7O7[M+H]+: 835.37, found 836.44, UPLC retention time: 2.89 min, purity >95%.

[0576] Example 101

[0577] Procedure: Formula 13 (30.00 mg, 0.05 mmol), 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (14.7 mg, 0.05 mmol)

[0578] Dissolved in 1 mL DMF, DIPEA (39.31 mg, 0.30 mmol) was added, and the reaction mixture was stirred at 100 °C for 5 h. After the reaction, the mixture was purified by preparative liquid chromatography with a mobile phase of methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-12 (9.8 mg, yield: 22.8%). LC-MS calculated C0... 49 H 49 N7O7[M+H]+: 847.37, found 848.36, UPLC retention time: 2.82 min, purity >95%.

[0579] Example 102

[0580] Procedure: Formula 14 (30.00 mg, 0.05 mmol), 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (15.38 mg, 0.06 mmol) were dissolved in 1 mL of DMF, and DIPEA (41.12 mg, 0.32 mmol) was added. The reaction system was stirred at 100 °C for 5 h. After the reaction was completed, the mixture was purified by preparative liquid chromatography. The mobile phase was methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column yielded a yellow solid LD-331-13 (1.3 mg, yield: 3.0%). LC-MS calculated C0... 47 H 47 N7O7[M+H]+: 821.35, found 822.40, UPLC retention time: 2.76 min, purity >95%.

[0581] Example 103

[0582] Procedure: Dissolve Formula 1 (30.00 mg, 0.06 mmol) and Formula 15 (24.13 mg, 0.07 mmol) in 1 mL of DMF. Add 2–3 drops of DIPEA, then add HATU (30.67 mg, 0.08 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction is complete, purify the mixture using preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TMRP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-14 (10.60 mg, yield: 20.4%). LC-MS calculated C0.05... 48 H 49 N7O7[M+H]+: 835.37, found 836.38, UPLC retention time: 2.80 min, purity >95%.

[0583] Example 104

[0584] Procedure: Dissolve Formula 1 (30.00 mg, 0.06 mmol) and Formula 16 (24.91 mg, 0.07 mmol) in 1 mL of DMF. Add 2–3 drops of DIPEA, then add HATU (30.67 mg, 0.08 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction, purify the mixture using preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-15 (14.50 mg, yield: 27.6%). LC-MS calculated C0... 49 H 49 N7O7[M+H]+: 847.37, found 848.38, UPLC retention time: 2.86 min, purity >95%.

[0585] Example 105

[0586] Procedure: Dissolve Formula 1 (30.00 mg, 0.06 mmol) and Formula 17 (25.89 mg, 0.07 mmol) in 1 mL of DMF. Add 2–3 drops of DIPEA, then add HATU (30.67 mg, 0.08 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction, purify the mixture using preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-16 (24.90 mg, yield: 46.5%). LC-MS calculated C0... 49 H 50 N8O7[M+H]+: 862.38, found 863.40, UPLC retention time: 2.76 min, purity >95%.

[0587] Example 106

[0588] Procedure: Formula 18 (30.00 mg, 0.05 mmol), 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (14.34 mg, 0.05 mmol) were dissolved in 1 mL of DMF, and DIPEA (38.34 mg, 0.30 mmol) was added. The reaction system was stirred at 100 °C for 5 h. After the reaction was completed, the mixture was purified by preparative liquid chromatography. The mobile phase was methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-17 (9.00 mg, yield: 21.1%). LC-MS calculated C0... 49 H 50 N8O7[M+H]+: 862.38, found 863.39, UPLC retention time: 2.71 min, purity >95%.

[0589] Example 107

[0590] Procedure: Formula 19 (30.00 mg, 0.05 mmol), 2-(2,6-dioxadiazin-3-yl)-5-fluoroisoindoline-1,3-dione (15.01 mg, 0.05 mmol) were dissolved in 1 mL of DMF, and DIPEA (40.13 mg, 0.31 mmol) was added. The reaction system was stirred at 100 °C for 5 h. After the reaction was completed, the mixture was purified by preparative liquid chromatography. The mobile phase was methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 column chromatography (5 μm, 19 × 150 mm) yielded a yellow solid LD-331-18 (23.50 mg, yield: 54.3%). LC-MS calculated C0... 48 H 49 N7O7[M+H]+: 835.37, found 836.47, UPLC retention time: 2.87 min, purity >95%.

[0591] Example 108

[0592] Procedure: Dissolve Formula 1 (30.00 mg, 0.06 mmol) and Formula 20 (21.39 mg, 0.07 mmol) in 1 mL of DMF. Add 2–3 drops of DIPEA, then add HATU (28.31 mg, 0.07 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction is complete, purify the mixture using preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TMRP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-19 (8.30 mg, yield: 15.8%). LC-MS calculated C0... 45 H 43 N7O7[M+H]+: 793.32, found 794.35, UPLC retention time: 2.57 min, purity >90%.

[0593] Example 109

[0594] Procedure: Dissolve Formula 1 (30.00 mg, 0.06 mmol) and Formula 21 (21.30 mg, 0.07 mmol) in 1 mL of DMF. Add 2–3 drops of DIPEA, then add HATU (28.31 mg, 0.07 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction is complete, purify the mixture using preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-20 (8.80 mg, yield: 16.0%). LC-MS calculated C0... 46 H 45 N7O7[M+H]+: 807.34, found 808.33, UPLC retention time: 2.58 min, purity >90%.

[0595] Example 110

[0596] Procedure: Dissolve Formula 1 (30.00 mg, 0.06 mmol) and Formula 22 (24.00 mg, 0.07 mmol) in 1 mL of DMF. Add 2–3 drops of DIPEA, then add HATU (28.31 mg, 0.07 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction is complete, purify the mixture using preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column yielded a yellow solid LD-331-21 (2.10 mg, yield: 4.0%). LC-MS calculated C0... 48 H 47 N7O7[M+H]+: 833.35, found 834.35, UPLC retention time: 3.10 min, purity >90%.

[0597] Example 111

[0598] Procedure: Dissolve Formula 1 (30.00 mg, 0.06 mmol) and Formula 23 (25.04 mg, 0.07 mmol) in 1 mL of DMF. Add 2–3 drops of DIPEA, then add HATU (28.31 mg, 0.07 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction, purify the mixture using preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-23 (14.40 mg, yield: 26.1%). LC-MS calculated C0... 49 H 51 N7O7[M+H]+: 849.38, found 850.51, UPLC retention time: 3.13 min, purity >95%.

[0599] Example 112

[0600] Procedure: Dissolve Formula 1 (30.00 mg, 0.06 mmol) and Formula 24 (25.3 mg, 0.07 mmol) in 1 mL of DMF. Add 2–3 drops of DIPEA, then add HATU (28.31 mg, 0.07 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction is complete, purify the mixture using preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-24 (4.50 mg, yield: 8.0%). LC-MS calculated C0... 48 H 48 N7O7[M+H]+: 853.36, found 854.47, UPLC retention time: 3.10 min, purity >90%.

[0601] Example 113

[0602] Procedure: Formula 25 (23.00 mg, 0.04 mmol), 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (11.16 mg, 0.04 mmol) were dissolved in 1 mL of DMF, and DIPEA (29.84 mg, 0.23 mmol) was added. The reaction mixture was stirred at 100 °C for 5 h. After the reaction, the mixture was purified by preparative liquid chromatography. The mobile phase was methanol / water = 70 / 30. Column type: SymmetryShield. TMRP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-25 (0.90 mg, yield: 2.5%). LC-MS calculated C0... 48 H 48 N7O7[M+H]+: 853.36, found 854.47, UPLC retention time: 3.09 min, purity >90%.

[0603] Example 114

[0604] Procedure: Formula 26 (0.2 mmol) and TX-16 (0.18 mmol) were dissolved in 1 mL of dichloromethane. 1–2 drops of acetic acid were added, and the reaction mixture was stirred for 1 h. Then, sodium triacetoxyborohydride (0.2 mmol) was added, and the reaction mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was purified by preparative liquid chromatography (PCLC) with a mobile phase of methanol / water = 70 / 30. The resulting yellow solid was LD-331-26. Column type: Prep C18 OBD TM A 5 μm, 50 × 100 mm column chromatography yielded a yellow solid LD-331-25 (yield: 40%). LC-MS calculations showed that C0... 50 H 51 N7O7[M+H]+: 861.38, found 861.48, UPLC retention time: 3.4 min, purity >95%.

[0605] Example 115

[0606] Procedure: Formula 27 (0.2 mmol) and TX-16 (0.18 mmol) were dissolved in 1 mL of dichloromethane. 1–2 drops of acetic acid were added, and the reaction mixture was stirred for 1 h. Then, sodium triacetoxyborohydride (0.2 mmol) was added, and the reaction mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was purified by preparative liquid chromatography (PCC) with a mobile phase of methanol / water = 70 / 30. The resulting product was a yellow solid, LD-331-26. Column type: Prep C18 OBD TM A 5 μm, 50 × 100 mm column chromatography yielded a yellow solid LD-331-27 (yield: 45%). LC-MS calculations showed that C0... 46 H 49 N7O7[M+H]+: 847.37, found 847.62, UPLC retention time: 3.6 min, purity >95%.

[0607] Example 116

[0608] Procedure: Dissolve Formula 1 (30.00 mg, 0.06 mmol) and Formula 28 (25.04 mg, 0.07 mmol) in 1 mL of DMF. Add 2–3 drops of DIPEA, then add HATU (28.31 mg, 0.07 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction, purify the mixture using preparative HPLC. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-28 (0.5 mg, yield: 1.0%). LC-MS calculated C0.0... 49 H 51 N7O7[M+H]+: 849.38, found 850.44, UPLC retention time: 2.82 min, purity >95%.

[0609] Example 117

[0610] Procedure: Formula 29 (0.1 mmol), 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.1 mmol) was dissolved in 1 mL DMF, and DIPEA (0.5 mmol) was added. The reaction system was stirred at 100 °C for 5 h. After the reaction, the mixture was purified by preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: Prep C18 OBD TM A 5 μm, 50 × 100 mm column chromatography yielded a yellow solid LD-331-29 (yield: 35%). LC-MS calculations showed that C0... 46 H 47 N7O7[M+H]+: 809.35, found 809.62. UPLC retention time: 7.8 min, purity >95%.

[0611] Example 118

[0612] Procedure: Dissolve Formula 1 (30.00 mg, 0.06 mmol) and Formula 30 (26.47 mg, 0.07 mmol) in 1 mL of DMF. Add 2–3 drops of DIPEA, then add HATU (28.31 mg, 0.07 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction is complete, purify the mixture using preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column, yielded a yellow solid LD-331-30 (0.8 mg, yield: 1.4%). LC-MS calculated C 48H 47 F2N7O7[M+H]+: 871.35, found 872.45. UPLC retention time: 3.19 min, purity >95%.

[0613] Example 119

[0614] Procedure: Formula 31 (30.00 mg, 0.05 mmol), 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (15.05 mg, 0.05 mmol) were dissolved in 1 mL of DMF, and DIPEA (37.78 mg, 0.30 mmol) was added. The reaction system was stirred at 100 °C for 5 h. After the reaction was completed, the mixture was purified by preparative liquid chromatography. The mobile phase was methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-31 (0.70 mg, yield: 1.6%). LC-MS calculated C0.05... 48 H 48 F3N7O7[M+H]+: 889.34, found 890.31, UPLC retention time: 2.91 min, purity >95%.

[0615] Example 120

[0616] Procedure: Dissolve Formula 1 (30.00 mg, 0.06 mmol) and Formula 32 (25.04 mg, 0.07 mmol) in 1 mL of DMF. Add 2–3 drops of DIPEA, then add HATU (28.31 mg, 0.07 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction, purify the mixture using preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-32 (1.00 mg, yield: 1.7%). LC-MS calculated C0... 49 H 51 N7O7[M+H]+: 849.38, found 850.55, UPLC retention time: 3.29 min, purity >90%.

[0617] Example 121

[0618] Procedure: Formula 33 (0.1 mmol), 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.1 mmol) was dissolved in 1 mL DMF, and DIPEA (0.5 mmol) was added. The reaction system was stirred at 100 °C for 5 h. After the reaction, the mixture was purified by preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-33 (yield: 46%). LC-MS calculations showed C0... 49 H 51 N7O7[M+H]+: 849.38, found 849.48, UPLC retention time: 5.2 min, purity >95%.

[0619] Example 122

[0620] Procedure: Dissolve Formula 1 (0.1 mmol) and Formula 34 (0.1 mmol) in 1 mL of DMF, add 2–3 drops of DIPEA, then add HATU (0.12 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction, purify the mixture using preparative liquid chromatography (PCLC). Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5μm, 19×150mm column chromatography yielded a yellow solid LD-331-34 (yield: 65%). LC-MS calculations showed C0... 49 H 47 F2N7O7[M+H]+: 883.35, found 883.39, UPLC retention time: 5.2 min, purity >95%.

[0621] Example 123

[0622] Procedure: Formula 35 (0.1 mmol), 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.1 mmol) was dissolved in 1 mL DMF, and DIPEA (0.5 mmol) was added. The reaction system was stirred at 100 °C for 5 h. After the reaction, the mixture was purified by preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5μm, 19×150mm column chromatography yielded a yellow solid LD-331-35 (yield: 54%). LC-MS calculations showed C0... 49 H 47F2N7O7[M+H]+: 883.35, found 883.49, UPLC retention time: 4.3 min, purity >95%.

[0623] Example 124

[0624] Procedure: Formula 36 (0.1 mmol), 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.1 mmol) was dissolved in 1 mL DMF, and DIPEA (0.5 mmol) was added. The reaction system was stirred at 100 °C for 5 h. After the reaction, the mixture was purified by preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5μm, 19×150mm column chromatography yielded a yellow solid LD-331-36 (yield: 55%). LC-MS calculations showed C0... 50 H 51 N7O7[M+H]+: 861.38, found 861.52, UPLC retention time: 4.9 min, purity >95%.

[0625] Example 125

[0626] Procedure: Dissolve Formula 1 (30.00 mg, 0.06 mmol) and Formula 37 (25.82 mg, 0.07 mmol) in 1 mL of DMF. Add 2–3 drops of DIPEA, then add HATU (28.31 mg, 0.07 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction, purify the mixture using preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-37 (8.5 mg, yield: 15.3%). LC-MS calculated C0... 50 H 51 N7O7[M+H]+: 861.38, found 862.47, UPLC retention time: 3.20 min, purity >95%.

[0627] Example 126

[0628] Procedure: Formula 38 (0.1 mmol), 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.1 mmol) was dissolved in 1 mL DMF, and DIPEA (0.5 mmol) was added. The reaction system was stirred at 100 °C for 5 h. After the reaction, the mixture was purified by preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-38 (yield: 46%). LC-MS calculations showed C0... 47 H 45 F2N7O7[M+H]+: 857.33, found 857.62, UPLC retention time: 5.6 min, purity >95%.

[0629] Example 127

[0630] Procedure: Dissolve Formula 1 (0.1 mmol) and Formula 39 (0.11 mmol) in 1 mL of DMF, add 2–3 drops of DIPEA, then add HATU (0.15 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction is complete, purify the mixture using preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5μm, 19×150mm column chromatography yielded a yellow solid LD-331-39 (yield: 75%). LC-MS calculations showed C0... 47 H 45 F2N7O7[M+H]+: 857.3, found 857.5, UPLC retention time: 5.9 min, purity >95%.

[0631] Example 128

[0632] Procedure: Formula 40 (0.12 mmol) and TX-16 (0.1 mmol) were dissolved in 1 mL of dichloromethane. 1–2 drops of acetic acid were added, and the reaction mixture was stirred for 1 h. Then, sodium triacetoxyborohydride (0.15 mmol) was added, and the reaction mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was purified by preparative liquid chromatography. The mobile phase was methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5μm, 19×150mm column chromatography yielded a yellow solid LD-331-40 (yield: 45%). LC-MS calculations showed C0... 52 H 53N7O7[M+H]+: 887.40, found 887.56, UPLC retention time: 5.8 min, purity >95%.

[0633] Example 129

[0634] Procedure: Formula 41 (0.12 mmol) and TX-16 (0.1 mmol) were dissolved in 1 mL of dichloromethane. 1–2 drops of acetic acid were added, and the reaction mixture was stirred for 1 h. Then, sodium triacetoxyborohydride (0.15 mmol) was added, and the reaction mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was purified by preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5μm, 19×150mm column chromatography yielded a yellow solid LD-331-41 (yield: 59%). LC-MS calculations showed C0... 52 H 53 N7O7[M+H]+: 887.40, found 887.56, UPLC retention time: 6.8 min, purity >95%.

[0635] Example 130

[0636] Procedure: Formula 42 (0.12 mmol) and TX-16 (0.1 mmol) were dissolved in 1 mL of dichloromethane. 1–2 drops of acetic acid were added, and the reaction mixture was stirred for 1 h. Then, sodium triacetoxyborohydride (0.15 mmol) was added, and the reaction mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was purified by preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5μm, 19×150mm column chromatography yielded a yellow solid LD-331-42 (yield: 55%). LC-MS calculations showed C0... 50 H 49 N7O7[M+H]+: 859.37, found 859.49, UPLC retention time: 4.1 min, purity >95%.

[0637] Example 131

[0638] Procedure: Dissolve Formula 1 (0.1 mmol) and Formula 43 (0.1 mmol) in 1 mL of DMF, add 2–3 drops of DIPEA, then add HATU (0.15 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction, purify the mixture using preparative liquid chromatography (PCLC). Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TMRP18 5μm, 19×150mm column chromatography yielded a yellow solid LD-331-43 (yield: 69%). LC-MS calculations showed C0... 49 H 50 FN7O7[M+H]+: 867.38, found 867.56, UPLC retention time: 6.5 min, purity > 95%.

[0639] Example 132

[0640] Procedure: Dissolve Formula 1 (0.12 mmol) and Formula 44 (0.1 mmol) in 1 mL of DMF, add 2–3 drops of DIPEA, then add HATU (0.15 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction, purify the mixture using preparative liquid chromatography (PCLC). Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-44 (yield: 65%). LC-MS calculations showed C0... 49 H 46 F3N7O7[M+H]+: 901.34, found 901.65. UPLC retention time: 6.5 min, purity >95%.

[0641] Example 133

[0642] Procedure: Formula 45 (0.1 mmol), 2-(2,6-dioxopiperidin-3-yl)-5,6-difluoroisoindoline-1,3-dione (0.1 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.5 mmol) was added. The reaction system was stirred at 100 °C for 5 h. After the reaction, the mixture was purified by preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5 μm, 19 × 150 mm column chromatography yielded a yellow solid LD-331-45 (yield: 45%). LC-MS calculations showed C0... 49 H 46 F3N7O7[M+H]+: 901.34, found 901.58, UPLC retention time: 5.9 min, purity >95%.

[0643] Example 134

[0644] Procedure: Formula 46 (0.1 mmol), 2-(2,6-dioxopiperidin-3-yl)-5,6-difluoroisoindoline-1,3-dione (0.1 mmol) was dissolved in 1 mL DMF, and DIPEA (0.5 mmol) was added. The reaction system was stirred at 100 °C for 5 h. After the reaction, the mixture was purified by preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5μm, 19×150mm column chromatography yielded a yellow solid LD-331-46 (yield: 50%). LC-MS calculations showed C0... 50 H 50 FN7O7[M+H]+: 879.38, found 879.54, UPLC retention time: 7.1 min, purity >95%.

[0645] Example 135

[0646] Procedure: Dissolve Formula 1 (0.12 mmol) and Formula 47 (0.1 mmol) in 1 mL of DMF, add 2–3 drops of DIPEA, then add HATU (0.15 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction, purify the mixture using preparative liquid chromatography (PCLC). Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5μm, 19×150mm column chromatography yielded a yellow solid LD-331-47 (yield: 76%). LC-MS calculations showed C0... 50 H 50 FN7O7[M+H]+: 879.38, found 879.51, UPLC retention time: 7.0 min, purity >95%.

[0647] Example 136

[0648] Procedure: Dissolve Formula 1 (0.12 mmol) and Formula 48 (0.1 mmol) in 1 mL of DMF, add 2–3 drops of DIPEA, then add HATU (0.15 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction, purify the mixture using preparative liquid chromatography (PCLC). Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5μm, 19×150mm column chromatography yielded a yellow solid LD-331-48 (yield: 72%). LC-MS calculations showed C0... 48 H 46F3N7O7[M+H]+: 889.34, found 889.42, UPLC retention time: 6.2 min, purity >95%.

[0649] Example 137

[0650] Procedure: Formula 49 (0.1 mmol), 2-(2,6-dioxopiperidin-3-yl)-5,6-difluoroisoindoline-1,3-dione (0.1 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.5 mmol) was added. The reaction system was stirred at 100 °C for 5 h. After the reaction, the mixture was purified by preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5μm, 19×150mm column chromatography yielded a yellow solid LD-331-49 (yield: 42%). LC-MS calculations showed C0... 47 H 44 F3N7O7[M+H]+: 875.33, found 875.42, UPLC retention time: 5.6 min, purity >95%.

[0651] Example 138

[0652] Procedure: Dissolve Formula 1 (0.12 mmol) and Formula 50 (0.1 mmol) in 1 mL of DMF, add 2–3 drops of DIPEA, then add HATU (0.15 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction, purify the mixture using preparative liquid chromatography (PCLC). Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5μm, 19×150mm column chromatography yielded a yellow solid LD-331-50 (yield: 74%). LC-MS calculations showed C0... 47 H 44 F3N7O7[M+H]+: 875.33 found 875.39, UPLC retention time: 5.8 min, purity >95%.

[0653] Example 139

[0654] Procedure: Dissolve Formula 1 (0.12 mmol) and Formula 51 (0.1 mmol) in 1 mL of DMF, add 2–3 drops of DIPEA, then add HATU (0.15 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction, purify the mixture using preparative liquid chromatography (PCLC). Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TMRP18 5μm, 19×150mm column chromatography yielded a yellow solid LD-331-51 (yield: 65%). LC-MS calculations showed C0... 48 H 46 FN7O7[M+H]+: 851.34 found 851.41, UPLC retention time: 5.3 min, purity >95%.

[0655] Example 140

[0656] Procedure: Dissolve Formula 1 (0.12 mmol) and Formula 52 (0.1 mmol) in 1 mL of DMF, add 2–3 drops of DIPEA, then add HATU (0.5 mmol). Stir the reaction mixture at room temperature for 30 min. After the reaction, purify the mixture using preparative liquid chromatography (PCLC). Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5μm, 19×150mm column chromatography yielded a yellow solid LD-331-52 (yield: 65%). LC-MS calculations showed C0... 49 H 50 FN7O7[M+H]+: 867.38 found 867.51, UPLC retention time: 6.4 min, purity >95%.

[0657] Example 141

[0658] Procedure: Formula 53 (0.1 mmol), 2-(2,6-dioxopiperidin-3-yl)-5,6-difluoroisoindoline-1,3-dione (0.1 mmol) were dissolved in 1 mL of DMF, and DIPEA (0.5 mmol) was added. The reaction system was stirred at 100 °C for 5 h. After the reaction, the mixture was purified by preparative liquid chromatography. Mobile phase: methanol / water = 70 / 30. Column type: SymmetryShield. TM RP18 5μm, 19×150mm column chromatography yielded a yellow solid LD-331-53 (yield: 35%). LC-MS calculations showed C0... 48 H 46 FN7O7[M+H]+: 851.34 found 851.42, UPLC retention time: 5.6 min, purity >90%.

[0659] Example 142 Bioactivity Test

[0660] I. Western blotting was used to verify the degradation effects on histone demethylases, deubiquitinases, and C-myc, and the accumulation of H3K4me2 and H3K4me3:

[0661] 1. Cell culture:

[0662] Human acute myeloid leukemia cells (Molm-13) and human acute promyelocytic leukemia cells (HL-60) were cultured in 1640 medium (Gibco) containing 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin (Gibco). Human Burkitt's lymphoma cells (Daudi) were cultured in 1640 medium (Gibco) containing 20% ​​FBS and 1% penicillin / streptomycin (Gibco). Human myeloid monocytic leukemia cells (MV4.11) were cultured in IMDM (Gibco) containing 10% FBS and 1% penicillin / streptomycin (Gibco). All cells were placed in a cell culture incubator at 37°C with 5% CO2, and the medium was changed every 2-3 days. Cells were passaged when the cell density reached approximately 90%. All of the above cell types are commercially available.

[0663] 2. Cell treatment:

[0664] Day 1:

[0665] Depending on the cell growth rate, cells were passaged into 6-well plates at ratios of 1:2 to 1:5, with 2 ml of the corresponding culture medium added to each well. After 4-6 hours of cell seeding, the appropriate dosage of the compound was added for incubation according to the experimental plan.

[0666] the next day:

[0667] Cells were collected at different time points within 6 to 24 hours after drug incubation, and proteins were extracted for Western blotting experiments.

[0668] 3. Extraction of total cellular proteins:

[0669] Cell collection:

[0670] Transfer the treated cells to a 15ml centrifuge tube using a 1ml pipette, centrifuge at 1200rpm for 3min, discard the supernatant, wash the collected cells once with 1ml of pre-cooled PBS, and discard the supernatant.

[0671] Cell lysis:

[0672] Add 30 μl of RIPA lysis buffer (Beyotime) to each sample and mix well by pipetting. Place on ice for lysis for 30 min, mixing with a vortex mixer every 10 min during the process. Then centrifuge at 14000 rpm for 30 min at 4 °C and collect the supernatant into a new EP tube. Store at -80 °C or perform protein quantification.

[0673] Protein quantification:

[0674] Take 2 μl of protein sample and dilute it with 28 μl of H2O. Add 4 μl of the diluted protein sample to a 96-well plate, then add 200 μl of Bradford staining solution (Bio-Rad), incubate at room temperature in the dark for 8 min, and then use a microplate reader to measure the absorbance at 595 nM. Calculate the protein concentration according to the standard curve.

[0675] Western Blot analysis:

[0676] After protein quantification, an equal volume of protein extract was transferred to a new EP tube, and an equal volume of 2×SDS loading buffer was added. The tube was then boiled at 100°C for 10 minutes to denature the protein.

[0677] The 2×SDS loading buffer consists of: 0.5% SDS, 3% β-mercaptoethanol, 15% glycerol, and an appropriate amount of bromophenol blue.

[0678] Based on the size of the target protein to be detected, select an appropriate concentration of polyacrylamide protein separating gel, choose β-actin as the internal control protein, perform Western blotting, and use ECL chromogenic solution (Bio-Rad) for development. The antibodies used are shown in the table below.

[0679] Table A

[0680] II. Validation of the degradation effect on LSD1 and the accumulation of H3K4me1 and H3K4me2 by Western blotting:

[0681] 1. Cell culture:

[0682] H520, BT474, and BT549 cells were cultured in Gibco 1640 medium containing 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin (Gibco). A549, H1299, MDA-MB-231, MDA-MB-453, and MCF-7 cells were cultured in Gibco DMEM containing 10% FBS and 1% penicillin / streptomycin (Gibco). All cells were cultured in a 37°C, 5% CO2 incubator, with medium changed every 2 days. Cells were passaged when the cell density reached approximately 90%. All of these cell lines are commercially available.

[0683] 2. Cell treatment:

[0684] Day 1:

[0685] Depending on the cell growth rate, cells were passaged into 6-well plates at ratios of 1:2 to 1:5, with 2 ml of the corresponding culture medium added to each well.

[0686] the next day:

[0687] After 24 hours of cell seeding, 2 ml of medium was changed in each well, and the appropriate dose of compound was added according to the experimental plan for incubation.

[0688] Day 4:

[0689] After 48 hours of drug incubation, cells were collected and proteins were extracted for Western blotting experiments.

[0690] 3. Extraction of total cellular proteins:

[0691] Cell collection:

[0692] After processing, scrape off the cells with a cell scraper, centrifuge at 1200 rpm for 3 min, discard the supernatant, wash the collected cells once with 1 ml of pre-cooled PBS, and discard the supernatant.

[0693] Cell lysis:

[0694] Add 40 μl of RIPA lysis buffer (Beyotime) to each sample and mix well by pipetting. Place on ice for lysis for 30 min, mixing with a vortex mixer every 10 min during the process. Then centrifuge at 14000 rpm for 30 min at 4 °C and collect the supernatant into a new EP tube. Store at -80 °C or perform protein quantification.

[0695] Protein quantification:

[0696] Take 2 μl of protein sample and dilute it with 18 μl of H2O. Add 4 μl of the diluted protein sample to a 96-well plate, then add 200 μl of Bradford staining solution (Bio-Rad), incubate at room temperature in the dark for 8 min, and then use a microplate reader to measure the absorbance at 595 nM. Calculate the protein concentration according to the standard curve.

[0697] Western Blot analysis:

[0698] After protein quantification, an equal volume of protein extract was transferred to a new EP tube, and an equal volume of 2×SDS loading buffer was added. The tube was then boiled at 100°C for 10 minutes to denature the protein.

[0699] The 2×SDS loading buffer consists of: 0.5% SDS, 3% β-mercaptoethanol, 15% glycerol, and an appropriate amount of bromophenol blue.

[0700] Based on the size of the target protein to be detected, select an appropriate concentration of polyacrylamide protein separating gel, choose β-actin as the internal control protein, perform Western blotting, and use ECL chromogenic solution (Bio-Rad) for development. The antibodies used are shown in Table 1.

[0701] Table 1.

[0702] III. Cell Inhibition Experiment 1:

[0703] 1. Cell culture:

[0704] Molm-13 and HL-60 cells were cultured in Gibco 1640 medium containing 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin (Gibco). Daudi cells (MV4) were cultured in Gibco 1640 medium containing 20% ​​FBS and 1% penicillin / streptomycin (Gibco). 11 cells were cultured in Gibco IMDM medium containing 10% FBS and 1% penicillin / streptomycin (Gibco). All cells were cultured in a 37°C, 5% CO2 incubator, with medium changes every 2-3 days. Cells were passaged when the cell density reached approximately 90%. All of these cell lines are commercially available.

[0705] 2.IC 50 Detection:

[0706] Day 1: When the cells reach a density of 80%–90%, gently mix and pipette 1 ml into a 15 ml centrifuge tube. Centrifuge at 1000 rpm for 3 min, then resuspend in the appropriate culture medium to a concentration of 1×10⁻⁶. 4 / ml, add 100μl of cell suspension to each 96-well plate;

[0707] For LD-331-LLH, in Molm-13, HL-60, Daudi, and MV4;11 cells, the compound was prepared into a 60 μM solution using the corresponding culture medium, and then serially diluted 3-fold with the corresponding culture medium. Three replicates were set up for each concentration gradient, and 100 μl of the compound solution was added to each replicate well, resulting in a final concentration of 1–30000 nM. Three wells without the compound were set up as negative controls, and 200 μl of the corresponding culture medium was added to each well. The cells were incubated at 37°C in a cell culture incubator with a CO2 concentration of 5%. The detection methods for other compounds were the same as described above.

[0708] Day 4: After incubating the cells with the compound for 72 hours, add 20 μl of CCK8 stock solution (MCE) to each well, gently shake to mix, and then incubate in a cell culture incubator at 37°C and 5% CO2 for 2-4 hours. Use a microplate reader to detect the absorbance at 450 nM, calculate cell viability, and plot a curve.

[0709] IV. Cell Inhibition Experiment 2:

[0710] 1. Cell culture:

[0711] H520, BT474, and BT549 cells were cultured in Gibco 1640 medium containing 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin (Gibco). A549, H1299, MDA-MB-231, MDA-MB-453, and MCF-7 cells were cultured in Gibco DMEM containing 10% FBS and 1% penicillin / streptomycin (Gibco). All cells were cultured in a 37°C, 5% CO2 incubator, with medium changed every 2 days. Cells were passaged when the cell density reached approximately 90%. All of these cell lines are commercially available.

[0712] 2.IC 50 Detection:

[0713] Day 1: When the cells reach a density of 80%–90%, digest them with 0.05% trypsin (Gibco) and resuspend them in the appropriate culture medium to a concentration of 2 × 10⁻⁶. 4 / ml, add 100μl of cell suspension to each 96-well plate;

[0714] Day 2: For the LSD1 inhibitor LI-1 used in the degradation agent of this invention, the compound was prepared into a 64 μM solution using the corresponding culture medium, and then serially diluted 2-fold with the corresponding culture medium. Three replicates were set up for each concentration gradient. 100 μl of LI-1 solution was added to each replicate well, that is, the final concentration was 0.125-32 μM. At the same time, three wells without LI-1 were set up as negative controls. 100 μl of the corresponding culture medium was added to each well.

[0715] For other compounds, in H520, A549, and H1299 cells, the compounds were prepared into 60 μM solutions using the corresponding culture media, and then serially diluted 3-fold with the corresponding culture media. Three replicates were set up for each concentration gradient, and 100 μl of the compound solution was added to each replicate well, resulting in a final concentration of 1–30000 nM. Three wells without the compound were set up as negative controls, with 100 μl of the corresponding culture media added to each well. In MDA-MB-231, MCF-7, and BT474 cells, the compounds were prepared into 12.5 μM solutions using the corresponding culture media, and then serially diluted 5-fold with the corresponding culture media. Three replicates were set up for each concentration gradient, and 100 μl of the compound solution was added to each replicate well, resulting in a final concentration of 0.0032–6250 nM. Three wells without the compound were set up as negative controls, with 100 μl of the corresponding culture media added to each well. The cells were incubated at 37°C in a cell culture incubator with 5% CO2.

[0716] Day 5: Add 100 μl of CCK8 stock solution (MCE) to 1 ml of the corresponding complete culture medium to prepare CCK8 working solution. After incubating the cells with the compound for 72 h, discard the original culture medium and add 100 μl of CCK8 working solution to each well. Then, incubate the cells in a cell culture incubator at 37°C and 5% CO2 for 2 h. Use a microplate reader to detect the absorbance at 450 nM, calculate cell viability, and plot the curve.

[0717] V. Animal Experiment 1:

[0718] For HL-60 cells, pipette 1 ml of the cells into a 15 ml centrifuge tube, centrifuge at 1000 rpm for 3 min, discard the supernatant, wash twice with PBS, count the cells, and resuspend the cells in PBS to a concentration of 5 × 10⁻⁶ cells / mL. 7 5 × 10⁶ / ml of cell suspension was subcutaneously injected into the groin of 4-5 week old female BALB / c nude mice. 6 One cell;

[0719] For MV4;11 cells, pipette 1 ml of the cells into a 15 ml centrifuge tube, centrifuge at 1000 rpm for 3 min, discard the supernatant, wash twice with PBS, count the cells, and resuspend the cells in PBS to a concentration of 8 × 10⁻⁶. 7 8 × 10⁸ cells / ml were subcutaneously injected into the groin of 4-5 week old female BALB / c nude mice using a cell suspension. 6 One cell;

[0720] When the tumor grows to a size of 50-100 mm 3Mice were divided into three groups according to tumor size. LD-331-LLH was dissolved in 50% PEG200 to a concentration of 40 mg / ml. Each group of mice was injected intraperitoneally with 0.1 mg / kg of LD-110, 1 mg / kg of LD-110, or the same volume of solvent. The administration was once a day for five days a week, followed by a two-day break. The total administration period was 3-4 weeks. During this period, the body weight and tumor volume of the mice were measured three times a week.

[0721] VI. Animal Experiment 2:

[0722] For A549 cells, the cells were digested with 0.05% trypsin (Gibco), neutralized with an equal volume of complete culture medium, centrifuged at 800 rpm for 3 min, the supernatant was discarded, the cells were washed twice with PBS, and after counting, the cells were resuspended in PBS to a concentration of 1×10⁻⁶ cells / mL. 8 1 × 10⁶ / ml of cell suspension was administered subcutaneously to the groin of 4-5 week old male BALB / c nude mice. 7 One cell;

[0723] For MDA-MB-231 cells, the cells were digested with 0.05% trypsin (Gibco), neutralized with an equal volume of complete culture medium, centrifuged at 800 rpm for 3 min, the supernatant was discarded, the cells were washed twice with PBS, and after counting, the cells were resuspended in PBS to a concentration of 5 × 10⁻⁶ cells / mL. 7 5 × 10⁶ / ml of cell suspension was subcutaneously injected into the groin of 4-5 week old female BALB / c nude mice. 6 One cell;

[0724] When the tumor grows to a size of 50-100 mm 3 Mice were divided into three groups according to tumor size. LD-110 and LI-1 were dissolved in 50% PEG200 to a concentration of 10 mg / ml. Each group of mice was injected intraperitoneally with 50 mg / kg of LD-110, LI-1 or the same volume of solvent. The drugs were administered once a day for five days a week, followed by a two-day break. The total administration period was 3-4 weeks. During this period, the body weight and tumor volume of the mice were measured three times a week.

[0725] Example 143 Animal Pharmacokinetic (PK) Study

[0726] 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.

[0727] Sample collection and analysis:

[0728] Blood volume collected: 0.03 mL

[0729] Blood collection method: orbital venous plexus

[0730] Anticoagulant: EDTA-K2

[0731] Blood collection time:

[0732] IV: 5min, 15min, 30min, 1h, 2h, 4h, 7h, 24h

[0733] PO: 15min, 30min, 1h, 2h, 4h, 7h, 24h

[0734] 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 freezer at -75℃±15℃.

[0735] Sample pretreatment was performed according to the Standard PK procedure.

[0736] The obtained data were used to calculate pharmacokinetic parameters using WinNonlin software. IV pharmacokinetic parameters include 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.

[0737] As shown in Figure 1, in solid tumor cell breast cancer cell lines, the compounds of the present invention can dose-dependently degrade LSD1, and in particular, can efficiently and dose-dependently downregulate the level of c-MYC protein.

[0738] As shown in Figure 2, in multiple hematologic malignancy cell lines, the compounds of the present invention can efficiently and dose-dependently degrade demethylase protein targets, such as KDM5A, and can efficiently and dose-dependently downregulate the level of c-MYC protein.

[0739] As shown in Figure 3, in multiple hematologic malignancy cell lines, the compound of this invention can efficiently and time-dependently degrade histone demethylase KDM5A, while simultaneously efficiently and time-dependently downregulating the level of c-MYC protein.

[0740] As shown in Figure 4, the compounds of the present invention can effectively inhibit the growth of tumor cells in multiple hematologic malignancy cell lines.

[0741] As shown in Figure 5, in multiple hematologic malignancies, the compounds of the present invention can effectively degrade the levels of histone demethylases and deubiquitinases.

[0742] As shown in Figure 6, in multiple solid tumor cells, the compounds of the present invention can effectively degrade the level of pathogenic proteins and effectively inhibit cell proliferation.

[0743] As shown in Figures 7-9, the compounds exhibited different levels of degradation activity against LSD1 protein in lung cancer cell lines.

[0744] As shown in Figures 10 and 11, compound LD-110 exhibits superior inhibitory activity against lung cancer cells compared to its corresponding LSD1 inhibitor LI-1.

[0745] As shown in Figure 12, LD-110 has a good degradation effect on LSD1 protein in breast cancer cells, while significantly accumulating the substrates H3K4Me1 and H3K4Me2.

[0746] As shown in Figure 13, LD-110 can effectively inhibit the growth of breast cancer cells.

[0747] As shown in Figure 14, LD-110 can effectively degrade LSD1 protein and inhibit cell proliferation in esophageal cancer cells.

[0748] As shown in Figure 15, the mechanism study showed that the LSD1 inhibitor LI-1, the CRBN E3 ligase ligand thalidomide, the mimetic inhibitor MLN4924, and the proteasome inhibitor MG132 can effectively block the degradation of LSD1 protein by LD-110, indicating that this series of compounds exert their effects through the ubiquitination proteasome pathway.

[0749] As shown in Figure 16, LD-110 can effectively inhibit the growth of A549 lung cancer xenografts without affecting the weight of mice.

[0750] As shown in Figure 17, LD-110 can effectively inhibit the growth of the MDA-MB-231 breast cancer model xenograft without affecting the weight of mice.

[0751] As shown in Figure 18, LD331 and its derivatives LD331-14 and LD331-18 can significantly reduce the levels of KDM5A, c-MYC, and RNF6 in the acute myeloid leukemia (AML) cell line HL-60.

[0752] As shown in Figure 19, LD331 and its derivatives LD331-14 and LD331-18 significantly reduced the expression of KDM5A, as well as c-MYC and RNF6 in the acute myeloid leukemia (AML) cell line MV4;11.

[0753] As shown in Figure 20, LD331 and its derivatives LD331-14 and LD331-23 significantly reduced the expression of KDM5A, c-MYC, and RNF6 in various solid malignant tumor cell lines.

[0754] As shown in Tables 1 and 2, the compounds of the present invention exhibit significant inhibitory activity against a variety of hematologic malignancies and solid tumor cell lines.

[0755] Table 1. IC50 of the compounds of the present invention against various hematologic malignancy cell lines 50 value

[0756] Table 2. Half-maximal inhibitory concentrations (IC50) of the compounds of the present invention against various solid tumor cell lines. 50 )

[0757] As shown in Table 3 below, the compounds of the present invention exhibit good pharmacokinetic parameters in mice.

[0758] Table 3

[0759] 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

A compound of general formula (I) or its stereoisomers, enantiomers, transisomers, optical isomers, racemates, tautomers or pharmaceutically acceptable salts thereof, their hydrates, or their isotopically labeled compounds: SLD(I) in, S is the target protein binding ligand, which has the structure shown below: Ring A is selected from: substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-10 membered heterocyclic group, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-10 membered heteroaryl; R is selected from: chemical bond, non-existent, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 alkylamino, C3-C8 cycloalkyl, 4-10 heterocyclic, C6-C10 aryl, 5-10 heteroaryl; Z is selected from: chemical bond, CO; R m Selected from: chemical bonds, H, halogens, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 alkylamino, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-10 heterocyclic groups, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-10 heteroaryl; L is -L1-(L2) p -L3-, which connects S and D via covalent bonds; L1 and L3 are each independently selected from the following groups: chemical bond, absent, CO, -O-, -CONH-, substituted or unsubstituted CH2, substituted or unsubstituted NH, S(O), S(O)2, ethynyl, -CH2CH2O-, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-10 heterocyclic group, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-10 heteroaryl; Each L2 is independently selected from the following groups: chemical bond, absent, CO, -O-, -CONH-, substituted or unsubstituted CH2, substituted or unsubstituted NH, S(O), S(O)2, ethynyl, -CH2CH2O-, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-10 heterocyclic group, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-10 heteroaryl; The substitution refers to having one or more substituents selected from the group consisting of: halogen, amino, hydroxyl, cyano, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 alkylamino, C3-C8 cycloalkyl, C6-C10 aryl, 4-10 heterocyclic, and 5-10 heteroaryl. p is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20; D is the E3 ubiquitin ligase-binding ligand. The compound of claim 1, wherein, D has a structure selected from the following group: Z 1 R1 is CH or N; R1 is selected from halogen, amino, hydroxyl, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 alkylamino, C3-C8 cycloalkyl, C6-C10 aryl, 4-10 heterocyclic, 5-10 heteroaryl; among which, This indicates the site covalently connected to L. The compound of claim 1, wherein, The compound has the structure shown in formula (II): Among them, L, D, Z, R m A is as defined in claim 1. The compound of claim 1, wherein, The compound has the structure shown in formula (III): Among them, L, Z, R m A is as defined in claim 1. The compound of claim 1, wherein, The compound has the structure shown in formula (IV): Among them, L, Z, R m As defined in claim 1. The compound of claim 1, wherein, L is selected from the following group: m, n, and q are each independently selected from: 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. The compound of claim 1, wherein, The compound has a structure selected from the group consisting of: A pharmaceutical composition comprising the compound of any one of claims 1-7 or a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof, and a pharmaceutically acceptable carrier. Use of the compound of any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating histone demethylases, deubiquitinases, c-MYC and / or RNF6-mediated diseases or conditions; wherein the histone demethylases, deubiquitinases, c-MYC and / or RNF6-mediated diseases or conditions include oncological diseases. The use as described in claim 9, wherein, The tumor diseases mentioned are selected from the following group: breast cancer, leukemia, lymphoma, lung cancer, colon cancer, liver cancer, ovarian cancer, pancreatic cancer, colorectal cancer, thyroid cancer, skin cancer, bone cancer, melanoma, salivary gland tumors, neuroendocrine tumors, brain tumors, neuroblastoma, mesothelioma, esophageal cancer, pulmonary sarcoma, medulloblastoma, glioblastoma, retinoblastoma, kidney cancer, bladder cancer, osteosarcoma, gastric cancer, uterine cancer, vulvar cancer, small intestine cancer, prostate cancer, bile duct cancer, ureteral cancer, adrenocortical cancer, head and neck cancer, colorectal cancer, nasopharyngeal carcinoma, myeloma, ovarian cancer, and cervical cancer.

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