Cyclic BRD4 protein degradation agent and application thereof

Abstract

The invention provides a BRD4 protein degradation agent and application thereof, and particularly provides a compound with a structure as shown in a formula (I) or pharmaceutically acceptable salt thereof, or a stereoisomer or a prodrug molecule thereof. The compound can degrade BRD4 protein in a targeted manner through a ubiquitin-proteasome way, so that the compound can be used for treating indications mediated by abnormal expression of the BRD4 protein. Wherein each group in the formula (I) is defined in the specification.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of pharmaceutical chemistry, in particular to a class of cyclic compounds targeting ubiquitination degradation of BRD4 protein and pharmaceutical compositions and uses thereof. BACKGROUND

[0002] Bromodomain family proteins contain 4 members, respectively BRD2, BRD3, BRD4 and BRDT. BET family proteins recognize acetylated chromatin through their bromodomains and are involved in regulating gene expression. As one of the BET family members, BRD4 contains two bromodomains, BD1 and BD2. BDs are chromatin "readers": by interacting with acetylated lysines in the histone tail, they recruit chromatin-modulating proteins at the promoter region to regulate gene expression and repression. Bromodomain-containing protein 4 (BRD4) is a chromatin reading protein that can recognize and bind to acetylated lysine residues in histones and plays a key role in the epigenetic memory transmission of cell division and transcriptional regulation. BRD4 has become a potential therapeutic target for various human malignancies, including various solid tumors and hematological malignancies.

[0003] Target protein degradation technology is currently widely used in drug research and development and is considered a major breakthrough in the field of small molecule drugs. Among them, the research of protein-targeted chimeric degradation (PROTAC) is the most mature. PROTAC molecular structure is composed of target protein recognition ligand, linker and ubiquitination E3 ligase recognition ligand. PROTAC can effectively induce ubiquitination labeling of target proteins and effectively degrade them through the proteasome pathway. Compared with traditional small molecule inhibitors, PROTAC molecules have the advantages of small dosage, less drug resistance, high safety, etc. At present, a number of PROTAC molecules have entered clinical research. SUMMARY

[0004] The present application provides a class of BRD4 protein degrading agents and uses thereof. Specifically, the present application provides a compound having a structure shown in formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer or prodrug molecule thereof. The compound can target the degradation of BRD4 protein through the ubiquitin-proteasome pathway, and thus can be used for treating indications mediated by abnormal expression of BRD4 protein.

[0005] In the first aspect of the present application, a compound as shown in formula (I) or a stereoisomer thereof is provided,

[0006]

[0007] wherein,

[0008] L 1, L 2 each independently selected from a structure having a formula:

[0009]

[0010] W1, W2, W3, W4, W5are each independently selected from the group consisting of -O-, -S-, -NH-, -CH2-, -CONH-, -NHCO-, -C≡C-, -CH=CH-, -C(O)-, -P(=O)-, -S(O)2-, -S(O)-, -P(O)2(OH)-, -NH-S(O)-NH-, -C(O)O-, -OC(O)-, -NHCONH-, 3-12 membered ring having 0-4 heteroatoms; n1, n2, n3, n4are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10; wherein the 3-12 membered ring can be monocyclic, spirocyclic, annular or bridged structure, and can each independently be optionally substituted with one or more substituents selected from oxo, halogen, C1-6alkyl, C1-6alkoxy and C1-6haloalkyl.

[0011] R 1 is selected from the group consisting of no substitution or the following groups: D, CN, NO2, halogen, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C3-C6cycloalkyl, 3-12 membered heterocyclyl containing 1-4 heteroatoms selected from the group consisting of oxygen, sulfur and nitrogen, aryl, 5-14 membered heteroaryl containing 1-5 heteroatoms selected from the group consisting of oxygen, sulfur and nitrogen;

[0012] B is selected from the following formula:

[0013]

[0014] wherein R 2 may be mono- or polysubstituted, each R 2 is the same or different, independently of one another, selected from the group consisting of H, OH, CN, NO2, halogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C6cycloalkyl, 5-14 membered heteroaryl containing 1-5 heteroatoms selected from the group consisting of oxygen, sulfur and nitrogen or 5-14 membered heteroaryl containing 1-5 heteroatoms selected from the group consisting of oxygen, sulfur and nitrogen optionally substituted by one, two or more R 3 is the same or different, independently of one another, selected from the group consisting of H, OH, CN, NO2, halogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C6cycloalkyl, 5-14 membered heteroaryl containing 1-5 heteroatoms selected from the group consisting of oxygen, sulfur and nitrogen or 5-14 membered heteroaryl containing 1-5 heteroatoms selected from the group consisting of oxygen, sulfur and nitrogen optionally substituted by one, two or more R 3 is selected from the group consisting of halogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C6cycloalkyl, halogen mono-, di- or polysubstituted C1-C6alkyl;

[0015] In some embodiments, R 1selected from the group consisting of C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C1-C6alkoxy, C3-C6cycloalkyl, 3- to 12-membered heterocyclyl containing from 1 to 4 heteroatoms selected from oxygen, sulfur, and nitrogen, aryl, 5- to 14-membered heteroaryl containing from 1 to 5 heteroatoms selected from oxygen, sulfur, and nitrogen;

[0016] Preferably, R 1 is selected from the group consisting of H, methyl, ethyl, isopropyl, tert-butyl, cyclopropyl.

[0017] Preferably, R 1 is tert-butyl.

[0018] In some embodiments, L 1 , L 2 has a structure selected from the group consisting of:

[0019]

[0020] wherein each of W1, W2, W3, W4, W5is independently selected from the group consisting of -O-, -S-, -NH-, -CH2-, -CONH-, -NHCO-, -C(O)-, -C(O)O-, -OC(O)-, -NHCONH-, 3- to 10-membered ring having from 0 to 4 heteroatoms; each of n1, n2, n3, n4is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10.

[0021] In some embodiments, L 1 , L 2 has a structure selected from the group consisting of:

[0022]

[0023]

[0024] wherein each X and Y is independently selected from the group consisting of -O-, -S-, -CO-, -NH-, -CONH-, -NHCO-, -C(O)O-, and -OC(O)-; each n and m is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0025] In some embodiments, B is selected from the group consisting of:

[0026]

[0027] Preferably, R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10the same or different, are each independently selected from the group consisting of H, Cl, F, Br;

[0028] Preferably, R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 is H or F.

[0029] Preferably, R 2 is selected from a 5-14 membered heteroaryl containing 1-5 heteroatoms selected from oxygen, sulfur and nitrogen, or a 5-14 membered heteroaryl optionally substituted with one, two or more R 3 substituted with one, two or more R

[0030] Preferably, R 3 are the same or different, and are each independently selected from the group consisting of C1-C6 alkyl, C3-C6 cycloalkyl, halogen mono-, di- or poly-substituted C1-C6 alkyl;

[0031] Preferably, B is selected from:

[0032]

[0033]

[0034] In some embodiments, the compound is selected from the following structures:

[0035]

[0036]

[0037]

[0038]

[0039]

[0040]

[0041]

[0042]

[0043] In a second aspect of the present application, there is provided a pharmaceutical composition for treating and / or preventing a tumor, comprising:

[0044] (1) a compound, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, or a prodrug molecule thereof as described in the first aspect of the present application;

[0045] optionally (2) a pharmaceutically acceptable carrier.

[0046] In a third aspect of the present application, there is provided a use of a compound of the first aspect of the present application, a pharmaceutically acceptable salt thereof, a stereoisomer thereof or a prodrug molecule thereof, or a pharmaceutical composition of the second aspect of the present application, for the manufacture of a medicament for treating and / or preventing a disease or disorder associated with abnormal expression of BRD4.

[0047] In another preferred embodiment, the disease or disorder is selected from the group consisting of a tumor, an autoimmune disease, a viral infection, a neurodegenerative disease.

[0048] In some embodiments, the disease or disorder is selected from the group consisting of a hematological tumor, a gastrointestinal stromal tumor, a histiocytic lymphoma, a non-small cell lung cancer, a small cell lung cancer, a lung adenocarcinoma, a lung squamous carcinoma, a pancreatic cancer, a breast cancer, a prostate cancer, a liver cancer, a skin cancer, an epithelial cell cancer, a colorectal cancer, a kidney cancer, a stomach cancer, a head and neck cancer, or a nasopharyngeal cancer.

[0049] In a fourth aspect of the present application, there is provided a method of degrading BRD4 in a subject in need thereof, by administering to the subject a compound of the first aspect of the present application, a pharmaceutically acceptable salt thereof, a stereoisomer thereof or a prodrug molecule thereof, or a pharmaceutical composition of the second aspect of the present application.

[0050] In another preferred embodiment, the subject is a human.

[0051] It should be understood that, within the scope of the present application, the above technical features of the present application and the technical features specifically described hereinafter (e.g. in the examples) can be combined with each other to form new or preferred technical solutions. Due to the limited space, they are not listed one by one here.

[0052] Definitions of terms and explanations

[0053] Unless otherwise specified, the definitions of groups and terms in the present application specification and claims, including the definitions of examples, exemplary definitions, preferred definitions, definitions in tables, definitions of specific compounds in examples, etc., can be combined and integrated with each other in any manner. The group definitions and compound structures after such combination and integration should be understood as within the scope of the present application specification and / or claims.

[0054] In the compounds of the present application, when any variable (e.g. R 1 , R 2If more than one occurrence of a component (e.g., R1, R2, etc.) is present in any group, then each occurrence of that component is defined independently of the other occurrences. Also, combinations of substituents and variables are permissible only if such combinations result in chemically sensible compounds. A line drawn to a ring system from a substituent denotes that the bond can be attached to any available carbon atom of the ring system. If the ring system is polycyclic, it means that the bond is attached only to any appropriate carbon atom of the adjacent ring. It is understood that one of ordinary skill in the art can select substituents and substitution patterns for the compounds of the application that result in chemically stable and synthetically readily accessible compounds from readily available starting materials, using techniques known in the art and methods presented below. If a substituent is itself substituted with more than one group, it is understood that the groups can be on the same carbon atom or on different carbon atoms, as long as the structure is stable.

[0055] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

[0056] The following terms are used to describe the present application. In instances in which a term is not specifically defined herein, the person of ordinary skill in the art with the context in which the term is used to describe the present application applies the recognized meaning of the term in the art.

[0057] The term "ubiquitin ligase" as used herein refers to a family of proteins that facilitate the transfer of ubiquitin to a particular substrate protein, thereby targeting the substrate protein for degradation. For example, the Von Hippel-Lindau E3 ubiquitin ligase is a protein that, alone or in combination with an E2 ubiquitin conjugating enzyme, results in the attachment of ubiquitin to a lysine on a target protein and subsequent targeting of the particular protein substrate for degradation by the proteasome. Thus, the E3 ubiquitin ligase, alone or in complex with an E2 ubiquitin conjugating enzyme, is responsible for the transfer of ubiquitin to the target protein. In general, the ubiquitin ligase is involved in polyubiquitination, whereby a second ubiquitin is attached to the first ubiquitin; a third ubiquitin is attached to the second ubiquitin, and so on. Polyubiquitination marks the protein for degradation by the proteasome. However, there are some ubiquitination events that are limited to mono-ubiquitination, in which only a single ubiquitin is added to the substrate molecule by the ubiquitin ligase. Mono-ubiquitinated proteins are not targeted to the proteasome for degradation, but rather can be altered in their cellular location or function, for example, by binding to other proteins that have domains capable of binding ubiquitin. Adding further complexity, different lysines on the ubiquitin can be targeted by the E3 to form a chain. The most common lysine is Lys48 on the ubiquitin chain. This is the lysine that is used to make the polyubiquitin that is recognized by the proteasome.

[0058] It is to be understood that in describing one, two, or more items, "more" shall mean greater than 2, e.g., an integer greater than or equal to 3, such as 3, 4, 5, 6, 7, 8, 9, or 10.

[0059] The term "optional" (or "optionally", "option") means the occurrence of the situation in which zero, one or more substituents are present, e.g., "optionally substituted with one, two or more R" means that there can be no substitution (no substituents) or there can be one, two or more substitutions (one, two or more substituents).

[0060] The term "halogen" denotes fluorine, chlorine, bromine and iodine.

[0061] The term "C 1- C 12 "Alkyl" is understood to mean a straight-chain or branched saturated monovalent hydrocarbon radical having from 1 to 12 carbon atoms. For example, "C 1- C 10 "Alkyl" means a straight-chain and branched alkyl group having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, "C 1- "C8-alkyl" means a straight-chain and branched alkyl group having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms, "C 1- "C6-alkyl" means a straight-chain and branched alkyl group having 1, 2, 3, 4, 5 or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1 -methylbutyl, 1 -ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1 -dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1 -methylpentyl, 2-ethylbutyl, 1 -ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1 -dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl and the like or isomers thereof.

[0062] The term "C 2- "C6-alkenyl" is understood to mean preferably a straight-chain or branched monovalent hydrocarbon group which contains one or more double bonds and has 2 to 6 carbon atoms. For example, having 2 or 3 carbon atoms (i.e., C 2-C3alkenyl). It is to be understood that where said alkenyl group contains more than one double bond, said double bonds can be separated from one another or conjugated. Said alkenyl group is, for example, ethenyl, allyl, (E)-2-methylethenyl, (Z)-2-methylethenyl, (E)-but-2-enyl, (Z)-but-2-enyl, (E)-but-l-enyl, (Z)-but-l-enyl, pent-4-enyl, (E)-pent-3-enyl, (Z)-pent-3-enyl, (E)-pent-2-enyl, (Z)-pent-2-enyl, (E)-pent-l-enyl, (Z)-pent-l-enyl, hex-5-enyl, (E)-hex-4-enyl, (Z)-hex-4-enyl, (E)-hex-3-enyl, (Z)-hex-3-enyl, (E)-hex-2-enyl, (Z)-hex-2-enyl, (E)-hex-l-enyl, (Z)-hex-l-enyl, isopropenyl, 2-methylprop-2-enyl, l-methylprop-2-enyl, 2-methylprop-l-enyl, (E)-l-methylprop-l-enyl, (Z)-l-methylprop-l-enyl, 3-methylbut-3-enyl, 2-methylbut-3-enyl, l-methylbut-3-enyl, 3-methylbut-2-enyl, (E)-2-methylbut-2-enyl, (Z)-2-methylbut-2-enyl, (E)-l-methylbut-2-enyl, (Z)-l-methylbut-2-enyl, (E)-3-methylbut-l-enyl, (Z)-3-methylbut-l-enyl, (E)-2-methylbut-l-enyl, (Z)-2-methylbut-l-enyl, (E)-l-methylbut-l-enyl, (Z)-l-methylbut-l-enyl, 1,1-dimethylprop-2-enyl, 1-ethylprop-l-enyl, 1-propylvinyl, 1-isopropylvinyl.

[0063] The term "C 2- C6alkynyl" is to be understood as meaning a straight-chain or branched one- valent hydrocarbon group which contains one or more triple bonds and which has 2 to 6 carbon atoms. For example, a "C2-C6alkynyl" group has 2, 3, 4, 5 or 6 carbon atoms. Said alkynyl group is, for example, ethynyl, propynyl, (E)-but-2-ynyl, (Z)-but-2-ynyl, (E)-pent-2-ynyl, (Z)-pent-2-ynyl, (E)-hex-2-ynyl, (Z)-hex-2-ynyl, (E)-hex-3-ynyl, (Z)-hex-3-ynyl, (E)-hex-4-ynyl, (Z)-hex-4-ynyl, (E)-hex-5-ynyl, (Z)-hex-5-ynyl, 1-methylprop-2-ynyl, 2-methylprop-2-ynyl, 1-ethylprop-2-ynyl, 1-propylvinyl, 1-isopropylvinyl, 1,1-dimethylprop-2-ynyl, 1-ethylprop-2-ynyl, 1-propylvinyl, 1-isopropylvinyl. 2-(C3 ynyl group). The ynyl group is, for example, ethynyl, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, but-2-ynyl, but-3-ynyl, pent-1-ynyl, pent-2-ynyl, pent-3-ynyl, pent-4-ynyl, hex-1-ynyl, hex-2-ynyl, hex-3-ynyl, hex-4-ynyl, hex-5-ynyl, 1-methylprop-2-ynyl, 2-methylbut-3-ynyl, 1-methylbut-3-ynyl, 1-methylbut-2-ynyl, 3-methylbut-1-ynyl, 1-ethylprop-2-ynyl, 3-methylpent-4-ynyl, 2-methylpent-4-ynyl, 1-methylpent- 4-Alynyl, 2-methylpentan-3-alkynyl, 1-methylpentan-3-alkynyl, 4-methylpentan-2-alkynyl, 1-methylpentan-2-alkynyl, 4-methylpentan-1-alkynyl, 3-methylpentan-1-alkynyl, 2-ethylbutan-3-alkynyl, 1-ethylbutan-3-alkynyl, 1-ethylbutan-2-alkynyl, 1-propylpropan-2-alkynyl, 1-isopropylpropan-2-alkynyl, 2,2-dimethylbutan-3-alkynyl, 1,1-dimethylbutan-3-alkynyl, 1,1-dimethylbutan-2-alkynyl, or 3,3-dimethylbutan-1-alkynyl. In particular, the alkynyl group is ethynyl, propan-1-alkynyl, or propan-2-alkynyl.

[0064] Term "C" 3- C 12 "Cycloalkyl" should be understood to refer to saturated monovalent monocyclic, bicyclic (such as fused ring, bridged ring, spiro ring) hydrocarbon rings or tricyclic alkanes, having 3 to 12 carbon atoms, preferably "C". 3- C 10 "Cycloalkyl", more preferably "C" 3- C8 cycloalkyl. The term "C8 cycloalkyl" 3- C 10 "Cycloalkyl" should be understood to refer to a saturated monovalent monocyclic, bicyclic (e.g., bridged, spirocyclic) hydrocarbon ring or tricyclic alkane having 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. The C... 3- C 10 Cycloalkyl groups can be monocyclic hydrocarbon groups, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or cyclodecyl; or bicyclic hydrocarbon groups, such as borneolyl, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, 6,6-dimethylbicyclo[3.1.1]heptyl, 2,6,6-trimethylbicyclo[3.1.1]heptyl, bicyclo[2.2.2]octyl, 2,7-diazaspiro[3,5]nonyl, 2,6-diazaspiro[3,4]octyl; or tricyclic hydrocarbon groups, such as adamantyl. For example: wait.

[0065] Term "C" 3- C12 "Cyclic hydrocarbon group" contains saturated C 3- C 12 cycloalkyl groups and unsaturated C 3- C 12 Cycloalkenyl, C 3- C 12 Cycloalkynyl, for example wait

[0066] Term "C" 1- C 12 "Alkoxy" refers to a compound with a carbonyl group. 1- C 12 Alkyl-O- groups, preferably "C" 1- C6 alkoxy", such as -OCH3, -OCH2CH3, -OCH2CH2CH3, -O-CH2CH(CH3)2, -OCH2CH2CH2CH3, -O-CH(CH3)2, etc.

[0067] The term "3-12 membered heterocyclic group" refers to a saturated or partially unsaturated monocyclic, bicyclic, or polycyclic cyclic substituent, wherein one or more ring atoms are selected from heteroatoms of N, O, or S(O)m (where m is an integer from 0 to 2), and the remaining ring atoms are carbon. Bicyclic or polycyclic groups include spirocyclic, fused-ring, and bridged-ring groups. Examples include: morpholinyl, piperidinyl, tetrahydropyrrolyl, pyrrolylalkyl, dihydroimidazolyl, dihydroisoxazolyl, dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl, dihydropyrazinyl, dihydropyrazoleyl, dihydropyridinyl, dihydropyrimidinyl, dihydropyrrolyl, dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl, dihydrothiopheneyl, dihydrotriazolyl, dihydroazacyclobutane, tetrahydrofuranyl, tetrahydrothiopheneyl, etc. And so on, and their N-oxides. The connection of heterocyclic substituents can be achieved through carbon atoms or through heteroatoms.

[0068] Term "C" 6- C 14 "Aryl" should preferably be understood to represent a monocyclic, bicyclic (such as fused ring, bridged ring, spiro ring), or tricyclic hydrocarbon ring having 6 to 14 carbon atoms and possessing monovalent aromaticity or partial aromaticity. It can be a monoaromatic ring or a polyaromatic ring fused together, preferably "C". 6- C 10 "Aromatic". The term "C" 6- C 10 "Aryl" should be understood to preferably represent a monovalent aromatic or partially aromatic monocyclic, bicyclic, or tricyclic hydrocarbon ring having 6, 7, 8, 9, or 10 carbon atoms, particularly a ring having 6 carbon atoms ("C6 aryl"), such as phenyl; or biphenyl, or a ring having 9 carbon atoms ("C9 aryl"), such as indenyl or indenyl, or a ring having 10 carbon atoms ("C9 aryl"). 10aryl") having 13 carbon atoms, e.g. tetrahydronaphthyl, dihydronaphthyl or naphthyl, or a ring ("C 13 aryl") having 13 carbon atoms, e.g. tetrahydronaphthyl, dihydronaphthyl or naphthyl, or a ring ("C 14 aryl") having 13 carbon atoms, e.g. tetrahydronaphthyl, dihydronaphthyl or naphthyl, or a ring ("C 6-20 When the aryl group is substituted, it can be mono- or polysubstituted. Also, the substitution sites thereof are not limited, and can be, for example, ortho-, para- or meta-substitution.

[0069] The term "5-14 membered heteroaryl" is to be understood as including a monovalent monocyclic, bicyclic (e.g. fused, bridged, spirocyclic) or tricyclic aromatic ring system having 5 to 14 ring atoms and comprising 1 to 5 heteroatoms independently selected from N, O and S, e.g. "5-10 membered heteroaryl". The term "5-10 membered heteroaryl" is to be understood as including a monovalent monocyclic, bicyclic or tricyclic aromatic ring system having 5, 6, 7, 8, 9 or 10 ring atoms, in particular 5 or 6 or 9 or 10 carbon atoms, and comprising 1 to 5, preferably 1 to 3, heteroatoms each independently selected from N, O and S and, additionally in each case, can be benzo-fused. "Heteroaryl" also refers to radicals in which the heteroaromatic ring is fused to one or more aryl, alicyclic or heterocyclyl rings, wherein the point of attachment is on the heteroaromatic ring. Non-limiting examples include indolizinyl, isoindolyl, indolyl, indazolyl, purinyl, quinolizinyl, quinolyl, isoquinolyl, phthalazinyl, naphthyridinyl, quinazolinyl, cinnolinyl, pteridinyl, carbazolyl, carbolinyl, phenanthridinyl, acridinyl, phenanthrolinyl, phenoxazinyl, phenothiazinyl, phenazinyl, benzoisoquinolyl, thieno[2,3-b]furanyl, pyrazino[2,3-c]carbazolyl, furano[3,2-b]-pyranyl, pyrido[2,3-d]-oxazinyl, pyrazolo[4,3-d]-oxazolyl, imidazo[4,5-d]thiazolyl, pyrazino[2,3-d]pyridazinyl, imidazo[2,1-b]thiazolyl, furano[3,4-c]cinnolinyl, pyrido[2,3-c]carbazolyl, imidazo[1,2-b][1,2,4]triazinyl, benzo[b]thiophenyl, benzimidazolyl, benzothiazolyl, benzoxapinyl, benzoxazinyl, pyrrolo[1,2-b][2]benzazapinyl.

[0070] The term "spirocyclic" refers to a ring system in which two rings share one ring atom, which can contain aliphatic, heterocyclic, aromatic or heteroaromatic rings as described above.

[0071] The term "fused ring" refers to a ring system in which two rings share two ring atoms, which can contain aliphatic, heterocyclic, aromatic or heteroaromatic rings as described above.

[0072] The term "bridged ring" refers to ring systems in which two rings share 3 or more ring atoms, which can contain aliphatic, heteroaliphatic, aromatic, or heteroaromatic rings as previously described.

[0073] pharmaceutically acceptable salt

[0074] The present application includes the free form of the compounds of Formula I, as well as pharmaceutically acceptable salts and stereoisomers thereof. Some of the specific exemplary compounds herein are protonated salts of amines. The term "free form" refers to the amine in non-salt form. The pharmaceutically acceptable salts which are included are not only those salts which are specifically mentioned herein, but also all typical pharmaceutically acceptable salts of the free form of the compounds of Formula I. The free form of a particular salt of a compound can be isolated using techniques known in the art. For example, the free form can be regenerated by treating the salt with a dilute aqueous solution of the appropriate base, such as NaOH, KOH, ammonium hydroxide, and NaHC03. The free form will often differ from the salt form useful for the purposes of the present application in some physical property, such as solubility in some solvent, but otherwise the two forms are equivalent for the purposes of the present application.

[0075] The pharmaceutically acceptable salts of the present application can be synthesized from the compounds of the present application which contain a basic or acidic moiety by conventional chemical methods. Generally, the salt will be prepared by dissolving the free base in a suitable solvent, such as water, ethanol or methanol, and passing a solution of the desired salt through the solution of the free base. The salts can be precipitated from solution by adding a solution of the free base in a suitable solvent, such as ether or hexane, to a solution of the desired salt in the same solvent. The salts can also be prepared by dissolving the free base in a suitable solvent, such as water or ethanol, and adding to the solution an excess of the desired salt, either in solid form or in a suitable solvent, such as ether or hexane.

[0076] Thus, pharmaceutically acceptable salts of the compounds of this application include the conventional nontoxic salts of the compounds of the present application as formed by reaction with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid and the like, and organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, p-aminobenzoic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethane disulfonic acid, oxalic acid, isethionic acid, trifluoroacetic acid and the like.

[0077] If the compound of the present application is acidic, suitable "pharmaceutically acceptable salts" refers to salts of pharmaceutically acceptable, non-toxic bases including inorganic bases and organic bases. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic, manganous, potassium, sodium, zinc, and the like. Particularly preferred are the ammonium, calcium, magnesium, potassium, and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethyldiamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydroxycobalamin, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, purines, procaine, pyrimidines, triethylamine, trimethylamine, tripropylamine, trolamine, and the like.

[0078] The preparation of the above-mentioned pharmaceutically acceptable salts and other typical pharmaceutically acceptable salts is described in more detail in Berg et al., "Pharmaceutical Salts," J. Pharm. Sci. 1977: 66: 1-19.

[0079] Pharmaceutical compositions and uses thereof

[0080] The present application also provides a pharmaceutical composition comprising a safe and effective amount of the active ingredient, and a pharmaceutically acceptable carrier or excipient. Since the active ingredient of the present application, the compound of Formula I and its pharmaceutically acceptable salts have a good inhibitory effect on the abnormal expression of BRD4 protein, the pharmaceutical composition can be used for treating cancer, autoimmune diseases, viral infections, neurodegenerative diseases and the like related to the abnormal expression of BRD4 protein in humans or other mammals.

[0081] The present application provides a class of protein kinase degraders and uses thereof, in particular, the present application provides a compound having a structure shown in Formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer or a prodrug molecule thereof. The compound can degrade BRD4 protein through the ubiquitination pathway, and can be used for treating BRD4 abnormal expression related indications.

[0082] In one embodiment, the active ingredient of the present application, or the pharmaceutical composition containing the active ingredient can be used for preventing and / or treating prostate cancer, non-small cell lung cancer, malignant melanoma, renal cancer, bladder cancer, ovarian cancer, colon cancer, rectal cancer, breast cancer, cervical cancer, lung cancer, laryngeal cancer, nasopharyngeal cancer, pancreatic cancer, multiple myeloma, B lymphoma, leukemia and the like tumors, or for preventing tumor recurrence after surgery.

[0083] The "active ingredient" according to the present application refers to the compound of Formula I according to the present application or a pharmaceutically acceptable salt thereof or a stereoisomer thereof or a prodrug thereof.

[0084] The "active ingredient" and the pharmaceutical composition according to the present application can be used as a BRD4 protein degrading agent and can be used for the preparation of a medicament for the prevention and / or treatment of cancer, autoimmune diseases, viral infection, neurodegenerative diseases, etc.

[0085] The "safe and effective amount" refers to an amount of the active ingredient sufficient to significantly improve the condition without causing serious side effects. Generally, the pharmaceutical composition contains 1-2000 mg of the active ingredient per dose, more preferably, 10-200 mg of the active ingredient per dose. Preferably, the "one dose" is one tablet.

[0086] The "pharmaceutically acceptable carrier or excipient" refers to one or more compatible solid or liquid filler or gel materials that are suitable for human use and must have sufficient purity and sufficiently low toxicity.

[0087] "Compatibility" here refers to the ability of the components of the composition to be incorporated with the active ingredient of the present application and to each other without significantly diminishing the efficacy of the active ingredient.

[0088] Examples of the pharmaceutically acceptable carrier or excipient include cellulose and its derivatives (e.g., sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g., stearic acid, magnesium stearate), calcium sulfate, vegetable oils (e.g., soybean oil, sesame oil, peanut oil, olive oil, etc.), polyhydric alcohols (e.g., propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers wetting agents (e.g., sodium dodecyl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0089] In another preferred embodiment, the compound of Formula I according to the present application can form a complex with a macromolecular compound or a polymer through non-covalent interaction. In another preferred embodiment, the compound of Formula I according to the present application as a small molecule can also be linked to a macromolecular compound or a polymer through a chemical bond. The macromolecular compound can be a biological macromolecule such as a polysaccharide, a protein, a nucleic acid, a polypeptide, etc.

[0090] The administration of the active ingredient or the pharmaceutical composition according to the present application is not particularly limited, and representative administration methods include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular, or subcutaneous), etc.

[0091] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.

[0092] In solid dosage forms of the invention, the active drug compound is admixed with at least one inert pharmaceutically acceptable excipient or carrier (or both) such as sodium citrate or dicalcium phosphate and / or a) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and silicic acids;

[0093] (a) fillers or bulking agents, such as starch, lactose, sucrose, glucose, mannitol, and silicic acid;

[0094] (b) binders, such as hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia;

[0095] (c) humectants, such as glycerol;

[0096] (d) disintegrants, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate;

[0097] (e) agents for retarding dissolution, such as paraffin;

[0098] (f) absorption accelerators, such as quaternary ammonium compounds;

[0099] (g) agents for retarding dissolution, such as paraffin;

[0100] (h) adsorbents, such as kaolin;

[0101] (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets, and pills, the dosage forms can also comprise buffering agents.

[0102] Solid dosage forms of the invention can also be prepared with coatings and shells, such as enteric coatings and other coatings known in the art. They can contain opacifying agents, and can be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the digestive tract. Examples of embedding compositions that can be used are polymeric substances and waxes.

[0103] Liquid dosage forms for oral administration of the compounds of the invention include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient, the liquid dosage forms can contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and the like, either with or without the addition of

[0104] In addition to the active ingredient, the suspensions can contain suspending agents, as for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide gel, and agar-agar, or mixtures of these substances, and the like.

[0105] The compositions for parenteral injection can contain physiologically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols and suitable mixtures thereof.

[0106] The compounds of the present application can be administered alone or in combination with other therapeutic agents.

[0107] The pharmaceutical compositions are administered in a safe and effective amount, which is an amount that is sufficient to treat the condition for which it is administered, but low enough to avoid serious side effects. The specific dose level and frequency of dosage will vary depending on the nature of the condition being treated, the age, weight and general health condition of the patient, and other factors known to those skilled in the art. In general, the dose per day for an average-sized adult human is in the range of 1 to 2000 mg, preferably 20 to 500 mg, of the compound of the present application. Of course, the specific dose will depend on the route of administration as well as the patient's needs. DETAILED DESCRIPTION

[0108] The technical solutions of the present application will be further described in detail below in combination with specific examples. It should be understood that the following examples are only illustrative and explanatory of the present application, and should not be interpreted as limiting the scope of protection of the present application. Any technology realized based on the above description of the present application is covered within the scope of the present application.

[0109] Unless otherwise indicated, the starting materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0110] Example 1: Synthesis of intermediates 1-7

[0111]

[0112] Step 1:

[0113] In a round bottom flask with stir bar, (2S,4R)-4-hydroxypyrrolidine-2-carboxylic acid methyl ester hydrochloride (10 g, 55.2 mmol), (S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanoic acid (12.75 g, 55.2 mmol) were dissolved in DMF (80 mL), then DIEA (30 mL, 172 mmol) was added, followed by transfer to an ice bath at 0 °C, HATU (22 g, 58 mmol) was added, then the reaction was allowed to warm to room temperature. LC-MS monitoring showed the reaction was complete, 200 mL water and 200 mL ethyl acetate were added and extracted 3 times, the organic layer was washed with 5% citric acid solution once, then saturated sodium bicarbonate solution once, and finally water once. Spinning to dryness gave 17.8 g of compound 1-2 crude product. LC-MS: (M+H) + : 359.2.

[0114] Step 2:

[0115] In a round bottom flask with stir bar, compound 1-2 (15 g, 41.9 mmol) from Step 1 was dissolved in THF / H2O (2:1) 120 mL, then lithium hydroxide monohydrate (5 g, 125 mmol) was added and the reaction was allowed to warm to room temperature. LC-MS monitoring showed the reaction was complete, the THF was spun to dryness, the pH was adjusted to weakly acidic, and a white solid precipitated, which was filtered to give 13.8 g of compound 1-3 as a white solid. LC-MS: (M+H) + : 345.1.

[0116] Step 3:

[0117] In a round bottom flask with stir bar, 4-bromo-2-hydroxybenzaldehyde (10 g, 49.7 mmol), tert-butyl carbamate (8.74 g, 74.6 mmol) were dissolved in dichloromethane / acetonitrile (3:1) (120 mL), then Et3SiH (24 mL, 149 mmol), TFA (8 mL, 99 mmol) were added and the reaction was allowed to warm to room temperature. LC-MS monitoring showed the reaction was complete, 400 mL saturated sodium bicarbonate solution was added to quench the reaction, and 200 mL ethyl acetate was added and extracted 3 times, the organic phase was spun to dryness and slurried in dichloromethane / petroleum ether. This gave 10.3 g of compound 1-5 crude product. LC-MS: (M+H) + : 303.2.

[0118] Step 4:

[0119] To a solution of compound 1-5 (10 g, 33.1 mmol) from Step 3 in a 250 mL Schleck tube, potassium acetate (6.49 g, 66.2 mmol), palladium acetate (300 mg, 13.2 mmol), 4-methylthiazole (6 mL, 66.2 mmol) were added, respectively. After three times of argon gas exchange, 60 mL DMF was added. The reaction was carried out at 90 °C overnight. LC-MS was used to monitor the reaction completion. 200 mL water was added and the mixture was extracted with 200 mL ethyl acetate for three times. The organic phase was dried and column chromatography was used to separate the product (eluent: petroleum ether / ethyl acetate = 1:1) to give compound 1-6 as a light yellow solid. LC-MS: (M+H) + : 321.4.

[0120] Step 5:

[0121] After deprotection of compound 1-6 (5 g, 15.6 mmol) from Step 4 with TFA / DCM, compound 1-3 (5.4 g, 15.7 mmol) from Step 2 was dissolved in DMF (30 mL). DIEA (28 mL, 156 mmol) was added, followed by moving the reaction to an ice bath at 0 °C. HATU (6 g, 17.2 mmol), HOAt (400 mg, 3 mmol) were added. The reaction was moved to room temperature. LC-MS was used to monitor the reaction completion. 100 mL water was added and the mixture was extracted with 100 mL ethyl acetate for three times. The organic phase was washed with 5% citric acid solution once, saturated sodium bicarbonate solution once and water once. The crude product was obtained after drying as compound 1-7, 8.7 g. LC-MS: (M+H) + : 547.3.

[0122] Example 2: Synthesis of compound 15a

[0123]

[0124] Step 1:

[0125] In a Schlenk tube with stir bar, (6S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H- thieno[3,2-F][l,2,4]thiazolo[4,3-A][l,4]diazepine-6-acetic acid (2-1) (500 mg, 1.3 mmol), KOH (280 mg, 5.0 mmol) and t-butyl BrettPhos (484 mg, 1.0 mmol) were dissolved in 1,4-dioxane / water (20 mL, v / v = 4 / 1) under nitrogen protection, then Pd2(dba)3(220 mg, 0.25 mmol) was added and the reaction was carried out at 100 °C. LC-MS was used to monitor the reaction completion, then 200 mL water and 200 mL ethyl acetate were added to extract 3 times, the organic phase was discarded, the water layer was adjusted to pH 4-5 with 2 M hydrochloric acid solution, and then dried and redissolved in dichloromethane / methanol (20:1). After drying, 410 mg of crude product compound 2-2 was obtained. LC-MS: (M+H) + : 383.2.

[0126] Step 2:

[0127] In a round-bottom reaction bottle with a stir bar, compound 2-2 (382 mg, 1 mmol) obtained in step 1 was dissolved in 15 mL of methanol, and thionyl chloride (177 mg, 107 μL, 1.5 mmol) was added dropwise at 0 °C and then the reaction was carried out at room temperature. LC-MS was used to monitor the reaction completion, and the methanol was dried and separated by column chromatography (eluent: dichloromethane / methanol). 376 mg of yellow solid compound 2-3 was obtained. LC-MS: (M+H) + : 397.1.

[0128] Step 3:

[0129] In a round-bottom reaction bottle with a stir bar, compound 2-3 (350 mg, 0.9 mmol) obtained in step 2, tert-butyl 5-bromopentanoate (320 mg, 1.4 mmol) and K2CO3(372 mg, 2.7 mmol) were dissolved in acetonitrile (15 mL) and the reaction was carried out at 80 °C. LC-MS was used to monitor the reaction completion, then filtered, the filter cake was washed with dichloromethane, and the organic phase was dried and separated by column chromatography (eluent: dichloromethane / methanol). 432 mg of compound 2-4 was obtained. LC-MS: (M+H) + : 553.3.

[0130] Step 4:

[0131] In a round bottom flask with stir bar, compound 2-4 (275 mg, 0.5 mmol) and the previously obtained intermediate 1-7 (270 mg, 0.5 mmol) from step 3 were dissolved in DMF (15 mL) after deprotection with TFA / DCM, DIEA (1.4 mL, 7.3 mmol) was added, followed by HATU (228 mg, 0.6 mmol). The reaction was monitored by LC-MS until completion. The reaction mixture was diluted with 50 mL water and extracted with 50 mL ethyl acetate three times. The organic phase was dried and purified by column chromatography (eluent: dichloromethane / methanol) to give 358 mg of compound 2-5 as a solid. LC-MS: (M+H) + : 925.1.

[0132] Step 5:

[0133] Compound 2-5 (370 mg, 0.4 mmol) from step 4, N-BOC-6-bromohexylamine (84 mg, 0.7 mmol) and K2CO3 (165 mg, 1.2 mmol) were dissolved in acetonitrile (15 mL) and reacted at 80 °C. The reaction was monitored by LC-MS until completion. The reaction mixture was filtered, the filter cake was washed with dichloromethane, the organic phase was dried and dissolved in tetrahydrofuran / water (2:1) 30 mL, followed by the addition of 30 mg LiOH and reaction at room temperature. The reaction was monitored by LC-MS until completion. The tetrahydrofuran was removed by rotary evaporation, the pH was adjusted to 5-6 by the addition of 2 M hydrochloric acid, the reaction mixture was diluted with 50 mL water and extracted with 50 mL ethyl acetate three times. The organic phase was dried and purified by column chromatography (eluent: dichloromethane / methanol) to give 308 mg of compound 2-6 as a white solid. LC-MS: (M+H) + : 1110.4.

[0134] Step 6:

[0135] Compound 2-6 (110 mg, 0.1 mmol) from step 5 was dissolved in DMF (30 mL) after deprotection with TFA / DCM, followed by the addition of DIEA (63 μL, 0.36 mmol) and HATU (57 mg, 0.15 mmol) and reaction at room temperature. The reaction was monitored by LC-MS until completion. The solvent was removed by rotary evaporation and the product was purified by column chromatography (eluent: dichloromethane / methanol) to give compound 15a as a white solid. 1H NMR (trans / cis, 600 MHz, Methanol-d4) δ 8.85 (s, 0.7H), 8.82 (s, 0.3H), 7.39 (d, J = 8.3 Hz, 1.4H), 7.34 - 7.28 (m, 1.4H), 7.02 - 6.92 (m, 3.3H), 6.86 (d, J = 1.5 Hz, 0.3H), 6.82 (d, J = 9.0 Hz, 0.6H), 4.64 - 4.53 (m, 3.3H), 4.50 (s, 0.3H), 4.48 (s, 1.3H), 4.46 - 4.42 (m, 0.3H), 4.38 (d, J = 14.8 Hz, 0.7H), 4.35 (d, J = 16.0 Hz, 0.3H), 4.11 (t, J = 6.3 Hz, 1.4H), 4.08 - 3.98 (m, 2.1H), 3.95 - 3.88 (m, 0.6H), 3.84 (d, J = 11.1 Hz, 0.7H), 3.74 (dd, J = 11.0, 4.1 Hz, 0.7H), 3.71 - 3.44 (m, 2.8H), 3.11 (dt, J = 13.6, 6.9 Hz, 0.7H), 3.04 (dd, J = 14.5, 2.9 Hz, 0.7H), 3.02 - 2.97 (m, 0.6H), 2.69 (d, J = 1.4 Hz, 3.0H), 2.47 (s, 2.1H), 2.44 (s, 2.1H), 2.41 (d, J = 2.4 Hz, 2.1H), 2.37 - 2.17 (m, 2.0H), 2.17 - 2.11 (m, 1.4H), 1.97 - 1.87 (m, 2.0H), 1.86 - 1.73 (m, 4.0H), 1.73 - 1.60 (m, 7.0H), 1.60 - 1.51 (m, 2.1H), 1.01 (s, 2.5H), 0.97 (s, 6.5H). 13CNMR (trans / cis, 150 MHz, Methanol-d4) δ 175.92, 175.04, 173.87, 173.81, 173.21, 173.01, 172.63, 172.48, 166.72, 166.69, 162.92, 162.79, 158.52, 157.87, 157.54, 157.51, 152.89, 152.75, 152.19, 152.15, 149.17, 149.10, 133.62, 133.61, 133.26, 133.09, 132.99, 132.90, 132.76, 132.68, 132.66, 132.53, 132.51, 131.90, 131.78, 131.48, 131.31, 130.74, 129.36, 128.03, 128.00, 122.46, 122.41, 115.56, 115.33, 113.43, 113.30, 71.10, 69.52, 69.48, 69.15, 69.08, 68.81, 61.30, 60.71, 59.48, 58.33, 57.89, 55.89, 54.98, 54.93, 41.33, 40.68, 40.52, 39.48, 39.15, 39.10, 38.93, 38.38, 37.46, 36.34, 35.99, 30.63, 30.50, 30.31, 29.46, 28.89, 28.05, 27.85, 27.17, 27.11, 27.06, 27.05, 24.01, 23.57, 15.96, 14.51, 14.47, 12.98, 11.68, 11.62. HRMS (ESI) calcd for C 52 H 65 N9O7S2[M+H] + 992.4527, found 992.4522. HPLC purity 98.87%.

[0136] Example 3: Synthesis of compound 10c

[0137]

[0138] Step 1:

[0139] In a round-bottom flask with a stir bar, the previously obtained intermediate 2-4 (553 mg, 1 mmol), LiOH (60 mg, 3.0 mmol) were dissolved in tetrahydrofuran / water (2:1) 30 mL, reaction at room temperature, LC-MS monitoring reaction complete, spin off tetrahydrofuran, add 2M hydrochloric acid to adjust pH to 5-6, add 50 mL water and 50 mL ethyl acetate extraction 3 times, spin dry organic phase, column chromatography separation (eluent: dichloromethane / methanol). 510 mg of compound 3-0 was obtained. LC-MS: (M+H) + : 539.2.

[0140] Step 2:

[0141] In a round-bottom flask with a stir bar, compound 3-1 (synthesis reference https: / / doi.org / 10.1038 / s41467-022-33430-6) (388 mg, 1 mmol) was dissolved in super dry DMF / toluene (20 mL, v / v = 1 / 10), then DBU (380 mg, 348 μL, 2.5 mmol) and DPPA (687 mg, 2.5 mmol) were added, and the reaction was carried out at 50°C. LC-MS monitoring reaction complete, add 100 mL water and 100 mL ethyl acetate extraction 3 times, column chromatography separation (eluent: petroleum ether / ethyl acetate), 336 mg of oily compound 3-2 was obtained. LC-MS: (M+H) + : 416.3.

[0142] Step 3:

[0143] In a round-bottom flask with a stir bar, compound 3-2 (210 mg, 0.5 mmol) obtained in step 2 was deprotected with TFA / DCM, then the previously obtained intermediate 1-3 (172 mg, 0.5 mmol) was dissolved in DMF (20 mL), then DIEA (1 mL, 5 mmol) was added, and HATU (228 mg, 0.6 mmol) was added, and the reaction was carried out at room temperature. LC-MS monitoring reaction complete, add 50 mL water and 50 mL ethyl acetate extraction 3 times, spin dry organic phase, column chromatography separation (eluent: dichloromethane / methanol). 204 mg of compound 3-3 was obtained. LC-MS: (M+H) + : 642.3.

[0144] Step 4:

[0145] In a round bottom flask with stir bar, compound 3-3 (193 mg, 0.3 mmol) from Step 3 was dissolved in 1,4-dioxane / water (20 mL, v / v = 5 / 1), and triphenylphosphine (257 mg, 1 mmol) was added. The reaction was stirred at room temperature for 1 h, and then ammonia (2 mL) was added. The reaction was monitored by LC-MS until completion. The organic phase was evaporated, and the product was isolated by column chromatography (eluent: 0.5% triethylamine in dichloromethane / methanol) to give 182 mg of compound 3-4 as a solid. LC-MS: (M+H) + : 616.2.

[0146] Step 5:

[0147] Compound 3-4 (182 mg, 0.3 mmol) from Step 4 and compound 3-0 (162 mg, 0.3 mmol) from Step 1 were dissolved in DMF (15 mL), and DIEA (0.6 mL, 3 mmol) was added. HATU (152 mg, 0.4 mmol) was added, and the reaction was stirred at room temperature. The reaction was monitored by LC-MS until completion. The reaction was extracted with 50 mL water and 50 mL ethyl acetate three times. The organic phase was evaporated, and the product was isolated by column chromatography (eluent: dichloromethane / methanol) to give 160 mg of compound 3-5 as a solid. LC-MS: (M+H) + : 1136.5.

[0148] Step 6:

[0149] Compound 3-5 (113 mg, 0.1 mmol) from Step 5 was deprotected with TFA / DCM and dissolved in DMF (30 mL). DIEA (63 μL, 0.36 mmol) and HATU (57 mg, 0.15 mmol) were added, and the reaction was stirred at room temperature. The reaction was monitored by LC-MS until completion. The solvent was evaporated, and the product was isolated by column chromatography (eluent: dichloromethane / methanol) to give compound 10c as a white solid. 1H NMR (trans, 600 MHz, Methanol-d4) δ 8.87 (s, 1H), 7.50 - 7.33 (m, 6H), 7.07 (d, J = 9.0 Hz, 2H), 4.88 (dd, J = 9.7, 4.8 Hz, 1H), 4.67 (s, 1H), 4.62 - 4.56 (m, 2H), 4.43 (s, 1H), 4.13 - 4.02 (m, 2H), 3.87 (d, J = 11.1 Hz, 1H), 3.74 (dd, J = 11.1, 3.7 Hz, 1H), 3.67 - 3.60 (m, 1H), 3.49 (dd, J = 14.3, 11.6 Hz, 1H), 3.02 (dd, J = 14.3, 2.4 Hz, 1H), 2.95 (dt, J = 13.7, 7.0 Hz, 1H), 2.70 (s, 3H), 2.48 (s, 3H), 2.45 (s, 3H), 2.44 - 2.41 (m, 1H), 2.33 - 2.27 (m, 1H), 2.20 - 2.15 (m, 1H), 1.93 - 1.74 (m, 8H), 1.72 (s, 3H), 1.70 - 1.63 (m, 2H), 1.55 - 1.47 (m, 3H), 1.05 (s, 9H). 13 C NMR (trans, 150 MHz, Methanol-d4) δ 175.73, 173.81, 172.96, 172.02, 166.72, 162.77, 157.54, 152.89, 152.16, 149.11, 145.45, 133.43, 133.11, 132.98, 132.74, 132.50, 132.11, 131.51, 131.43, 130.50, 127.92, 115.90, 71.09, 69.16, 60.63, 59.04, 58.17, 55.00, 54.91, 49.61, 40.80, 39.03, 38.80, 37.95, 36.69, 35.72, 30.55, 28.15, 28.13, 27.49, 27.14, 24.24, 15.85, 14.51, 12.98, 11.63. HRMS (ESI) calcd for C 51 H 63 N9O6S2[M+H] + 962.4421, experimental 962.4416. HPLC purity 99.05%.

[0150] Example 4: Synthesis of compound 15b

[0151]

[0152] Preparation Method refers to the synthesis of 15a. 1 H NMR (trans / cis, 600 MHz, Methanol-d4) δ 8.87 (s, 0.7H), 8.86 (s, 0.3H), 7.39 (d, J = 8.3 Hz, 2H), 7.34 (d, J = 7.7 Hz, 0.7H), 7.23 (d, J = 7.8 Hz, 0.3H), 7.02 (dd, J = 7.7, 1.5 Hz, 0.3H), 6.99 (d, J = 1.3 Hz, 0.7H), 6.98 (dd, J = 7.7, 1.5 Hz, 0.7H), 6.95 - 6.89 (m, 1.7H), 6.86 (d, J = 9.0 Hz, 0.6H), 4.65 - 4.60 (m, 0.6H), 4.59 - 4.52 (m, 2.4H), 4.49 - 4.43 (m, 1.7H), 4.38 (d, J = 14.5 Hz, 0.7H), 4.30 (d, J = 16.7 Hz, 0.3H), 4.11 - 3.97 (m, 3.3H), 3.97 - 3.89 (m, 0.7H), 3.86 (d, J = 10.9 Hz, 0.7H), 3.75 (dd, J = 11.0, 4.1 Hz, 0.7H), 3.71 - 3.65 (m, 0.3H), 3.65 - 3.59 (m, 0.6H), 3.56 - 3.42 (m, 1.7H), 3.21 (q, J = 7.3 Hz, 0.3H), 3.13 - 3.06 (m, 0.7H), 3.05 - 3.02 (m, 0.7H), 2.99 (dd, J = 14.0, 2.8 Hz, 0.3H), 2.96 - 2.91 (m, 0.3H), 2.69 (s, 3.0H), 2.49 (s, 2.1H), 2.46 (s, 0.9H), 2.43 (s, 2.1H), 2.43 - 2.38 (m, 2.1H), 2.36 - 2.20 (m, 1.7H), 2.12 (dd, J = 8.3, 4.0 Hz, 1.4H), 1.95 - 1.89 (m, 1.4H), 1.86 - 1.72 (m, 5.0H), 1.70 (s, 2.1H), 1.68 (s, 0.9H), 1.67 - 1.61 (m, 2.1H), 1.60 - 1.55 (m, 1.4H), 1.51 - 1.45 (m, 4.6H), 1.01 (s, 2.5H), 0.99 (s, 6.5H). 13C NMR (trans / cis, 150 MHz, Methanol-d4) δ 175.97, 174.82, 173.94, 173.81, 173.13, 172.98, 172.60, 172.52, 166.65, 166.52, 162.94, 162.81, 158.60, 157.83, 157.52, 157.49, 152.91, 152.81, 152.13, 149.17, 149.10, 133.63, 133.59, 133.37, 133.11, 133.07, 133.01, 132.99, 132.71, 132.69, 132.54, 132.49, 132.42, 131.86, 131.64, 131.48, 131.15, 128.29, 128.20, 127.89, 122.44, 122.39, 115.43, 115.24, 113.37, 113.16, 71.08, 69.50, 69.47, 69.16, 69.03, 68.81, 61.36, 60.65, 59.48, 58.26, 57.86, 55.95, 55.14, 55.02, 42.62, 41.43, 40.64, 40.43, 39.55, 39.14, 39.03, 38.36, 37.48, 37.44, 36.24, 36.11, 36.03, 32.79, 31.17, 30.76, 30.69, 30.50, 30.38, 30.37, 30.20, 29.71, 29.32, 29.16, 28.90, 28.17, 28.16, 28.02, 27.19, 27.10, 27.03, 26.90, 24.00, 23.74, 23.46, 23.03, 21.55, 19.87, 19.18, 15.99, 15.97, 14.67, 14.59, 14.51, 14.47, 12.99, 12.98, 11.80, 11.62. HRMS (ESI) calcd for C 53 H 67 N9O7S2[M+H] + 1006.4683, experimental value 1006.4683. HPLC purity 95.35%.

[0153] Example 5: Synthesis of compound 15c

[0154]

[0155] Preparation method refers to the synthesis of 15a. 1H NMR (trans / cis, 600 MHz, Methanol-d4) δ 8.86 (s, 0.7H), 8.85 (s, 0.3H), 7.41 - 7.34 (m, 2H), 7.32 (d, J = 7.6 Hz, 0.7H), 7.25 (d, J = 7.8 Hz, 0.3H), 7.01 (dd, J = 7.8, 1.5 Hz, 0.3H), 6.99 - 6.95 (m, 1.4H), 6.95 - 6.89 (m, 1.7H), 6.85 (d, J = 9.0 Hz, 0.6H), 4.65 (d, J = 16.2 Hz, 0.3H), 4.61 (d, J = 9.3 Hz, 0.6H), 4.59 - 4.55 (m, 1.4H), 4.53 (s, 0.7H), 4.50 - 4.42 (m, 1.7H), 4.38 (d, J = 15.0 Hz, 0.7H), 4.28 (d, J = 16.2 Hz, 0.3H), 4.12 - 4.06 (m, 1.4H), 4.05 - 3.92 (m, 2.7H), 3.87 (d, J = 10.7 Hz, 0.7H), 3.75 (dd, J = 10.9, 4.1 Hz, 0.7H), 3.66 - 3.61 (m, 0.9H), 3.53 - 3.44 (m, 2.3H), 3.11 - 3.04 (m, 1.4H), 3.02 - 2.96 (m, 0.6H), 2.69 (s, 0.9H), 2.69 (s, 2.1H), 2.48 (s, 2.1H), 2.45 (s, 0.9H), 2.43 (s, 2.1H), 2.42 - 2.41 (m, 1.2H), 2.40 - 2.24 (m, 1.7H), 2.22 - 2.17 (m, 0.3H), 2.15 (dd, J = 8.2, 3.7 Hz, 1.4H), 1.92 - 1.85 (m, 1.7H), 1.85 - 1.74 (m, 4.3H), 1.71 (s, 2.1H), 1.67 (s, 0.9H), 1.64 - 1.55 (m, 3.4H), 1.52 - 1.39 (m, 6.6H), 1.01 (s, 2.7H), 0.98 (s, 6.3H). 13C NMR (trans / cis, 150 MHz, Methanol-d4) δ 176.03, 174.87, 173.95, 173.87, 173.15, 172.95, 172.65, 172.53, 166.81, 166.65, 162.94, 162.84, 158.48, 157.97, 157.52, 157.51, 152.89, 152.81, 152.16, 152.14, 149.15, 149.13, 133.63, 133.17, 133.11, 133.08, 133.00, 132.75, 132.72, 132.69, 132.52, 132.46, 131.85, 131.73, 131.48, 131.46, 130.53, 128.80, 128.22, 128.03, 122.40, 122.38, 115.44, 115.29, 113.38, 113.27, 71.12, 69.57, 69.41, 69.18, 69.09, 68.78, 66.96, 61.33, 60.70, 59.58, 58.30, 57.89, 57.66, 57.52, 57.38, 56.03, 55.07, 54.98, 41.40, 40.71, 40.60, 39.45, 39.19, 39.14, 38.98, 38.41, 37.46, 36.18, 36.10, 31.06, 30.89, 30.77, 30.71, 30.67, 30.59, 30.53, 30.39, 29.74, 29.25, 28.34, 28.24, 27.48, 27.14, 27.10, 27.03, 23.91, 23.58, 15.96, 15.49, 14.52, 14.49, 12.97, 11.64, 11.62. HRMS (ESI) calcd for C 54 H 69 N9O7S2[M+H] + 1020.4840, found 1020.4834. HPLC purity 99.47%.

[0156] Example 6: Synthesis of compound 10a

[0157]

[0158] Preparation method refers to the synthesis of 10c. 1H NMR (600 MHz, Methanol-d4) δ 8.87 (s, 1H), 7.53 - 7.42 (m, 4H), 7.41 (d, J = 8.3 Hz, 1H), 7.17 (d, J = 8.5 Hz, 1H), 4.84 - 4.81 (m, 1H), 4.77 (s, 1H), 4.70 - 4.64 (m, 1H), 4.61 (dd, J = 11.6, 2.3 Hz, 1H), 4.38 (s, 1H), 4.10 - 4.04 (m, 1H), 4.04 - 3.99 (m, 1H), 3.84 (d, J = 11.2 Hz, 1H), 3.71 (dd, J = 11.1, 3.8 Hz, 1H), 3.69 - 3.64 (m, 1H), 3.51 (dd, J = 14.4, 11.6 Hz, 1H), 3.04 (dd, J = 14.3, 2.4 Hz, 1H), 2.98 - 2.89 (m, 1H), 2.70 (s, 3H), 2.48 (s, 3H), 2.45 (s, 3H), 2.45 - 2.40 (m, 1H), 2.37 - 2.30 (m, 1H), 2.25 - 2.19 (m, 1H), 2.00 - 1.82 (m, 5H), 1.82 - 1.75 (m, 3H), 1.74 (s, 3H), 1.72 - 1.64 (m, 1H), 1.04 (s, 9H). 13 C NMR (150 MHz, Methanol-d4) δ 175.33, 173.94, 173.02, 171.74, 166.57, 162.72, 157.61, 152.91, 152.14, 149.11, 145.65, 133.42, 133.03, 133.00, 132.83, 132.44, 131.96, 131.59, 131.48, 130.51 (2C), 127.94 (2C), 115.99, 70.92, 69.55, 60.34, 58.67, 58.14, 55.80, 54.79, 49.61, 40.77, 39.06, 38.87, 37.06 (2C), 35.52, 35.26, 28.50, 27.64 (3C), 27.13, 24.48, 15.85, 14.53, 12.99, 11.62. HRMS (ESI) calcd for C 49 H 59 N9O6S2[M+H] + 934.4105, found 934.4104. HPLC purity 97.46%

[0159] Example 7: Synthesis of compound 10b

[0160]

[0161] Preparation method refers to the synthesis of 10c. 1 H NMR (600 MHz, Methanol-d4) δ 8.87 (s, 1H), 7.47 - 7.40 (m, 6H), 7.15 (d, J = 8.9 Hz, 2H), 4.92 (t, J = 7.5 Hz, 1H), 4.70 (s, 1H), 4.61 - 4.53 (m, 2H), 4.45 (s, 1H), 4.18 - 4.12 (m, 1H), 4.11 - 4.04 (m, 1H), 3.84 (d, J = 11.1 Hz, 1H), 3.76 (dd, J = 11.1, 3.7 Hz, 1H), 3.60 - 3.52 (m, 1H), 3.48 (dd, J = 13.9, 11.6 Hz, 1H), 3.03 - 2.94 (m, 2H), 2.70 (s, 3H), 2.48 (s, 3H), 2.45 (s, 3H), 2.43 - 2.37 (m, 2H), 2.16 (dd, J = 13.1, 7.3 Hz, 1H), 1.97 - 1.82 (m, 7H), 1.80 - 1.74 (m, 1H), 1.73 (s, 3H), 1.71 - 1.62 (m, 2H), 1.56 - 1.48 (m, 1H), 1.06 (s, 9H). 13 C NMR (150 MHz, Methanol-d4) δ 175.39, 175.31, 173.59, 173.17, 171.88, 171.85, 166.62, 162.69, 157.50, 152.92, 152.17, 149.15, 144.58, 133.39, 133.16, 132.98, 132.69, 132.49, 132.21, 131.71, 131.55, 130.55, 128.29, 115.80, 71.17, 69.85, 60.86, 59.16, 59.07, 58.26, 55.17, 54.56, 49.61, 40.10, 39.12, 38.79, 37.24, 37.22, 36.23, 30.88, 28.16, 27.23, 24.68, 24.47, 15.86, 14.55, 12.98, 11.64. HRMS (ESI) calcd for C 50 H 61 N9O6S2[M+H] + 948.4264, found 948.4263. HPLC purity 97.21%.

[0162] Example 8: Synthesis of compound 10d

[0163]

[0164] Preparation method refers to the synthesis of 10c. 1 H NMR (trans, 600 MHz, Methanol-d4) δ 8.87 (s, 1H), 7.47 - 7.36 (m, 6H), 7.02 (d, J = 9.0 Hz, 2H), 4.94 - 4.88 (m, 1H), 4.64 (s, 1H), 4.61 - 4.54 (m, 2H), 4.44 (s, 1H), 4.14 - 4.06 (m, 2H), 3.84 (d, J = 11.2 Hz, 1H), 3.74 (dd, J = 11.1, 3.7 Hz, 1H), 3.57 - 3.48 (m, 2H), 3.07 - 3.01 (m, 1H), 2.99 (dd, J = 14.1, 3.0 Hz, 1H), 2.70 (s, 3H), 2.48 (s, 3H), 2.44 (s, 3H), 2.39 (t, J = 6.8 Hz, 3H), 2.19 - 2.12 (m, 1H), 1.98 - 1.89 (m, 2H), 1.89 - 1.78 (m, 6H), 1.71 (s, 3H), 1.68 - 1.58 (m, 2H), 1.53 - 1.44 (m, 5H), 1.05 (s, 9H). 13 C NMR (trans, 150 MHz, Methanol-d4) δ 175.49, 173.73, 173.14, 172.00, 166.57, 162.78, 157.52, 152.89, 152.17, 149.11, 145.06, 133.42, 133.17, 132.99, 132.66, 132.49, 132.04, 131.53, 130.50, 128.04, 115.69, 71.20, 69.08, 60.93, 59.18, 58.18, 55.07, 54.42, 40.55, 39.03, 38.73, 37.87, 37.07, 36.14, 30.95, 30.25, 28.48, 27.67, 27.20, 27.08, 24.16, 15.85, 14.55, 12.98, 11.64. HRMS (ESI) calcd for C 52 H 65 N9O6S2[M+H] + 976.4577, found 976.4574. HPLC purity 95.97%.

[0165] Example 9: Synthesis of compound 10e

[0166]

[0167] The preparation method refers to the synthesis of 10c. 1 H NMR (trans, 600 MHz, Methanol-d4) d 8.86 (s, 1H), 7.46 - 7.35 (m, 6H), 6.98 (d, J = 9.1 Hz, 1H), 4.90 (dd, J = 10.2, 4.2 Hz, 1H), 4.62 (s, 1H), 4.60 - 4.54 (m, 2H), 4.44 (s, 1H), 4.11 - 4.01 (m, 2H), 3.87 (d, J = 11.2 Hz, 1H), 3.75 (dd, J = 11.1, 3.7 Hz, 1H), 3.65 - 3.59 (m, 1H), 3.48 (dd, J = 14.1, 11.6 Hz, 1H), 3.00 (dd, J = 14.0, 2.6 Hz, 1H), 2.98 - 2.91 (m, 1H), 2.69 (s, 3H), 2.48 (s, 3H), 2.44 (s, 3H), 2.43 - 2.39 (m, 1H), 2.38 - 2.32 (m, 1H), 2.18 - 2.12 (m, 1H), 1.97 - 1.90 (m, 1H), 1.88 - 1.78 (m, 6H), 1.72 (s, 3H), 1.66 - 1.59 (m, 2H), 1.48 - 1.41 (m, 8H), 1.07 (s, 9H). 13 C NMR (trans, 150 MHz, Methanol-d4) d 175.74, 173.76, 172.91, 172.10, 166.62, 162.80, 157.49, 152.87, 152.14, 149.08, 145.37, 133.40, 133.12, 132.96, 132.66, 132.47, 131.97, 131.44, 130.47, 127.88, 115.50, 71.16, 68.96, 60.80, 59.29, 58.16, 54.99, 54.69, 40.80, 39.10, 38.77, 38.30, 36.65, 36.02, 31.09, 31.04, 30.84, 28.63, 27.90, 27.19, 27.13, 24.20, 15.86, 15.84, 14.52, 12.99, 11.63. HRMS (ESI) calcd for C 53 H 67 N9O6S2[M+H] + 990.4734, found 990.4732. HPLC purity 98.31%.

[0168] Example 10: Synthesis of compound 10f

[0169]

[0170] Preparation method refers to the synthesis of 10c. 1 H NMR (trans, 600 MHz, Methanol-d4) δ 8.86 (s, 1H), 7.44 - 7.38 (m, 6H), 6.96 (d, J = 8.8 Hz, 2H), 4.90 (dd, J = 10.0, 4.6 Hz, 1H), 4.60 (s, 1H), 4.59 - 4.55 (m, 2H), 4.43 (s, 1H), 4.10 - 4.03 (m, 2H), 3.86 (d, J = 11.2 Hz, 1H), 3.73 (dd, J = 11.1, 3.8 Hz, 1H), 3.60 - 3.54 (m, 1H), 3.51 - 3.45 (m, 1H), 3.04 - 2.97 (m, 2H), 2.69 (s, 3H), 2.48 (s, 3H), 2.44 (s, 3H), 2.41 - 2.36 (m, 2H), 2.19 - 2.13 (m, 1H), 1.97 - 1.88 (m, 1H), 1.87 - 1.77 (m, 5H), 1.71 (s, 3H), 1.65 - 1.60 (m, 2H), 1.47 - 1.38 (m, 10H), 1.05 (s, 9H). 13 C NMR (trans, 150 MHz, Methanol-d4) δ 175.72, 173.77, 173.11, 172.13, 166.64, 162.84, 157.52, 152.87, 152.14, 149.09, 145.31, 133.43, 133.11, 132.99, 132.71, 132.47, 131.91, 131.47, 131.44, 130.47, 127.93, 115.46, 71.17, 68.81, 60.83, 58.12, 55.05, 54.56, 40.64, 39.06, 38.78, 38.20, 37.34, 36.69, 36.14, 31.20, 30.85, 30.71, 30.20, 28.85, 28.09, 27.49, 27.22, 27.09, 24.09, 23.03, 15.85, 14.54, 12.98, 11.62. HRMS (ESI) calcd for C 54 H 69 N9O6S2[M+H] + 1004.4890, found 1004.4887. HPLC purity 96.47%.

[0171] Example 11: Synthesis of 10g of compound

[0172]

[0173] The preparation method is the same as that used in the synthesis of 10c. 1 H NMR(trans,600MHz,Methanol-d4)δ8.87(s,1H),7.45–7.35(m,6H),7.05(d,J=8.9Hz,2H),4.93(dd,J=10.0,4.7Hz,1H),4.61–4.5 5(m,2H),4.44(s,1H),4.39(s,1H),4.15–4.08(m,1H),4.07–3.98(m,1H),3.86(d,J=11.1Hz,1H),3.71(dd,J=10.9,3.8Hz,1H),3. 64–3.56(m,1H),3.53–3.46(m,1H),3.06–2.98(m,2H),2.70(s,3H),2.55–2.47(m,1H),2.48(s,3H),2.47–2.41(m,4H),2.16–2.11 (m,2H),2.10–2.03(m,1H),1.94–1.87(m,1H),1.85–1.75(m,2H),1.69(s,3H),1.65–1.58(m,2H),1.55–1.44(m,6H),1.00(s,9H). 13 C NMR(trans,150MHz,Methanol-d4)δ175.52,173.93,173.11,171.94,166.79,162.69,157.50,152 .88,149.10,145.26,133.44,133.16,132.97,132.69,132.48,132.32,131.49,130.49,127.97,1 15.94,71.21,68.42,60.83,60.79,57.96,55.02,54.05,40.18,39.02,38.82,37.73,36.25,33.2 7,30.67,29.74,27.39,27.21,27.09,26.99,26.47,15.85,14.51,12.97,11.65.HRMS(ESI)calcd for C 51 H 63 N9O6S2[M+H] + 962.4421, experimental value 962.4418. HPLC purity 99.49%.

[0174] Example 12: Synthesis of compound 10h

[0175]

[0176] Preparation method refers to the synthesis of 10c. 1 H NMR (trans, 600 MHz, Methanol-d4) d 8.87 (s, 1H), 7.47 - 7.35 (m, 6H), 6.96 (d, J = 9.0 Hz, 2H), 4.91 - 4.87 (m, 1H), 4.58 - 4.53 (m, 3H), 4.43 (s, 1H), 4.10 - 4.03 (m, 2H), 3.87 (d, J = 11.0 Hz, 1H), 3.75 (dd, J = 11.0, 3.9 Hz, 1H), 3.54 - 3.45 (m, 2H), 3.08 - 3.02 (m, 1H), 3.00 (dd, J = 14.0, 2.8 Hz, 1H), 2.69 (s, 3H), 2.48 (s, 3H), 2.44 (s, 3H), 2.37 - 2.31 (m, 1H), 2.28 - 2.24 (m, 1H), 2.16 - 2.11 (m, 1H), 1.96 - 1.90 (m, 1H), 1.87 - 1.73 (m, 5H), 1.70 (s, 3H), 1.65 - 1.61 (m, 2H), 1.54 (p, J = 7.0 Hz, 3H), 1.49 - 1.41 (m, 6H), 1.06 (s, 9H). 13 C NMR (trans, 150 MHz, Methanol-d4) d 176.29, 176.20, 173.74, 173.66, 173.10, 172.16, 172.14, 166.62, 163.01, 157.53, 152.90, 152.16, 149.12, 145.02, 145.00, 133.42, 133.12, 133.03, 132.73, 132.45, 131.79, 131.55, 131.49, 130.50, 128.08, 115.64, 71.17, 69.32, 60.82, 60.77, 59.70, 59.61, 58.09, 55.14, 54.42, 54.32, 40.32, 39.08, 38.73, 37.67, 37.64, 36.90, 36.86, 36.57, 30.98, 30.02, 29.59, 27.57, 27.22, 27.10, 27.03, 26.87, 26.50, 15.85, 14.51, 12.98, 12.97, 11.62. HRMS (ESI) calcd for C 53 H67 N9O6S2C 51 H 63 N9O6S2[M+H] + 990.4734, found 990.4725. HPLC purity 98.62%.

[0177] Example 13: Synthesis of compound 10i

[0178]

[0179] Preparation method refers to the synthesis of 10c. 1 H NMR (trans, 600 MHz, Methanol-d4) δ 8.87 (s, 1H), 7.44 (d, J = 8.3 Hz, 2H), 7.41 - 7.38 (m, 4H), 6.94 (d, J = 9.0 Hz, 2H), 4.92 (dd, J = 8.9, 5.8 Hz, 1H), 4.67 (s, 1H), 4.59 - 4.52 (m, 2H), 4.44 (s, 1H), 4.10 - 4.00 (m, 2H), 3.84 (d, J = 11.2 Hz, 1H), 3.75 (dd, J = 11.1, 3.7 Hz, 1H), 3.54 - 3.45 (m, 2H), 3.06 (dt, J = 13.2, 7.3 Hz, 1H), 3.00 (dd, J = 14.0, 2.9 Hz, 1H), 2.69 (s, 3H), 2.48 (s, 3H), 2.44 (s, 3H), 2.33 - 2.28 (m, 2H), 2.16 - 2.11 (m, 1H), 1.98 - 1.91 (m, 1H), 1.88 - 1.72 (m, 5H), 1.70 (s, 3H), 1.67 - 1.55 (m, 5H), 1.51 - 1.37 (m, 8H), 1.07 (s, 9H). 13C NMR(trans,150MHz,Methanol-d4)δ175.85,173.66,173.07,172.08,166.56,163.02,157.53,152.90,15 2.16,149.12,144.94,133.40,133.14,132.99,132.69,132.48,131.79,131.57,131.50,130.51,128.10, 115.49,71.15,69.14,60.79,58.95,58.17,55.08,54.23,40.51,39.05,38.70,37.67,36.99,36.02,31. 05,30.12,30.03,29.03,27.78,27.21,27.10,26.53,26.50,15.86,14.52,12.98,11.63.HRMS(ESI)calcd for C 54 H 69 N9O6S2[M+H] + 1004.4890, experimental value 1004.4889. HPLC purity 98.77%.

[0180] Example 14: Degradation of BRD4 protein in PC-3 tumor cells by the compound

[0181] Method: At a density of 2×10 6 Cells were seeded at 10 nM / ml in 6-well plates, followed by the addition of 10 nM of the drug. Cells were treated at 37°C for 24 hours, and the supernatant was collected and washed twice with PBS. Cells were lysed using SDS lysis buffer (62.5 mM Tris, 2% (w / v) SDS, 10% glycerol, 50 mM DTT, 0.01% (w / v) bromophenol blue, pH 6.8). All cell clusters were then disrupted on ice using an ultrasonic cell disruptor for 10 seconds, heated at 100°C for 10 minutes, and centrifuged at 12000 RPM for 5 minutes at 4°C. The denaturing lysis buffer was separated by SDS-PAGE and then electrophoretically transferred to a polyvinylidene fluoride membrane. The membrane was then incubated in blocking buffer at room temperature for 1 hour. It was then incubated overnight at 4°C with primary antibody, followed by incubation with secondary antibody for 1 hour. Chemiluminescence detection was performed using Western Lightning Plus ECL. Data analysis and statistical analysis were performed using GraphPad Prism 7.0.

[0182] Table 1. Activity of compounds on BRD4 protein degradation in PC-3 cells.

[0183]

[0184]

[0185]

[0186]

[0187] The above has exemplarily described the embodiments of the technical scheme of the present application. It should be understood that the protection scope of the present application is not limited to the above embodiments. Any modification, equivalent replacement, improvement, etc. made by those skilled in the art within the spirit and principle of the present application shall be included in the protection scope of the claims of the present application.

Claims

1. A compound as shown in formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof or a prodrug molecule thereof: ###0001### (I) wherein, B is selected from the group consisting of: ###0002### wherein, each of W1, W2, W3, W4, W5 is independently selected from the group consisting of -0-, -S-, -NH-, -CH2-, -CONH-, -NHCO-, -C(O)-, -C(O)O-, -OC(O)-, -NHCONH-, a phenyl ring, a 3-10 membered ring having 0-4 heteroatoms, a 5-14 membered heteroaromatic ring having 1-5 heteroatoms, said heteroatoms are selected from N, S or O; each of n1, n2, n3, n4 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10; and each of X and Y is independently selected from the group consisting of -0-, -S-, -CO-, -NH-, -CONH-, -NHCO-, -C(O)O-, and -OC(O)-; each of n and m is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10. L 1 、L 2 each independently selected from a structure having a formula: wherein, W1, W2, W3, W4, W5are each independently selected from the group consisting of -0-, -S-, -NH-, -CH2-, -CONH-, -NHCO-, -C≡C-, -CH=CH-, -C(O)-, -P(=O)-, -S(O)2-, -S(O)-, -P(O)2(OH)-, -NH-S(O)-NH-, -C(O)O-, -OC(O)-, -NHCONH-, a phenyl ring, a 3-12 membered heterocyclic ring having 0-4 heteroatoms, a 5-14 membered heteroaromatic ring having 1-5 heteroatoms, said heteroatoms being selected from N, S or O; n1, n2, n3, n4are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10; wherein said 3-12 membered ring can be a monocyclic, spirocyclic, fused or bridged ring structure, and can each independently be optionally substituted with one or more substituents selected from oxo, halogen, C 1-6 alkyl, C 1-6 alkoxy and C 1-6 haloalkyl; said 3-12 membered heterocyclic ring is a saturated or partially unsaturated ring, but does not include an aromatic structure; R 1 selected from the group consisting of H, D, CN, NO2, halogen, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C2-C6alkenyl, substituted or unsubstituted C2-C6alkynyl, substituted or unsubstituted C1-C6alkoxy, substituted or unsubstituted C3-C6cycloalkyl, 3-12 membered heterocyclyl containing from 1-4 heteroatoms selected from the group consisting of oxygen, sulfur and nitrogen, aryl, 5-14 membered heteroaryl containing from 1-5 heteroatoms selected from the group consisting of oxygen, sulfur and nitrogen; Preferably, W1 is selected from the group consisting of O or -NHCO-. wherein R 2 may be one or more substituents on the ring, and when said substituents are more than one, each R 2 are the same or different, and each is independently selected from the group consisting of H, OH, CN, NO2, halogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C6cycloalkyl, optionally substituted with one, two or more R 3 substituted 5-14 membered heteroaryl containing 1-5 heteroatoms selected from oxygen, sulfur and nitrogen; R 3 is selected from halogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C6cycloalkyl, or halogen mono-, di- or poly-substituted C1-C6alkyl.

2. The compound or pharmaceutically acceptable salt thereof, or stereoisomer or prodrug molecule thereof, according to claim 1, wherein R 1 selected from the group consisting of C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C1-C6alkoxy, C3-C6cycloalkyl, 3- to 12-membered heterocyclyl containing from 1 to 4 heteroatoms selected from oxygen, sulfur, and nitrogen, aryl, 5- to 14-membered heteroaryl containing from 1 to 5 heteroatoms selected from oxygen, sulfur, and nitrogen; Preferably, R 1 is selected from H, methyl, ethyl, isopropyl, tert-butyl, cyclopropyl; Preferably, R 1 is tert-butyl.

3. The compound of claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer or a prodrug molecule thereof, wherein L 1 、L 2 having the structure shown in the following formula: W2 is selected from the group consisting of -NHCO-, O or CH2. wherein each of X and Y is independently selected from the group consisting of -0-, -S-, -CO-, -NH-, -CONH-, -NHCO-, -C(O)O-, and -OC(O)-; each of n and m is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10. B is selected from the group consisting of:

4. The compound of claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer or a prodrug molecule thereof, wherein, The L 1 , L 2 each independently has a structure selected from the group consisting of Preferably, B is selected from the group consisting of:

5. The compound of claim 1, or a stereoisomer thereof, wherein said compound is selected from the group consisting of the following structures: R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 are identical or different, each independently selected from the group consisting of H, Cl, F, Br; Preferably, R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 is H or F. Preferably, R 2 Selected from 5-14 heteroaryl groups containing 1-5 heteroatoms selected from oxygen, sulfur, and nitrogen, or optionally surrounded by one, two, or more R atoms. 3 Substituted 5-14 membered heteroaryl groups containing 1-5 heteroatoms selected from oxygen, sulfur, and nitrogen; Preferably, R 3 They may be the same or different, and are independently selected from: C1-C6 alkyl, C3-C6 cycloalkyl, halogen monosubstituted, disubstituted or polysubstituted C1-C6 alkyl; 7. A pharmaceutical composition for treating and / or preventing a tumor, comprising:

6. The compound according to any one of claims 1 to 5, or a stereoisomer thereof, wherein (1) a compound as claimed in any one of claims 1-6, a pharmaceutically acceptable salt thereof, a stereoisomer thereof or a prodrug molecule thereof; optionally (2) a pharmaceutically acceptable carrier. for use in the preparation of a medicament for treating and / or preventing a disease or disorder associated with abnormal expression of BRD4. said disease or disorder is selected from the group consisting of a hematological tumor, a gastrointestinal stromal tumor, a histiocytic lymphoma, a non-small cell lung cancer, a small cell lung cancer, a lung adenocarcinoma, a lung squamous carcinoma, a pancreatic cancer, a breast cancer, a prostate cancer, a liver cancer, a skin cancer, an epithelial cell cancer, a colorectal cancer, a kidney cancer, a stomach cancer, a head and neck cancer, or a nasopharyngeal carcinoma.

8. The use of a compound, pharmaceutically acceptable salt, stereoisomer, or prodrug molecule thereof of any one of claims 1-6, or the pharmaceutical composition of claim 7, wherein, administering to a subject a compound as claimed in any one of claims 1-6, a pharmaceutically acceptable salt thereof, a stereoisomer thereof or a prodrug molecule thereof, or a pharmaceutical composition as claimed in claim 7.

9. The use according to claim 8, characterized in that, In another preferred embodiment, the subject is a human.

10. A method of degrading BRD4 in a subject in need thereof, other than for diagnosis or treatment, comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-9. ​ ​

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