Compound for degrading USP7 protein by means of targeted ubiquitination, pharmaceutical composition thereof and use thereof

Abstract

A compound for degrading USP7 protein by means of targeted ubiquitination, a pharmaceutical composition thereof and the use thereof. The structural formula thereof is shown as formula (I), wherein A is a specific protein ligand in an E3 ubiquitin ligase complex, and L is a divalent linker group between a small molecule ligand of the USP7 protein and the specific protein ligand in the E3 ubiquitin ligase complex. The protein degradation chimera has both USP7 protein inhibitory activity and USP7 protein-degrading activity, can effectively inhibit malignant proliferation of acute lymphoblastic leukemia cells, and thus can be used for diseases associated with abnormal USP7 protein expression.

Description

A compound targeting ubiquitination degradation of USP7 protein, a pharmaceutical composition thereof and application TECHNICAL FIELD

[0001] The present application relates to the field of medicinal chemistry, in particular to a kind of compound targeting ubiquitination degradation of USP7 protein, its preparation method and pharmaceutical composition and application. BACKGROUND

[0002] Normal cellular homeostasis requires a balance between protein synthesis and degradation. Ubiquitination and deubiquitination, a protein post-translational modification, is an important regulatory mechanism for protein stability and degradation. There are about 1000 enzymes encoded in the human genome involved in ubiquitination regulation. Under the participation of these enzymes, the ubiquitination modification process of proteins undergoes an enzymatic cascade reaction and finally adds a ubiquitin tag to the lysine residue on the substrate protein. In addition, ubiquitination is a highly dynamic and reversible process, and the covalently linked ubiquitin can be removed from the ubiquitin-labeled protein by proteolysis mediated by deubiquitinating enzymes (DUBs), thereby adding another layer of regulation. The ubiquitin-proteasome system (UPS) plays a very important role in maintaining cell growth, survival and homeostasis. The imbalance of UPS related enzymes will cause the imbalance of protein homeostasis, leading to the occurrence of many diseases such as tumors, cardiovascular diseases, Alzheimer's disease and so on. Therefore, a lot of efforts have been devoted to the development of drugs targeting the UPS system. Currently, the human DUBs identified include 99 enzymes, which can be divided into 7 subfamilies according to the sequence similarity of their respective catalytic domains. The USPs family is the largest subfamily of DUBs, with about 60 members. Among the USPs family, ubiquitin-specific protease USP7 is the most thoroughly studied deubiquitinating enzyme. The human USP7 protein has a molecular weight of about 135 kDa, containing 1102 amino acid residues, which can be divided into four domains: the N-terminal polyglutamine extension fragment (poly Q), the tumor necrosis factor receptor-associated factor (TRAF) domain, the catalytic core domain and the C-terminal tandem of five ubiquitin-like (UBL) domains. USP7 regulates the abundance or function of multiple disease-related proteins through its deubiquitinating enzyme activity and is involved in various pathological processes, especially in various carcinogenic pathways. USP7 plays a multidimensional role in many cancers such as prostate cancer, bladder cancer, lung cancer, liver cancer, brain cancer, colon cancer, breast cancer, epithelial ovarian carcinoma (EOC), liver cancer, leukemia and so on. Mechanistically, USP7 stabilizes MDM2 through its deubiquitination to promote the degradation of the tumor suppressor protein p53, thereby inhibiting the apoptosis of tumor cells. Designing small molecule inhibitors to target the activation of the USP7-MDM2-p53 signaling pathway is an important strategy in anti-tumor therapy. More and more evidence supports the key role of USP7 in various diseases, and therefore USP7 is a very promising target for drug development. Currently reported USP7 inhibitors have poor physicochemical properties, no in vivo data, and no small molecule inhibitors targeting USP7 approved by FDA. The PROTAC strategy can not only block the catalytic function of proteins, but also remove proteins as a whole, which has a good application prospect of overcoming drug resistance. Therefore, the present application provides an important reference for the feasibility of USP7 as a drug development target, leukemia treatment and the development and application of PROTAC technology by designing and synthesizing a series of USP7 degradation agents. SUMMARY

[0003] To solve the above problems, one of the purposes of the present application is to provide a novel compound or its pharmaceutically acceptable salt, hydrate or solvate for targeting ubiquitination degradation of USP7 protein.

[0004] According to one embodiment of the present application, a compound represented by formula (I) or a pharmaceutically acceptable salt, stereoisomer, enantiomer, diastereomer, atropisomer, racemate, or isotopically-labeled compound thereof is provided:

[0005] wherein,

[0006] Y1is selected from the group consisting of hydrogen, deuterium, C 1-6 alkyl, halogenated C 1-6 alkyl, C 3-8 cycloalkyl, and 3-8 membered heterocycloalkyl;

[0007] B ring is selected from the group consisting of C 6-10 aromatic ring, C 3-10 saturated or unsaturated alicyclic ring, and 5-10 membered heteroaromatic ring;

[0008] Y2is selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, halogenated C 1-6 alkoxy, -CN, -NO2, and C 1-6 alkylsulfonyl;

[0009] Y3is selected from the group consisting of C 1-6 alkylene, -CH2NY 3a -, -C(O)NY 3a -, -NY 3a -, -O-C 2-6 alkylene-NY 3a -, and 3-8 membered heterocycloalkylene, wherein Y 3a is hydrogen, C 1-6 alkyl, halogenated C 1-6 alkyl, or C 3-8 cycloalkyl;

[0010] Y4is selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 alkyl, and halogenated C 1-6 alkoxy;

[0011] C ring is selected from the group consisting of C 6-10aromatic ring, C 3-10 saturated or unsaturated alicyclic, and 5-10 membered heteroaromatic ring;

[0012] Y5is selected from the group consisting of: hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 alkyl, halogenated C 1-6 alkoxy, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, -OH, -C(O)OY 5a , -C(O)NHY 5a , -C(O)NHOH, and -NH(CO)Y 5a , wherein Y 5a is hydrogen, C 1-6 alkyl or C 3-8 cycloalkyl;

[0013] A is the following general structure A1:

[0014] wherein,

[0015] R1is selected from the group consisting of: hydrogen, substituted 5-10 membered heteroaryl, C 1-6 alkyl, substituted C 6-10 aryl, substituted C 1-6 alkoxy, and substituted 3-8 membered heterocyclyl; when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 alkyl, halogenated C 1-6 alkoxy, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, and -OH;

[0016] R2is selected from the group consisting of: hydrogen, halogen, -CN, substituted C 1-6 alkyl, or substituted C 1-6 alkoxy, and substituted C 3-8 cycloalkyl; when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, and -OH;

[0017] R 3a and R 3b are each independently selected from the group consisting of: hydrogen, substituted C 3-8 cycloalkyl, substituted C 1-6 alkyl, and substituted 3-8 membered heterocycloalkyl; when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, C1-6 alkylsulfonyl, -CN, -NO2, -NH2, and -OH;

[0018] R4is selected from the group consisting of: hydrogen, substituted C 1-6 alkyl, and substituted C 3-8 cycloalkyl; and when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, -CN, -NO2, -NH2, and -OH;

[0019] R5and R6are independently selected from the group consisting of hydrogen, -OH, -SH, and substituted C 1-6 alkyl, or R5, R6and the carbon atom to which they are attached form a carbonyl group; and when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, -CN, -NO2, -NH2, and -OH;

[0020] R7is selected from the group consisting of: hydrogen and substituted C 1-6 alkyl; and when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, -CN, -NO2, -NH2, and -OH;

[0021] R8is selected from the group consisting of: hydrogen and OH;

[0022] R 9a and R 9b are each independently selected from the group consisting of: hydrogen and substituted C 1-6 alkyl; or R 9a and R 9b groups can together form an oxo or substituted C 3-8 cycloalkyl; and when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, -CN, -NO2, -NH2, and -OH;

[0023] R 10 and R 11 are selected from the group consisting of: hydrogen, substituted C 1-6 alkyl, substituted C 3-8 cycloalkyl, substituted C 1-6 alkoxy, substituted C 1-6 alkylthio, and substituted 5-10 membered heteroaryl; or R 10 , R 11 and the carbon atom to which they are attached form a substituted C 3-8 cycloalkyl; and when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, and -OH;

[0024] R 12 is selected from the group consisting of: substituted 3-8 membered heterocyclylene, substituted 5-10 membered heteroarylene, substituted C 6-10 arylene, and In the substitutions, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 alkyl, halogenated C 1-6 alkoxy, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, and -OH;

[0025] wherein R 13 is selected from the group consisting of hydrogen, deuterium, substituted C 1-6 alkyl, and substituted C 3-8 cycloalkyl; in the substitutions, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, and -OH;

[0026] The L has the structure of L 1 -L 2 -L 3 wherein L 1 , L 2 , and L 3 may be present simultaneously or one or both of them are present; wherein, when L 1 and / or L 3 are present, they are independently selected from the following structures: and

[0027] wherein n = 0-20, preferably 0-5, more preferably 0-2;

[0028] R 15 is selected from the group consisting of H, C 1-10 alkyl, and C 3-10 cycloalkyl;

[0029] R 16 is selected from the group consisting of H, C 1-10 alkyl, and C 3-10 cycloalkyl;

[0030] When L 2 is present, it is selected from the group consisting of:

[0031] and

[0032] wherein m = 0-20, preferably 0-12.

[0033] According to one embodiment of the present application, wherein the compound is selected from the group consisting of compounds of formula (II):

[0034] wherein,

[0035] Y1is selected from the group consisting of: hydrogen, deuterium, C 1-6 alkyl, and C 3-8 cycloalkyl;

[0036] B ring is selected from the group consisting of: C 6-10 aromatic ring, C 3-10 saturated or unsaturated alicyclic, and 5-10 membered heteroaromatic ring;

[0037] Y2is selected from the group consisting of: hydrogen, deuterium, halogen, C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, halogenated C 1-6 alkyl, CN, NO2, and C 1-6 alkylsulfonyl;

[0038] Y3is selected from the group consisting of: C 1-6 alkylene, -CH2NY 3a -, and -NY 3a -; wherein Y 3a is hydrogen, C 1-6 alkyl, halogenated C 1-6 alkyl, or C 3-8 cycloalkyl;

[0039] Y4is selected from the group consisting of: C 1-6 alkyl, C 3-8 cycloalkyl, and halogenated C 1-6 alkyl;

[0040] C ring is selected from the group consisting of: C 6-10 aromatic ring, C 3-10 cycloalkyl, and 5-10 membered heteroaromatic ring;

[0041] Y5is selected from the group consisting of: hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 alkyl, and halogenated C 1-6 alkoxy;

[0042] R1is selected from the group consisting of: substituted 5-10 membered heteroaryl, substituted C 6-10 aryl, and substituted 3-8 membered heterocyclyl; when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8cycloalkyl, haloC 1-6 alkyl, haloC 1-6 alkoxy, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, and -OH;

[0043] R 3a and R 3b are each independently selected from the group consisting of hydrogen, substituted C 1-6 alkyl, substituted C 3-8 cycloalkyl, and substituted 3-8 membered heterocycloalkyl; and wherein said substituents, when present, are selected from the group consisting of hydrogen, deuterium, and halogen;

[0044] R5and R6are each independently selected from the group consisting of hydrogen, -OH, -SH, and substituted C 1-6 alkyl, or R5, R6and the carbon atom to which they are attached form a carbonyl group; and wherein said substituents, when present, are selected from the group consisting of hydrogen, deuterium, halogen;

[0045] R 10 and R 11 are selected from the group consisting of hydrogen, substituted C 1-6 alkyl, substituted C 3-8 cycloalkyl, substituted C 1-6 alkoxy, substituted C 1-6 alkylthio, and substituted 5-10 membered heteroaryl; or R 10 , R 11 and the carbon atom to which they are attached form a substituted C 3-8 cycloalkyl; and wherein said substituents, when present, are selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, haloC 1-6 alkyl, haloC 1-6 alkoxy, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, and -OH;

[0046] R 12 is selected from the group consisting of substituted 5-10 membered heteroarylene, substituted C 6-10 arylene, and and wherein said substituents, when present, are selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, haloC 1-6 alkyl, haloC 1-6 alkoxy, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, and -OH;

[0047] R 13selected from the group consisting of hydrogen and substituted C 1-6 alkyl; when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, and -OH;

[0048] said L has the structure of L 1 -L 2 -L 3 wherein L 1 , L 2 , L 3 may be present simultaneously or one or both of them is present; wherein, when L 1 and / or L 3 is present, it is independently selected from the following structures:

[0049] wherein n = 0-20, preferably 0-5, more preferably 0-2;

[0050] R 15 is selected from the group consisting of H, C 1-10 alkyl and C 3-10 cycloalkyl;

[0051] when L 2 is present, it is selected from the group consisting of:

[0052] and

[0053] wherein m = 0-20, preferably 0-12.

[0054] According to one embodiment of the present application, wherein the compound is selected from the group consisting of compounds represented by formula (III):

[0055] wherein,

[0056] Y1is selected from the group consisting of C 1-6 alkyl and C 3-8 cycloalkyl;

[0057] the B ring is selected from the group consisting of C 6-10 aromatic ring, C 3-10 saturated or unsaturated alicyclic ring, and 5-10 membered heteroaromatic ring;

[0058] Y2is selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, and halogenated C 1-6Alkoxy;

[0059] Y3 is selected from: C 1-3 Alkyl groups, -CH2NH-, and -NH-;

[0060] Y5 is selected from: hydrogen, deuterium, halogens, and carbon. 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 Alkyl groups and halogenated C 1-6 Alkoxy;

[0061] R1 is selected from:

[0062] Among them, R 17 It is hydrogen or a substituted C 1-6 Alkyl group; when substituted, the substituent is selected from hydrogen, deuterium, halogen, -CN, -NO2, -NH2, and -OH;

[0063] R 3a and R 3b Selected independently from: hydrogen and the substituted C 1-6 Alkyl group; when substituted, the substituent is selected from hydrogen, deuterium, or halogen;

[0064] R 10 and R 11 Selected from: hydrogen, substituted C 1-6 Alkyl groups, and substituted C groups 3-8 cycloalkyl, or R 10 R 11 The carbon atoms they are attached to form substituted C atoms. 3-8 Cycloalkyl; when substituted, the substituent is selected from hydrogen, deuterium, or halogen;

[0065] R 12 Selected from: substituted 5-10 quinone heteroaryl groups, substituted C 6-10 Aspartic acid, and When substituted, the substituent is selected from hydrogen, deuterium, halogen, C. 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 Alkyl groups and halogenated C 1-6 Alkoxy;

[0066] R 13 Selected from: Hydrogen and C 1-6 alkyl;

[0067] The L has L 1 -L 2 -L 3 The structure, where L1 , L 2 , L 3 , L 1 and / or L 3 , when present, is independently selected from the following structures:

[0068] wherein n = 0-20, preferably 0-5, more preferably 0-2;

[0069] R 15 is selected from the group consisting of H, C 1-10 alkyl and C 3-10 cycloalkyl;

[0070] L 2 , when present, is selected from the group consisting of:

[0071]

[0072] wherein m = 0-20, preferably 0-12.

[0073] According to one embodiment of the present application, the compound is selected from the group consisting of compounds of formula (IV):

[0074] wherein,

[0075] Y2and Y5are each selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, and halogenated C 1-6 alkoxy;

[0076] R 3a and R 3b are each independently selected from the group consisting of hydrogen and substituted C 1-6 alkyl; when substituted, the substituents are selected from the group consisting of hydrogen, deuterium and halogen;

[0077] R 10 and R 11 are selected from the group consisting of hydrogen, substituted C 1-6 alkyl, and substituted C 3-8 cycloalkyl; when substituted, the substituents are selected from the group consisting of hydrogen, deuterium and halogen;

[0078] R 12 is selected from the group consisting of substituted 5-6 membered heteroarylene and ​​​​​​​​​​​​​​​​​​​​​​​​In the substituents, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 alkyl, and halogenated C 1-6 alkyl;

[0079] R 13 is selected from the group consisting of hydrogen or C 1-6 alkyl;

[0080] The L has the structure of L 1 -L 2 -L 3 , wherein L 1 , L 2 , L 3 may exist simultaneously or one or both of them exist; wherein, when L 1 and / or L 3 exist, they are independently selected from the following structures: and

[0081] wherein n = 0-20, preferably 0-5, more preferably 0-2;

[0082] R 15 is selected from the group consisting of H, C 1-10 alkyl and C 3-10 cycloalkyl;

[0083] When L 2 exists, it is selected from the group consisting of: and

[0084] wherein m = 0-20, preferably 0-12.

[0085] According to one embodiment of the present application, wherein the compound is selected from the group consisting of compounds shown in formula (V):

[0086] wherein,

[0087] Y1is selected from the group consisting of C 1-6 alkyl;

[0088] Y2is selected from the group consisting of hydrogen, halogen, C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, halogenated C 1-6 alkyl;

[0089] Y5is selected from: hydrogen, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 alkyl, halogenated C 1-6 alkoxy;

[0090] said A3moiety is selected from:

[0091] said L is the same as defined above.

[0092] According to one embodiment of the present application, wherein,

[0093] said L is selected from:

[0094] wherein m = 1-10.

[0095] According to one embodiment of the present application, wherein, the compound is selected from:

[0096] In another aspect, the present application provides a pharmaceutical composition comprising a therapeutically effective amount of one or more of the above-mentioned compounds, pharmaceutically acceptable salts, stereoisomers, enantiomers, diastereomers, atropisomers, optical isomers, racemates, polymorphs, solvates and isotopically-labeled compounds thereof, and optionally a pharmaceutically acceptable carrier or excipient.

[0097] In another aspect, the present application provides the use of the above-mentioned compounds, or pharmaceutically acceptable salts, stereoisomers, enantiomers, diastereomers, atropisomers, optical isomers, racemates, polymorphs, solvates or isotopically-labeled compounds thereof, for the preparation of a medicament for the prevention and / or treatment of a disease or disorder or disease state mediated by USP7.

[0098] In another aspect, in the disease or disorder or disease state mediated by USP7, the disease related to abnormal expression of USP7 protein activity includes a tumor, a hemangion, a leukemia, an ovarian cancer, a breast cancer, a lung cancer, a pancreatic cancer, a kidney cancer, a melanoma, a liver cancer, a colon cancer, a sarcoma, a brain cancer, a prostate cancer, a lymphoma or a multiple myeloma, etc. and a non-tumor related to expression includes hyperglycemia, diabetes, obesity, hyperlipidemia, hypercholesterolemia, hyperlipoproteinemia, hypertriglyceridemia, hypertension, hyperinsulinemia, hyperuricemia, Parkinson's disease, Alzheimer's disease, etc. Advantages

[0099] The compound provided by the application can effectively inhibit USP7 protein and has USP7 target protein degradation function. The protein degradation mechanism is to combine USP7 protein and E3 ubiquitin ligase at the same time, and then ubiquitinate and degrade the target protein. The compound can significantly inhibit the malignant proliferation of tumor cells such as leukemia cells, and can provide an important reference for the research and development of USP7 as a drug development target, the treatment of leukemia, and the development and application feasibility of PROTAC technology. BRIEF DESCRIPTION OF DRAWINGS

[0100] Figure 1 is a comparison of the tumor growth in mice and the change in body weight of mice, and the content of USP7 protein, p53 protein and p21 protein in tumor tissue of mice after administration of blank control, azacitidine group and different concentrations of compound s18 of the application in RS4; 11 xenotransplant tumor model mice. DETAILED DESCRIPTION

[0101] In order to enable those skilled in the art to understand the characteristics and effects of the present application, the following is a general description and definition of the terms and phrases mentioned in the specification and the scope of the application. Unless otherwise specified, all technical and scientific words used in the text have the usual meaning understood by those skilled in the art for the present application, and in case of conflict, the definition in the specification shall prevail.

[0102] In this text, the words "include", "include", "have", "contain" or any other similar words are open-ended transitional phrases, which are intended to cover non-exclusive inclusion. For example, a composition or article containing a plurality of elements is not limited to the elements listed herein, but can also include other elements not explicitly listed but inherent in the composition or article. In addition, unless specifically stated otherwise, the word "or" means inclusive "or", not exclusive "or". For example, any of the following conditions satisfies the condition "A or B": A is true (or exists) and B is false (or does not exist), A is false (or does not exist) and B is true (or exists), A and B are both true (or exist). In addition, in this text, the interpretation of the words "include", "include", "have", "contain" should be considered as having been specifically disclosed and at the same time covering closed or semi-closed transitional words such as "consisting of" and "consisting essentially of".

[0103] In this document, all features or conditions that are described in terms of a numerical range or a percentage range are only intended to simplify the description and the convenience. Accordingly, the description of a numerical range or a percentage range is to be considered as having encompassed and specifically disclosed all possible subranges and individual numerical values within the range, in particular integer values. For example, a range description of "1 to 8" is to be considered as having specifically disclosed all subranges, in particular subranges defined by all integer values, such as 1 to 7, 2 to 8, 2 to 6, 3 to 6, 4 to 8, 3 to 8, etc. and as having specifically disclosed individual numerical values within the range, such as 1, 2, 3, 4, 5, 6, 7, 8, etc. The aforementioned interpretation method applies to all content of the present application, regardless of the extent of the range, unless otherwise indicated.

[0104] If a numerical value or other numerical value or parameter is expressed in a range, a preferred range or a series of upper and lower limits, it is to be understood that all ranges formed by any pair of the upper or preferred limit of the range and the lower or preferred limit of the range have been specifically disclosed herein, whether or not these ranges have been separately disclosed. Furthermore, if a range of values is mentioned herein, the range should include its endpoints and all integers and fractions within the range, unless otherwise stated.

[0105] In this document, numerical values are to be understood as having the precision of the number of significant figures of the numerical value, provided that the object of the invention can be achieved. For example, the number 40.0 is to be understood as encompassing the range from 39.50 to 40.49.

[0106] In this document, in the case where Markush groups or alternative expressions are used to describe features or examples of the invention, the person skilled in the art will understand that subgroups of all the elements of the Markush group or any combination of individual elements can also be used to describe the invention. For example, if X is described as "selected from the group consisting of X1, X2 and X3", it is also to be understood that the claim has been fully described in which X is X1 and in which X is X1 and / or X2. Furthermore, in the case where Markush groups or alternative expressions are used to describe features or examples of the invention, the person skilled in the art will understand that subgroups of all the elements of the Markush group or any combination of individual elements can also be used to describe the invention. Accordingly, for example, if X is described as "selected from the group consisting of X1, X2 and X3" and Y is described as "selected from the group consisting of Y1, Y2 and Y3", it is to be understood that the claim has been fully described in which X is X1 or X2 or X3 and Y is Y1 or Y2 or Y3.

[0107] In the present invention, the aromatic, alicyclic or heteroaromatic ring can be monocyclic or annelated.

[0108] The following detailed description is merely illustrative in nature and is not intended to limit the application or the use of embodiments of the application. Furthermore, there is no intention to be bound by any theory of operation described or implied herein.

[0109] The application is further described by the following examples with reference to the accompanying drawings.

[0110] Example 1:

[0111] First Step: Synthesis of compound m1-3:

[0112] Compound m1-1 (3 g, 13.1 mmol), compound m1-2 (5.59 g, 26.2 mmol) and cesium carbonate (4.69 g, 14.4 mmol) were added to N,N-dimethylformamide (50 mL), the resulting suspension was heated to 80 °C for 18 hours. The reaction was cooled to room temperature, quenched with saturated ammonium chloride (200 mL), then extracted with ethyl acetate (50 mL) for three times. The organic phase was combined and dried over column to give compound m1-3 (4.2 g). LC-MS: m / z = 442, 444 [M+H] + . 1 H NMR (500 MHz, CDC13): δ 7.93 (s, 1H), 4.18-3.98 (m, 2H), 4.12 (s, 3H), 3.95-3.75 (m, 2H), 3.22-3.05 (m, 2H), 1.68-1.48 (m, 4H), 1.43 (s, 9H).

[0113] Second Step: Synthesis of compound m1-4:

[0114] Trifluoroacetic acid (20 mL) was added to a solution of m1-3 (10 g, 22.6 mmol) in dichloromethane, the resulting reaction was stirred at 0 °C for 2 hours. The reaction was dried to give m1-4 (7.6 g) which was used directly in the next step. LC-MS: m / z = 342, 344 [M+H] + .

[0115] Third Step: Synthesis of compound m1-5:

[0116] N,N-diisopropylethylamine (4.08 mL, 23.4 mmol) was added to a solution of m1-4 (2 g, 5.84 mmol), (R)-3-phenylbutyric acid (1.06 g, 6.43 mmol) and HATU (2.45 g, 6.43 mmol) in dichloromethane (70 mL). The resulting reaction was stirred at room temperature for 2 hours, quenched with water and extracted with dichloromethane (100 mL) three times. The combined organic phase was dried and concentrated to give compound m1-5 (2.4 g). LC-MS: m / z = 488, 490 [M+H] + . 1 H NMR (500 MHz, CDC13): δ 8.07 - 7.97 (m, 1H), 7.34 - 7.08 (m, 5H), 4.90 - 4.80 (m, 1H), 4.08 (s, 3H), 4.08 - 3.84 (m, 3H), 3.69 - 3.58 (m, 1H), 3.26 - 3.10 (m, 2H), 2.91 - 2.81 (m, 1H), 2.65 - 2.52 (m, 2H), 1.54 - 1.22 (m, 4H), 1.20 (d, J = 7.0 Hz, 3H).

[0117] Fourth Step: Synthesis of compound m1:

[0118] Compound m1-5 (1.56 g, 3.19 mmol), 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (78 mg, 0.2 mmol), palladium acetate (21.5 mg, 0.1 mmol), 4-(N-BOC- aminomethyl)phenylboronic acid (1.2 g, 4.8 mmol) and potassium phosphate (1.36 g, 6.38 mmol) were added to n-butanol (20 mL), purged with argon and then reacted at 100 °C for 2 h. The reaction was cooled to room temperature, concentrated and purified by column to give (1.2 g). LC-MS: m / z = 615 [M+H] + ; the resulting compound was dissolved in dichloromethane (10 mL) and trifluoroacetic acid (5 mL), stirred at room temperature for 2 h and concentrated to give compound m1 (900 mg). LC-MS: m / z = 515 [M+H] + . 1H NMR (500 MHz, CDC13): δ 8.03 - 7.92 (m, 1H), 7.69 - 7.43 (m, 4H), 7.30 - 7.21 (m, 4H), 7.19 - 7.12 (m, 1H), 4.87 (s, 1H), 4.10 (s, 3H), 4.07 - 3.86 (m, 3H), 3.81 (s, 2H), 3.70 - 3.61 (m, 1H), 3.28 - 3.12 (m, 2H), 2.93 - 2.83 (m, 1H), 2.66 - 2.52 (m, 2H), 2.31 (br. s, 2H), 1.57 - 1.23 (m, 4H), 1.21 (d, J = 6.9 Hz, 3H).

[0119] Example 2:

[0120] Synthesis of compound m2:

[0121] Compound m1-5 (1.56 g, 3.19 mmol), 2-dicyclohexylphosphino-2',6'- dimethoxybiphenyl (78 mg, 0.2 mmol), palladium acetate (21.5 mg, 0.1 mmol), 4- formylphenylboronic acid (720 mg, 4.8 mmol) and potassium phosphate (1.36 g, 6.38 mmol) were added to n-butanol (20 mL), purged with argon and then reacted at 100 °C for 2 hours. After the reaction solution was cooled to room temperature, it was spin dried and columned to give m2 (1.4 g). LC-MS: m / z = 514 [M+H] + .

[0122] Synthesis of compound m2-1: Potassium carbonate (93.8 mg, 0.680 mmol) was added to a solution of m2 (70.0 mg, 0.136 mmol) and dimethyl (1-diazo-2-oxopropyl)phosphonate (78.4 mg, 0.408 mmol) in methanol (10 mL), and the resulting solution was reacted at room temperature for 12 hours, diluted with ethyl acetate (50 mL), and then extracted with water (2 x 15 mL). After the organic phase was dried, filtered, and spin dried, it was columned to give m2-1 (55 mg, 79% yield). UPLC-MS: [M+H] = 510.20 found. +

[0123] Example 3:

[0124] ​Synthesis of compound m3-2: Triethylamine (252 g, 2.50 mol) was added to a solution of compound m3-1 (200 g, 1.92 mol), benzyl alcohol (200 g, 1.92 mol) in dichloromethane (500 mL). The resulting reaction was stirred at room temperature for 4 h. After the reaction was complete, the reaction was poured into water (1000 mL) and extracted with dichloromethane (3 x 250 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to yield m3-2 (272 g) after column purification.

[0125] Synthesis of compound m3-3: A solution of compound m3-2 (150 g, 0.852 mol) in tetrahydrofuran (200 mL) was slowly added to 511 mL of lithium diisopropylamide (1.0 M) in tetrahydrofuran at -78 °C. The resulting reaction was stirred at -78 °C for 1 h. Then 2-iodopropane (174 g, 1.02 mol) was added. The reaction was continued at -78 °C for 1 h. After the reaction was complete, it was quenched with 100 mL of 5% hydrochloric acid. It was extracted with diethyl ether (3 x 250 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to yield m3-3 (134 g) after column purification.

[0126] Synthesis of compound m3-4: Sodium bicarbonate (688 mg, 6.88 mol) was added to a solution of m3-3 (1.00 g, 4.59 mmol), 1,1-dibromoformoxime (954 mg, 4.59 mol) in N,N-dimethylformamide (20 mL) at 0 °C. The resulting reaction was stirred at room temperature for 12 h. After the reaction was complete, the reaction was poured into water (100 mL) and extracted with ethyl acetate (3 x 25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to yield m3-4 (856 mg) after column purification.

[0127] Synthesis of compound m3-5: Sodium carbonate (779 mg, 7.35 mol) was added to a solution of m3-4 (1.00 g, 2.94 mmol), 4-(dimethoxymethyl)-piperidine (561 mg, 3.53 mol) in n-butanol (20 mL) and the resulting mixture was reacted at 120 °C for 12 h. After the reaction was complete, the reaction was poured into water (100 mL) and extracted with ethyl acetate (3 x 25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to yield m3-5 (553 mg) after column purification.

[0128] Synthesis of compound m3-6: Compound iodine (17.3 g, 68 mmol) was added to a solution of m3-5 (18.9 g, 45.3 mmol) and imidazole (9.25 g, 136 mmol) in toluene (200 mL). The resulting reaction was heated at 110 °C for 12 h. After the reaction was complete, the reaction was poured into water (1000 mL) and extracted with ethyl acetate (3 x 250 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo and purified by column to give m3-6 (4.72 g).

[0129] Synthesis of compound m3: Compound m3-6 (3.74 g, 9.01 mmol) and palladium on carbon (10%, 374 mg) were added to methanol (40 mL). The resulting reaction was hydrogenated at room temperature for 12 h. After the reaction was complete, the palladium on carbon was filtered and concentrated in vacuo to give compound m3 (2.65 g).

[0130] Example 4:

[0131] Synthesis of compound m4-1: Sodium carbonate (779 mg, 7.35 mmol) was added to a solution of m3-4 (1.00 g, 2.94 mmol), 4-(dimethoxymethyl)-piperidine (504 mg, 3.53 mmol) in n-butanol (20 mL) and the resulting mixture was heated at 120 °C for 12 h. After the reaction was complete, the reaction was poured into water (100 mL) and extracted with ethyl acetate (3 x 25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo and purified by column to give m4-1 (530 mg).

[0132] Synthesis of compound m4-2: Compound iodine (17.3 g, 68 mmol) was added to a solution of m4-1 (18.1 g, 45 mmol) and imidazole (9.25 g, 136 mmol) in toluene (200 mL). The resulting reaction was heated at 110 °C for 12 h. After the reaction was complete, the reaction was poured into water (1000 mL) and extracted with ethyl acetate (3 x 250 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo and purified by column to give m4-2 (4.32 g).

[0133] Synthesis of compound m4: Compound m4-2 (4.02 g, 10 mmol) and palladium on carbon (10%, 400 mg) were added to methanol (40 mL). The resulting reaction was hydrogenated at room temperature for 12 h. After the reaction was complete, the palladium on carbon was filtered and concentrated in vacuo to give compound m4 (2.45 g).

[0134] Example 5:

[0135] Synthesis of compound m5: refer to the synthesis of compound m4, replace 4-(dimethoxymethyl)-piperidine with tert-butyl piperazine-1-carboxylate to undergo substitution reaction with compound m3-4, and then prepare compound m5 through 2 steps.

[0136] Example 6:

[0137] First step: synthesis of compound m6-3:

[0138] Compound m6-1 (3 g, 12.4 mmol, which was prepared with reference to patent CN110088096A, Example 120), compound m1-2 (5.28 g, 24.8 mmol) and cesium carbonate (4.44 g, 13.6 mmol) were added to N,N-dimethylformamide (50 mL), and the obtained suspension was heated to 80°C and reacted for 18 hours. After the reaction solution was cooled to room temperature, it was quenched with saturated ammonium chloride (200 mL), and then extracted with ethyl acetate (50 mL) three times. The combined organic phase was rotary evaporated and column chromatographed to obtain compound m6-3 (4.0 g). LC-MS: m / z = 456.42 [M+H] + .

[0139] Second step: synthesis of compound m6-4:

[0140] Trifluoroacetic acid (20 mL) was added to a dichloromethane solution of m6-3 (4 g, 22.6 mmol), and the obtained reaction solution was reacted at 0°C for 2 hours. The reaction solution was rotary evaporated to obtain m6-4 (2.8 g) which was directly used in the next step. LC-MS: m / z = 357.344 [M+H] + .

[0141] Third step: synthesis of compound m6-5:

[0142] N,N-diisopropylethylamine (4.08 mL, 23.4 mmol) was added to a dichloromethane (100 mL) solution of m6-4 (2.8 g, 7.84 mmol), (R)-4,4,4-trifluoro-3-phenylbutanoic acid (2.05 g, 9.41 mmol) and HATU (3.57 g, 9.41 mmol). After the obtained reaction solution was stirred at room temperature for 2 hours, it was quenched with water, and then extracted with dichloromethane (100 mL) three times. The combined organic phase was rotary evaporated and column chromatographed to obtain compound m6-5 (2.9 g). LC-MS: m / z = 556.490 [M+H] + .

[0143] Fourth step: synthesis of compound m6b:

[0144] Compound m6-5 (2.9 g, 5.21 mmol), 2-dicyclohexylphosphino-2',6'- dimethoxybiphenyl (78 mg, 0.2 mmol), palladium acetate (21.5 mg, 0.1 mmol), 4- (N-BOC-aminomethyl)phenylboronic acid (2.6 g, 7.8 mmol) and potassium phosphate (1.65 g, 7.8 mmol) were added to n-butanol (40 mL), purged with argon and reacted at 100 °C for 2 hours. After the reaction solution was cooled to room temperature, it was spin-dried and columned to give (3 g). LC-MS: m / z = 683.26 [M+H] + The resulting compound was dissolved in dichloromethane (10 mL) and trifluoroacetic acid (5 mL), stirred at room temperature for 2 hours and spin-dried to give compound m6b (900 mg). LC-MS: m / z = 583 [M+H] + .

[0145] Fifth step (parallel): synthesis of compound m6a:

[0146] Compound m6-5 (2.9 g, 5.21 mmol), 2-dicyclohexylphosphino-2',6'- dimethoxybiphenyl (78 mg, 0.2 mmol), palladium acetate (21.5 mg, 0.1 mmol), 4- (N-BOC-aminomethyl)phenylboronic acid (2.6 g, 7.8 mmol) and potassium phosphate (1.65 g, 7.8 mmol) were added to n-butanol (40 mL), purged with argon and reacted at 100 °C for 2 hours. After the reaction solution was cooled to room temperature, it was spin-dried and columned to give (3 g). LC-MS: m / z = 683.26 [M+H] + .

[0147] Example 7:

[0148] Synthesis of compounds m7a and m7b: refer to the synthesis of compounds m6a and m6b. Replace 4-formylphenylboronic acid with 3-fluoro-4-formylphenylboronic acid, and compound m1-5 is coupled to obtain compound m7a, LC-MS: m / z = 532.2 [M+H] + ; replace 4-(N-BOC-aminomethyl)phenylboronic acid with 4-((tert- butyloxycarbonylamino)methyl)-3-fluorophenylboronic acid, and compound m1-5 is coupled to obtain compound m7b, LC-MS: m / z = 533.1 [M+H] + .

[0149] Example 8:

[0150] Synthesis of compound m9a and m9b: Refer to the synthesis of compound m1. Replace (R)-3-phenylbutyric acid with (R)-4,4,4-trifluoro-3-phenylbutyric acid to prepare m9-1, LC-MS: m / z = 542.1 [M+H] + ; and compound m9a, LC-MS: m / z = 568.2 [M+H] was obtained by coupling reaction of compound m9-1 with 4-formylphenylboronic acid + ; and compound m9b, LC-MS: m / z = 569.2 [M+H] was obtained by coupling reaction of m9-1 with 4-(N-BOC-aminomethyl)phenylboronic acid + .

[0151] Example 9:

[0152] Synthesis of compound m10a and m10b: Refer to the synthesis of compound m1. Replace (R)-3-phenylbutyric acid with (R)-4,4-difluoro-3-phenylbutyric acid to prepare m10-1, LC-MS: m / z = 524.2 [M+H] + ; and compound m10a, LC-MS: m / z = 550.2 [M+H] was obtained by coupling reaction of compound 10-1 with 4-formylphenylboronic acid + ; and compound m10b, LC-MS: m / z = 551.1 [M+H] was obtained by coupling reaction of m10-1 with 4-(N-BOC-aminomethyl)phenylboronic acid + .

[0153] Example 10:

[0154] Synthesis of compound m11a and m11b: Refer to the synthesis of compound m1. Replace (R)-3-phenylbutyric acid with (R)-4,4,4-trifluoro-3-(4-fluorophenyl)butyric acid to prepare m11-1, LC-MS: m / z = 560.08 [M+H] + ; and compound m11a, LC-MS: m / z = 586.2 [M+H] was obtained by coupling reaction of compound m11-1 with 4-formylphenylboronic acid + ; and compound m11b, LC-MS: m / z = 587.23 [M+H] was obtained by coupling reaction of m11-1 with 4-(N-BOC-aminomethyl)phenylboronic acid + .

[0155] Example 11:

[0156] Synthesis of compound m12a and m12b: Refer to the synthesis of compound m6a and m6b, replace 4-formylphenylboronic acid with 4-formyl-3-trifluoromethylphenylboronic acid, and compound m12a is obtained by coupling reaction with m9-1, LC-MS: m / z = 636.3 [M+H] + ; replace 4-(N-BOC-aminomethyl)phenylboronic acid with 4-((tert-butoxycarbonylamino)methyl)-3-trifluoromethylboronic acid, and compound m12b is obtained by coupling reaction, LC-MS: m / z = 637.2 [M+H] + .

[0157] Example 12:

[0158] Synthesis of compound m13a and m13b: Refer to the synthesis of compound m1. Replace (R)-3-phenylbutyric acid with 3,3-dicyclopropylpropionic acid to prepare m13-1, LC-MS: m / z = 478.11 [M+H] + ; compound m13a is obtained by coupling reaction of compound m13-1 with 4-formylphenylboronic acid, LC-MS: m / z = 504.2 [M+H] + ; compound m13b is obtained by coupling reaction with 4-(N-BOC-aminomethyl)phenylboronic acid, LC-MS: m / z = 505.12 [M+H] + .

[0159] Example 13:

[0160] Synthesis of compound m14a and m14b: Refer to the synthesis of compound m1. Replace (R)-3-phenylbutyric acid with (R)-4,4,4-trifluoro-3-(5-methylthiophen-2-yl)butyric acid to prepare m14-1, LC-MS: m / z = 488.31 [M+H] + ; compound m14a is obtained by coupling reaction of compound m14-1 with 4-formylphenylboronic acid, LC-MS: m / z = 588.7 [M+H] + ; compound m14b is obtained by coupling reaction with 4-(N-BOC-aminomethyl)phenylboronic acid, LC-MS: m / z = 589.3 [M+H] + .

[0161] Example 14:

[0162] Synthesis of compound m14a-2: NaBD4 (185 mg, 4.88 mmol) was added to a solution of m14a-1 (500 mg, 2.32 mmol) in tetrahydrofuran / deuterated methanol (10 mL / 1 mL) and the reaction was stirred at 50 °C for 12 h. After the reaction was complete, the reaction was poured into water (30 mL) and extracted with ethyl acetate (3 x 25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo before purification by column to give m14a-2 (450 mg).

[0163] Synthesis of compound m14a-3: Compound m14a-2 (450 mg, 2.39 mmol), bis(pinacolato)diboron (1.22 g, 4.78 mmol), 1,1'-bis(diphenylphosphino)ferrocene palladium(II) dichloride (174 mg, 0.239 mmol), potassium acetate (703 mg, 7.17 mmol) were added to 1.4-dioxane (5 mL), purged with argon and then stirred at 100 °C for 2 h. After the reaction was cooled to room temperature, it was concentrated in vacuo and purified by column to give m14a-3 (505 mg), 1 H NMR (600 MHz, Chloroform-d) δ 7.82 (d, J = 8.0 Hz, 2H), 7.38 (d, J = 8.0 Hz, 2H), 3.71 (s, 1H), 1.36 (s, 12H).

[0164] Synthesis of compound m14c: Refer to the synthesis of compound m6a, replace 4- formylphenylboronic acid with m14a-3, and couple with m9-1 to give compound m14c, LC-MS: m / z = 572.2 [M+H] + .

[0165] Example 15:

[0166] Synthesis of compound m15a: Refer to the synthesis of compound m6a, replace 4- formylphenylboronic acid with 4-formyl-3-methoxyphenylboronic acid, and couple with m9-1 to give compound m15a, LC-MS: m / z = 598.3 [M+H] + .

[0167] Example 16:

[0168] Synthesis of compound m16a: Refer to the synthesis of compound m6a, replace 4- formylphenylboronic acid with 4-formyl-3-methoxyphenylboronic acid pinacol ester, and couple with m1-5 to give compound m16a, LC-MS: m / z = 544.3 [M+H] + .

[0169] Example 17:

[0170] Synthesis of compound m17a: Refer to the synthesis of compound m6a, replace 4- formylphenylboronic acid with 4-formyl-3-deuteriomethoxyphenylboronic acid pinacol ester, and compound m17a was obtained by coupling reaction with m9-1, LC-MS: m / z = 601.3 [M+H] + .

[0171] Example 18:

[0172] Synthesis of compound m19a: Refer to the synthesis of compound m6a, replace 4- formylphenylboronic acid with 4-formylmethylphenylboronic acid, and compound m19a was obtained by coupling reaction with m1-5, LC-MS: m / z = 528.3 [M+H] + .

[0173] Example 19:

[0174] Synthesis of compound m23a: Refer to the synthesis of compound m1. Replace (R)-3- phenylbutyric acid with (R)-3-phenylbutyl-4,4,4-d3 acid to prepare m24a-1, LC-MS: m / z = 491.1, 493.1 [M+H] + ; and refer to the synthesis of compound m6a, compound m23a was obtained by coupling reaction of compound m23a-1 with 4-formylphenylboronic acid, LC-MS: m / z = 517.3 [M+H] + .

[0175] Example 20:

[0176] Synthesis of compound m24a: Refer to the synthesis of compound m6a, replace 4- formylphenylboronic acid with 4-formyl-3-methoxyphenylboronic acid pinacol ester, and compound m24a was obtained by coupling reaction with m23a-1, LC-MS: m / z = 547.3 [M+H] + .

[0177] Example 21:

[0178] Synthesis of compound m25a: Refer to the synthesis of compound m6a, replace 4- formylphenylboronic acid with 5-fluoro-4-formyl-3-methoxyphenylboronic acid, and compound m25a was obtained by coupling reaction with m9-1, LC-MS: m / z = 616.2 [M+H] + .

[0179] Example 22:

[0180] First step: synthesis of compound m26a-2: m26a-1 (513 mg, 3 mmol) and potassium carbonate (830 mg, 6 mmol) were dissolved in acetone (70 mL), and deuterated methyl iodide (373 μL, 6 mmol) was added. The resulting mixture was heated at 70 °C for 4 h, and the reaction was monitored by TLC. After the reaction was completed, the organic solvent was evaporated under reduced pressure. Water (6 mL) was added to the residue, which was extracted with ethyl acetate (7 mL x 3). The combined organic phase was washed with saturated aqueous sodium chloride solution (10 mL) and dried over anhydrous sodium sulfate. The organic solvent was evaporated under reduced pressure, and the residue was purified by column chromatography on silica gel (petroleum ether / ethyl acetate = 2:1) to give compound m26a-2 (150 mg). LC-MS: m / z = 211.0 [M+Na] + .

[0181] Second step: synthesis of compound m26a-3: Compound m26a-2 (2.54 g, 13.7 mmol) was dissolved in methanol (50 mL), and 10% Pd / C (0.5 g) was added. The resulting mixture was stirred at room temperature under a hydrogen atmosphere overnight. The reaction was monitored by TLC, and the reaction solution was filtered through celite. The filtrate was concentrated under reduced pressure to give purple solid of compound m26a-3 (2.06 g). The crude product was used directly in the next step without purification. LC-MS: m / z = 181.1 [M+Na] + .

[0182] Third step: synthesis of compound m26a-4: The crude product of compound m26a-3 (1.96 g, 12.6 mmol) was dissolved in N,N-diisopropylethylamine (10.7 mL) and methanol (10.7 mL). Formamidinium acetate (1.45 g, 13.9 mmol) was added to the mixture portionwise slowly. The resulting solution was reacted at 110 °C for 1 h. The reaction was monitored by TLC, and the reaction solution was cooled to room temperature. White solid was precipitated from the system. The solid was collected by filtration, and the filter cake was washed with diethyl ether and dried under reduced pressure to give compound m26a-4 (1.77 g). LC-MS: m / z = 170.1 [M+H] + .

[0183] Fourth Step: Synthesis of compound m26a-5: Compound m26a-4 (3.24 g, 21.6 mmol) was placed in a sealed tube, acetic acid (21.6 mL) and liquid bromine (3.34 mL, 64.7 mmol) were added, and the reaction was stirred at 95 °C for 24 h. The reaction was monitored by TLC and was complete. Saturated sodium thiosulfate solution (50 mL) was added and a solid precipitated. The solid was collected by filtration, the filter cake was washed with water, and dried under vacuum to give compound m26a-5 as a yellow solid (1.81 g). LC-MS: m / z = 248.0 / 250.0 [M+H] + .

[0184] Fifth Step: Synthesis of compound m26a-6: Compound m26a-5 (3 g, 13.0 mmol), 1-oxa-6- azaspiro[2-4]octane-6-carboxylic acid tert-butyl ester (5.59 g, 26.2 mmol), and cesium carbonate (4.69 g, 14.4 mmol) were added to N,N-dimethylformamide (50 mL), and the resulting suspension was heated to 80 °C for 18 h. The reaction was cooled to room temperature, quenched with saturated ammonium chloride (200 mL), and extracted with ethyl acetate (50 mL) three times. The organic phases were combined, dried, and concentrated to give compound m26a-6 (4.0 g). LC-MS: m / z = 445, 447 [M+H] + .

[0185] Sixth Step: Synthesis of compound m26a-7: Trifluoroacetic acid (20 mL) was added to a solution of m26a-6 (10 g, 22.6 mmol) in dichloromethane, and the resulting reaction was stirred at 0 °C for 2 h. The reaction was concentrated to give m26a-7 (7.6 g) which was used directly in the next step. LC-MS: m / z = 345, 347 [M+H] + .

[0186] Seventh Step: Synthesis of compound m26a-8: N,N-diisopropylethylamine (4.08 mL, 23.4 mmol) was added to a solution of m37a-7 (2 g, 5.84 mmol), (R)-4,4,4-trifluoro-3- phenylbutanoic acid (1.06 g, 6.43 mmol), and HATU (2.45 g, 6.43 mmol) in dichloromethane (70 mL). The resulting reaction was stirred at room temperature for 2 h, quenched with water, and extracted with dichloromethane (100 mL) three times. The organic phases were combined, dried, and concentrated to give compound m26a-8 (2.4 g). LC-MS: m / z = 545.1, 547.1 [M+H] + .

[0187] Step 8: Synthesis of compound m26a: Refer to the synthesis of compound m6a, replace m6-5 with m26a-8, and 4-formylphenylboronic acid to undergo coupling reaction to obtain compound m26a, LC-MS: m / z = 568.3 [M+H] + .

[0188] Example 23:

[0189] Synthesis of compound m27a: Refer to the synthesis of compound m6a, replace 4- formylphenylboronic acid with 4-formylmethylphenylboronic acid, and m9-1 to undergo coupling reaction to obtain compound m27a, LC-MS: m / z = 582.3 [M+H] + .

[0190] Example 24:

[0191] Synthesis of compound s1-2: s1-1 (1.00 g, 2.25 mmol) was added to a solution of 6- ((tert-butoxycarbonyl)amino)hexanoic acid (850 mg, 2.70 mmol), HATU (855 mg, 2.25 mmol) and N,N-diisopropylethylamine (435 mg, 3.38 mmol) in N,N- dimethylformamide (20 mL) at 0 °C. The reaction was stirred at 0 °C for 2 h. After the reaction was completed, the reaction was poured into water (100 mL) and extracted with ethyl acetate (3 x 25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give s1-2 (910 mg) after column purification. UPLC-MS: [M+H] = 658.53 found. + .

[0192] Synthesis of compound s1-3: s1-2 (910 mg, 1.38 mmol) was added to a mixture of dichloromethane (10 mL) and trifluoroacetic acid (2.5 mL) at room temperature. The reaction was stirred at room temperature for 1 h and concentrated to give s1-3 (755 mg). UPLC-MS: [M+H]+ = 558.5 found.

[0193] Synthesis of compound S1: Sodium cyanoborohydride (25.8 mg, 0.410 mmol) was added to a dichloromethane / methanol (5 mL / 1 mL) solution containing S1-3 (100 mg, 0.180 mmol), M2 (105 mg, 0.205 mmol), sodium acetate (84.0 mg, 1.025 mmol), and acetic acid (1.27 mg, 0.0205 mmol). The reaction solution was reacted at room temperature for 12 h. After the reaction was complete, the reaction solution was poured into water (100 mL) and extracted with dichloromethane (3 × 25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Compound S1 (30 mg) was prepared by HPLC. UPLC-MS: [M+H] + =1055.87 (actual measurement); 1 H NMR (500MHz, methanol-d4) δ8.84(d,J=21.2Hz,1H),7.99(dd,J=25.2,7.9Hz,1H),7.82–7.58(m,4H),7.48–7.37(m,4H),7.34–7.12 (m,5H),5.00(q,J=6.8Hz,1H),4.63–4.53(m,2H),4.44–4.40(m,1H),4.23(s,2H),4.15–3.82(m,6H),3.78–3.53(m,2H),3.30 –3.14(m,3H),3.10–2.87(m,3H),2.76(ddd,J=32.2,14.3,8.1Hz,1H),2.63–2.41(m,4H),2.38–2.15(m,3H),2.05–1.92(m,1 H),1.72(p,J=8.6,8.1Hz,2H),1.68–1.57(m,3H),1.50(d,J=7.0Hz,3H),1.31(dt,J=19.2,5.6Hz,8H),1.03(d,J=9.9Hz,9H); 13C NMR (126 MHz, Methanol-d4) δ 173.63, 171.20, 170.72, 170.66, 170.35, 156.77, 156.69, 150.87, 147.11, 147.03, 146.98, 145.20, 145.07, 143.63, 135.69, 131.34, 129.53, 129.31, 128.87, 128.63, 128.51, 127.72, 127.52, 126.17, 126.00, 125.74, 125.64, 125.48, 125.41, 68.98, 58.63, 57.12, 56.01, 53.04, 50.34, 48.15, 41.18, 40.20, 37.96, 36.96, 36.86, 36.75, 36.63, 36.50, 34.49, 34.29, 34.13, 34.00, 33.57, 33.34, 25.47, 25.23, 25.07, 24.23, 20.38, 20.28, 20.08, 13.82. HPLC >95%; HRMS (ESI-TOF) m / z: [M+H] + Calculated C 58 H 75 N 10 O7S+, 1055.5535; Found, 1055.5535.

[0194] Example 25:

[0195] Synthesis of compound s2: Refer to the synthesis of compound s1. Replace 6-((tert- butoxycarbonyl)amino)hexanoic acid with BOC-8-aminooctanoic acid (CAS No.: 30100-16-4) to prepare compound s2-1, then follow similar procedures to obtain compound s2. UPLC-MS: [M+H]+= 1083.81 found;1H NMR (500 MHz, Methanol-d4) δ 8.85 (d, J = 8.5 Hz, 1H), 7.97 (d, J = 25.9 Hz, 1H), 7.65 - 7.50 (m, 4H), 7.45 - 7.14 (m, 9H), 4.99 (q, J = 7.0 Hz, 1H), 4.62 - 4.53 (m, 2H), 4.42 (dt, J = 4.8, 3.0 Hz, 1H), 4.34 - 3.82 (m, 9H), 3.78 - 3.57 (m, 2H), 3.29 - 3.14 (m, 2H), 3.07 - 2.87 (m, 1H), 2.75 (ddd, J = 33.0, 14.2, 8.1 Hz, 1H), 2.67 - 2.42 (m, 6H), 2.33 - 2.13 (m, 3H), 2.02 - 1.91 (m, 1H), 1.67 - 1.54 (m, 5H), 1.50 (d, J = 7.0 Hz, 3H), 1.42 - 1.27 (m, 14H), 1.03 (s, 9H).13C NMR (126 MHz, Methanol-d4) δ 176.03, 173.26, 172.72, 172.66, 172.33, 158.85, 158.77, 152.86, 149.10, 148.77, 148.72, 147.20, 147.07, 145.68, 138.42, 136.07, 135.92, 133.35, 131.54, 131.04, 130.50, 130.30, 129.72, 129.52, 128.16, 128.00, 127.75, 127.64, 127.48, 70.97, 60.60, 59.00, 58.04, 55.07, 54.90, 53.92, 50.14, 43.18, 42.20, 39.87, 38.95, 38.78, 38.63, 38.50, 36.60, 36.49, 36.26, 35.98, 35.60, 35.37, 30.24, 30.14, 30.09, 28.21, 27.07, 26.93, 22.40, 22.27, 22.08, 15.81. HPLC > 95%; HRMS (ESI-TOF) m / z: [M+H] + Calculated C60H79N10O7S+, 1083.5848; found, 1083.5848.

[0196] Example 26:

[0197] Synthesis of compound s3: Refer to the synthesis of compound s1. Replace 6-((tert- butoxycarbonyl)amino)hexanoic acid with 9-((tert-butoxycarbonyl)amino)nonanoic acid (CAS No.: 173435-78-4) to prepare compound s3-1, then follow similar procedures to obtain compound s3. UPLC-MS: [M+H]+=1097.57 found; HPLC >95%;1H NMR (500 MHz, Methanol-d4): d 8.97 (d, J = 13.2 Hz, 1H), 8.01 (d, J = 25.1 Hz, 1H), 7.79 - 7.66 (m, 4H), 7.49 - 7.36 (m, 4H), 7.33 - 7.13 (m, 5H), 5.00 (q, J = 7.0 Hz, 1H), 4.62 (s, 1H), 4.60 - 4.54 (m, 1H), 4.43 (tt, J = 4.4, 2.0 Hz, 1H), 4.30 (d, J = 14.3 Hz, 2H), 4.24 - 4.11 (m, 4H), 4.10 - 3.95 (m, 1H), 3.90 - 3.85 (m, 1H), 3.77 - 3.54 (m, 2H), 3.30 - 3.15 (m, 3H), 3.13 - 3.06 (m, 2H), 2.97 (dtd, J = 48.9, 13.2, 12.5, 3.6 Hz, 1H), 2.76 (ddd, J = 31.9, 14.3, 8.1 Hz, 1H), 2.62 - 2.45 (m, 4H), 2.35 - 2.16 (m, 3H), 2.03 - 1.91 (m, 1H), 1.74 (p, J = 7.5 Hz, 2H), 1.60 (ddt, J = 17.2, 11.1, 5.1 Hz, 3H), 1.50 (d, J = 7.0 Hz, 3H), 1.35 (ddd, J = 28.0, 16.3, 6.2 Hz, 14H), 1.03 (s, 9H).13C NMR (126 MHz, Methanol-d4): d 174.57, 171.84, 171.29, 171.22, 170.91, 160.11, 157.34, 157.26, 151.87, 147.68, 147.63, 146.97, 145.77, 145.64, 144.50, 136.09, 132.49, 132.38, 130.22, 130.17, 129.75, 129.10, 128.54, 128.31, 128.11, 126.76, 126.58, 126.34, 126.28, 126.08, 69.56, 59.21, 57.59, 56.61, 53.63, 50.47, 48.75, 41.76, 40.78, 38.58, 37.54, 37.42, 37.33, 37.22, 37.08, 35.19, 35.12, 34.87, 34.58, 34.15, 33.91, 28.72, 28.58, 26.11, 25.77, 25.66, 25.49, 21.00, 20.88, 20.68, 14.12. HPLC >95%; HRMS (ESI-TOF) m / z: [M+H]. + Calculated C61H81N10O7S+, 1097.6005; found, 1097.6006.

[0198] Example 27:

[0199] Synthesis of compound s4: Refer to the synthesis of compound s1. Replace 6-((tert- butoxycarbonyl)amino)hexanoic acid with 10-((tert-butoxycarbonyl)amino)decanoic acid (CAS No.: 173606-50-3) to prepare compound s4-1, then follow similar procedures to obtain compound s4. UPLC-MS: [M+H]+= 1112.05 found; 1 HNMR (500 MHz, Methanol-d4) δ 9.07 (s, 1H), 8.01 (d, J = 25.1 Hz, 1H), 7.79 - 7.67 (m, 4H), 7.47 - 7.37 (m, 4H), 7.33 - 7.14 (m, 5H), 5.00 (q, J = 7.0 Hz, 1H), 4.62 (s, 1H), 4.56 (dd, J = 9.6, 7.1 Hz, 1H), 4.42 (dd, J = 4.8, 2.6 Hz, 1H), 4.31 (s, 2H), 4.14 (d, J = 3.5 Hz, 4H), 4.11 - 3.97 (m, 1H), 3.88 (dd, J = 13.3, 11.0 Hz, 1H), 3.78 - 3.54 (m, 2H), 3.30 - 3.15 (m, 3H), 3.12 - 3.05 (m, 2H), 3.05 - 2.89 (m, 1H), 2.76 (ddd, J = 31.9, 14.3, 8.1 Hz, 1H), 2.62 - 2.46 (m, 4H), 2.33 - 2.16 (m, 3H), 2.04 - 1.92 (m, 1H), 1.74 (dq, J = 15.8, 7.9, 7.2 Hz, 2H), 1.59 (dt, J = 15.0, 6.9 Hz, 3H), 1.50 (d, J = 7.0 Hz, 3H), 1.43 - 1.28 (m, 16H), 1.02 (d, J = 11.6 Hz, 9H). 13C NMR (126 MHz, Methanol-d4) δ 176.02, 173.26, 172.73, 172.66, 172.35, 160.54, 158.76, 158.68, 153.57, 149.11, 149.05, 147.85, 147.19, 147.07, 146.10, 137.52, 136.15, 134.15, 133.89, 131.65, 131.57, 130.88, 130.53, 129.99, 129.72, 129.52, 128.17, 127.99, 127.75, 127.57, 127.49, 70.98, 60.62, 59.01, 58.01, 55.05, 54.88, 51.90, 50.17, 43.18, 42.20, 39.97, 38.96, 38.82, 38.64, 38.51, 36.66, 36.54, 36.29, 36.00, 35.57, 35.33, 30.26, 30.11, 27.57, 27.20, 27.07, 26.97, 22.38, 22.28, 22.08, 15.28. HPLC > 95%; HRMS (ESI-TOF) m / z: [M+H] + Calculated for C62H83N10O7S+, 1111.6161; found, 1111.6158.

[0200] Example 28:

[0201] Synthesis of compound s5: Refer to the synthesis of compound s1. Replace 6- ((tert-butoxycarbonyl)amino)hexanoic acid with tert-butoxycarbonyl-11-aminoundecanoic acid (CAS Number: 10436-25-6) to prepare compound s5-1, then follow similar procedures to obtain compound s5. UPLC-MS: [M+H]+= 1125.58 found; 1HNMR (600 MHz, Methanol-d4) δ 8.88: (d, J = 7.1 Hz, 1H), 7.99 (d, J = 30.1 Hz, 1H), 7.81 - 7.59 (m, 4H), 7.45 - 7.34 (m, 4H), 7.32 - 7.11 (m, 5H), 4.98 (q, J = 7.0 Hz, 1H), 4.61 (s, 1H), 4.57 - 4.53 (m, 1H), 4.41 (tt, J = 4.3, 2.1 Hz, 1H), 4.29 (d, J = 7.3 Hz, 2H), 4.23 - 4.09 (m, 4H), 4.09 - 3.95 (m, 1H), 3.89 - 3.83 (m, 1H), 3.75 - 3.43 (m, 2H), 3.29 - 3.14 (m, 3H), 3.11 - 2.82 (m, 3H), 2.82 - 2.42 (m, 5H), 2.31 - 2.14 (m, 3H), 2.02 - 1.90 (m, 1H), 1.72 (ddd, J = 15.1, 10.4, 6.6 Hz, 2H), 1.63 - 1.54 (m, 3H), 1.49 (d, J = 7.0 Hz, 3H), 1.41 - 1.25 (m, 18H), 1.01 (d, J = 14.2 Hz, 9H). 13 CNMR (151 MHz, Methanol-d4): δ 174.61, 171.83, 171.29, 171.22, 170.90, 157.34, 157.26, 151.60, 147.68, 147.63, 147.44, 145.78, 145.65, 144.33, 136.09, 132.48, 132.09, 130.22, 130.16, 130.00, 129.09, 128.56, 128.31, 128.11, 126.76, 126.58, 126.33, 126.24, 126.07, 69.56, 59.19, 57.58, 56.59, 53.62, 50.48, 48.73, 41.76, 41.67, 40.78, 38.57, 37.54, 37.41, 37.33, 37.21, 37.08, 35.26, 35.12, 34.87, 34.58, 34.15, 33.91, 29.02, 28.98, 28.89, 28.76, 26.18, 25.81, 25.66, 25.59, 20.99, 20.88, 20.67, 14.31. HPLC >95%; HRMS (ESI-TOF) m / z: [M+H] + Calculated C63H85N10O7S+, 1125.6318; Found, 1125.6320.

[0202] Example 29:

[0203] Synthesis of compound s6: Refer to the synthesis of compound s1. Replace 6-((tert-butoxycarbonyl)amino)hexanoic acid with 12-(BOC-amino)dodecanoic acid (CAS No.: 18934-81-1) to prepare compound s6-1, then follow similar procedures to obtain compound s6. UPLC-MS: [M+H]+= 1139.95 found; 1 HNMR (600 MHz, Methanol-d4) δ 8.88: (d, J = 7.1 Hz, 1H), 7.99 (d, J = 30.1 Hz, 1H), 7.81 - 7.59 (m, 4H), 7.45 - 7.34 (m, 4H), 7.32 - 7.11 (m, 5H), 4.98 (q, J = 7.0 Hz, 1H), 4.61 (s, 1H), 4.57 - 4.53 (m, 1H), 4.41 (tt, J = 4.3, 2.1 Hz, 1H), 4.29 (d, J = 7.3 Hz, 2H), 4.23 - 4.09 (m, 4H), 4.09 - 3.95 (m, 1H), 3.89 - 3.83 (m, 1H), 3.75 - 3.43 (m, 2H), 3.29 - 3.14 (m, 3H), 3.11 - 2.82 (m, 3H), 2.82 - 2.42 (m, 5H), 2.31 - 2.14 (m, 3H), 2.02 - 1.90 (m, 1H), 1.72 (ddd, J = 15.1, 10.4, 6.6 Hz, 2H), 1.63 - 1.54 (m, 3H), 1.49 (d, J = 7.0 Hz, 3H), 1.41 - 1.25 (m, 18H), 1.01 (d, J = 14.2 Hz, 9H). 13CNMR (151 MHz, Methanol-d4): δ 174.61, 171.83, 171.29, 171.22, 170.90, 157.34, 157.26, 151.60, 147.68, 147.63, 147.44, 145.78, 145.65, 144.33, 136.09, 132.48, 132.09, 130.22, 130.16, 130.00, 129.09, 128.56, 128.31, 128.11, 126.76, 126.58, 126.33, 126.24, 126.07, 69.56, 59.19, 57.58, 56.59, 53.62, 50.48, 48.73, 41.76, 41.67, 40.78, 38.57, 37.54, 37.41, 37.33, 37.21, 37.08, 35.26, 35.12, 34.87, 34.58, 34.15, 33.91, 29.02, 28.98, 28.89, 28.76, 26.18, 25.81, 25.66, 25.59, 20.99, 20.88, 20.67, 14.31. HPLC >95%; HRMS (ESI-TOF) m / z: [M+H] + Calculated C64H87N10O7S+, 1139.6474; found, 1139.6477.

[0204] Example 30:

[0205] Synthesis of compound s7: Refer to the synthesis of compound s1. Replace 6-((tert- butoxycarbonyl)amino)hexanoic acid with N-tert-butoxycarbonyl-diglycol-carboxylic acid (CAS number: 1365655-91-9) to obtain compound s7-1, then follow similar procedures to obtain compound s7. UPLC-MS: [M+H]+=1101.13 found;1H NMR (600 MHz, Methanol-d4) δ 8.86 (d, J = 1.0 Hz, 1H), 7.97 (d, J = 30.8 Hz, 1H), 7.67 - 7.56 (m, 4H), 7.41 (q, J = 8.3 Hz, 4H), 7.31 (t, J = 7.6 Hz, 1H), 7.27 - 7.13 (m, 4H), 5.00 - 4.95 (m, 1H), 4.63 (s, 1H), 4.58 - 4.53 (m, 1H), 4.41 (dp, J = 4.3, 1.9 Hz, 1H), 4.25 - 4.09 (m, 4H), 4.06 - 3.81 (m, 5H), 3.74 (dddd, J = 16.6, 7.6, 6.1, 3.8 Hz, 3H), 3.67 - 3.59 (m, 6H), 3.30 - 3.13 (m, 3H), 3.05 - 2.98 (m, 1H), 2.92 (td, J = 13.0, 12.5, 3.5 Hz, 1H), 2.87 - 2.82 (m, 2H), 2.75 (ddd, J = 40.4, 14.3, 8.1 Hz, 1H), 2.60 - 2.53 (m, 1H), 2.50 - 2.43 (m, 4H), 2.18 (ddt, J = 13.1, 7.7, 2.0 Hz, 1H), 1.94 (ddd, J = 13.3, 9.0, 4.5 Hz, 1H), 1.65 - 1.26 (m, 10H), 1.01 (d, J = 10.4 Hz, 9H). 13C NMR (151 MHz, methanol-d4) δ 172.26, 171.83, 171.26, 171.18, 170.68, 151.45, 147.62, 147.32, 145.74, 145.61, 144.29, 136.99, 131.96, 130.06, 129.64, 129.08, 128.87, 128.31, 128.11, 126.71, 126.54, 126.38, 126.21, 126.05, 69.97, 69.53, 69.29, 66.89, 59.16, 57.52, 56.58, 53.64, 52.40, 48.72, 41.76, 40.78, 38.51, 37.41, 37.08, 35.91, 35.29, 34.55, 33.95, 25.65, 21.01, 20.87, 20.66, 14.42. HPLC >95%; HRMS (ESI-TOF) m / z: [M+H] + Calculated C59H77N10O9S+, 1101.5590; found, 1101.5591.

[0206] Example 31:

[0207] Synthesis of compound s8: Refer to the synthesis of compound s1. Replace 6-((tert- butoxycarbonyl)amino)hexanoic acid with N-tert-butoxy carbonyl-glyceryl carboxylic acid (CAS Number: 1347750-75-7) to prepare compound s8-1, then follow similar procedures to obtain compound s8. UPLC-MS: [M+H] + = 1145.55 found; 1H NMR (500 MHz, Methanol-d4) δ 8.99 (s, 1H), 8.01 (d, J = 24.6 Hz, 1H), 7.79 - 7.69 (m, 4H), 7.45 - 7.35 (m, 4H), 7.30 (t, J = 7.5 Hz, 1H), 7.26 - 7.11 (m, 4H), 4.99 (q, J = 7.0 Hz, 1H), 4.61 (s, 1H), 4.57 (dd, J = 9.2, 7.5 Hz, 1H), 4.42 (dd, J = 4.6, 2.5 Hz, 1H), 4.38 (d, J = 4.7 Hz, 2H), 4.24 - 3.95 (m, 5H), 3.91 - 3.79 (m, 3H), 3.76 - 3.72 (m, 1H), 3.71 - 3.55 (m, 11H), 3.33 (t, J = 5.0 Hz, 2H), 3.31 - 3.14 (m, 3H), 3.05 - 2.86 (m, 1H), 2.75 (ddd, J = 32.2, 14.3, 8.1 Hz, 1H), 2.61 - 2.39 (m, 6H), 2.20 (ddt, J = 13.3, 7.8, 2.0 Hz, 1H), 1.95 (ddd, J = 13.3, 9.2, 4.5 Hz, 1H), 1.66 - 1.25 (m, 10H), 1.02 (d, J = 9.9 Hz, 9H). 13 C NMR (126 MHz, Methanol-d4) δ 172.31, 171.84, 171.27, 171.20, 170.72, 151.93, 147.63, 146.89, 145.79, 145.66, 144.53, 136.11, 132.43, 130.39, 130.21, 129.70, 129.10, 128.57, 128.32, 128.13, 126.76, 126.59, 126.34, 126.08, 70.08, 69.86, 69.79, 69.57, 66.88, 65.34, 59.26, 57.66, 56.63, 53.62, 50.27, 48.76, 41.77, 40.82, 38.66, 37.55, 35.79, 35.26, 34.59, 33.92, 25.68, 21.04, 20.90, 20.69.14.17. HPLC > 95%; HRMS (ESI-TOF) m / z: [M+H] + Calculated C61H81N10O10S+, 1145.5852; found, 1145.5854.

[0208] Example 32:

[0209] Synthesis of compound s9: Refer to the synthesis of compound s1. Replace 6-((tert- butoxycarbonyl)amino)hexanoic acid with 5,8,11,14-tetraoxa-2-azaheneicosanedioic acid 1-tert-butyl ester (CAS No.: 756525-91-4) to prepare compound s9-1, then follow similar procedures to obtain compound s9. UPLC-MS: [M+H] + = 1189.99 found; 1 H NMR (500 MHz, methanol-d4) δ 8.86 (s, 1H), 8.00 (d, J = 25.1 Hz, 1H), 7.80 - 7.67 (m, 4H), 7.45 - 7.12 (m, 9H), 4.99 (t, J = 6.9 Hz, 1H), 4.61 (s, 1H), 4.57 - 4.53 (m, 1H), 4.42 (s, 1H), 4.31 (s, 2H), 4.24 - 4.03 (m, 5H), 4.00 - 3.87 (m, 1H), 3.83 (d, J = 11.0 Hz, 1H), 3.81 - 3.77 (m, 2H), 3.75 - 3.55 (m, 16H), 3.29 - 3.12 (m, 4H), 3.07 - 2.89 (m, 1H), 2.75 (ddd, J = 32.8, 14.3, 8.2 Hz, 1H), 2.63 - 2.49 (m, 2H), 2.46 (d, J = 6.2 Hz, 4H), 2.18 (ddd, J = 12.5, 7.5, 3.6 Hz, 1H), 1.95 (ddd, J = 13.4, 9.0, 4.6 Hz, 1H), 1.66 - 1.58 (m, 1H), 1.49 (d, J = 7.0 Hz, 3H), 1.36 - 1.24 (m, 6H), 1.01 (d, J = 10.6 Hz, 9H). 13C NMR (126 MHz, Methanol-d4) δ 173.70, 173.22, 172.72, 172.65, 172.14, 152.87, 149.11, 148.96, 147.23, 147.07, 145.64, 137.66, 133.33, 131.55, 130.51, 129.72, 129.52, 128.17, 128.00, 127.74, 127.63, 127.44, 71.48, 71.45, 71.38, 71.26, 71.11, 70.97, 68.31, 67.33, 60.63, 59.04, 58.01, 55.14, 52.05, 50.15, 43.18, 42.24, 40.01, 38.89, 38.63, 38.50, 37.28, 36.68, 27.08, 22.41, 22.28, 22.08, 15.82. HPLC >95%; HRMS (ESI-TOF) m / z: [M+H] + Calculated C63H85N10O11S+, 1189.6115; found, 1189.6114.

[0210] Example 33:

[0211] Synthesis of compounds s10-3s and s10-3r: s10-1 (1.02 g, 3.07 mmol) was added to a solution of m3 (1.00 g, 3.07 mmol), HATU (1.17 g, 3.07 mmol), and N,N- diisopropylethylamine (594 mg, 4.60 mmol) in N,N-dimethylformamide (20 mL) at 0 °C. The reaction was stirred at 0 °C for 2 h. After the reaction was complete, the reaction was poured into water (100 mL) and extracted with ethyl acetate (3 x 25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo before purification by column to give a mixture of non-corresponding isomers (1.49 g). UPLC-MS: [M+H] + = 640.32 Found. Chiral HPLC resolution gave s10-3r (0.49 g) and s10-3s (0.52 g). s10-3r: 21.8 min at room temperature. s10-3s: 23.9 min at room temperature. 1HNMR (600 MHz, Methanol-d4) δ 8.89 (s, 1H), 7.50 - 7.37 (m, 4H), 6.11 (s, 1H), 5.05 (q, J = 6.7 Hz, 1H), 4.53 (t, J = 8.1 Hz, 1H), 4.48 - 4.41 (m, 1H), 4.10 (d, J = 6.6 Hz, 1H), 3.86 (dd, J = 10.8, 4.2 Hz, 1H), 3.71 (dt, J = 12.8, 3.2 Hz, 2H), 3.66 - 3.59 (m, 2H), 3.38 (s, 6H), 2.82 (tt, J = 12.6, 2.7 Hz, 2H), 2.50 (s, 3H), 2.39 (dp, J = 9.5, 6.7 Hz, 1H), 2.20 (ddt, J = 12.3, 7.8, 2.1 Hz, 1H), 1.98 (ddd, J = 13.2, 8.7, 4.7 Hz, 1H), 1.85 - 1.75 (m, 3H), 1.57 (dd, J = 37.8, 7.0 Hz, 3H), 1.35 (tdd, J = 20.5, 18.9, 10.6, 3.4 Hz, 2H), 1.07 (dd, J = 6.7, 2.7 Hz, 3H), 0.92 (dd, J = 18.1, 6.7 Hz, 3H).

[0212] Synthesis of compound s10-4: 1M sulfuric acid (5 mL) was added to a solution of s10-3r (490 mg, 0.767 mmol) in tetrahydrofuran (5 mL). The resulting reaction was stirred at 60 °C for 3 h. After the reaction was completed, it was quenched with saturated sodium bicarbonate solution. It was extracted with ethyl acetate (3 x 25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated and purified by column to give s10-4 (368 mg). UPLC-MS: [M+H] = 594.56 found. +

[0213] Synthesis of compound s10-5: Sodium cyanoborohydride (78.2 mg, 1.24 mmol) was added to a solution of s10-4 (368 mg, 0.621 mmol), 4-(dimethoxymethyl)-piperidine (98.7 mg, 0.621 mmol), sodium acetate (192 mg, 3.10 mmol) and acetic acid (3.73 mg, 0.0621 mmol) in dichloromethane / methanol (5 mL / 1 mL). The resulting reaction was stirred at room temperature for 12 h. After the reaction was completed, the reaction was poured into water (100 mL) and extracted with dichloromethane (3 x 25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated and purified by column to give s10-5 (265 mg). UPLC-MS: [M+H] = 737.4 found. +

[0214] ​​Synthesis of compound s10-6: 1 M sulfuric acid (5 mL) was added to a solution of s10-5 (265 mg, 0.360 mmol) in tetrahydrofuran (5 mL). The resulting reaction was stirred at 60 °C for 3 h. After the reaction was complete, it was quenched with saturated sodium bicarbonate solution. It was extracted with ethyl acetate (3 x 25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column to give s10-6 (216 mg). UPLC-MS: [M+H] = 691.32 found. +

[0215] Synthesis of compound s10: Sodium cyanoborohydride (39.4 mg, 0.626 mmol) was added to a solution of s10-6 (216 mg, 0.313 mmol), m2 (161 mg, 0.313 mmol), sodium acetate (128 mg, 1.56 mmol), and acetic acid (1.88 mg, 0.0313 mmol) in dichloromethane / methanol (5 mL / 1 mL). The resulting reaction was stirred at room temperature for 12 h. After the reaction was complete, the reaction was poured into water (100 mL) and extracted with dichloromethane (3 x 25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by HPLC to give s10 (85.5 mg). UPLC-MS: [M+H] = 1189.51 found. + 1 ​​H NMR (500 MHz, Methanol-d4) δ 8.97 (d, J = 6.1 Hz, 1H), 8.01 (d, J = 24.9 Hz, 1H), 7.80 - 7.70 (m, 4H), 7.50 - 7.37 (m, 4H), 7.33 - 7.13 (m, 5H), 6.08 (d, J = 27.6 Hz, 1H), 5.03 (q, J = 7.0 Hz, 1H), 4.51 (t, J = 8.2 Hz, 1H), 4.44 (dp, J = 4.4, 2.3 Hz, 1H), 4.36 (s, 2H), 4.14 (d, J = 3.4 Hz, 4H), 4.11 - 4.03 (m, 1H), 4.01 - 3.80 (m, 2H), 3.75 - 3.58 (m, 7H), 3.55 - 3.37 (m, 1H), 3.22 (qd, J = 14.6, 9.5 Hz, 2H), 3.12 - 2.98 (m, 5H), 2.93 - 2.85 (m, 2H), 2.76 (ddd, J = 31.0, 14.2, 8.1 Hz, 1H), 2.65 (s, 1H), 2.63 - 2.44 (m, 4H), 2.42 - 2.29 (m, 1H), 2.22 - 1.93 (m, 6H), 1.83 (d, J = 12.7 Hz, 2H), 1.74 - 1.55 (m, 3H), 1.52 (d, J = 7.1 Hz, 3H), 1.43 - 1.25 (m, 9H), 1.05 (dd, J = 6.7, 3.8 Hz, 3H), 0.91 - 0.85 (m, 3H). 13C NMR (126 MHz, Methanol-d4) δ 171.74, 171.29, 169.81, 169.56, 166.82, 157.35, 157.27, 151.84, 147.66, 147.61, 147.04, 145.77, 145.65, 144.46, 136.09, 134.75, 134.71, 132.14, 130.37, 130.17, 129.78, 129.16, 129.10, 128.62, 128.31, 128.11, 126.75, 126.58, 126.33, 126.28, 126.18, 126.07, 93.49, 69.69, 69.56, 69.36, 62.22, 59.39, 56.00, 53.63, 52.36, 51.51, 51.13, 50.63, 48.71, 46.37, 41.76, 40.77, 38.57, 37.54, 37.33, 37.22, 37.08, 34.58, 33.91, 31.65, 31.08, 30.77, 28.61, 26.69, 20.94, 20.89, 20.68, 20.03, 19.07, 14.13. HPLC > 95%; HRMS (ESI-TOF) m / z: [M+H] + Calculated C66H85N12O7S+, 1189.6379; found, 1189.6375.

[0216] Example 34:

[0217] Synthesis of compound sll: Refer to the synthesis of compound slO. Compound sll-2 was used to react with compound m7b via reductive amination to produce compound sll, UPLC-MS: [M+H]+= 1161.18 found; 1H NMR (500 MHz, Methanol-d4) δ 8.95 (d, J = 4.9 Hz, 1H), 8.01 (dd, J = 25.1, 1.9 Hz, 1H), 7.79 - 7.65 (m, 4H), 7.49 - 7.36 (m, 4H), 7.34 - 7.15 (m, 5H), 6.08 (d, J = 26.7 Hz, 1H), 5.06 - 4.99 (m, 1H), 4.55 - 4.18 (m, 7H), 4.17 - 4.03 (m, 5H), 3.99 (d, J = 14.0 Hz, 1H), 3.92 - 3.79 (m, 1H), 3.77 - 3.33 (m, 8H), 3.28 - 3.15 (m, 3H), 3.06 - 2.69 (m, 4H), 2.68 - 2.54 (m, 1H), 2.48 (d, J = 2.8 Hz, 3H), 2.42 - 2.27 (m, 1H), 2.26 - 2.15 (m, 1H), 2.07 - 1.91 (m, 1H), 1.90 - 1.80 (m, 1H), 1.75 (d, J = 12.8 Hz, 2H), 1.67 - 1.56 (m, 1H), 1.52 (d, J = 7.0 Hz, 3H), 1.47 - 1.24 (m, 10H), 1.05 (dd, J = 6.7, 3.6 Hz, 3H), 0.89 (dd, J = 15.7, 6.7 Hz, 3H). 13 C NMR (126 MHz, Methanol-d4) δ 171.74, 171.29, 169.83, 169.56, 166.78, 157.36, 157.28, 151.75, 147.65, 147.61, 147.19, 145.78, 145.65, 144.40, 136.05, 134.77, 134.72, 132.07, 130.22, 130.09, 129.86, 129.15, 129.09, 128.68, 128.31, 128.11, 126.75, 126.58, 126.33, 126.27, 126.17, 126.07, 93.48, 69.69, 69.57, 69.36, 59.40, 55.99, 53.63, 53.47, 50.84, 50.63, 48.70, 48.52, 46.35, 46.33, 41.76, 40.77, 38.58, 37.53, 37.33, 37.22, 37.08, 34.58, 33.91, 31.91, 31.65, 28.12, 20.94, 20.88, 20.68, 20.03, 19.07, 14.20. HPLC > 95%; HRMS (ESI-TOF) m / z: [M+H] +C64H81N12O7S+, 1161.6066; found, 1161.6063.

[0218] Example 35:

[0219] Synthesis of compound s12: Refer to the synthesis of compound s10. Compound s12 was prepared by reductive amination of compound s12-2 with compound m7b. UPLC-MS: [M+H]+= 1147.55 found; 1 H NMR (500 MHz, Methanol-d4) δ 8.97 (d, J = 6.4 Hz, 1H), 8.03 (d, J = 25.3 Hz, 1H), 7.77 (q, J = 7.9 Hz, 4H), 7.44 (tq, J = 16.3, 8.3, 7.6 Hz, 4H), 7.35 - 7.15 (m, 5H), 6.14 (d, J = 9.3 Hz, 1H), 5.09 - 5.00 (m, 1H), 4.63 - 4.40 (m, 6H), 4.35 (s, 2H), 4.28 - 4.05 (m, 5H), 4.04 - 3.82 (m, 2H), 3.78 - 3.59 (m, 5H), 3.30 - 3.17 (m, 3H), 3.08 - 2.70 (m, 4H), 2.65 - 2.45 (m, 4H), 2.38 (ddt, J = 14.4, 11.3, 5.9 Hz, 1H), 2.28 - 2.17 (m, 1H), 2.10 - 1.83 (m, 2H), 1.83 - 1.70 (m, 2H), 1.63 (dq, J = 24.9, 7.6, 5.7 Hz, 2H), 1.53 (dd, J = 14.3, 7.1 Hz, 3H), 1.48 - 1.27 (m, 10H), 1.07 (dd, J = 6.6, 2.4 Hz, 3H), 0.94 - 0.87 (m, 3H). 13C NMR (126 MHz, Methanol-d4) δ 171.25, 170.81, 169.32, 169.07, 166.31, 156.87, 156.78, 151.26, 147.13, 146.69, 145.27, 145.15, 143.90, 143.85, 135.59, 134.23, 134.21, 132.12, 131.75, 129.81, 129.68, 128.66, 128.60, 128.10, 127.82, 127.62, 126.26, 126.09, 125.84, 125.78, 125.66, 125.58, 93.01, 69.20, 69.08, 68.87, 60.25, 58.91, 55.50, 53.15, 50.13, 48.66, 45.82, 41.27, 40.28, 38.09, 37.03, 36.73, 36.58, 34.07, 33.42, 31.42, 31.15, 28.93, 28.42, 27.60, 20.46, 20.40, 20.19, 19.53, 18.59, 18.38, 13.77. HPLC > 95%; HRMS (ESI-TOF) m / z: [M+H] + Calculated C63H79N12O7S+, 1147.5910; found, 1147.5907.

[0220] Example 36:

[0221] Synthesis of compound s13: Refer to the synthesis of compound s10. Compound s13 was prepared by reductive amination reaction of compound s13-2 with compound m7b. UPLC-MS: [M+H] + = 1175.89 found; 1H NMR (500 MHz, Methanol-d4) δ 9.00 (d, J = 6.9 Hz, 1H), 8.01 (d, J = 24.7 Hz, 1H), 7.78 - 7.70 (m, 4H), 7.47 - 7.37 (m, 4H), 7.32 - 7.13 (m, 5H), 6.09 (d, J = 28.5 Hz, 1H), 5.03 (q, J = 7.1 Hz, 1H), 4.52 (t, J = 8.2 Hz, 1H), 4.43 (d, J = 11.9 Hz, 3H), 4.24 - 4.06 (m, 5H), 3.99 (d, J = 13.9 Hz, 1H), 3.92 - 3.78 (m, 3H), 3.77 - 3.58 (m, 6H), 3.29 - 2.97 (m, 6H), 2.91 (qd, J = 12.6, 12.1, 3.1 Hz, 2H), 2.75 (ddd, J = 29.1, 14.3, 8.1 Hz, 1H), 2.67 - 2.45 (m, 7H), 2.43 - 2.29 (m, 1H), 2.27 - 1.80 (m, 7H), 1.55 (dd, J = 36.2, 7.0 Hz, 4H), 1.48 - 1.26 (m, 9H), 1.05 (dd, J = 6.6, 4.1 Hz, 3H), 0.89 (dd, J = 16.7, 6.8 Hz, 3H). 13 C NMR (126 MHz, Methanol-d4) δ 171.76, 171.28, 171.21, 169.78, 169.57, 166.82, 157.37, 157.28, 151.95, 147.66, 147.61, 146.81, 145.78, 145.66, 144.53, 136.06, 134.76, 134.71, 132.48, 132.21, 130.23, 130.20, 129.67, 129.16, 129.10, 128.62, 128.31, 128.12, 126.76, 126.58, 126.31, 126.21, 126.08, 93.55, 69.69, 69.57, 69.37, 59.42, 56.01, 53.64, 53.48, 50.64, 48.72, 46.38, 41.76, 41.67, 40.81, 40.78, 39.05, 38.61, 37.56, 37.52, 37.34, 37.23, 37.07, 34.59, 33.91, 31.61, 30.93, 28.61, 20.96, 20.90, 20.70, 20.05, 19.09, 14.06. HPLC > 95%; HRMS (ESI-TOF) m / z: [M+H]+calcd for C65H83N12O7S+, 1175.6223; found, 1175.6222.

[0222] Example 37:

[0223] Synthesis of compound s14-2: s10-1 (1.02 g, 3.07 mmol) was added to a solution of m4 (1.00 g, 3.07 mmol, prepared according to example 4), HATU (1.17 g, 3.07 mmol), and N,N-diisopropylethylamine (594 mg, 4.60 mmol) in N,N-dimethylformamide (20 mL) at 0 °C. The reaction was stirred at 0 °C for 2 h. After completion of the reaction, the reaction was poured into water (100 mL) and extracted with ethyl acetate (3 x 25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo and purified by column to give a mixture of non-corresponding isomers s14-2 (1.55 g). UPLC-MS: [M+H]+= 624.7 [M+H] + found.

[0224] 1 M sulfuric acid (5 mL) was added to a solution of s14-2 (500 mg, 0.767 mmol) in tetrahydrofuran (5 mL). The resulting reaction was stirred at 60 °C for 3 h. After completion of the reaction, it was quenched with saturated sodium bicarbonate solution. It was extracted with ethyl acetate (3 x 25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo and purified by column to give s14-3 (368 mg). UPLC-MS: [M+H]+= 580.2 [M+H] + found.

[0225] Chiral HPLC resolution to give s14-4r and s14-4s. s14-4r: UPLC-MS: [M+H] + = 580.25 found; 1H NMR (600 MHz, Methanol-d4) δ 8.89 (s, 1H), 7.49 - 7.38 (m, 4H), 6.26 - 6.05 (m, 1H), 5.09 - 5.00 (m, 1H), 4.54 (td, J = 8.2, 2.4 Hz, 1H), 4.46 (dq, J = 4.7, 2.3 Hz, 1H), 3.89 - 3.83 (m, 1H), 3.66 (tt, J = 12.9, 6.3 Hz, 4H), 2.56 - 2.48 (m, 6H), 2.45 - 2.34 (m, 1H), 2.20 (ddd, J = 12.0, 7.9, 3.4 Hz, 1H), 1.98 (ddd, J = 13.3, 8.8, 4.7 Hz, 1H), 1.80 (tt, J = 13.7, 6.9 Hz, 1H), 1.61 (t, J = 6.6 Hz, 1H), 1.54 (d, J = 7.0 Hz, 3H), 1.38 - 1.28 (m, 1H), 1.17 (d, J = 6.1 Hz, 1H), 1.08 (t, J = 7.3 Hz, 3H), 0.96 - 0.89 (m, 3H).

[0226] Synthesis of compound s14: Refer to the synthesis of compound s10. Compound s14 was prepared by reductive amination reaction of compound s14-6 with compound m7b. UPLC-MS: [M+H] + = 1175.51 found.

[0227] Example 38:

[0228] Synthesis of compound s15: Refer to the synthesis of compound s10. Compound s15 was prepared by reductive amination reaction of compound s15-6 with compound m7b. UPLC-MS: [M+H] + = 1161.53 found; 1H NMR (500 MHz, Methanol-d4) δ 8.91 (d, J = 4.0 Hz, 1H), 8.01 (d, J = 25.2 Hz, 1H), 7.81 - 7.69 (m, 4H), 7.50 - 7.37 (m, 4H), 7.33 - 7.14 (m, 5H), 6.13 (d, J = 27.9 Hz, 1H), 5.03 (td, J = 7.3, 5.5 Hz, 1H), 4.51 (t, J = 8.2 Hz, 1H), 4.47 - 4.38 (m, 2H), 4.22 (d, J = 13.2 Hz, 1H), 4.17 - 3.95 (m, 5H), 3.93 - 3.43 (m, 10H), 3.27 - 3.14 (m, 3H), 3.06 - 2.87 (m, 3H), 2.76 (ddd, J = 31.7, 14.2, 8.1 Hz, 1H), 2.67 - 2.45 (m, 6H), 2.43 - 2.27 (m, 1H), 2.25 - 1.71 (m, 8H), 1.56 (dd, J = 37.1, 7.0 Hz, 4H), 1.48 - 1.25 (m, 9H), 1.05 (dd, J = 6.5, 3.2 Hz, 3H), 0.90 (dd, J = 13.9, 6.6 Hz, 3H). 13 CNMR (126 MHz, Methanol-d4) δ 173.22, 173.14, 172.73, 171.68, 170.94, 167.68, 158.68, 153.06, 149.11, 149.06, 148.82, 147.20, 147.07, 145.76, 137.46, 136.19, 136.15, 133.55, 131.70, 131.59, 131.51, 131.41, 130.59, 130.52, 129.73, 129.53, 128.18, 128.00, 127.75, 127.67, 127.55, 127.49, 94.93, 71.11, 70.99, 70.79, 60.81, 57.41, 55.04, 54.87, 52.04, 50.23, 50.12, 47.13, 47.08, 43.18, 42.19, 39.98, 39.00, 38.97, 38.74, 38.63, 38.51, 36.00, 35.32, 33.12, 26.81, 22.38, 22.35, 22.29, 22.09, 21.43, 20.49, 15.69. HPLC >95%; HRMS (ESI-TOF) m / z: [M+H] + Calculated C64H81N12O7S+, 1161.6066; found, 1161.6067.

[0229] Example 39:

[0230] Synthesis of compound s16: Refer to the synthesis of compound s10. Compound s16 was prepared by reductive amination of compound s16-6 with compound m7b. UPLC-MS: [M+H] + = 1133.50 found; 1 H NMR (500 MHz, Methanol-d4) δ 8.93 (d, J = 7.2 Hz, 1H), 8.03 (d, J = 25.5 Hz, 1H), 7.80 - 7.66 (m, 4H), 7.52 - 7.38 (m, 4H), 7.36 - 7.16 (m, 5H), 6.15 (d, J = 16.3 Hz, 1H), 5.00 (d, J = 6.5 Hz, 1H), 4.60 (t, J = 8.1 Hz, 1H), 4.54 - 4.40 (m, 3H), 4.39 - 4.20 (m, 5H), 4.19 - 4.04 (m, 4H), 4.04 - 3.89 (m, 1H), 3.88 - 3.57 (m, 6H), 3.44 (t, J = 11.9 Hz, 1H), 3.32 - 3.16 (m, 2H), 3.09 - 2.71 (m, 4H), 2.66 - 2.44 (m, 4H), 2.44 - 2.34 (m, 1H), 2.23 (td, J = 14.4, 12.5, 7.4 Hz, 1H), 2.12 - 1.91 (m, 3H), 1.56 (dd, J = 44.3, 7.1 Hz, 6H), 1.44 - 1.28 (m, 8H), 1.02 (dd, J = 55.3, 6.6 Hz, 3H), 0.92 (dd, J = 7.0, 4.8 Hz, 3H). 13C NMR (126 MHz, Methanol-d4) δ 171.36, 170.82, 169.55, 168.88, 165.80, 156.87, 156.79, 151.10, 147.13, 147.08, 146.98, 145.27, 145.14, 143.79, 135.76, 134.18, 133.89, 131.56, 129.68, 129.47, 129.32, 128.96, 128.73, 128.58, 127.82, 127.62, 126.26, 126.08, 125.91, 125.84, 125.63, 125.58, 93.27, 69.20, 69.08, 68.88, 61.70, 58.70, 55.18, 53.14, 49.65, 48.88, 44.59, 41.27, 40.28, 38.05, 37.04, 36.72, 36.59, 34.07, 33.41, 30.59, 28.93, 28.42, 26.20, 24.69, 20.53, 20.38, 20.18, 19.59, 18.34, 13.85. HPLC > 95%; HRMS (ESI-TOF) m / z: [M+H]+calcd for C62H77N12O7S+, 1133.5753; found, 1133.5751.

[0231] Example 40:

[0232] Synthesis of compound s17: Refer to the synthesis of compound s10. Compound s17 was prepared by reductive amination of compound s17-6 with compound m7b, UPLC-MS: [M+H]+= 1147.91 found; 1H NMR (500 MHz, Methanol-d4) δ 8.98 (d, J = 7.0 Hz, 1H), 8.00 (d, J = 24.6 Hz, 1H), 7.79 - 7.69 (m, 4H), 7.47 - 7.36 (m, 4H), 7.32 - 7.14 (m, 5H), 6.12 (d, J = 25.2 Hz, 1H), 5.03 (q, J = 6.9 Hz, 1H), 4.51 (t, J = 8.2 Hz, 1H), 4.47 - 4.29 (m, 5H), 4.25 - 4.03 (m, 7H), 4.03 - 3.78 (m, 4H), 3.77 - 3.57 (m, 3H), 3.57 - 3.39 (m, 3H), 3.30 - 3.14 (m, 2H), 3.08 - 2.84 (m, 3H), 2.75 (ddd, J = 29.7, 14.3, 8.2 Hz, 1H), 2.67 - 2.54 (m, 1H), 2.48 (d, J = 2.9 Hz, 4H), 2.43 - 2.14 (m, 2H), 2.11 - 1.87 (m, 3H), 1.78 - 1.55 (m, 2H), 1.55 - 1.25 (m, 12H), 1.05 (dd, J = 6.6, 3.4 Hz, 3H), 0.88 (t, J = 7.5 Hz, 3H). 13 C NMR (126 MHz, Methanol-d4) δ 171.74, 171.29, 170.17, 169.50, 166.29, 157.37, 157.29, 151.89, 147.59, 146.93, 145.78, 145.66, 144.49, 136.05, 134.77, 134.71, 132.41, 132.08, 130.23, 130.21, 129.73, 129.17, 129.10, 128.67, 128.31, 128.12, 126.76, 126.58, 126.33, 126.29, 126.17, 126.07, 93.58, 69.69, 69.57, 69.36, 59.40, 55.99, 53.64, 50.85, 50.62, 48.71, 48.49, 45.10, 41.76, 40.77, 38.60, 37.56, 37.52, 37.34, 37.23, 37.07, 34.58, 33.91, 31.65, 26.37, 25.02, 20.95, 20.90, 20.69, 20.03, 19.07, 14.10. HPLC >95%; HRMS (ESI-TOF) m / z: [M+H] + Calculated C63H79N12O7S+, 1147.5910; Found, 1147.5912.

[0233] Example 41:

[0234] Synthesis of compound s18-2: s10-1 (744 mg, 2.25 mmol) was added to a solution of m5 (953 mg, 2.70 mmol, prepared according to Example 5), HATU (855 mg, 2.25 mmol), and N,N-diisopropylethylamine (580 mg, 4.50 mmol) in N,N-dimethylformamide (20 mL) at 0 °C. The reaction was stirred at 0 °C for 2 h. After the reaction was complete, the reaction was poured into water (100 mL) and extracted with ethyl acetate (3 x 25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to dryness before purification by column to give a mixture of non-corresponding isomers (929 mg). UPLC-MS: [M+H]+= 667.32 [M+H]+ found.

[0235] Synthesis of compounds s18-3R and s18-3S: Compound s18-2 (929 mg, 1.39 mmol) was dissolved in dichloromethane (10 mL) and trifluoroacetic acid (2.5 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 h and then concentrated to dryness to give 790 mg of a mixture of non-corresponding isomers. UPLC-MS: [M+H]+= 567.31 [M+H]+ found. s18-3R (0.316 g) and s18-3S (0.300 g) were obtained by chiral resolution.

[0236] s18-3R: 1H NMR (600 MHz, Methanol-d4) δ 8.89 (s, 1H), 7.45 (q, J = 8.4 Hz, 4H), 6.13 (s, 1H), 5.06 (q, J = 6.8 Hz, 1H), 4.53 (t, J = 8.2 Hz, 1H), 4.46 (dp, J = 4.6, 2.2 Hz, 1H), 3.86 (dd, J = 10.9, 4.2 Hz, 1H), 3.68 - 3.55 (m, 2H), 3.30 - 3.24 (m, 4H), 2.99 - 2.92 (m, 4H), 2.50 (s, 3H), 2.43 - 2.34 (m, 1H), 2.23 - 2.17 (m, 1H), 1.98 (ddd, J = 13.2, 8.7, 4.7 Hz, 1H), 1.57 (dd, J = 40.7, 7.0 Hz, 3H), 1.07 (dd, J = 6.7, 2.5 Hz, 3H), 0.91 (d, J = 6.7 Hz, 3H).

[0237] Synthesis of compound s18: Sodium cyanoborohydride (25.8 mg, 0.410 mmol) was added to a solution of s18-3R (116 mg, 0.205 mmol), m2 (105 mg, 0.205 mmol), sodium acetate (84.0 mg, 1.025 mmol) and acetic acid (1.27 mg, 0.0205 mmol) in dichloromethane / methanol (5 mL / 1 mL). The resulting reaction was stirred at room temperature for 12 h. After the reaction was complete, the reaction was poured into water (100 mL) and extracted with dichloromethane (3 x 25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. s18 (39.2 mg) was obtained by HPLC purification. UPLC-MS: [M+H] += 1064.46 Found,1H NMR (600 MHz, Methanol-d4) δ 8.89 (s, 1H), 8.00 (d, J = 31.5 Hz, 1H), 7.70 - 7.58 (m, 4H), 7.45 (q, J = 8.2 Hz, 4H), 7.36 - 7.16 (m, 5H), 6.10 (d, J = 32.1 Hz, 1H), 5.05 (p, J = 6.8 Hz, 1H), 4.53 (t, J = 8.2 Hz, 1H), 4.48 - 4.40 (m, 1H), 4.28 - 4.15 (m, 4H), 4.14 - 4.05 (m, 1H), 4.02 - 3.89 (m, 1H), 3.86 (dd, J = 10.9, 4.2 Hz, 1H), 3.78 - 3.60 (m, 5H), 3.32 - 3.17 (m, 6H), 3.08 - 2.91 (m, 1H), 2.79 (ddd, J = 40.1, 14.2, 8.1 Hz, 1H), 2.70 - 2.58 (m, 5H), 2.50 (s, 3H), 2.43 - 2.33 (m, 1H), 2.20 (t, J = 10.0 Hz, 1H), 2.07 - 1.94 (m, 1H), 1.69 - 1.58 (m, 1H), 1.54 (d, J = 7.1 Hz, 3H), 1.48 - 1.29 (m, 6H), 1.07 (d, J = 6.5 Hz, 3H), 0.93 - 0.87 (m, 3H).13C NMR (151 MHz, Methanol-d4) δ 171.78, 171.20, 170.62, 169.39, 165.96, 157.33, 157.26, 152.69, 147.68, 145.83, 145.73, 145.53, 144.97, 135.85, 134.82, 134.72, 133.27, 131.73, 130.21, 129.76, 129.19, 129.13, 128.98, 128.35, 128.17, 126.79, 126.62, 126.46, 126.33, 126.11, 93.73, 69.72, 69.60, 69.41, 59.68, 59.46, 56.08, 50.63, 50.55, 48.76, 44.12, 41.80, 40.82, 38.85, 37.68, 37.44, 37.02, 34.66, 33.97, 31.63, 21.08, 21.02, 20.80, 20.09, 19.19, 13.66. HPLC > 95%; HRMS (ESI-TOF) m / z: [M + H] + Calculated C58H70N11O7S+, 1064.5175; found, 1064.5179.

[0238] Example 42:

[0239] Synthesis of compound s19: Sodium cyanoborohydride (39.4 mg, 0.626 mmol) was added to a solution of s10-4 (185 mg, 0.313 mmol), m1 (161 mg, 0.313 mmol, prepared according to reference example 1), sodium acetate (128 mg, 1.56 mmol) and acetic acid (1.88 mg, 0.0313 mmol) in dichloromethane / methanol (5 mL / 1 mL). The resulting reaction was stirred at room temperature for 12 h. After the completion of the reaction, the reaction was poured into water (100 mL) and extracted with dichloromethane (3 x 25 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give s18 (95.5 mg) which was purified by HPLC. UPLC-MS: [M+H] + = 1092.52 found, 1 H NMR (600 MHz, Methanol-d4) δ 8.88 (d, J = 5.5 Hz, 1H), 7.99 (dd, J = 31.0, 3.5 Hz, 1H), 7.66 - 7.57 (m, 4H), 7.47 - 7.42 (m, 4H), 7.34 - 7.15 (m, 5H), 6.08 (d, J = 29.8 Hz, 1H), 5.08 - 5.01 (m, 1H), 4.54 (t, J = 8.1 Hz, 1H), 4.47 - 4.40 (m, 1H), 4.29 - 4.09 (m, 5H), 4.09 - 3.82 (m, 5H), 3.77 - 3.56 (m, 5H), 3.31 - 3.17 (m, 2H), 2.99 (dtd, J = 58.2, 12.9, 12.4, 3.4 Hz, 1H), 2.90 - 2.71 (m, 3H), 2.57 - 2.45 (m, 6H), 2.43 - 2.33 (m, 1H), 2.19 (ddt, J = 12.7, 8.1, 1.9 Hz, 1H), 1.98 (ddd, J = 13.2, 8.7, 4.7 Hz, 1H), 1.83 (dd, J = 13.7, 3.5 Hz, 2H), 1.74 (ttt, J = 10.6, 6.7, 3.6 Hz, 1H), 1.57 (dd, J = 35.3, 7.1 Hz, 4H), 1.48 - 1.21 (m, 8H), 1.06 (dd, J = 6.6, 2.3 Hz, 3H), 0.93 - 0.88 (m, 3H). 13C NMR (151 MHz, methanol-d4) δ 171.76, 171.28, 169.65, 169.59, 167.03, 157.34, 151.46, 147.68, 147.33, 147.28, 145.80, 145.66, 144.26, 141.14, 137.05, 134.63, 134.46, 131.94, 130.12, 129.55, 129.13, 129.09, 128.72, 128.31, 128.11, 126.76, 126.59, 126.34, 126.24, 126.17, 126.07, 125.99, 93.58, 69.69, 69.57, 69.36, 59.38, 55.98, 54.25, 53.66, 52.81, 50.66, 48.70, 41.77, 40.79, 38.48, 37.52, 37.22, 37.08, 35.59, 34.56, 33.96, 31.67, 29.41, 20.97, 20.88, 20.68, 20.07, 19.10, 14.43. HPLC >95%; HRMS (ESI-TOF) m / z: [M+H] + Calculated C60H74N11O7S+, 1092.5488; found, 1092.5488.

[0240] Example 43:

[0241] Synthesis of compound s21: Refer to the synthesis of compound s18. Compound s21 was prepared by reductive amination reaction of compound s18-3R with compound m7a, UPLC-MS: [M+H] + = 1082.64 found.

[0242] Example 44:

[0243] Synthesis of compound s22: Refer to the synthesis of compound s18. Compound s22 was prepared by reductive amination reaction of compound s18-3R with compound m9a, UPLC-MS: [M+H] + = 1118.32 found. 1H NMR (600 MHz, DMSO-d6) δ 8.99 (s, 1H), 8.41 (d, J = 7.3 Hz, 1H), 7.99 (d, J = 13.6 Hz, 1H), 7.74 - 7.66 (m, 2H), 7.60 - 7.49 (m, 2H), 7.48 - 7.39 (m, 4H), 7.39 - 7.29 (m, 5H), 6.15 (s, 1H), 5.12 (q, J = 3.2 Hz, 1H), 4.41 - 4.34 (m, 1H), 4.29 (s, 1H), 4.17 - 4.08 (m, 4H), 4.03 - 3.90 (m, 3H), 3.80 - 3.74 (m, 1H), 3.72 (dd, J = 10.8, 4.2 Hz, 1H), 3.64 - 3.57 (m, 2H), 3.43 (d, J = 11.1 Hz, 1H), 3.30 - 3.14 (m, 6H), 3.06 - 2.75 (m, 3H), 2.61 - 2.48 (m, 4H), 2.46 (d, J = 2.5 Hz, 3H), 2.22 (tdd, J = 23.4, 12.1, 7.7 Hz, 1H), 2.03 (t, J = 10.6 Hz, 1H), 1.79 (td, J = 13.8, 12.4, 8.6 Hz, 1H), 1.48 - 1.30 (m, 6H), 1.27 - 1.16 (m, 1H), 0.97 (dd, J = 10.0, 6.6 Hz, 3H), 0.80 (d, J = 6.7 Hz, 3H).

[0244] Example 45:

[0245] Synthesis of compound s23: Refer to the synthesis of compound s18. Compound s23 was prepared by reductive amination reaction of compound s18-3R with compound m10a, UPLC-MS: [M+H] + = 1110.49 found.

[0246] Example 46:

[0247] Synthesis of compound s24: Refer to the synthesis of compound s18. Compound s24 was prepared by reductive amination reaction of compound s18-3R with compound m11a, UPLC-MS: [M+H] + = 1136.73 found.

[0248] Example 47:

[0249] Synthesis of compound s25: Refer to the synthesis of compound s18. Compound s25 was prepared by reductive amination reaction of compound s18-3R with compound m12a, UPLC-MS: [M+H] = 1186.55 found. +

[0250] Example 48:

[0251] Synthesis of compound s26: Refer to the synthesis of compound s18. Compound s26 was prepared by reductive amination reaction of compound s18-3R with compound m6a, UPLC-MS: [M+H] = 1132.81 found. +

[0252] Example 49:

[0253] Synthesis of compound s27: Refer to the synthesis of compound s18. Compound s27 was prepared by reductive amination reaction of compound s18-3R with compound m13a, UPLC-MS: [M+H] = 1054.21 found. +

[0254] Example 50:

[0255] Synthesis of compound s29: Refer to the synthesis of compound s19. Compound s29 was prepared by reductive amination reaction of compound s10-4 with compound m9b, UPLC-MS: [M+H] = 1146.32 found. +

[0256] Example 51:

[0257] Synthesis of compound s30: Refer to the synthesis of compound s19. Compound s30 was prepared by reductive amination reaction of compound s10-4 with compound m10b, UPLC-MS: [M+H] = 1128.11 found. +

[0258] Example 52:

[0259] Synthesis of compound s31: Refer to the synthesis of compound s19. Compound s31 was prepared by reductive amination reaction of compound s10-4 with compound m11b, UPLC-MS: [M+H] = 1164.45 found. +

[0260] Example 53: ​​​​​​

[0261] Synthesis of compound s32: Refer to the synthesis of compound s19. Compound s32 was prepared by reductive amination reaction of compound s10-4 with compound m12b, UPLC-MS: [M+H] + = 1214.68 found.

[0262] Example 54:

[0263] Synthesis of compound s33: Refer to the synthesis of compound s19. Compound s33 was prepared by reductive amination reaction of compound s10-4 with compound m6b, UPLC-MS: [M+H] + = 1160.89 found.

[0264] Example 55:

[0265] Synthesis of compound s34: Refer to the synthesis of compound s19. Compound s34 was prepared by reductive amination reaction of compound s10-4 with compound m13b, UPLC-MS: [M+H] + = 1082.18 found.

[0266] Example 56:

[0267] Synthesis of compound s36: Refer to the synthesis of compound s19. Compound s36 was prepared by reductive amination reaction of compound s10-4 with compound m7b, UPLC-MS: [M+H] + = 1110.47 found.

[0268] Example 57:

[0269] Synthesis of compound s37: Refer to the synthesis of compound s18. Compound s37 was prepared by reductive amination reaction of compound s18-3R with compound m14a, UPLC-MS: [M+H] + = 1084.34 found.

[0270] Example 58:

[0271] Synthesis of compound s38: Refer to the synthesis of compound s19. Compound s38 was prepared by reductive amination reaction of compound s10-4 with compound m14b, UPLC-MS: [M+H] + = 1112.62 found.

[0272] Example 59:

[0273] Synthesis of compound m14a-4: MsCI (45.2 mg, 0.393 mmol) was added to a solution of compound m14c (150 mg, 0.262 mmol) and triethylamine (79.4 mg, 0.786 mmol) in dichloromethane (3 mL) and the reaction was allowed to proceed at room temperature for 1 h. After the reaction was complete, the reaction was quenched with water (10 mL) and extracted with dichloromethane (3 x 5 mL). The organic phase was dried, filtered and concentrated in vacuo and the residue was purified by column chromatography to give m14a-4 (130 mg). UPLC-MS: [M+H] = 572.2 found. +

[0274] Synthesis of compound s39: s18-3R (113.4 mg, 0.200 mmol) was added to a solution of m14a-4 (130 mg, 0.200 mmol), potassium carbonate (83 mg, 0.600 mmol) and potassium iodide (33 mg, 0.200 mmol) in acetonitrile (3 mL) and the resulting suspension was heated to 90 °C for 2 h. The reaction was cooled to room temperature and quenched with water (10 mL) and extracted with ethyl acetate (5 mL) three times. The organic phases were combined, dried, filtered and concentrated in vacuo and the residue was purified by column chromatography to give compound m1-3 (42 mg), UPLC-MS: [M+H] = 1120.5 found. + 1 ​​HNMR (600 MHz, Methanol-d4) δ 8.89 (s, 1H), 8.00 (d, J = 28.6 Hz, 1H), 7.66 (dt, J = 8.2, 1.7 Hz, 2H), 7.61 (dt, J = 8.2, 1.8 Hz, 2H), 7.47 - 7.31 (m, 9H), 6.11 (s, 1H), 5.04 (dq, J = 14.3, 7.2 Hz, 1H), 4.53 (t, J = 8.2 Hz, 1H), 4.46 (dp, J = 4.7, 2.3 Hz, 1H), 4.19 - 4.13 (m, 4H), 4.10 - 4.05 (m, 1H), 3.97 (q, J = 14.1 Hz, 1H), 3.86 (dd, J = 10.9, 4.2 Hz, 1H), 3.83 - 3.74 (m, 1H), 3.65 (d, J = 9.9 Hz, 1H), 3.62 (dt, J = 10.9, 1.8 Hz, 1H), 3.32 - 3.18 (m, 6H), 3.06 - 2.88 (m, 2H), 2.68 - 2.55 (m, 4H), 2.49 (d, J = 2.0 Hz, 3H), 2.44 - 2.31 (m, 1H), 2.23 - 2.16 (m, 1H), 2.01 - 1.94 (m, 1H), 1.67 - 1.42 (m, 6H), 1.26 - 1.17 (m, 1H), 1.07 (dd, J = 6.6, 2.6 Hz, 3H), 0.92 (dd, J = 17.5, 6.7 Hz, 3H).

[0275] Example 60:

[0276] Synthesis of compound s40: Refer to the synthesis of compound s18. Compound s40 was prepared by reductive amination of compound s18-3R with compound m15a. UPLC-MS: [M+H] = 1048.5 found. + = 1048.5 found. 1H NMR (600 MHz, Methanol-d4) δ 8.89 (d, J = 1.4 Hz, 1H), 8.08 - 8.03 (m, 1H), 7.68 (dd, J = 7.8, 1.5 Hz, 1H), 7.49 - 7.30 (m, 11H), 6.23 (s, 1H), 5.05 (q, J = 7.0 Hz, 1H), 4.55 - 4.40 (m, 4H), 4.20 (d, J = 4.4 Hz, 3H), 4.17 - 4.05 (m, 3H), 4.03 (s, 3H), 3.99 (d, J = 6.7 Hz, 1H), 3.88 - 3.74 (m, 3H), 3.70 (d, J = 9.9 Hz, 1H), 3.64 (dt, J = 10.9, 1.8 Hz, 1H), 3.58 - 3.35 (m, 6H), 3.29 - 3.15 (m, 2H), 3.06 - 2.90 (m, 2H), 2.49 (d, J = 4.8 Hz, 3H), 2.40 (d h, J = 9.4, 6.5 Hz, 1H), 2.25 - 2.17 (m, 1H), 1.98 (ddd, J = 13.2, 8.8, 4.6 Hz, 1H), 1.67 - 1.59 (m, 1H), 1.54 (d, J = 7.0 Hz, 3H), 1.45 (t, J = 10.2 Hz, 1H), 1.36 - 1.30 (m, 1H), 1.22 (td, J = 12.8, 11.7, 4.5 Hz, 1H), 1.08 (dd, J = 6.6, 2.4 Hz, 3H), 0.92 (dd, J = 15.0, 6.7 Hz, 3H).

[0277] Example 61:

[0278] Synthesis of compound s41: Refer to the synthesis of compound s18. Compound s41 was prepared by reductive amination reaction of compound s18-3R with compound m16a, UPLC-MS: [M+H] + = 1174.5 found.

[0279] Example 62:

[0280] Synthesis of compound s42: Refer to the synthesis of compound s18. Compound s42 was prepared by reductive amination reaction of compound s18-3R with compound m17a, UPLC-MS: [M+H] + = 1151.5 found.

[0281] Example 63:

[0282] Synthesis of compound s47: Refer to the synthesis of compound s18. Compound s47 was prepared by reductive amination reaction of compound s18-3R with compound m25a, UPLC-MS: [M+H] = 1166.5 found. + = 1166.5 found.

[0283] Example 64:

[0284] Synthesis of compound s48: Refer to the synthesis of compound s18. Compound s48 was prepared by reductive amination reaction of compound s18-3R with compound m26a, UPLC-MS: [M+H] = 1121.5 found. + = 1121.5 found.

[0285] Example 65:

[0286] Synthesis of compound s49: Refer to the synthesis of compound s18. Compound s49 was prepared by reductive amination reaction of compound s18-3R with compound m19a, UPLC-MS: [M+H] = 1078.6 found. + = 1078.6 found.

[0287] Example 66:

[0288] Synthesis of compound s50: Refer to the synthesis of compound s18. Compound s50 was prepared by reductive amination reaction of compound s18-3R with compound m27a, UPLC-MS: [M+H] = 1132.6 found. + = 1132.6 found.

[0289] Example 67:

[0290] Synthesis of compound s51: Refer to the synthesis of compound s18. Compound s51 was prepared by reductive amination reaction of compound s18-3R with compound m23a, UPLC-MS: [M+H] = 1067.5 found. + = 1067.5 found.

[0291] Example 68:

[0292] Synthesis of compound s52: Refer to the synthesis of compound s18. Compound s52 was prepared by reductive amination reaction of compound s18-3R with compound m24a, UPLC-MS: [M+H] = 1097.5 found. + = 1097.5 found.

[0293] Example 69: Protein degradation experiment

[0294] Cells in logarithmic growth phase were inoculated in 12-well plates, 1*10^6 cells per well, with 1 mL culture solution containing different concentrations of compounds. After 12 hours of treatment, the cells were collected by centrifugation. The cells were washed twice with pre-cooled PBS, then centrifuged to remove PBS, 100 μL of loading buffer was added, the cells were resuspended, and then the sample was boiled at 100°C for 10 minutes. Take an appropriate amount of protein sample for Tricine-SDS-PAGE electrophoresis. After electrophoresis, under the action of stable current, the protein was transferred to a nitrocellulose membrane plate, and then 5% BSA was used for blocking for 2 hours. After blocking, TBST was used for washing three times, each for 10 minutes. An appropriate amount of solution containing the corresponding primary antibody was added to the incubation box at 4°C, and the nitrocellulose membrane was incubated overnight, the next day the secondary antibody was incubated, and finally the developing solution was added for exposure. Image Lab 6.0 was used for gray scale analysis. GraphPad Prism 8.0 was used to calculate the IC 50 (degradation agent concentration at which 50% USP7 degradation is induced). U7D-1 was used as a reference compound (see Angew. Chem., Int. Ed. 2022, 61, e202204395), and representative data are shown in Tables 1 and 2.

[0295] Table 1

[0296] For representative examples in Table 1, the USP7 degradation activity is classified as follows:

[0297] Table 2

[0298] These representative compounds have very strong USP7 protein degradation ability, far superior to the control compound U7D-1.

[0299] Example 70: Assay for the activity of compounds in inhibiting tumor cell proliferation

[0300] Cells in logarithmic growth phase were inoculated in 96-well culture plates, and an appropriate number of cells were inoculated according to the growth rate of the cells. Each well contains 100 μL of culture solution and the corresponding concentration of compounds. After incubating for the corresponding time, 20 μL of PMS-MTS mixture (Promega) was added to each well. After 3 hours of incubation, the absorbance value was read using a SpectraMaxR 340PC384 (Molecular Device) microplate reader. The absorbance difference between 490 nm and 690 nm was taken as the final data. Finally, the IC 50 value of the compound was calculated using the non-linear regression method of GraphPad 8.0. The IC 50The data results come from three independent repeated experiments. U7D-1 is used as a reference compound (see Angew. Chem., Int. Ed. 2022, 61, e202204395), and representative data are shown in Table 3.

[0301] Table 3

[0302] The proliferation inhibition activity of the example compounds on RS4;11 cells is significantly better than that of the control compound U7D-1

[0303] Example 71: In vivo pharmacokinetic experiment of the compound

[0304] After the compound is coated with sulfobutyl cyclodextrin and dissolved in physiological saline to form a clear solution, it is administered intravenously. After ICR mice are administered 20 mg / kg of the drug intravenously, 0.2 mL of blood is continuously taken from the orbital plexus at time points of 2 min, 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, and 24 h, and placed in a heparin anticoagulation tube. After the sample is centrifuged at 12,000 r at 4°C for 2 min, 0.1 mL of the upper plasma is taken and stored at -80°C, and then subjected to LC-MS / MS analysis. The data are analyzed by WinNolin non-compartment model to obtain key pharmacokinetic parameters.

[0305] The data in Table 4 show that the representative compounds s18, s22, s39, and s40 have good plasma exposure after intravenous administration, and the half-lives of s22, s39, and s40 are significantly prolonged.

[0306] Table 4

[0307] Example 72: In vivo anti-tumor effect of the compound

[0308] (1) Model establishment

[0309] RS4;11 xenograft tumor model: female NOD / SCID mice were inoculated with RS4;11 cells (5*10^6 per mouse) subcutaneously near the back, and tumors were formed about 20 days later

[0310] (2) Evaluation of the in vivo anti-tumor effect of s18

[0311] Experimental grouping: I) blank group; II) s18 group 5mg / kg; III) s18 group 10mg / kg; IV) s18 group 20mg / kg; V) azacitidine group 2.5mg / kg. Administration method: intraperitoneal injection (azacitidine administration method is subcutaneous administration). Drug dissolution method: 5%_DMSO, 50%_PEG300, 5%_ethanol, 40%_normal saline. Evaluation index: a. When the tumor volume≈2000mm 3 As the endpoint, the mouse tumor was dissected out, and the tumor size of each group was compared; b. The health status, diet, drinking water and body weight changes of the mice during the experiment were recorded, and the side effects were compared. Figure 1 shows that s18 significantly inhibits the growth of RS4; 11CDX mouse tumors.

[0312] The above only describes the preferred embodiments of the present application, and it should be noted that for those skilled in the art, without departing from the method of the present application, several improvements and supplements can also be made, which should also be considered as the protection scope of the present application.

Claims

A compound of Formula (I), or a pharmaceutically acceptable salt, stereoisomer, enantiomer, diastereomer, atropisomer, racemate, or isotopically labeled compound thereof: wherein, Y1is selected from the group consisting of: hydrogen, deuterium, C 1-6 alkyl, halo-substituted C 1-6 alkyl, C 3-8 cycloalkyl, and 3-8 membered heterocycloalkyl; B ring is selected from: C 6-10 aromatic ring, C 3-10 saturated or unsaturated alicyclic, and 5-10 membered heteroaromatic ring; Y2is selected from the group consisting of: hydrogen, deuterium, halogen, C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, halogenated C 1-6 alkoxy, -CN, -NO2, and C 1-6 alkylsulfonyl; Y3is selected from: C 1-6 alkylene, -CH2NY 3a -, -C(O)NY 3a -, -NY 3a -, -O-C 2-6 alkylene-NY 3a -, and 3-8 membered heterocycloalkylene, wherein Y 3a is hydrogen, C 1-6 alkyl, halogenated C 1-6 alkyl, or C 3-8 cycloalkyl; Y4is selected from the group consisting of: hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 alkyl, and halogenated C 1-6 alkoxy; C ring is selected from: C 6-10 aromatic ring, C 3-10 saturated or unsaturated alicyclic, and 5-10 membered heteroaromatic ring; Y5is selected from the group consisting of: hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 alkyl, halogenated C 1-6 alkoxy, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, -OH, -C(O)OY 5a , -C(O)NHY 5a , -C(O)NHOH, and -NH(CO)Y 5a , wherein Y 5a is hydrogen, C 1-6 alkyl or C 3-8 cycloalkyl; A is the following general structure A1: wherein, R1is selected from the group consisting of: hydrogen, substituted 5-10 membered heteroaryl, C 1-6 alkyl, substituted C 6-10 aryl, substituted C 1-6 alkoxy, and substituted 3-8 membered heterocyclyl; when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 alkyl, halogenated C 1-6 alkoxy, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, and -OH; R2is selected from the group consisting of: hydrogen, halogen, -CN, substituted C 1-6 alkyl, or substituted C 1-6 alkoxy, and substituted C 3-8 cycloalkyl; when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, and -OH; R 3a and R 3b are each independently selected from the group consisting of hydrogen, substituted or unsubstituted C 3-8 cycloalkyl, substituted or unsubstituted C 1-6 alkyl, and substituted or unsubstituted 3-8 membered heterocycloalkyl; and when substituted, said substituents are selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, and -OH; R4is selected from the group consisting of: hydrogen, substituted C 1-6 alkyl, and substituted C 3-8 cycloalkyl; when substituted, said substituent is selected from the group consisting of hydrogen, deuterium, halogen, -CN, -NO2, -NH2, and -OH; R5and R6are independently selected from the group consisting of hydrogen, -OH, -SH, and substituted C 1-6 alkyl, or R5, R6and the carbon atom to which they are attached form a carbonyl group; and when substituted, said substituents are selected from the group consisting of hydrogen, deuterium, halogen, -CN, -NO2, -NH2, and -OH; R7is selected from the group consisting of: hydrogen and substituted C 1-6 alkyl; when substituted, the substituent is selected from the group consisting of hydrogen, deuterium, halogen, -CN, -NO2, -NH2, and -OH; R8is selected from the group consisting of: hydrogen and OH; R 9a and R 9b are each independently selected from the group consisting of hydrogen and substituted C 1-6 alkyl; or R 9a and R 9b groups can together form an oxo or substituted C 3-8 cycloalkyl; and when substituted, said substituents are selected from the group consisting of hydrogen, deuterium, halogen, -CN, -NO2, -NH2, and -OH; R 10 and R 11 are selected from the group consisting of hydrogen, substituted C 1-6 alkyl, substituted C 3-8 cycloalkyl, substituted C 1-6 alkoxy, substituted C 1-6 alkylthio, and substituted 5-10 membered heteroaryl; or R 10 , R 11 and the carbon atom to which they are attached form a substituted C 3-8 cycloalkyl; and when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, and -OH; R 12 is selected from: substituted 3-8 membered heterocyclylene, substituted 5-10 membered heteroarylene, substituted C 6-10 arylene, and In the substituents, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 alkyl, halogenated C 1-6 alkoxy, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, and -OH; Among them, R 13 Selected from: hydrogen, deuterium, substituted C 1-6 Alkyl groups, and substituted C groups 3-8 Cycloalkyl; when substituted, the substituent is selected from hydrogen, deuterium, halogen, C 1-6 Alkyl sulfonyl, -CN, -NO2, -NH2, and -OH; L has the structure of L 1 -L 2 -L 3 L 1 L 2 L 3 may be present simultaneously or one or both of them is present; wherein, in L 1 and / or L 3 is present, it is independently selected from the following structures: and wherein n = 0-20, preferably 0-5, more preferably 0-2; R 15 selected from the group consisting of H, C 1-10 alkyl and C 3-10 cycloalkyl; R 16 selected from the group consisting of H, C 1-10 alkyl and C 3-10 cycloalkyl; In L 2 when present, is selected from the group consisting of: and wherein, m = 0-20, preferably 0-12. The compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer, enantiomer, diastereomer, atropisomer, racemic, or isotopically labeled compound thereof, wherein, The compound is selected from the group consisting of compounds of formula (II): wherein, Y1is selected from the group consisting of: hydrogen, deuterium, C 1-6 alkyl, and C 3-8 cycloalkyl; B ring is selected from: C 6-10 aromatic ring, C 3-10 saturated or unsaturated alicyclic, and 5-10 membered heteroaromatic ring; Y2is selected from the group consisting of: hydrogen, deuterium, halogen, C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, halogenated C 1-6 alkoxy, CN, NO2, and C 1-6 alkylsulfonyl; Y3is selected from: C 1-6 alkylene, -CH2NY 3a -, and -NY 3a -; wherein Y 3a is hydrogen, C 1-6 alkyl, halogenated C 1-6 alkyl, or C 3-8 cycloalkyl; Y4is selected from: C 1-6 alkyl, C 3-8 cycloalkyl, and haloC 1-6 alkyl; C ring is selected from: C 6-10 aromatic ring, C 3-10 cycloalkyl, and 5-10 membered heteroaromatic ring; Y5is selected from the group consisting of: hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 alkyl, and halogenated C 1-6 alkoxy; R1is selected from: substituted 5-10 membered heteroaryl, substituted C 6-10 aryl, and substituted 3-8 membered heterocyclyl; when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkyl, C 3-8 cycloalkyl, halogenated C 1-6 alkyl, halogenated C 1-6 alkyl, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, and -OH; R 3a and R 3b are each independently selected from the group consisting of hydrogen, substituted C 1-6 alkyl, substituted C 3-8 cycloalkyl, and substituted 3-8 membered heterocycloalkyl; and when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, and halogen; R5and R6are each independently selected from the group consisting of hydrogen, -OH, -SH, and substituted C 1-6 alkyl, or R5, R6and the carbon atom to which they are attached form a carbonyl group; and when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen; R 10 and R 11 are selected from the group consisting of hydrogen, substituted C 1-6 alkyl, substituted C 3-8 cycloalkyl, substituted C 1-6 alkoxy, substituted C 1-6 alkylthio, and substituted 5-10 membered heteroaryl; or R 10 , R 11 and the carbon atom to which they are attached form a substituted C 3-8 cycloalkyl; and when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 alkyl, halogenated C 1-6 alkoxy, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, and -OH; R 12 selected from: substituted 5-10 membered heteroaryl, substituted 5-10 membered heteroarylene, substituted C 6-10 arylene, and In the substituents, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 alkyl, halogenated C 1-6 alkoxy, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, and -OH; R 13 selected from the group consisting of hydrogen and substituted C 1-6 alkyl; when substituted, said substituent is selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkylsulfonyl, -CN, -NO2, -NH2, and -OH; L has the structure of L 1 -L 2 -L 3 L 1 L 2 L 3 may be present simultaneously or one or both of them is present; wherein, in L 1 and / or L 3 is independently selected from the following structures: and wherein n = 0-20, preferably 0-5, more preferably 0-2; R 15 selected from the group consisting of H, C 1-10 alkyl and C 3-10 cycloalkyl; In L 2 when present, is selected from the group consisting of: and wherein, m = 0-20, preferably 0-12. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt, stereoisomer, enantiomer, diastereomer, atropisomer, racemate, or isotopically labeled compound thereof, wherein, The compound is selected from the group consisting of compounds of formula (III): wherein, Y1is selected from: C 1-6 alkyl and C 3-8 cycloalkyl; B ring is selected from: C 6-10 aromatic ring, C 3-10 saturated or unsaturated alicyclic, and 5-10 membered heteroaromatic ring; Y2is selected from the group consisting of: hydrogen, deuterium, halogen, C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, and halogenated C 1-6 alkoxy; Y3is selected from: C 1-3 alkylene, -CH2NH-, and -NH-; Y5is selected from the group consisting of: hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 alkyl, and halogenated C 1-6 alkoxy; R1is selected from: wherein R is hydrogen or substituted C1-C6alkyl; when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, -CN, -NO2, -NH2, and -OH; 17 1-6 alkyl; when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, -CN, -NO2, -NH2, and -OH;​ R 3a and R 3b are each independently selected from the group consisting of hydrogen and substituted C 1-6 alkyl; and when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen; R 10 and R 11 is selected from the group consisting of hydrogen, substituted C 1-6 alkyl, and substituted C 3-8 cycloalkyl, or R 10 , R 11 and the carbon atom to which they are attached form a substituted C 3-8 cycloalkyl; and when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, halogen; R 12 selected from: substituted 5-10 membered heteroaryl, substituted C 6-10 arylene, and In the substituents, the substituents are selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 alkyl, and halogenated C 1-6 alkoxy; R 13 selected from: hydrogen and C 1-6 alkyl; L has the structure of L 1 -L 2 -L 3 L has the structure of L 1 L has the structure of L 2 L has the structure of L 3 L can be present simultaneously or one or both of L 1 and / or L 3 are independently selected from the following structures: and wherein n = 0-20, preferably 0-5, more preferably 0-2; R 15 selected from: H, C 1-10 alkyl and C 3-10 cycloalkyl; In L 2 when present, is selected from: and wherein, m = 0-20, preferably 0-12. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt, stereoisomer, enantiomer, diastereomer, atropisomer, racemate, or isotopically labeled compound thereof, wherein, The compound is selected from the group consisting of compounds of formula (IV): wherein, Y2and Y5are each selected from the group consisting of: hydrogen, deuterium, halogen, C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, and halogenated C 1-6 alkoxy; R 3a and R 3b are each independently selected from the group consisting of hydrogen and substituted C 1-6 alkyl; and when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, and halogen; R 10 and R 11 is selected from the group consisting of hydrogen, substituted C 1-6 alkyl, and substituted C 3-8 cycloalkyl; and when substituted, the substituents are selected from the group consisting of hydrogen, deuterium, and halogen; R 12 selected from: substituted 5-6 membered heteroaryl and In the substituents, the substituents are selected from hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 alkyl, and halogenated C 1-6 alkoxy; R 13 selected from: hydrogen or C 1-6 alkyl; L has the structure of L 1 -L 2 -L 3 L 1 L 2 L 3 may be present simultaneously or one or both of them is present; wherein, in L 1 and / or L 3 is present, it is independently selected from the following structures: and wherein n = 0-20, preferably 0-5, more preferably 0-2; R 15 selected from the group consisting of H, C 1-10 alkyl and C 3-10 cycloalkyl; In L 2 when present, is selected from: and wherein, m = 0-20, preferably 0-12. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt, stereoisomer, enantiomer, diastereomer, atropisomer, racemate, or isotopically labeled compound thereof, wherein, The compound is selected from the group consisting of compounds of formula (V): wherein, Y1is selected from: C 1-6 alkyl; Y2is selected from: hydrogen, halogen, C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, halogenated C 1-6 alkoxy; Y5is selected from: hydrogen, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 alkyl, halogenated C 1-6 alkoxy; the A3 moiety is selected from: said L is the same as defined in claim 4. The compound according to claim 5, or a pharmaceutically acceptable salt, stereoisomer, enantiomer, diastereomer, atropisomer, racemate, or isotopically-labeled compound thereof, wherein, said A3, Y1, Y2and Y5are the same as defined in claim 5, L is selected from: wherein m = 1-10. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt, stereoisomer, enantiomer, diastereomer, atropisomer, racemate, or isotopically labeled compound thereof, wherein, The compound is selected from: A pharmaceutical composition comprising one or more compounds selected from the group consisting of a compound according to any one of claims 1-7, a pharmaceutically acceptable salt, stereoisomer, enantiomer, diastereomer, atropisomer, optical isomer, racemate, polymorph, solvate, and isotopically-labeled compound thereof, and optionally a pharmaceutically acceptable carrier or excipient. Use of a compound according to any one of claims 1-7, or a pharmaceutically acceptable salt, stereoisomer, enantiomer, diastereomer, atropisomer, racemate, polymorph, solvate, or isotopically-labeled compound thereof, in the manufacture of a medicament for the prevention and / or treatment of a disease or disorder or disease state mediated by USP7. The use of claim 9, wherein the disease or disorder or disease state mediated by USP7 comprises a tumor, a hemangiosarcoma, a leukemia, an ovarian cancer, a breast cancer, a lung cancer, a pancreatic cancer, a renal cancer, a melanoma, a liver cancer, a colon cancer, a sarcoma, a brain cancer, a prostate cancer, a lymphoma, a multiple myeloma, hyperglycemia, diabetes, obesity, hyperlipidemia, hypercholesterolemia, hyperlipoproteinemia, hypertriglyceridemia, hypertension, hyperinsulinemia, hyperuricemia, Parkinson's disease, and / or Alzheimer's disease.

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