BRD4 protein degradation agent compound as well as preparation method and application thereof

Abstract

The invention provides a compound as shown in a formula (I) and a racemate, a stereoisomer, a tautomer, an isotope label, a nitrogen oxide, a solvate, a polymorphic substance, a metabolite, ester, a prodrug or pharmaceutically acceptable salt thereof, which have better BRD4 inhibition and degradation effects as a drug, and can be used for preparing drugs related to BRD4 inhibition and degradation. And preventing and / or treating related diseases. # imgabs0 #

Description

[0001] The present invention claims:

[0002] Priority to the prior application, patent application number 2023112633767, filed with the State Intellectual Property Office of China on September 27, 2023, entitled “A class of BRD4 protein degrader compounds, preparation methods and applications thereof”;

[0003] Priority to the prior application No. 18 / 475,989, filed with the U.S. Patent Office on September 27, 2023, entitled “BRD4 ​​protein degrader compounds, methods for their preparation and use”;

[0004] The entirety of said prior application is incorporated herein by reference. Technical Field

[0005] The present invention belongs to the field of medicine, and specifically relates to a class of BRD4 protein degradation agent compounds and a preparation method and application thereof. Background Art

[0006] Bromodomain (BRD) is a class of highly conserved protein domains that can recognize and bind to acetylated lysine residues of histones. It consists of four anti-parallel α-helices (αZ, αA, αB, αC) and two hydrophobic loops (ZA and BC) formed by about 110 amino acids. In the human genome, there are 61 bromodomains distributed in 46 bromodomain proteins. Based on the similarity of structure or sequence, bromodomain proteins are divided into eight families, among which the bromodomain and extraterminal domain (BET) protein family has been studied most intensively and extensively.

[0007] BET family proteins include BRD2 (bromodomain-containing protein 2), BRD3, BRD4 and testis-specific BRDT (Testis-specific bromodomain-containing protein). Their structures are similar, including two tandem bromodomains (BD1 and BD2) at the N-terminus and an extra-terminal domain (ET). In addition, BRD4 and BRDT also contain an extended C-terminal domain (CTM). BET proteins are important epigenetic "readers" that can specifically bind to N-ε-acetyl-lysine (Kac) residues in histones H3, H4 or non-histone proteins through BD1 and BD2, and participate in transcriptional regulation and chromatin remodeling.

[0008] BRD4 has three characteristic proteins with different C-terminal amino acid residues: a long form (BRD-L) and two short forms (BRD-S), the short forms BRD-S (a) and BRD-S (b) are generated by splicing of other mRNAs. The amino acids 1-719 of the three BRD4s are similar, including two highly conserved N-terminal tandem bromodomains (BD1 and BD2), an N-terminal phosphorylation site cluster (NPS), a basic residue-rich interaction domain (BID), an extra-terminal domain (ET), and a C-terminal phosphorylation site cluster (CPS).

[0009] Mutations in bromodomain-containing proteins have been observed to be associated with cancer and immune and neurological dysfunction. Moreover, recent findings have demonstrated that small molecule inhibition of BRD4 bromodomains may have clinical utility in a variety of human diseases ranging from autoimmunity to cardiac hypertrophy. This is possible because the underlying mechanism lies in transcriptional regulation. Therefore, selective inhibition of bromodomains in this family creates various opportunities as novel therapeutic agents in human dysfunction. Therefore, there is a need for treatment of cancer, immunological disorders and other bromodomain-related diseases, and a need for compounds that can inhibit or degrade bromodomains (such as BRD4). Summary of the invention

[0010] In order to solve the problems existing in the prior art, the present invention provides a compound represented by formula (I) and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt:

[0011]

[0012] in,

[0013] Represents a single bond or a double bond; Indicates that the ring in which it exists is aromatic; m is 1 or 2;

[0014] Ring A is selected from unsubstituted or optionally substituted with one, two or more R a Substituted with the following groups: C 6-10 Aryl, 5-10 membered heteroaryl; each R a are the same or different and are independently selected from halogen, oxo (=O), C 1-12 Alkyl, C 1-12 Alkoxy;

[0015] R1 is selected from unsubstituted or optionally substituted with one, two or more R b Substituted groups: 3-10 membered heterocyclic group, 5-10 membered heteroaryl group, -C(O)OR 11 、-C(O)N(R 12)(R 13 ) or -OC(O)R 14 ; R 11 , R 12 , R 13 , R 14 The same or different, independently selected from H, OH, NH2, C 1-12 Alkyl, C 3-12 Cycloalkyl, 3-10 membered heterocyclyl, 5-10 membered heteroaryl, C 6-10 Aryl, 3-10 membered heterocyclic-C 1-12 Alkyl; each R b The same or different, independently selected from OH, NH2, C 1-12 Alkyl, C 2-12 Alkynyl;

[0016] R2 is selected from C 1-12 Alkyl, C 1-12 Alkoxy, halogenated C 1-12 Alkyl, halogenated C 1-12 Alkoxy;

[0017] R3 and R4 are the same or different and are independently selected from H, C 1-12 Alkyl, C 1-12 Alkoxy, halogenated C 1-12 Alkyl, halogenated C 1-12 Alkoxy;

[0018] when middle When it is a double bond, R5 is selected from =NR 54 ,

[0019] when middle When it is a single bond, R5 is selected from H, halogen, unsubstituted or optionally substituted by one, two or more R c Substituted with the following groups: NH2, C 1-12 Alkyl, -C 1-12 Alkyl-N(R 51 )(R 52 )、-C(O)N(R 51 )(R 52 ),-C 1-12 Alkyl-OR 53 、-C(O)OR 53 , -C 1-12 Alkyl-NH-C(=NR 54 )(N(R 51 )(R 52 ))、-CH=N(R 54 )、-S(O)2R 55、-C(O)R 55 ; R 51 , R 52 , R 53 , R 54 , R 55 The same or different, independently selected from H, OH, NH2, C 1-12 Alkyl, C 1-12 Alkoxy, C 1-12 Alkyl-C(O)-, C 2-12 Alkenyl-C(O)-, C 1-12 Alkyl-S(O)2-; each R c The same or different, independently selected from H, C 1-12 Alkyl-C(O)-, Boc, Boc-O-;

[0020] X is selected from C or N;

[0021] X1 is selected from S or C(R6); R6 is selected from H, C 1-12 Alkyl, C 1-12 Alkoxy;

[0022] X2 is absent or is C(R7); R7 is selected from H, C 1-12 Alkyl, C 1-12 Alkoxy;

[0023] X3 is selected from N or C(R8); R8 is selected from H, C 1-12 Alkyl, C 1-12 Alkoxy;

[0024] L is absent or selected from halogen, NH, unsubstituted or optionally substituted with one, two or more R L Substituted with the following groups: 1-12 Alkyl, C 2-12 Alkenyl, C 2-12 Alkynyl, C 3-10 Cycloalkyl, 3-10 heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl; each R L are the same or different and are independently selected from H, halogen, oxo (=O), C 1-12 Alkyl, C 1-12 Alkoxy.

[0025] According to an embodiment of the present invention, Ring A is selected from unsubstituted or optionally substituted with one, two or more R a Substituted with the following groups: phenyl or

[0026] According to an embodiment of the present invention, ring A is selected from

[0027] According to an embodiment of the present invention, R1 is selected from unsubstituted or optionally substituted with one, two or more R b Substituted with the following groups: C 1-6 Alkyl, 5-8 membered heterocyclic group, 5-6 membered heteroaryl (e.g., thiazolyl, oxazolyl, oxadiazolyl, triazolyl), -C(O)OR 11 、-C(O)N(R 12 )(R 13 ) or -OC(O)R 14 ; R 11 , R 12 , R 13 , R 14 The same or different, independently selected from H, OH, NH2, C 1-6 Alkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, 5-6 membered heteroaryl, C 6-10 Aryl, 3-8 membered heterocyclic group-C 1-6 Alkyl; each R b The same or different, independently selected from OH, NH2, C 1-6 Alkyl, C 2-6 Alkynyl or -OC(O)R 14 ;

[0028] According to an embodiment of the present invention, R 11 , R 12 , R 13 , R 14 are the same or different and are independently selected from H, OH, NH2, methyl, ethyl, isopropyl, tert-butyl, hydroxyphenyl, propynyl, hydroxyethyl, methylcyclopropyl,

[0029] According to an embodiment of the present invention, R1 is selected from

[0030] According to an embodiment of the present invention, R2 is selected from C 1-6 An alkyl group, such as a methyl group.

[0031] According to an embodiment of the present invention, R3 and R4 are the same or different and are independently selected from H, C 1-6 Alkyl, C 1-6 Alkoxy, for example H, methyl, methoxy.

[0032] According to an embodiment of the present invention, when middle When it is a double bond, R5 is selected from =NR 54 ,

[0033] when middle When it is a single bond, R5 is selected from H, halogen, unsubstituted or optionally substituted by one, two or more R c Substituted with the following groups: NH2, C 1-6 Alkyl, -C 1-6 Alkyl-N(R 51 )(R 52 )、-C(O)N(R 51 )(R 52 ),-C 1-6 Alkyl-OR 53 、-C(O)OR 53 , -C 1-6 Alkyl-NH-C(=NR 54 )(N(R 51 )(R 52 ))、-CH=N(R 54 )、-S(O)2R 55 、-C(O)R 55 ; R 51 , R 52 , R 53 , R 54 , R 55 The same or different, independently selected from H, OH, NH2, C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Alkyl-C(O)-, C 2-6 Alkenyl-C(O)-, C 1-6 Alkyl-S(O)2-; each R c The same or different, independently selected from H, C 1-6 Alkyl-C(O)-, Boc, Boc-O-;

[0034] According to an embodiment of the present invention, when middle When it is a double bond, R5 is selected from

[0035] when middle When it is a single bond, R5 is selected from H, Cl, NH2, methyl,

[0036] According to an embodiment of the present invention, X1 is selected from S or C(R6); R6 is selected from H, C 1-6 Alkyl, C 1-6 Alkoxy;

[0037] X2 is absent or is C(R7); R7 is selected from H, C1-6 Alkyl, C 1-6 Alkoxy;

[0038] When X2 does not exist, middle is a single bond, X1 is S; when X2 is C(R7), middle is a double bond, and X1 is C(R6).

[0039] According to an embodiment of the present invention, X1 is selected from S or CH, X2 is absent or is CH; when X2 is absent, middle is a single bond, X1 is S; when X2 is CH, middle is a double bond, and X1 is CH.

[0040] According to an embodiment of the present invention, X3 is selected from N or CH.

[0041] According to an embodiment of the present invention, the Selected from

[0042] According to an embodiment of the present invention, the Selected from

[0043] According to an embodiment of the present invention, the Selected from

[0044] According to an embodiment of the present invention, L is absent or selected from halogen, NH, unsubstituted or optionally substituted by one, two or more R L Substituted with the following groups: 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, 3-8 heterocyclic group, C 6-10 Aryl, 5-6 membered heteroaryl; each R L are the same or different and are independently selected from H, halogen, oxo (=O), C 1-6 Alkyl, C 1-6 Alkoxy;

[0045] According to an embodiment of the present invention, L is absent or selected from Cl, NH, unsubstituted or optionally substituted with one, two or more R L Substituted groups of the following: vinylene, ethynylene, 2,3-dihydroisoxazolylene, pyrazolylene, thiazolylene, furylene, azetidinylene, pyrrolylene, 2-azaspiro[3.3]heptanylene, thienylene, phenylene, cyclopropylene, cyclohexenylene, cyclopentenylene;

[0046] According to an embodiment of the present invention, L is absent or selected from Cl, NH,

[0047] According to an embodiment of the present invention, the compound represented by formula (I) is selected from the following structures:

[0048]

[0049] in, Represents a single bond or a double bond; It means that the ring it exists in is aromatic; Ring A, R1, R2, R3, R4, R5, X1, X2, L, and m have the meanings as described above.

[0050] According to an embodiment of the present invention, the compound represented by formula (I) is selected from the following structures:

[0051]

[0052] Wherein, Ring A, R1, R2, R3, R4, R5, and L have the definitions as described above.

[0053] The present invention also provides a compound represented by formula (III) and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt:

[0054]

[0055] wherein ring A, R1, R2, R3, R4, R5, L, m have the definitions given above, and Y is selected from -(CH2) p -[O-(CH2) q ] r , p is selected from 1, 2, 3, 4 or 5, q is selected from 1, 2, 3, 4 or 5, and r is selected from 0, 1, 2, 3, 4 or 5.

[0056] According to an embodiment of the present invention, exemplary specific compounds of the compound represented by formula (I) are as follows:

[0057]

[0058]

[0059]

[0060]

[0061]

[0062]

[0063]

[0064]

[0065]

[0066]

[0067]

[0068] According to an embodiment of the present invention, exemplary specific compounds of the compound represented by formula (I) are as follows:

[0069]

[0070]

[0071]

[0072]

[0073] According to an embodiment of the present invention, exemplary specific compounds of the compound represented by formula (I) are as follows:

[0074]

[0075]

[0076] According to an embodiment of the present invention, exemplary specific compounds of the compound represented by formula (III) are as follows:

[0077]

[0078] The present invention also provides a pharmaceutical composition, comprising a compound represented by formula (I) and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or a pharmaceutically acceptable salt thereof.

[0079] According to an embodiment of the present invention, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

[0080] According to an embodiment of the present invention, the pharmaceutical composition further comprises an additional biologically active agent.

[0081] The present invention also provides the use of the compound represented by formula (I) and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof and a pharmaceutical composition containing the same in the preparation of a drug for preventing and / or treating a bromodomain-mediated disease or condition.

[0082] According to an embodiment of the present invention, the bromodomain protein is bromodomain protein 4 (BRD4).

[0083] According to an embodiment of the present invention, the disease is cancer.

[0084] According to an embodiment of the present invention, the cancer is selected from the group consisting of acoustic neuroma, acute leukemia, acute lymphocytic leukemia, acute myeloid leukemia, acute T-cell leukemia, basal cell carcinoma, bile duct cancer, bladder cancer, brain cancer, breast cancer, bronchial cancer, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic myeloid leukemia, colon cancer, colorectal cancer, craniopharyngioma, cystic leukemia, Adenocarcinoma, diffuse large B-cell lymphoma, abnormal proliferation, embryonal carcinoma, endometrial carcinoma, endothelial sarcoma, ependymoma, epithelial carcinoma, erythroleukemia, esophageal cancer, estrogen receptor-positive breast cancer, essential thrombocythemia, Ewing's tumor, fibrosarcoma, follicular lymphoma, germ cell testicular cancer, glioma, glioblastoma, gliosarcoma, heavy chain disease, hemangioblastoma, hepatoma, hepatocellular carcinoma, hormone-insensitive prostate cancer, leiomyosarcoma, leukemia, liposarcoma, lung cancer, lymphoma endothelial sarcoma, lymphangiosarcoma, lymphoblastic leukemia, lymphomas (Hodgkin and non-Hodgkin), malignancies and hyperproliferative disorders of the bladder, breast, colon, lung, ovary, pancreas, prostate, skin, and uterus, lymphoid malignancies of T-cell or B-cell origin, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, multiple myeloma, myeloid leukemia, myeloma, myxosarcoma, neuroblastoma, NUT midline carcinoma (NMC), non-small cell Lung cancer, oligodendroglioma, oral cancer, osteogenic sarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pinealoma, polycythemia vera, prostate cancer, colorectal cancer, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, small cell lung cancer, solid tumors (carcinomas and sarcomas), small cell lung cancer, gastric cancer, squamous cell carcinoma, synovioma, sweat gland carcinoma, thyroid cancer, Waldenstrom's macroglobulinemia, testicular tumors, uterine cancer, and Wilms' tumor.

[0085] According to an embodiment of the present invention, the disease is an autoimmune disease or inflammatory disease selected from the group consisting of Addison's disease, acute gout, ankylosing spondylitis, asthma, atherosclerosis, Behcet's disease, bullous dermatosis, chronic obstructive pulmonary disease (COPD), Crohn's disease, dermatitis, eczema, giant cell arteritis, glomerulonephritis, hepatitis, hypophysitis, inflammatory bowel disease, Kawasaki disease, lupus nephritis, multiple sclerosis, myocarditis, myositis, nephritis, organ transplant rejection, osteoarthritis, pancreatitis, pericarditis, polyarteritis nodosa, pneumonia, primary biliary cirrhosis, psoriasis, psoriatic arthritis, rheumatoid arthritis, scleritis, sclerosing cholangitis, sepsis, systemic lupus erythematosus, Takayasu's arteritis, toxic shock, thyroiditis, type I diabetes, ulcerative colitis, uveitis, vitiligo, vasculitis and Wegener's granulomatosis.

[0086] According to an embodiment of the present invention, the disease or condition is selected from the group consisting of: AIDS; chronic kidney disease, diabetic nephropathy, hypertensive nephropathy, HIV-related nephropathy, glomerulonephritis, lupus nephritis, IgA nephropathy, focal segmental glomerulosclerosis, membranous glomerulonephritis, minimal change nephropathy, polycystic kidney disease and tubulointerstitial nephritis; acute kidney injury or disease or condition; obesity; dyslipidemia; hypercholesterolemia; Alzheimer's disease; metabolic syndrome; hepatic steatosis; type II diabetes; insulin resistance; and diabetic retinopathy.

[0087] The present invention also provides a method for degrading a bromodomain-containing protein in a cell, comprising exposing the cell to an effective amount of a compound represented by formula (I) and its racemate, stereoisomer, tautomer, isotope label, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising the same, wherein the compound achieves degradation of the bromodomain-containing protein.

[0088] According to an embodiment of the present invention, the bromodomain-containing protein is bromodomain-containing protein 4 (BRD4).

[0089] The present invention also provides a method for preventing and / or treating BRD4-mediated diseases or conditions, which comprises administering to a patient in need thereof a therapeutically effective amount of a compound represented by formula (I) and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising the same.

[0090] The present invention also provides a compound of formula (I) for preventing and / or treating BRD4-mediated diseases or conditions and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof.

[0091] Beneficial Effects

[0092] The present invention provides a compound represented by formula (I), which has good BRD4 protein degradation effect as a drug and can be used to prepare drugs related to BRD4 inhibition and protein degradation, as well as to prevent and / or treat related diseases.

[0093] Definition and explanation of terms

[0094] Unless otherwise specified, the definitions of groups and terms recorded in the specification and claims of this application, including their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, definitions of specific compounds in examples, etc., can be arbitrarily combined and combined with each other. The definitions of groups and compound structures after such combinations and combinations should be understood to be within the scope of the specification and / or claims of this application.

[0095] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 12 carbon atoms (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12), preferably an alkyl group containing 1 to 6 carbon atoms (C 1-6 Alkyl). Non-limiting examples of alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and various branched chain isomers thereof, etc. The alkyl group may be substituted or unsubstituted.

[0096] The term "alkoxy" refers to -O-(alkyl), wherein alkyl is as defined herein. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, and butoxy. Alkoxy groups can be substituted or unsubstituted.

[0097] The term "alkenyl" is understood to mean a linear or branched monovalent hydrocarbon radical containing from 2 to 12 carbon atoms (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12), preferably an alkyl radical containing from 2 to 6 carbon atoms (C 2-6 It is understood that where the alkenyl group contains more than one double bond, the double bonds may be separated from one another or conjugated. The alkenyl group is, for example, vinyl, allyl, (E)-2-methylvinyl, (Z)-2-methylvinyl, (E)-but-2-enyl, (Z)-but-2-enyl, (E)-but-1-enyl, (Z)-but-1-enyl, pent-4-enyl, (E)-pent-3-enyl, (Z)-pent-3-enyl, (E)-pent-2-enyl, (Z)-pent-2-enyl, (E)- Pent-1-enyl, (Z)-pent-1-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-1-enyl, (Z)-hex-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2-enyl , 2-methylprop-1-enyl, (E)-1-methylprop-1-enyl, (Z)-1-methylprop-1-enyl, 3-methylbut-3-enyl, 2-methylbut-3-enyl, 1-methylbut-3-enyl, 3-methylbut-2-enyl, (E)-2-methylbut-2-enyl, (Z)-2-methylbut-2-enyl, (E)-1-methylbut-2-enyl, (Z)-1-methyl but-2-enyl, (E)-3-methylbut-1-enyl, (Z)-3-methylbut-1-enyl, (E)-2-methylbut-1-enyl, (Z)-2-methylbut-1-enyl, (E)-1-methylbut-1-enyl, (Z)-1-methylbut-1-enyl, 1,1-dimethylprop-2-enyl, 1-ethylprop-1-enyl, 1-propylvinyl, 1-isopropylvinyl.

[0098] The term "alkynyl" is understood to mean a linear or branched monovalent hydrocarbon radical containing one or more triple bonds and having 2 to 12 carbon atoms, preferably "C 2-10 The term "C 2-10 "Alkynyl" is understood to mean preferably a straight or branched monovalent hydrocarbon radical containing one or more triple bonds and having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, for example having 2, 3, 4, 5, 6, 7 or 8 carbon atoms (i.e., "C 2-8 alkynyl”), having 2, 3, 4, 5 or 6 carbon atoms (i.e., “C 2-6 Alkynyl”), having 2 or 3 carbon atoms (“C 2-3The alkynyl 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 In some embodiments, the alkynyl group is ethynyl, prop-1-ynyl or prop-2-ynyl.

[0099] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, and the cycloalkyl ring preferably contains 3 to 12 or 3 to 8 (e.g., 3, 4, 5, 6, 7, and 8) carbon atoms, more preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc.; polycyclic cycloalkyls include spirocyclic, fused ring, and bridged ring cycloalkyls.

[0100] The cycloalkyl ring includes a cycloalkyl group (including monocyclic, spirocyclic, fused and bridged rings) as described herein fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the ring connected to the parent structure is a cycloalkyl group, non-limiting examples include etc.; preferred and The cycloalkyl group may be substituted or unsubstituted.

[0101] The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic substituent containing 3 to 14 ring atoms, one or more of which are heteroatoms selected from nitrogen, oxygen and sulfur, the sulfur being optionally oxo-substituted (i.e. forming a sulfoxide or sulfone), but excluding the ring part of -OO-, -OS- or -SS-, and the remaining ring atoms being carbon. Preferably, it contains 3 to 12 (e.g. 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) ring atoms, of which 1 to 4 (e.g. 1, 2, 3 and 4) are heteroatoms; more preferably, it contains 3 to 8 ring atoms (e.g. 3, 4, 5, 6, 7 and 8), of which 1-3 (e.g. 1, 2 and 3) are heteroatoms; more preferably, it contains 3 to 6 ring atoms, of which 1-3 are heteroatoms; most preferably, it contains 5 or 6 ring atoms, of which 1-3 are heteroatoms. Non-limiting examples of monocyclic heterocyclyls include pyrrolidinyl, tetrahydropyranyl, 1,2,3,6-tetrahydropyridinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc. Polycyclic heterocyclyls include spiro, fused and bridged heterocyclyls.

[0102] The heterocyclyl ring includes a heterocyclyl as described herein (including monocyclic, spiro heterocyclic, fused heterocyclic and bridged heterocyclic) fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring attached to the parent structure is a heterocyclyl, non-limiting examples of which include:

[0103] The heterocyclic group may be substituted or unsubstituted.

[0104] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (fused polycyclic is a ring that shares adjacent carbon atom pairs) group with a conjugated π electron system, preferably 6- to 10-membered, such as phenyl and naphthyl. The aryl ring includes an aryl ring as described herein fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring, and non-limiting examples thereof include:

[0105]

[0106] Aryl groups can be substituted or unsubstituted.

[0107] The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 (e.g., 1, 2, 3, and 4) heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur, and nitrogen. Heteroaryl is preferably 5 to 10 yuan (e.g., 5, 6, 7, 8, 9, or 10 yuan), more preferably 5 yuan or 6 yuan, such as furanyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidyl, pyrazinyl, pyridazinyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, etc. The heteroaryl ring includes a heteroaryl as described herein fused to an aryl, heterocyclyl, or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, non-limiting examples of which include:

[0108]

[0109] A heteroaryl group can be substituted or unsubstituted.

[0110] Those skilled in the art will appreciate that the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl groups include residues derived from a parent ring atom by removing one hydrogen atom, or residues derived from the same or two different ring atoms of the parent by removing two hydrogen atoms, i.e., "alkylene", "alkenylene", "alkynylene", "cycloalkylene", "heterocyclylene", "arylene" and "heteroarylene".

[0111] The term "halogen" refers to F, Cl, Br or I.

[0112] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs or does not occur. For example, "a heterocycloalkyl group optionally substituted with an alkyl group" means that an alkyl group may but need not be present, and the description includes instances where the heterocycloalkyl group is substituted with an alkyl group and instances where the heterocycloalkyl group is not substituted with an alkyl group.

[0113] The term "more" includes 3, 4, 5 and more.

[0114] The pharmaceutically acceptable salts of the compounds of the present invention may be inorganic or organic salts. If these compounds have a basic center, they may form acid addition salts; if these compounds have an acidic center, they may form base addition salts; if these compounds contain both an acidic center (e.g., a carboxyl group) and a basic center (e.g., an amino group), they may also form internal salts.

[0115] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. For example, cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, racemic mixtures and other mixtures, as well as mixtures enriched in enantiomers or diastereomers, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl. All of these isomers and their mixtures are included within the scope of the present invention.

[0116] The compounds and intermediates of the present invention can also exist in different tautomeric forms, and all such forms are included in the scope of the present invention. "Tautomer" refers to structural isomers of different energies that can interconvert via a low energy barrier. For example, proton tautomers (also referred to as prototransfer tautomers) include interconversion via proton migration, such as keto-enol isomerization, imine-enamine isomerization, and lactam-lactim isomerization. All tautomeric forms of all compounds in the present invention are within the scope of the present invention. The name of the compound named in a single manner does not exclude any tautomer.

[0117] The present invention also includes isotopically labeled compounds of the present invention having the same structure as described herein, but with one or more atoms replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as 2 H. 3 H. 11 C. 13 C. 14 C. 13 N. 15 N. 15 O. 17 O. 18 O. 31 P. 32 P. 35 S. 18 F. 123 I. 125 I and 36 All isotopic changes of the compounds of the present invention, whether radioactive or not, are included in the scope of the present invention.

[0118] Unless otherwise stated, when a position is specifically designated as deuterium (D), the position is understood to have deuterium (i.e., at least 10% deuterium incorporation) at least 1000 times greater than the natural abundance of deuterium (which is 0.015%). The natural abundance of the compound in the example may be at least 1000 times greater than deuterium, at least 2000 times greater than deuterium, at least 3000 times greater than deuterium, at least 4000 times greater than deuterium, at least 5000 times greater than deuterium, at least 6000 times greater than deuterium, or more. Each available hydrogen atom connected to a carbon atom may be independently replaced by a deuterium atom. Those skilled in the art can synthesize deuterated compounds with reference to the relevant literature. In preparing deuterated forms of the compounds, commercially available deuterated starting materials may be used, or they may be synthesized using conventional techniques using deuterated reagents including, but not limited to, deuterated borane, trideuterated borane in tetrahydrofuran, deuterated lithium aluminum hydride, deuterated iodoethane, deuterated iodomethane, and the like.

[0119] The term "prodrug" refers to a compound of the present invention that can be converted into a biologically active compound under physiological conditions or by solvolysis. For example, the prodrug of the present invention can be prepared by modifying the functional groups in the compound, and the modification can be removed by conventional operations or in vivo to obtain the parent compound. For example, the prodrug of the present invention includes a compound of the present invention as a drug payload for a conjugated drug (e.g., an ADC drug), nanoparticles, and hydrosols, thereby further preparing the conjugated drug, nanoparticles, and hydrosols.

[0120] The "therapeutically effective amount" of the present invention refers to the amount of active compound or drug that causes a biological or medical response in a tissue, system, animal, individual or human that is sought by researchers, veterinarians, physicians or other clinicians, and includes one or more of the following: (1) Prevention of disease: for example, prevention of disease, disorder or condition in an individual who is susceptible to the disease, disorder or condition but has not yet experienced or developed the disease pathology or symptoms. (2) Suppression of disease: for example, suppression of disease, disorder or condition in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder or condition (i.e., preventing further development of the pathology and / or symptoms). (3) Relief of disease: for example, relief of disease, disorder or condition in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder or condition (i.e., reversing the pathology and / or symptoms). For drugs or pharmacologically active agents, a "therapeutically effective amount" refers to a sufficient amount of a drug or agent that is non-toxic but can achieve the desired effect. The determination of the effective amount varies from person to person, depends on the age and general condition of the recipient, and also depends on the specific active substance. The appropriate effective amount in an individual case can be determined by a person skilled in the art based on routine experiments. DETAILED DESCRIPTION

[0121] The technical scheme of the present invention will be further described in detail below in conjunction with specific embodiments. It should be understood that the following embodiments are only exemplary descriptions and explanations of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are included in the scope that the present invention is intended to protect.

[0122] Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0123] Example 1-1

[0124]

[0125] Compound 4 (1.00 g, 0.44 mmol), compound 5 (0.60 g, 0.66 mmol), Pd2(dba)3 (0.20 g, 0.04 mmol), SPhos (0.09 g, 0.04 mmol) and Cs2CO3 (2.10 g, 1.31 mmol) were mixed in toluene (10 mL) and stirred at 110° C. under N2 for 16 hours. The solvent was removed under vacuum. The residue was purified by flash column chromatography (PE / EA=1 / 1) to obtain yellow oily compound 6 (1.3 g, 98.5%).

[0126] After compound 6 (1.30g, 2.16mmol) and HCl (26mL, 1M in THF) were mixed in THF (50mL), stirred at room temperature for 1 hour. The solvent was removed under vacuum, and the residue was diluted with water (30mL). The pH of the solution was adjusted to 7 by saturated NaHCO3 solution, extracted with EtOAc (30mL×4). The combined organic layers were dried with Na2SO4, filtered, concentrated and purified by chromatography (MeOH / DCM=5%) to obtain compound 7 (770.0mg, 81.6%) as a yellow solid.

[0127] To a solution of t-BuONO (75 mg, 0.74 mmol) in CH3CN (10 mL) was added KI (76 mg, 0.46 mmol) at room temperature. The mixture was stirred and heated at 65 ° C for 2 hours. Then a solution of compound 7 (100 mg, 0.23 mmol) in CH3CN (10 mL) was added, and the mixture was stirred for another 18 hours at 65 ° C. The reaction was quenched with water (15 mL), and the mixture was extracted with EtOAc (20 mL × 3). The organic layer was washed with brine (10 mL × 2), dried over Na2SO4, concentrated and purified by FCC (MeOH / DCM=5%) to give compound 8 (103 mg, 82%) as a yellow solid.

[0128] K2CO3 (1.08 g, 0.008 mol) was added to a DMF (10 mL) solution of compound 1 (1.4 g, 0.006 mol) and compound 2 (1.31 g, 0.011 mol), and the mixture was stirred at room temperature for 4 hours. The solution was diluted with water, and the aqueous phase was extracted with ethyl acetate. The organic layer was washed with brine, dried over MgSO4 and concentrated under reduced pressure to obtain a residue. The residue was purified by FCC (PE / EtOAc=95 / 5) to obtain compound 3 (1 g, 55% yield) as a colorless oil.

[0129] Under microwave irradiation, compound 3 (90 mg, 0.164 mmol), compound 8 (90 mg (0.328 mmol)), Cy2NMe (46 mg, 0.131 mmol), tri-tert-butylphosphine tetrafluoroborate (116 mg, 0.492 mmol) and Pd2(dba)3 (36 mg, 0.025 mmol) were dissolved in NMP (3 mL) solution and stirred at 80°C for 2 hours. The solution was filtered and the filtrate was collected, the solvent was removed under reduced pressure, and the residue was extracted with EA (30 mL). The crude product was purified by FCC (DCM / MeOH=20 / 1) to give yellow solid compound 9 (30 mg, 13.2% yield).

[0130] Compound 9 (30 mg, 0.043 mmol) was dissolved in a mixed solution of HCl in EA / MeOH (1 mL / 1 mL, 1 N). The mixture was stirred at room temperature for 6 hours. The solvent was removed under reduced pressure and the crude product was purified by preparative HPLC (column: Phenomenexluna C18 250 mm×100 mm×10 um; mobile phase: [H2O(0.1% FA)-ACN]; B%: 10%-55%, 40 minutes) to give 9011 (10 mg, 43.16% yield) as a yellow solid.

[0131] LCMS: [M+H] + =494.2;

[0132] 1 H NMR: (400MHz, d4-MeOD) δ7.458(d,J=8.4Hz,2H),7.404(d,J=8.4Hz,2H),6.641(d,J=16.0Hz,1H),6.433(dt,J=15.8,6. 4Hz,1H),4.542(m,1H),3.628(d,J=6.4Hz,2H),3.412(m,2H),2.698(s,3H),2.455(s,3H),1.704(s,3H),1.496(s,9H).

[0133] HPLC: 95.15% purity;

[0134] SFC: Rt = 13.319 min.

[0135] Example 1-2

[0136]

[0137] Compound 2 (75 mg, 1.367 mmol), CuI (17 mg, 0.091 mmol), Pd(PPh3)4 (105 mg, 0.09 mmol) and TEA (277 mg, 2.735 mmol) were added to DMF (7 mL) of compound 1 (500 mg, 0.912 mmol). The resulting mixture was stirred at room temperature under N2 for 2 hours. The mixture was filtered and concentrated under reduced pressure to give a residue. The crude product was directly purified by preparative HPLC (column: Phenomenex moon: C18 250 mm×100 mm×10 um; mobile phase: [ACN-H20 (0.1% FA)]; B%: 5%-45%, 40 minutes) to give GNE-0011 (280 mg, 61.3% yield) as a white solid.

[0138] LCMS: [M+H] + =476.1;

[0139] 1 H NMR: (400MHz, MeOD) δ7.469–7.417 (m, 4H), 4.555 (dd, J = 8.2, 6.2Hz, 1H), 3.722 (s, 2H),3.421–3.301(m,2H),2.693(s,3H),2.453(s,3H),1.694(s,3H),1.491(s,9H).

[0140] HPLC: 99.29% purity;

[0141] SFC: Rt = 2.395 min.

[0142] Examples 1-3

[0143]

[0144] Compound 1 (3 g, 0.007 mol), B2PiN2 (3.35 g, 0.013 mol), Cs2CO3 (6.45 g, 0.020 mol) and XPhos-Pd-G3 (1.12 g, 0.001 mol)) were dissolved in 1,4-dioxane (20 mL) and heated at 80 ° C under N2 for 2 hours. The solvent was removed under reduced pressure to obtain a residue, which was diluted with water (100 mL), extracted with DCM (10 mL×3), washed with brine (100 mL×3), dried over Na2SO4, filtered and concentrated. The crude product was then purified by flash chromatography (MeOH / DCM=7%) to give a yellow solid compound 2 (3.43 g, 5.6 mmol, 85% yield).

[0145] A solution of compound 2 (116 mg, 0.212 mmol), compound 3 (40 mg, 0.212 mmol), Cs2CO3 (138 mg, 0.423 mmol) and Pd(dppf)Cl2 (16 mg, 0.021 mmol) in 1.4-dioxane / H2O=5:1 (1 mL) was stirred at 80°C and N2 for 2 hours. The mixture was diluted with water (10 mL) and extracted with EA (100 mL×3). The organic layer was washed with saturated aqueous sodium chloride solution (50 mL), dried over Na2SO4 and concentrated under reduced pressure. The residue was purified by FCC (DCM / MeOH=20 / 1) to give compound 4 (60 mg, 50.8% yield) as a yellow oil.

[0146] A solution of compound 4 (35 mg, 0.066 mmol), AcOH (0.2 mL) and NH3·H2O (5.62 mg, 0.330 mmol) in MeOH (2 mL) was stirred at 50°C for 1 hour. Then NaBH3CN (21 mg, 0.330 mmol) was added and the resulting mixture was stirred at room temperature for 12 hours. The reaction mixture was concentrated under reduced pressure and the crude product was purified by preparative HPLC (column: Phenomenexluna: C18 250 mm×100 mm×10 um; mobile phase: [H2O (0.1% FA)-ACN]; B%: 20%-60%, 40 minutes) to give 9058 (8 mg, 0.015 mmol, 22.3% yield) as a white solid.

[0147] LCMS: [M+H] + =532.3.

[0148] 1H NMR: (400MHz, MeOD) δ8.541(s,0.6H),7.597(d,J=8.2Hz,2H),7.551(d,J=8.4Hz,2H),6.472(s,1H),4.595(d,J=8.4H z,1H),4.027(s,2H),3.878(s,3H),3.447(t,J=7.2Hz,2H),2.709(s,3H),2.470(s,3H),1.747(s,3H),1.504(s,9H).

[0149] HPLC: 98.95% purity.

[0150] SFC: Rt = 3.023 min.

[0151] Examples 1-4

[0152]

[0153] Under 0°C and N2 conditions, NaH (150 mg, 3.737 mmol) was added to a DMF (5 mL) solution of compound 2 (500 mg, 3.397 mmol). The mixture was stirred at 0°C for 0.5 hours, then compound 1 (444 mg, 3.737 mmol) was added dropwise and stirred for another hour at room temperature. The mixture was then diluted with water (30 mL), extracted with EtOAc (30 mL×2), washed with brine (50 mL×5), dried with Na2SO4 and concentrated under reduced pressure to give compound 3 (550 mg, crude product) as a colorless oil, which was used in the next step without further purification.

[0154] A DMF (2 mL) solution of compound 4 (100 mg, 0.182 mmol), compound 3 (68 mg, 0.365 mmol), CuI (10 mg, 0.055 mmol), Pd (PPh 3 ) 4 (21 mg, 0.018 mmol) and TEA (55 mg, 0.547 mmol) was stirred at room temperature for 1 hour. The solution was diluted with water (10 mL) and extracted with EA (10 mL×3). The organic layer was washed with brine (10 mL×5), dried over Na 2 SO 4 and concentrated. The crude product was purified by FCC (eluent MeOH in DCM=0-10%) to give compound 5 (60 mg, 0.079 mmol, 43.39% yield) as a yellow solid.

[0155] Compound 5 (60 mg, 0.082 mmol) in MeOH (1 mL) was added with 2N HCl / EA (1 ml) solution, and the mixture was stirred at room temperature for 2 hours. The solvent was then removed by N2. The residue was purified by preparative-HPLC (column: Phenomenexluna C18 250 mm×100 mm×10 um; mobile phase: [water (0.05% FA)-ACN]; B%: 5%-50%, 30 minutes) to give a white solid 9064 (13.94 mg, 0.028 mmol, 9.2% yield, 96.08% purity).

[0156] LCMS: [M+H] + =506.3;

[0157] 1 H NMR: (400MHz, CD3OD) δ7.434 (dd, J=8.4Hz, 4.0Hz, 4H), 4.570–4.535 (m, 1H), 3.833 (s, 2H), 3 .574(s,3H),3.441–3.408(m,2H),2.693(s,3H),2.452(s,3H),1.696(s,3H),1.491(s,9H).

[0158] HPLC: 96.08% purity;

[0159] SFC: Rt = 5.559 min.

[0160] Examples 1-5

[0161]

[0162] At 0°C, triethylamine (46 mg, 0.457 mmol) was added to a solution of compound 1 (150 mg, 0.305 mmol) in DCM (5 mL), and then compound 2 (46 mg, 0.335 mol) was added dropwise. The resulting solution was warmed to room temperature and stirred for 30 minutes. The mixture was diluted with DCM (20 mL), washed with brine (20 ml), dried over anhydrous Na2SO4, and concentrated under vacuum to give compound 3 (160 mg, crude product) as a yellow oil.

[0163] DIBAL-H (0.81 mmol) was added dropwise to a solution of compound 3 (160 mg, 0.270 mmol) in THF (3 mL) at -78 ° C under N2. The reaction mixture was stirred at -78 ° C for 0.5 hours, then warmed to 0 ° C and stirred for another 1.5 hours. Saturated MeOH aqueous solution (3 mL) was added. The solvent was removed under reduced pressure, and the residue was diluted with water (10 mL) and extracted with EA (10 mL×3). The organic layer was dried over Na2SO4 and concentrated under reduced pressure, and the residue was purified by FCC (DCM / MeOH=10 / 1) to obtain yellow oily compound 4 (120 mg, 65.0% yield).

[0164] Pyridine (30 mg, 0.376 mmol) and compound 5 (45 mg, 0.376 mm mol) were added to DCM (5 mL) of compound 4 (120 mg, 0.251 mmol) and added dropwise at 0 ° C under N2. The reaction mixture was stirred at 0 ° C for 3 hours and quenched with H2O (10 mL), and the solution was extracted with DCM (10 mL×2). The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The residue was purified by FCC (DCM / MeOH=20:1) to give compound 6 (70 mg, 44.6% yield) as a yellow oil.

[0165] At room temperature and N2 atmosphere, compound 6 (70 mg, 0.125 mmol), compound 7 (21 mg, 0.374 mmol), Pd (PPh3) 4 (29 mg, 0.025 mmol), TEA (38 mg, 0.374 mm) and CuI (5 mg, 0.025 mmol) were stirred in DMF (1 mL) for 1 hour. The mixture was filtered and the filtrate was collected and analyzed by preparative HPLC (column: Phenomenex moon C18 250 mm × 100 mm × 10 um; mobile phase: [H2O (0.1% FA) -ACN]; B%: 5%-40%, 40 minutes) to give a yellow solid 9091 (9 mg, 14.1% yield).

[0166] LCMS: [M+H] + =490.2;

[0167] 1 H NMR: (400MHz, MeOD) δ7.490(s,4H),4.692–4.623(m,1H),4.432–4.365(m,1H),4.192(dd,J=9.8,4.8 Hz,1H),3.862(s,2H),2.893–2.726(m,2H),2.693(s,3H),2.437(s,3H),1.670(s,3H),1.088(s,9H).

[0168] HPLC: 99.2% purity;

[0169] SFC: Rt = 2.755 min.

[0170] Examples 1-6

[0171]

[0172] At room temperature and N2 atmosphere, compound 1 (300 mg, 0.547 mmol) was dissolved in DCM / TFA=2:1 (3 mL) and stirred for 3 hours. The mixture was diluted with water (10 mL) and extracted with DCM (10 mL×3). The organic layer was collected, dried over Na2SO4 and concentrated to give brown solid compound 2 (250 mg, 0.46 mmol, 83.55% yield).

[0173] At room temperature and N2 atmosphere, compound 2 (300mg, 0.609mmol), EDCI (155mg, 0.810mmol) and DMAP (6mg, 0.049mmol) were stirred in DCM (3mL). After 10 minutes, compound 3 (51mg, 0.701mmol) was added dropwise. The reaction mixture was stirred at room temperature for 15 hours. The mixture was diluted with water (10mL) and extracted with DCM (10mL×3). The organic layer was collected, dried and concentrated with Na2SO4. The crude product was then purified by flash chromatography (PE / EA=70:30) to obtain yellow solid compound 4 (100mg, 0.16mmol, 27% yield).

[0174] Compound 4 (90 mg, 0.165 mmol), compound 5 (11 mg, 0.198 mmol), Pd(PPh3)4 (38 mg, 0.033 mmol), CuI (6 mg, 0.032 mmol) and TEA (50 mg, 0.494 mmol) were stirred in DMF (1 mL) at room temperature and N2 atmosphere for 1 hour. The mixture was diluted with MeOH, filtered and purified by preparative HPLC (column: Phenomenex moon C18 250 mm×100 mm×10 um; mobile phase: [H2O(0.1% FA)-ACN]; B%: 20%-60%, 30 minutes) to give 9103 (15 mg, 0.03 mmol, 18.3% yield) as a white solid.

[0175] LCMS: [M+H] + =474.2;

[0176] 1H NMR: (400MHz, CDCl3) δ7.391 (t, J=10.2Hz, 4H), 4.584 (t, J=7.2Hz, 1H), 3.939 (s, 2H), 3.550 (d, J= 6.8Hz,2H),2.665(s,3H),2.400(s,3H),1.654(s,3H),1.559(s,3H),0.914(s,2H),0.656(s,2H).

[0177] HPLC: 99.55% purity;

[0178] SFC: Rt = 7.007 min.

[0179] Examples 1-7

[0180]

[0181] At room temperature and N2 atmosphere, compound 1 (250 mg, 0.456 mmol) was stirred in DCM / TFA=2:1 (3 mL) for 2 hours. The mixture was diluted with water (10 mL) and extracted with DCM (10 mL×3). The organic layer was collected, dried over Na2SO4 and concentrated to give compound 2 (200 mg, crude product) as a yellow oil.

[0182] At 70 ° C and N2 atmosphere, compound 2 (200 mg, 0.406 mmol), compound 3 (51 mg, 0.701 mmol), Bu4NI (15 mg, 0.041 mmol) and K2CO3 (112 mg, 0.812 mmol) were stirred in DMF (3 mL) for 15 hours. The mixture was diluted with water (10 mL) and extracted with EA (10 mL × 3). The organic layer was collected, dried over Na2SO4 and concentrated. The crude product was then purified by flash chromatography (DCM / MeOH=10:1) to obtain yellow oily compound 4 (100 mg, 0.156 mmol, 38.4% yield).

[0183] Compound 4 (90 mg, 0.156 mmol), compound 5 (17 mg, 0.312 mmol), Pd(PPh3)4 (36 mg, 0.031 mmol), CuI (6 mg, 0.031 mmmol) and TEA (50 mg, 0.46 mmol) were stirred in DMF (1 mL) for 2 h at room temperature and in a N2 atmosphere. The mixture was diluted with MeOH (3 mL) and purified by preparative HPLC (column: Phenomenex moon C18 250 mm×100 mm×10 um; mobile phase: [H2O(0.1% FA)-ACN]; B%: 10%-50%, 40 min) to give a yellow solid 9112 (15 mg, 14.6% yield).

[0184] LCMS: [M+H] + =504.1;

[0185] 1 H NMR: (400MHz, MeOD) δ8.525(s,0.8H),7.520–7.438(m,4H),4.675(t,J=7.2Hz,1H),4.502(dd,J=14.4,6.0Hz,2H),4.345(dd,J =6.0,2.0Hz,2H),4.249(s,2H),3.915(s,2H),3.591(d,J=7.4Hz,2H),2.691(s,3H),2.450(s,3H),1.685(s,3H),1.323(s,3H).

[0186] HPLC: 98.7% purity;

[0187] SFC: Rt = 5.801 min.

[0188] To a solution of 9112 (50 mg, 0.099 mmol) in anhydrous DCM (2 mL) was added a solution of BF3·Et2O (0.1 mL, 46.5%) in anhydrous DCM (1 mL) dropwise at -10°C and N2 atmosphere, and the reaction mixture was stirred at -10°C for 3 hours. The reaction was then quenched with trimethylamine. The mixture was concentrated and dissolved with MeOH, and then purified by preparative HPLC (column: Phenomenex lunaC18 250 mm×100 mm×10 um; mobile phase: [H2O (0.1% TFA)-ACN]; B%: 5%-40%, 40 minutes) to obtain 9113 as a TFA salt. HCl (2 drops, 1 M) was then added to the solution to give M019113 (15 mg, 28.5% yield) as a yellow solid.

[0189] LCMS: [M+H]+ =504.1;

[0190] 1 H NMR: (400MHz, MeOD) δ7.565 (s, 4H), 4.865 (d, J = 6.4Hz, 1H), 4.171–4.052 (m, 4H), 3.653–3. 545(m,2H),3.492–3.413(m,4H),2.930(s,3H),2.507(s,3H),1.726(s,3H),0.911(s,3H).

[0191] HPLC: 95.1% purity;

[0192] SFC: Rt = 3.066 min.

[0193]

[0194] 9112Int 2 (400 mg, 0.813 mmol), NH4Cl (130 mg, 2.438 mmol), HATU (618 mg, 1.625 mmol) and DIEA (525 mg, 4.063 mmol) were stirred in DMF (10 mL) at room temperature for 2 hours. The mixture was diluted with water (10 mL) and extracted with EA (10 mL×3). The combined organic layers were dried over Na2SO4 and concentrated. The crude product was purified by FCC (DCM / MeOH=10:1) to give yellow solid compound 1 (300 mg, 0.550 mmol, 67.63% yield).

[0195] Compound 1 (300 mg, 0.611 mmol) and Lawesson's reagent (494 mg, 1.221 mmol) were stirred in THF (10 mL) and heated at 40 ° C for 2 hours. The mixture was then diluted with water (10 mL) and extracted with EtOAc (10 mL×3). The combined organic layers were dried over Na2SO4 and concentrated. The crude product was purified by FCC (DCM / MeOH=10:1) to give a yellow solid compound 2 (250 mg, 0.443 mmol, 72.62% yield).

[0196] Compound 2 (250 mg, 0.493 mmol), compound 3 (125 mg, 0.739 mmol) and TsOH (17 mg, 0.099 mmol) were stirred in AcOH (5 mL) and heated at 50 ° C for 2 hours. The mixture was then diluted with water (5 mL) and extracted with EtOAc (5 mL×3). The combined organic layers were dried over Na2SO4 and concentrated. The crude product was purified by FCC (DCM / MeOH=10:1) to give yellow solid compound 4 (150 mg, 0.254 mmol, 51.55% yield).

[0197] Compound 4 (30 mg, 0.057 mmol), compound 5 (9 mg, 0.170 mmol), Pd(PPh3)4 (13 mg, 0.011 mmol), CuI (2 mg, 0.011 mg) and TEA (17 mg, 0.170 mol) were stirred in DMF (3 mL) at room temperature and N2 atmosphere for 2 hours. The mixture was diluted with MeOH, filtered and purified by preparative HPLC (column: Phenomenex moon C18 250 mm×100 mm×10 um; mobile phase: [H2O(0.1% FA)-ACN]; B%: 20%-60%, 30 minutes) to give 9135 (5 mg, 0.010 mmol, 95% yield) as a white solid.

[0198] LCMS: [M+H] + =459.1;

[0199] 1 H NMR: (400MHz, MeOD) δ7.723(d,J=3.4Hz,1H),7.639(d,J=3.4Hz,1H),7.446(d,J=6.6Hz,4H),4.569(t, J=6.8Hz,1H),4.064(dd,J=13.6,10.4Hz,2H),3.384(s,2H),2.605(s,3H),2.399(s,3H),1.597(s,3H).

[0200] HPLC: 95.86% purity;

[0201] SFC: Rt = 3.550 min.

[0202] Examples 1-9

[0203]

[0204] A solution of compound 1 (1 g, 0.002 mol) in MeOH / H2SO4=20:1 (20 mL) was stirred at 60°C and N2 for 16 hours. The mixture was evaporated and purified by flash chromatography (DCM / MeOH=98 / 2) to give compound 2 (0.89 g, yield 83.33%) as a yellow solid.

[0205] To a solution of compound 2 (890 mg, 1.758 mmol) in MeOH (20 mL) was added NH2NH2·H2O (879 mg, 17.576 mmol) dropwise. The reaction mixture was stirred and heated at 50°C for 5 hours. The solution was evaporated and purified by flash chromatography (DCM / MeOH=95 / 5) to give compound 3 (800 mg, yield 85.39%) as a yellow solid.

[0206] Compound 4 (168 mg, 1.580 mmol) was added dropwise to a solution of compound 3 (800 mg, 1.580 mmol) and DIEA (408 mg, 3.160 mmol) in DCM (50 mL) at room temperature. The reaction mixture was stirred at room temperature for 2 hours. The solution was evaporated and purified by flash chromatography (DCM / MeOH=96 / 4) to give compound 5 (700 mg, 73.02% yield) as a yellow solid.

[0207] A solution of compound 5 (700 mg, 1.214 mmol) in PPA (60 mL) was heated to 110° C. and stirred at 110° C. for 2 hours. The reactant was quenched with H2O (50 mL) at 25° C., diluted with EA (300 mL), washed with water and brine, dried over Na2SO4, filtered and concentrated. The mixture was evaporated and purified by flash chromatography (DCM / MeOH=95 / 5) to give compound 6 (500 mg, 70.05% yield) as a yellow solid.

[0208] Compound 6 [50 mg, 0.089 mmol], compound 7 (6 mg, 0.107 mmol), Pd (PPh 3) 4 (21 mg, 0.018 mmol), TEA (18 mg, 0.179 mmol) and copper (I) iodide were stirred in DMF (10 mL) at room temperature for 3 hours. The mixture was directly purified by prep-HPLC (chromatographic column: Phenomenex luna C18 250 mm × 100 mm × 10 um; mobile phase: [H 2 O (0.1% FA) -ACN]; B%: 10% to 50%, 40 minutes) to obtain a white solid 9160 (25 mg, yield 54.64%).

[0209] LCMS: [M+H] + =486.2;

[0210] 1 H NMR: (400MHz, MeOD) δ7.481-7.460 (d, J=8.4Hz, 2H), 7.421-7.400 (d, J=8.4Hz, 2H), 4.819-4.783 (m, 1H), 4.056-3 .999(m,2H),3.887(s,2H),3.298-3.203(m,1H),2.692(s,3H),2.453(s,3H),1.679(s,3H),1.402-1.379(m,3H).

[0211] HPLC: 97.45% purity.

[0212] Examples 1-10

[0213]

[0214] 9135Int 4 (1.5 g, 0.003 mol), 9167-Int6 (1.79 g, 0.006 mol), Pd (dppf) Cl2 (0.41 g, 0.5 mmol) and TEA (0.85 g, 0.008 mol) were heated to 80 ° C in 1,4-dioxane / H2O = 4 / 1 (15 mL) and stirred at 80 ° C for 2 hours under N2. The mixture was diluted with H2O (10 mL), extracted with EtOAc (20 mL × 3), washed with brine (20 mL), and dried over anhydrous sodium sulfate. The organic layer was concentrated under reduced pressure to give a crude product. The residue was purified by flash chromatography (eluent MeOH in DCM = 0-5%) to give a yellow solid compound 6 (600 mg, 0.80 mmol, 30% yield).

[0215] At room temperature and N2 atmosphere, compound 1 (600 mg, 0.884 mmol) was stirred in EA / MeOH (10 mL, 2M) solution of HCl for 6 hours. The solvent was removed under N2 flow to obtain a crude product. The residue was purified by reverse phase column (eluent ACN in H2O=5-30%, 0.1% TFA) to obtain a yellow solid 9166 (300 mg, 0.598 mmol, 68% yield).

[0216] LCMS: [M+H] + =477.10.

[0217] 1H NMR: (400MHz, MeOD) δ7.755(d,J=3.6Hz,1H),7.562(d,J=3.6Hz,5H),7.525(d,J=8.4Hz,2H),7.475(d,J=8.4Hz,2H),6.931(d,J=16.0Hz,1H),6.421– 6.305(m,1H),4.661(dd,J=8.8,5.2Hz,1H),4.196(ddd,J=20.4,15.2,7.0 Hz, 2H), 4.018 (d, J = 7.2Hz, 2H), 2.711 (s, 3H), 2.439 (s, 3H), 1.659 (s, 3H).

[0218] HPLC: 95.6% purity.

[0219] Examples 1-11

[0220]

[0221] Compound 2 (178 mg, 0.985 mmol), Cs2CO3 (642 mg, 1.97) and XPhos Pd G3 (55.6 mg, 0.0657 mmol) were added to a solution of compound 1 (300 mg, 0.657 mmol) in 1,4-dioxane (10 mL). The reaction mixture was stirred at 100 ° C, N2 for 8 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with EtOAc in petroleum ether from 0% to 50% to give compound 3 (300 mg, yield 72%) as a yellow solid. LC-MS: (ESI) m / z [M+H] + 602.2.

[0222] 1M HCl (1.0 mL) was added dropwise to a solution of compound 3 (300 mg, 0.499 mmol) in THF (5.0 mL) at 25°C and stirred. The resulting mixture was stirred at 25°C for 30 minutes. The reactant was concentrated under reduced pressure and purified by reverse phase chromatography (ACN / water (0.1% NH3HCO3)) to give compound 4 (150 mg, yield 69%) as a white solid. LC-MS: (ESI) m / z [M+H] + 438.2.

[0223] To a solution of compound 4 (60.0 mg, 0.137 mmol) in DCM (6.0 mL) was added TEA (41.6 mg, 0.411 mmol) and prop-2-enoyl chloride (24.8 mg, 0.274 mmol). The reaction mixture was stirred at 25 ° C for 10 minutes. The mixture was concentrated under reduced pressure and the residue was purified by Prep-HPLC (CH3CN from 45% to 75% in 10 minutes) to give compound 9230 as a white solid.

[0224] 1 H NMR (400MHz, DMSO-d6) δ10.40(s,1H),7.71(d,J=8.4Hz,2H),7.39(d,J=8.4Hz,2H),6.50–6.40(m,1H),6.33–6.24(m,1H),5. 82–5.73(m,1H),4.42–4.36(m,1H),3.33–3.28(m,2H),2.60(s,3H),2.42(s,3H),1.67(s,3H),1.43(s,9H).LC-MS:m / z[M+H] + 492.2.

[0225] Examples 1-12

[0226]

[0227] To a solution of compound 1 (60.0 mg, 0.137 mmol) in DCM (6.0 mL) was added TEA (41.6 mg, 0.411 mmol) and 2-chloroethanesulfonyl chloride (44.7 mg, 0.274 mmol). The reaction mixture was stirred at 25 ° C for 10 minutes. The mixture was concentrated under reduced pressure and the residue was purified by Prep-HPLC (CH3CN from 45% to 75% in 10 minutes) to give compound 9231 as a white solid.

[0228] 1 H NMR (400MHz, DMSO-d6) δ10.40(s,1H),7.71(d,J=8.4Hz,2H),7.39(d,J=8.4Hz,2H),6.51–6.38(m,1H),6.33–6.22(m,1H),5. 82–5.74(m,1H),4.44–4.34(m,1H),3.33–3.28(m,2H),2.60(s,3H),2.43(s,3H),1.67(s,3H),1.43(s,9H).LC-MS:m / z[M+H] + 528.2.

[0229] Examples 1-13

[0230]

[0231] At 25°C, 1-bromo-2-propanone (4281 mg, 31.25 mmol) and CF3SO3Ag (3212 mg, 12.50 mmol) were added to a solution of compound 1 (2500 mg, 6.25 mmol) in toluene (10 mL). After addition, the mixture was stirred at 90°C for 2 hours. The reaction mixture was purified by prep-HPLC (CH3CN from 10% to 70% in 10 minutes) to give compound 3 (100 mg, yield 3%) as a yellow solid. LC-MS: (ESI) m / z [M+H] + 438.1.

[0232] At 25°C and N2, a solution of compound 3 (100 mg, 0.22 mmol), X-phos-Pd G3 (19 mg, 0.02 mmol), compound 4 (137 mg, 0.34 mmol) and K3PO4 (145 mg, 0.68 mmol) in dioxane (1 mL) and H2O (0.1 mL) was mixed and stirred. The mixture was heated to 100°C and stirred at this temperature for 1 hour. The reaction mixture was cooled to 25°C, diluted with water (10 mL) and extracted with EtOAc (20 mL×2). The combined organic phases were dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting from 0% to 80% with EtOAc / petroleum ether in 15 minutes to give compound 5 (50 mg, yield 29%) as a yellow solid. LC-MS: (ESI) m / z[M+H] + 675.2.

[0233] To a solution of compound 5 (50 mg, 0.07 mmol) in EA (1 mL) was added HCl / EA (1 mL) at 25° C. After addition, the mixture was stirred at 25° C. for 1 hour. The reaction mixture was purified by prep-HPLC (CH3CN from 10% to 70% in 10 minutes) to give compound 9234 (4.2 mg, yield 11%) as a white solid.

[0234] 1H NMR (400MHz, DMSO-d6) δ11.80(s,2H),10.97(s,1H),7.75(s,1H),7.55–7.39(m,4H),6.86–6.82(m,1H),6.42–6.40(m, 1H),4.77–4.473(m,1H),3.88–3.82(m,4H),2.68(s,3H),2.49(s,3H),2.03(s,3H),1.61(s,3H).LC-MS:(ESI)m / z[M+H] + 475.1.

[0235] Examples 1-14

[0236]

[0237] A solution of compound 1 (3.6 g, 8.2 mmol), compound 2 (4.9 g, 12.3 mmol), X-phos-Pd G3 (690 mg, 0.8 mmol) and K3PO4 (5.22 g, 24.6 mmol) in 1,4-dioxane / water (44 mL) was heated in a microwave at 100 ° C for 1 hour. LCMS showed that the starting material was consumed and the desired product was formed. The mixture was diluted with H2O (50 mL), extracted with EtOAc (50 mL×3), washed with brine (50 mL), and dried over anhydrous sodium sulfate. The organic layer was concentrated under reduced pressure to give a crude product. The residue was purified by flash chromatography (eluent MeOH in DCM = 0-10%) to give compound 9235 (2.5 g, 3.7 mmol, yield 45%) as a yellow solid.

[0238] 1 H NMR (400MHz, DMSO-d6) δ7.71–7.70(m,1H),7.63–7.62(m,1H),7.50–7.40(m,4H),6.63–6.59(m,1H),6.37–6.32(m,1H),4.54–4.53(m,1H) ),4.30(s,2H),4.08–4.06(m,2H),2.60(s,3H),2.39(s,3H),1.58(s,3H),1.41–1.38(m,18H),1.75–1.50(m,6H).LC-MS: (ESI)m / z[M+H] + 677.2.

[0239] Examples 1-15

[0240]

[0241] At 25 ° C, compound 1 (400 mg, 0.875 mmol) and compound 2 (292 mg, 1.31 mmol) were added to a solution of 1,4-dioxane (10 mL) and H2O (1 mL) with K2CO3 (363 mg, 2.63 mmol) and Xphos Pd G3 (74.1 mg, 0.0875 mmol). The reaction mixture was stirred at 100 ° C, N2 for 6 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with THF in petroleum ether from 0% to 65% to obtain compound 9241 (410 mg, yield 86%) as a yellow solid.

[0242] 1 H NMR (400MHz, DMSO-d6) δ7.74(d,J=8.4Hz,2H),7.48(d,J=8.4Hz,2H),6.42(s,1H),4.50–4.40(m,1H),3.35–3.30(m,2H),2.81– 2.71(m,2H),2.63(s,3H),2.44(s,3H),2.41–2.33(m,2H),2.12–2.00(m,2H),1.64(s,3H),1.44(s,9H).LC-MS:(ESI)m / z[M+H] + 517.2.

[0243] Hydroxylamine hydrochloride (12.1 mg, 0.174 mmol) and TEA (17.6 mg, 0.174 mmol) were added to a solution of EtOH (6.0 mL) of compound 9241 (60.0 mg, 0.116 mmol). After stirring at 80 ° C for 3 hours, sodium cyanoborohydride (36.5 mg, 0.581 mmol) was added at 25 ° C. The mixture was then stirred for 12 hours at 25 ° C. The mixture was quenched with water (20 mL) and then extracted with ethyl acetate (30 mL x 2). The combined organic phases were dried over sodium sulfate and filtered. The mixture was concentrated under reduced pressure, and the residue was purified by Prep-HPLC (CH3CN from 30% to 40% in 8 minutes) to obtain compound 9236 (3.80 mg, yield 6.0%) as a white solid.

[0244] 1H NMR (400MHz, DMSO-d6) δ10.86(s,1H),7.37(d,J=8.4Hz,2H),7.06(d,J=8.4Hz,2H),6.46(s,1H),5.31(s,1H),4.17(s,1H),3.26-3.17(m,1H ),3.09-3.96(m,1H),2.78-2.69(m,1H),2.49-2.41(m,7H),2.34(s,3H),1.87(s,3H),1.81-1.72(m,2H),1.42(s,9H).LC-MS:(ESI)m / z[M+H] + 543.2.

[0245] Examples 1-16

[0246]

[0247] At 25 ° C, compound 1 (400 mg, 0.875 mmol) and compound 2 (273 mg, 1.31 mmol) were added to a solution of 1,4-dioxane (10 mL) and H2O (1 mL) with K2CO3 (363 mg, 2.63 mmol) and Xphos Pd G3 (74.1 mg, 0.0875 mmol). The reaction mixture was stirred at 100 ° C, N2 for 6 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with THF in petroleum ether from 0% to 45% to obtain compound 9242 (390 mg, yield 84%) as a yellow solid.

[0248] 1 H NMR (400MHz, DMSO-d6) δ7.87(d,J=8.4Hz,2H),7.51(d,J=8.4Hz,2H),6.80(s,1H),4.52–4.38(m,1H),3.37–3.35(m,2 H),3.08–2.97(m,2H),2.62(s,3H),2.49–2.45(m,2H),2.43(s,3H),1.64(s,3H),1.44(s,9H).LC-MS:(ESI)m / z[M+H] + 503.2.

[0249] Hydroxylamine hydrochloride (22.1 mg, 0.318 mmol) and TEA (48.3 mg, 0.477 mmol) were added to a solution of EtOH (6.0 mL) of compound 9242 (80.0 mg, 0.159 mmol). After stirring for 3 hours at 80 ° C, NaCNBH (50.0 mg, 0.796 mmol) was added at 25 ° C. The mixture was then stirred for 12 hours at 25 ° C. The mixture was quenched with water (20 mL) and then extracted with ethyl acetate (30 mL x 2). The combined organic phases were dried over sodium sulfate and filtered. The mixture was concentrated under reduced pressure, and the residue was purified by Prep-HPLC (CH CN from 30% to 40% in 8 minutes) to obtain compound 9237 (14.0 mg, yield 17%), which was a white solid.

[0250] 1 H NMR (400MHz, DMSO-d6) δ10.48(s,1H),7.45(d,J=8.4Hz,2H),7.07(d,J=8.4Hz,2 H),6.72(s,1H),5.31(d,J=8.0Hz,1H),4.23–4.12(m,1H),3.31–3.24(m,1H),3. 05(dd,J=16.4,7.2Hz,1H),2.81–2.76(m,2H),2.76–2.69(m,1H),2.67–2.60(m, 2H),2.42(s,3H),2.35(s,3H),1.88(s,3H),1.41(s,9H).LC-MS:(ESI)m / z[M+H] + 520.2.

[0251] Examples 1-17

[0252]

[0253] To a solution of compound 1 (400 mg, 0.875 mmol) and compound 2 (359 mg, 1.40 mmol) in 1,4-dioxane (10 mL) was added H2O (1 mL) K3PO4 (557 mg, 2.63 mmol) and Xphos Pd G3 (74.1 mg, 0.0875 mmol) at 25 °C. The reaction mixture was stirred at 100 °C under N2 for 1 hour. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with THF in petroleum ether from 0% to 65% to give compound 3 (350 mg, yield 76%) as a yellow solid. LC-MS: (ESI) m / z [M+H] + 477.2.

[0254] N-methylhydroxylamine hydrochloride (28.1mg, 0.336mmol) and TEA (34.0mg, 0.336mmol) were added to a solution of MeOH (6.0mL) of compound 3 (80.0mg, 0.168mmol). After stirring at 25°C for 30 minutes, NaBH3CN (106mg, 1.68mmol) was added at 25°C. The mixture was then stirred for 12 hours at 25°C. The mixture was quenched with water (20mL) and then extracted with ethyl acetate (30mL x 2). The combined organic phases were dried over sodium sulfate and filtered. The mixture was concentrated under reduced pressure, and the residue was purified by Prep-HPLC (CH3CN from 30% to 40% in 8 minutes) to obtain compound 9240 (14.0mg, yield 16%), which was a white solid.

[0255] 1 H NMR(400MHz,DMSO-d6)δ7.95(s,1H),7.51–7.44(m,2H),7.42–7.33(m,2H),6.62–6.50(m,1H),6.47–6.38(m,1H),4.48– 4.31(m,1H),3.34–3.30(m,4H),2.60(s,3H),2.50(s,3H),2.42(s,3H),1.65(s,3H),1.43(s,9H).LC-MS:(ESI)m / z[M+H] + 508.2.

[0256] Examples 1-18

[0257]

[0258] Compound 1 (500 mg, 1.24 mmol), compound 2 (750 mg, 1.87 mmol), X-phos-Pd G3 (105 mg, 0.12 mmol) and K3PO4 (1323 mg, 6.23 mmol) were dissolved in 1,4-dioxane / H2O (11 mL) and heated at 100 °C for 1 hour. LCMS showed that the starting material was consumed and the desired product was formed. The mixture was diluted with H2O (50 mL), extracted with EtOAc (50 mL×3), washed with brine (50 mL), and dried over anhydrous sodium sulfate. The organic layer was concentrated under reduced pressure to give a crude product. The residue was purified by flash chromatography (eluent MeOH in DCM = 0-10%) to give compound 3 (350 mg, yield 39%) as a yellow solid. LC-MS: (ESI) m / z[M+H] + 638.3.

[0259] Compound 4 (78 mg, 1.0551 mmol), HATU (401 mg, 1.05 mmol) and DIEA (272 mg, 2.11 mmol) were added to a solution of compound 3 (450 mg, 0.70 mmol) in DCM (10 mL). After addition, the mixture was stirred at 25 ° C for 1 hour. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting from 0% to 90% with EtOAc / petroleum ether and EtOAc in 15 minutes to give compound 5 (400 mg, yield 73%) as a yellow solid. LC-MS: (ESI) m / z [M+H] + 694.3.

[0260] A solution of compound 5 (400 mg, 0.57 mmol) in toluene (10 mL) was stirred at 130 °C for 1 hour. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with EtOAc / petroleum ether and EtOAc from 0% to 90% in 15 minutes to give compound 6 (250 mg, yield 61%) as a yellow solid. LC-MS: (ESI) m / z [M+H] + 676.2.

[0261] A solution of compound 6 (130 mg, 0.19 mmol) in FA (2 mL) was stirred at 25 °C for 1 hour. The reaction mixture was concentrated under reduced pressure. The reaction mixture was purified by prep-HPLC (CH3CN from 0% to 100% in 10 minutes) to give compound 9243 (10 mg, yield 10%) as a white solid.

[0262] 1 H NMR (400MHz, DMSO-d6) δ7.46–7.44(m,2H),7.34–7.32(m,2H),6.54–6.50(m,1H),6.44–6.37(m,1H),4.73–4.69(m,1 H),4.01–3.94(m,2H),3.51–3.49(m,2H),2.62(s,3H),2.42(s,3H),2.31(s,3H),1.63(s,3H).LC-MS:(ESI)m / z[M+H] + 476.2.

[0263] Examples 1-19

[0264]

[0265] At 25°C, H2SO4 (20 mL) was added to a solution of compound 1 (1 g, 2.10 mmol) in MeOH (20 mL). After addition, the mixture was stirred at 60°C for 16 hours. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (100 mL×2). The combined organic phases were dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to give a yellow solid compound 2 (800 mg, yield 79%). LC-MS: (ESI) m / z[M+H] + 415.0.

[0266] At 25°C, N2H4.H2O (772 mg, 24.10 mmol) was added to a solution of compound 2 (1000 mg, 2.41 mmol) in MeOH (20 mL). After addition, the mixture was stirred at 50°C for 2 hours. The reaction mixture was diluted with water (30 mL) and extracted with EtOAc (50 mL×2). The combined organic phases were dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting from 0% to 90% with EtOAc / petroleum ether and EtOAc in 15 minutes to give compound 3 (800 mg, yield 76%) as a yellow solid. LC-MS: (ESI) m / z[M+H] + 415.1.

[0267] To a solution of compound 3 (500 mg, 1.20 mmol) in DCM (10 mL) was added acetyl chloride (104 mg, 1.32 mmol) and DIEA (311 mg, 2.41 mmol) at 25 ° C. After addition, the mixture was stirred at 25 ° C for 1 hour. The reaction mixture was diluted with water (10 mL) and extracted with DCM (20 mL × 2). The combined organic phases were dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting from 0% to 90% with EtOAc / petroleum ether and EtOAc in 15 minutes to give compound 5 (300 mg, yield 54%) as a yellow solid. LC-MS: (ESI) m / z[M+H] + 457.1.

[0268] A solution of compound 5 (300 mg, 0.61 mmol) in PPA (5 mL) was stirred at 110 ° C for 1 hour. The reaction mixture was diluted with water (10 mL) and extracted with EtOAc (20 mL×2). The combined organic phase was dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting from 0% to 90% with EtOAc / petroleum ether and EtOAc in 15 minutes to give compound 6 (150 mg, yield 52%) as a yellow solid. LC-MS: (ESI) m / z[M+H]+ 439.1.

[0269] A solution of compound 6 (200 mg, 0.45 mmol), X-phos-Pd G3 (38 mg, 0.04 mmol), compound 7 (310 mg, 0.77 mmol) and K3PO4 (290 mg, 1.36 mmol) in dioxane (2 mL) and H2O (0.2 mL) was stirred at 25 ° C under N2. The mixture was heated to 100 ° C and stirred at this temperature for 1 hour. The reaction mixture was cooled to 25 ° C, diluted with water (10 mL) and extracted with EtOAc (20 mL×2). The combined organic phases were dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting from 0% to 80% with EtOAc / petroleum ether and EtOAc in 15 minutes to give compound 8 (150 mg, yield 46%) as a yellow solid. LC-MS: (ESI) m / z[M+H] + 676.3.

[0270] A solution of compound 8 (150 mg, 0.22 mmol) in FA (5 mL) was stirred at 25 °C for 10 minutes. The reaction mixture was concentrated under reduced pressure. The reaction mixture was purified by preparative HPLC (CH3CN from 0% to 100% in 10 minutes) to give compound 9244 (6 mg, 5% yield) as a white solid.

[0271] 1 H NMR (400MHz, DMSO-d6) δ7.46–7.44(m,2H),7.35–7.33(m,2H),6.58–6.54(m,1H),6.44–6.39(m,1H),4.68–4.64(m,1 H),3.94–3.90(m,2H),3.55–3.52(m,2H),2.62(s,3H),2.49(s,3H),2.42(s,3H),1.63(s,3H).LC-MS:(ESI)m / z[M+H] + 476.1.

[0272] Examples 1-20

[0273]

[0274] DMF-DMA (1148 mg, 9.64 mmol) was added to a solution of compound 1 (800 mg, 1.92 mmol) in DMF (10 mL) at 25°C. After addition, the mixture was stirred at 110°C for 2 hours. The reaction mixture was concentrated under reduced pressure to give compound 2 (800 mg, yield 79%) as a yellow solid. LC-MS: (ESI) m / z [M+H] + 470.2.

[0275] Methylamine (231 mg, 7.44 mmol) was added to a solution of compound 2 (700 mg, 1.48 mmol) in EtOH (10 mL) at 25 ° C. After addition, the mixture was stirred at 110 ° C for 1 hour. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (CH3CN from 0% to 100% in 10 minutes) to give compound 3 (300 mg, 43% yield) as a white solid. LC-MS: (ESI) m / z [M+H] + 438.1.

[0276] A solution of compound 3 (120 mg, 0.27 mmol), X-phos-Pd G3 (23 mg, 0.02 mmol), compound 4 (186 mg, 0.46 mmol) and K3PO4 (174 mg, 0.82 mmol) in dioxane (2 mL) and H2O (0.2 mL) was stirred at 25 ° C under N2. The mixture was heated to 100 ° C and stirred at this temperature for 1 hour. The reaction mixture was cooled to 25 ° C, diluted with water (10 mL) and extracted with EtOAc (20 mL×2). The combined organic phases were dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting from 0% to 80% with EtOAc / petroleum ether and EtOAc in 15 minutes to give compound 5 (100 mg, yield 51%) as a yellow solid. LC-MS: (ESI) m / z[M+H] + 675.2.

[0277] A solution of compound 5 (120 mg, 0.17 mmol) in FA (2 mL) was stirred at 25 °C for 1 hour. The reaction mixture was concentrated under reduced pressure. The reaction mixture was purified by preparative HPLC (CH3CN from 0% to 90% in 10 minutes) to give compound 9245 (10 mg, 11% yield) as a white solid.

[0278] 1H NMR(400MHz,DMSO-d6)δ8.41(s,1H),7.46–7.44(m,2H),7.35–7.33(m,2H),6.59–6.55(m,1H),6.43–6.39(m,1H),4.71–4.67(m,1H),3 .94–3.90(m,1H),3.80(s,3H),3.77–3.74(m,1H),3.52–3.50(m,2H),2.60(s,3H),2.41(s,3H),1.63(s,3H).).LC-MS:(ESI)m / z[M+H] + 475.1.

[0279] Embodiment 1-21

[0280]

[0281] To a solution of compound 1 (300 mg, 0.72 mmol) in AcOH (5 mL) was added TsOH.H2O (411 mg, 2.16 mmol) and compound 2 (330 mg, 1.80 mmol) at 25 ° C. After addition, the mixture was stirred at 100 ° C for 1 hour. The reaction mixture was diluted with water (10 mL) and extracted with EtOAc (20 mL × 2). The combined organic phases were dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting from 0% to 90% with EtOAc / petroleum ether in 15 minutes to give compound 3 (200 mg, yield 54%) as a yellow solid. LC-MS: (ESI) m / z[M+H] + 454.1.

[0282] Compound 3 (200 mg, 0.44 mmol), X-phos-Pd G3 (37 mg, 0.04 mmol), compound 4 (265 mg, 0.66 mmol) and K3PO4 (280 mg, 1.32 mmol) were dissolved in dioxane (5 mL) and H2O (0.5 mL) solution and stirred at 25 ° C under N2. The mixture was heated to 100 ° C and stirred at this temperature for 2 hours. The reaction mixture was cooled to 25 ° C, diluted with water (10 mL) and extracted with EtOAc (20 mL×2). The combined organic phases were dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting from 0% to 80% with EtOAc / petroleum ether and EtOAc in 15 minutes to obtain compound 5 (150 mg, yield 44%) as a yellow solid. LC-MS: (ESI) m / z[M+H] + 691.2.

[0283] A solution of compound 5 (100 mg, 0.14 mmol) in FA (2 mL) was stirred at 25 °C for 1 hour. The reaction mixture was concentrated under reduced pressure. The reaction mixture was purified by preparative HPLC (CH3CN from 0% to 100% in 10 minutes) to give compound 9246 (10 mg, 13% yield) as a white solid.

[0284] 1 H NMR(400MHz,DMSO-d6)δ7.45–7.36(m,5H),6.59–6.55(m,1H),6.45–6.38(m,1H),4.52–4.81(m,1H),4.00– 3.97(m,2H),3.51–3.42(m,2H),2.61(s,3H),2.41(s,3H),2.40(s,3H),1.62(s,3H).LC-MS:(ESI)m / z[M+H] + 491.9.

[0285] Examples 1-22

[0286]

[0287] Compound 2 (178 mg, 0.985 mmol), Cs2CO3 (642 mg, 1.97) and Xphos Pd G3 (55.6 mg, 0.0657 mmol) were added to a solution of compound 1 (300 mg, 0.657 mmol) in 1,4-dioxane (10 mL). The reaction mixture was stirred at 100 ° C, N2 for 8 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with EtOAc in petroleum ether from 0% to 50% to give compound 3 (300 mg, yield 72%) as a yellow solid. LC-MS: (ESI) m / z [M+H] + 602.2.

[0288] 1M HCl (2.0 mL) was added dropwise to a solution of compound 3 (1.10 g, 1.80 mmol) in THF (10 mL) at 25 ° C and stirred. The resulting mixture was stirred at 25 ° C for 30 minutes. The mixture was quenched with NaHCO3 solution (30 mL) and then extracted with ethyl acetate (30 mL x 2). The combined organic phases were dried over sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluted with a petroleum ether solution of THF from 0% to 80%, to give compound 4 (0.730 g, yield 89%) as a white solid. LC-MS: (ESI) m / z[M+H] + 438.2.

[0289] To a mixed solution of compound 4 (730 mg, 1.67 mmol) in 1,4-dioxane (8.0 mL) and NaHCO3 solution (1 M, 8.0 mL) was added 9-fluorenylmethyl chloroformate (518 mg, 2.00 mmol). The reaction mixture was stirred at 25 ° C for 16 hours. The mixture was quenched with water (10 mL) and then extracted with ethyl acetate (30 mL x 2). The combined organic phases were dried over sodium sulfate and filtered. The residue was purified by silica gel chromatography, eluted with a petroleum ether solution of THF from 0% to 20% to give compound 5 (730 mg, yield 65%) as a yellow solid. LC-MS: (ESI) m / z[M+H] + 660.3.

[0290] To a solution of compound 5 (730 mg, 1.11 mmol) in DCM (6.0 mL) was added TFA (2.0 mL) at 25°C. The reaction mixture was stirred at 25°C for 2 hours. The filtrate was concentrated under reduced pressure to give compound 6 (660 mg, crude product), which was used directly in the next step without further purification. LC-MS: (ESI) m / z [M+H] + 604.2.

[0291] At 25 ° C, HATU (462 mg, 1.22 mmol), DIPEA (857 mg, 6.63 mmol) and NH4Cl (177 mg, 3.32 mmol) were added to a solution of compound 6 (667 mg, 1.11 mmol) in DCM (5.0 mL). The reaction mixture was stirred at 25 ° C for 2 hours. The reaction mixture was added to water (20 mL), the aqueous phase was extracted with ethyl acetate (30 mL × 2), the combined organic phase was washed with NaCl aqueous solution (20 mL × 3) and dried with Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluted with methanol in DCM from 0% to 6%, to give compound 7 (609 mg, yield 87%) as a yellow solid. LC-MS: (ESI) m / z[M+H] + 603.2.

[0292] A solution of compound 7 (609 mg, 1.01 mmol) and Lawesson's reagent (CAS: 19172-47-5, 450 mg, 1.11 mmol) in THF (10 mL) was stirred and heated at 100 ° C for 1 hour. The mixture was diluted with water (30 mL) and extracted with EA (30 mL x 3). The combined organic layers were dried over Na2SO4 and concentrated. The crude product was purified by FCC (DCM / MeOH=10:1) to give compound 8 (500 mg, yield 78%) as a yellow solid. LC-MS: (ESI) m / z[M+H]+ 619.2.

[0293] Compound 8 (500 mg, 0.808 mmol), compound 9 (273 mg, 1.62 mmol) and TsOH (347 mg, 2.02 mmol) were mixed and stirred in AcOH (8.0 mL) and heated at 60 ° C for 1 hour using microwave. The mixture was then diluted with water (20 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were dried over Na2SO4 and concentrated. The crude product was purified by FCC (DCM / MeOH=10:1) to give compound 10 (330 mg, yield 62%) as a white solid. LC-MS: (ESI) m / z[M+H] + 643.2.

[0294] To a solution of compound 10 (330 mg, 0.513 mol) in DCM (1.5 mL) was added dimethylamine (2 M, 1.5 mL). The mixture was stirred at 25 °C for 1 hour. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with MeOH in PE from 0% to 6% to give compound 11 (200 mg, yield 88%) as a yellow solid. LC-MS: (ESI) m / z [M+H] + 421.1.

[0295] To a solution of compound 11 (20.0 mg, 0.0476 mmol) in DCM (2.0 mL) was added propene-2-enoyl chloride (6.46 mg, 0.0714 mmol) and TEA (14.5 mg, 0.143 mmol). The reaction mixture was stirred at 25 ° C for 10 minutes. The mixture was concentrated. The residue was purified by Prep HPLC (chromatographic column: -Xbridge-C18 150x 19 mm, 5um mobile phase: ACN-H2O, 45-75% MeCN in 0.1% FA / water, 30 mL / min) to obtain compound 9248 (4.10 mg, yield 18%) as a white solid.

[0296] 1H NMR(400MHz,DMSO-d6)δ10.39(s,1H),7.76–7.69(m,3H),7.66–7.61(m,1H),7.43(d,J=8.8Hz,2 H),6.51–6.40(m,1H),6.27(dd,J=17.2,2.0Hz,1H),5.78(dd,J=10.0,2.0Hz,1H),4.56–4.45(m 1H),4.16–3.96(m,2H),2.61(s,3H),2.41(s,3H),1.64(s,3H).LC-MS:(ESI)m / z[M+H] + 475.1.

[0297] Examples 1-23

[0298]

[0299] To a solution of compound 1 (30.0 mg, 0.0587 mmol) in DCM (2.0 mL) was added 2-chloroethanesulfonyl chloride (14.6 mg, 0.0881 mmol) and TEA (17.8 mg, 0.176 mmol) at 25°C. The reaction mixture was stirred at 25°C for 10 minutes. LCMS showed that the starting material was consumed and the desired product was formed. The mixture was concentrated. The residue was purified by Prep HPLC (chromatographic column: -Xbridge-C18 150×19 mm, 5 um mobile phase: ACN-H2O, 45-75% MeCN in 0.1% FA / water, 30 mL / min) to give compound 9249 (4.10 mg, 13% yield) as a white solid.

[0300] 1 H NMR (400MHz, DMSO-d6) δ7.71(d,J=1.2Hz,1H),7.61(d,J=1.2Hz,1H),7.35(d,J=2.4Hz,2H),7.10(d,J=2.4Hz,2H),6.71(dd,J=6.4,2.4Hz,1H),6.10( d,J=16.4Hz,1H),5.97(d,J=9.6Hz,1H),4.52–4.46(m,1H),4.06–3.99(m, 2H),2.59(s,3H),2.40(s,3H),1.62(s,3H).LC-MS:(ESI)m / z[M+H]+511.1.

[0301] Examples 1-24

[0302]

[0303] To a solution of compound 1 (20.0 mg, 0.0476 mmol) in DCM (2.0 mL) was added 2-chloroacetyl chloride (8.06 mg, 0.0714 mmol) and TEA (14.5 mg, 0.143 mmol) at 25°C. The reaction mixture was stirred at 25°C for 10 minutes. LCMS showed that the starting material was consumed and the desired product was formed. The mixture was concentrated. The residue was purified by Prep HPLC (chromatographic column: -Xbridge-C18 150×19 mm, 5 um mobile phase: ACN-H2O, 45-75% MeCN in 0.1% FA / water, 30 mL / min) to give compound 9250 (4.20 mg, 18% yield) as a white solid.

[0304] 1 H NMR (400MHz, DMSO-d6) δ10.55(s,1H),7.72(s,1H),7.72–7.63(m,3H),7.43(d,J=8.8Hz,2H),4.55–4.50(m, 1H),4.27(s,2H),4.06–4.00(m,2H),2.51(s,3H),2.40(s,3H),1.62(s,3H).LC-MS:(ESI)m / z[M+H]+497.1.

[0305] Examples 1-25

[0306]

[0307] At 25 ° C, to a DMF (2.0 mL) solution of compound 1 (20.0 mg, 0.0476 mmol), butane-2-ynoic acid (6.00 mg, 0.0714 mmol), DIPEA (18.5 mg, 0 0.143 mmol) and HATU (27.2 mg, 0.0714 mmol) were added. The reaction mixture was stirred at 25 ° C for 1 hour. LCMS showed that the raw material was consumed and the desired product was formed. The mixture was quenched with water (20 mL), then extracted with ethyl acetate (20 mL x 2). The combined organic phases were washed with NaCl aqueous solution (10 mL x 2), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by Prep HPLC (column: -Xbridge-C18 150 x 19 mm, 5 um mobile phase: ACN-H2O, 45-75% MeCN in 0.1% FA / water, 30 mL / min) to afford compound 9253 (5.70 mg, yield 24%) as a white solid.

[0308] 1H NMR (400MHz, DMSO-d6) δ10.82(s,1H),7.72(s,1H),7.71–7.62(m,3H),7.40(d,J=8.8Hz,2H),4.54–4.51(m, 1H),4.07–4.00(m,2H),2.51(s,3H),2.40(s,3H),2.04(s,3H),1.62(s,3H).LC-MS:(ESI)m / z[M+H]+487.1.

[0309] Examples 1-26

[0310]

[0311] At 25 ° C, to a solution of compound 1 (20.0 mg, 0.0421 mmol) in DCM (3.0 mL), prop-2-enoyl chloride (5.72 mg, 0.0631 mmol) and TEA (12.8 mg, 0.126 mmol) were added. The mixture was then stirred at 25 ° C for 10 minutes. LCMS showed that the starting material was consumed and the desired product was detected. The mixture was quenched with water (20 mL) and then extracted with ethyl acetate (20 mL x 2). The combined organic phases were dried over sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep HPLC (chromatographic column: -Xbridge-C18 150×19 mm, 5um mobile phase: ACN-H2O, 45-75% MeCN in 0.1% FA / water, 30 mL / min) to obtain compound 9254 (7.60 mg, 34% yield) as a white solid.

[0312] 1 H NMR (400MHz, DMSO-d6) δ8.66(t,J=5.2Hz,1H),7.47(d,J=8.4Hz,2H),7.41(d,J=8.4Hz,2H),6.26–6.11(m,2H),5.63(d,J=2.4Hz,1H),4.44 –4.40(m,1H),2.22(d,J=5.3Hz,2H),2.31–2.26(m,2H),2.60(s,3H),2.41(s,3H),1.62(s,3H),1.42(s,9H).LC-MS:(ESI)m / z[M+H]+530.2.

[0313] Examples 1-27

[0314]

[0315] At 25 ℃, to DMF (3.0mL) solution of compound 1 (100mg, 0.210mmol), propane-2-enoic acid (30.2mg, 0.420mmol), HOAt (3.43mg, 0.0251mmol), HOPO (23.3mg, 0.210mmol) and N-ethylmorpholine (72.5mg, 0.629mmol) were added. After stirring for 5 minutes, EDCI (52.3mg, 0.273mmol) was added at 25 ℃. The mixture was then stirred for 55 minutes at 25 ℃. LCMS showed that the raw material was consumed and the desired product was detected. The mixture was quenched with water (20mL), then extracted with ethyl acetate (20mL x 2). The combined organic phase was dried over sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC (column: -Xbridge-C18 150×19 mm, 5 um mobile phase: ACN-H 2 O, 45-75% MeCN in 0.1% FA / water, 30 mL / min) to give compound 9255 (8.50 mg, yield 8%) as a white solid.

[0316] 1 H NMR(400MHz, DMSO-d6)δ9.69(d,J=7.6Hz,1H),7.73–7.72(m,3H),7.67–7.64(m,2H),7.52–7.50(m,2H),7.45–7.42(m,1H),7.41–7.39(m,1H ),6.92(dd,J=7.6,15.6Hz,1H),4.60–4.53(m,2H),4.10–4.07(m,3H),2.62(s,3H),2.41(s,3H),1.61(s,3H).LC-MS:(ESI)m / z[M+H]+531.1.

[0317] Examples 1-28

[0318]

[0319] At 25 ° C, compound 2 (50.2.0 mg, 0.232 mmol), DIPEA (59.9 mg, 0.232 mmol) and HATU (88.2 mg, 0.232 mmol) were added to a solution of compound 1 (40.0 mg, 0.464 mmol) in DMF (2.0 mL). The reaction mixture was stirred at 25 ° C for 16 hours. LCMS showed that the starting material was consumed and the desired product was formed. The mixture was quenched with water (20 mL) and then extracted with ethyl acetate (20 mL×2). The combined organic phase was washed with NaCl aqueous solution (10 mL×2), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by Prep-HPLC (column: -Xbridge-C18 150×19 mm, 5 um mobile phase: ACN-H 2 O, 45-75% MeCN in 0.1% FA / water, 30 mL / min) to give compound 9257 (10.5 mg, yield 22%) as a white solid.

[0320] 1 H NMR(400MHz,DMSO-d6)δ10.17(d,J=24.0Hz,1H),7.72(d,J=3.6Hz,1H),7.65–7 .62(m,3H),7.42–7.39(m,2H),6.94(br,0.4H),6.32(br,0.6H),5.12–4.95(m, 1H),4.50–4.49(m,1H),4.09–3.99(m,2H),3.25(s,1H),3.07–2.95(m,4H),2.5 1(s,3H),2.40(s,3H),1.62(s,3H),1.42(s,9H).LC-MS:(ESI)m / z[M+H]+618.2.

[0321] Examples 1-29

[0322]

[0323] To a solution of compound 1 (80 mg, 0.1828 mol) and TEA (73.99 mg, 0.7312 mmol) in DCM (2 mL) was added 2-chloroacetyl chloride (30.97 mg, 0.2742 mmol) at 0°C. The mixture was stirred at 25°C for 7 hours. LCMS showed that the starting material was consumed and the desired product mass was detected. The mixture was concentrated to dryness. The residue was purified by Prep HPLC (chromatographic column: WELCH Xtimate C18 21.2×250 mm 10 μm, mobile phase: ACN-H2O (0.1% FA), gradient: 50-50) to give compound 9258 (30.8 mg, yield 32%) as a white solid.

[0324] 1 H NMR (400MHz, DMSO-d6) δ10.52(s,1H),7.63(d,J=9.2Hz,2H),7.41(d,J=8.4Hz,2H),4.40–4.36(m,1H),4.27( s,2H),3.33–3.29(m,2H),2.51(s,3H),2.42(s,3H),1.66(s,3H),1.46(s,9H).LC-MS:(ESI)m / z[M+H]+514.1.

[0325] Examples 1-30

[0326]

[0327] Compound 1 (55 mg, 0.1257 mol), but-2-ynoic acid (15.85 mg, 0.18855 mmol) and pyridine (149.14 mg, 1.8855 mmol) were dissolved in DCM (2 mL) and POCl3 (38.55 mg, 0.2514 mmol) was added at 0°C. The mixture was stirred at 25°C for 1 hour. LCMS showed that the starting material was consumed and the desired product mass was detected. The mixture was concentrated to dryness. The residue was purified by Prep HPLC (chromatographic column: WELCH Xtimate C18 21.2×250 mm 10 um, mobile phase: ACN-H2O (0.1% FA) gradient: 45-55) to give compound 9259 (15.6 mg, yield 24%) as a white solid.

[0328] 1H NMR (400MHz, DMSO-d6) δ10.83(s,1H),7.63(d,J=8.8Hz,2H),7.37(d,J=8.8Hz,2H),4.42–4.37(m,1H),3.33– 3.29(m,2H),2.51(s,3H),2.42(s,3H),2.05(s,3H),1.65(s,3H),1.45(s,9H).LC-MS:(ESI)m / z[M+H]+504.2.

[0329] Example 1-31

[0330]

[0331] Compound 1 (60 mg, 0.1371 mol), propiolic acid (14.41 mg, 0.2056 mmol) and pyridine (162.67 mg, 2.0564 mmol) were dissolved in DCM (2 mL) and POCl (42.04 mg, 0.2742 mmol) was added dropwise at 0°C. The mixture was stirred at 25°C for 0.5 hours. LCMS showed that the starting material was consumed and the desired product mass was detected. The mixture was concentrated to dryness. The residue was purified by Prep HPLC (chromatographic column: WELCH Xtimate C18 21.2*250mm 10um, mobile phase: ACN-H2O (0.1% FA) gradient: 47-53) to give compound 9260 (14.2 mg, yield 21%) as a yellow solid.

[0332] 1 H NMR (400MHz, DMSO-d6) δ11.02(s,1H),7.63(d,J=8.8Hz,2H),7.38(d,J=8.4Hz,2H),4.46(s,1H),4.41–4.36( m,1H),3.33–3.28(m,2H),2.50(s,3H),2.42(s,3H),1.65(s,3H),1.48(s,9H).LC-MS: (ESI)m / z[M+H]+490.2.

[0333] Example 1-32

[0334]

[0335] Compound 2 (226.60 mg, 1.2503 mmol), Pd2(dba)3 (114.49 mg, 0.12503 mmol), SPhos (102.77 mg, 0.25006 mmol) and cesium carbonate (1222.12 mg, 3.75 mmol) were added to a solution of compound 1 (500 mg, 1.2503 mmol) in DMF (5 mL). The reaction mixture was stirred at 110 ° C for 16 hours. LCMS showed the formation of the desired product. The reaction mixture was quenched with H2O (10 mL), extracted with EtOAc (10 mL*3), the organic layer was dried over anhydrous Na2SO4, filtered and concentrated in vacuo, and the residue was purified by silica gel chromatography, eluting with methanol in DCM from 0% to 5% to give compound 3 (60 mg, 3%) as a yellow oil. LC-MS: (ESI) m / z[M+H] + 545.2.

[0336] Compound 3 (60 mg, 0.1102 mmol) was dissolved in THF (1 mL) and 2M / HCl (1 mL). The reaction mixture was stirred at 25 ° C for 1 hour. LCMS showed the formation of the desired product. The reaction mixture was quenched with NaHCO3 solution (5 mL), extracted with EtOAc (10 mL*3), and the organic layer was dried over anhydrous Na2SO4, filtered and concentrated in vacuo to give compound 4 (40 mg, 29%) as a yellow oil. It can be used directly without further purification. LC-MS: (ESI) m / z [M+H] + 381.1.

[0337] To a solution of compound 4 (40 mg, 0.1051 mmol), propionic acid (11.04 mg, 0.15765 mmol), pyridine (16.63 mg, 0.2102 mmol) in DCM (3 mL) was added POCl (32.23 mg, 0.2102 mmol). The reaction mixture was stirred at 25 ° C for 30 minutes. LCMS showed the formation of the desired product. The reaction mixture was concentrated in vacuo and the residue was purified by pre-HPLC (chromatographic column: -Xbridge-C18 150x 19 mm, 5um mobile phase: ACN-H2O (0.1% FA)) (0-100%) to obtain compound 9263 (4.6 mg, 10%), white.

[0338] 1H NMR (400MHz, DMSO-d6-25℃) δ11.03(s,1H),7.63(d,J=8.8Hz,3H),7.39(d,J=8.6Hz,2H),6.97 (s,1H),4.46(d,J=1.8Hz,2H),3.21(d,J=7.2Hz,2H),2.59(s,3H),2.41(s,3H),1.64(s,3H).

[0339] 1 H NMR (400MHz, DMSO-d6-80℃) δ10.72(s,1H),7.61(d,J=8.6Hz,2H),7.39(d,J=8.6Hz,2H),6.67(s,2H),4.47(t,J=6 .8Hz,1H),4.22(s,1H),3.33–3.17(m,2H),2.58(s,3H),2.41(s,3H),1.66(s,3H).LC-MS:(ESI)m / z[M+H]+433.1.

[0340] Examples 1-33

[0341]

[0342] To a solution of compound 1 (300 mg, 0.748 mmol) in DCM (5.0 mL) was added ethylamine hydrochloride (183 mg, 2.245 mmol), DIEA (483 mg, 3.742 mmol) and HATU (341 mg, 0.898 mmol). The reaction mixture was stirred at 25 ° C for 1 hour. The reaction mixture was quenched with water (10 mL), extracted with DCM (10 mL×3), the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography, eluted with methanol in DCM from 0% to 5%, and compound 2 (320 mg, 95% yield) was obtained as a white solid. LC-MS: (ESI) m / z[M+H] + 428.1.

[0343] Compound 3 (216 mg, 1.19 mmol), Cs2CO3 (776 mg, 2.38 mmol) and Xphos Pd G3 (67 mg, 0.0794 mmol) were added to a solution of compound 2 (340 mg, 0.795 mmol) in dioxane (3.0 mL). The reaction mixture was stirred at 110 ° C for 16 hours in a sealed tube under N2 atmosphere. The mixture was quenched with water (10 mL) and extracted with EtOAc (10 mL×3). The combined organic phases were dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography, eluting with methanol in DCM from 0% to 5% to give compound 4 (280 mg, yield 61%) as a yellow solid. LC-MS: (ESI) m / z[M+H] + 573.2.

[0344] 2M HCl (2.0 mL) was added to a solution of compound 4 (280 mg) in THF (2.0 mL). The reaction mixture was stirred at 25 ° C for 1 hour. The mixture was quenched with NaHCO3 solution (5 mL) and then extracted with DCM (10 mL×3). The combined organic phases were dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated in vacuo. The residue was purified by silica gel chromatography, eluting with EtOAc in petroleum ether from 0% to 80% to give compound 5 (120 mg, yield 60%) as a yellow solid. LC-MS: (ESI) m / z[M+H] + 409.2.

[0345] To a solution of compound 5 (120 mg, 0.294 mmol) in DCM (2.0 mL) was added prop-2-ynoic acid (31 mg, 0.441 mmol), pyridine (35 mg, 0.441 mmol) and POCl3 (90 mg, 0.587 mmol). The reaction mixture was stirred at 25 ° C for 30 minutes. The mixture was vacuumed. The residue was purified by Prep HPLC (chromatographic column: -Xbridge-C18 150×19 mm, 5 um mobile phase: ACN-H2O, 23-55% MeCN in 0.1% FA / water, 30 mL / min) to obtain compound 6 (28.5 mg, yield 21%) as a yellow solid.

[0346] 1H NMR (400MHz, DMSO-d6) δ11.01(s,1H),8.19(t,J=5.2Hz,1H),7.64(d,J=8.4Hz,2H),7.38(d,J=8.4Hz,2H),4.53-4. 38(m,2H),3.21-3.09(m,4H),2.59(s,3H),2.41(s,3H),1.64(s,3H),1.06(t,J=7.2Hz,3H).LC-MS:(ESI)m / z[M+H] + 461.2.

[0347] Examples 1-34

[0348]

[0349] To a solution of compound 1 (400 mg, 0.998 mmol) in DCM (5.0 mL) were added EDCI (248 mg, 1.30 mmol), DMAP (61 mg, 0.499 mmol) and compound 2 (142 mg, 1.99 mmol). The reaction mixture was stirred at 25 ° C for 16 hours. The mixture was concentrated in vacuo. The residue was purified by silica gel chromatography, eluted with 0-70% EtOAc in petroleum ether to give compound 3 (360 mg, yield: 79%) as a white solid. LC-MS: (ESI) m / z [M+H] + 455.1.

[0350] Compound 4 (119 mg, 0.660 mmol), Xphos Pd G3 (37 mg, 0.0440 mmol) and Cs2CO3 (286 mg, 0.879 mmol) were added to a solution of compound 3 (200 mg, 0.440 mmol) in dioxane (2.0 mL). The reaction mixture was stirred at 110 ° C. under N2 atmosphere for 16 hours. The mixture was concentrated in vacuo. The residue was purified by silica gel chromatography, eluted with 0-60% EtOAc in petroleum ether to give compound 5 (160 mg, yield: 36%) as a yellow oil. LC-MS: (ESI) m / z[M+H] + 600.2.

[0351] To a solution of compound 5 (160 mg, 0.267 mmol) in THF (2.0 mL) was added 1N HCl aqueous solution (2.0 ml). The reaction mixture was stirred at 25 ° C for 1 hour. The mixture was quenched with NaHCO3 solution (5 mL) and then extracted with DCM (10 mL×3). The combined organic phases were dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated in vacuo. The residue was purified by silica gel chromatography, eluted with EtOAc in petroleum ether from 0% to 65%, to give compound 6 (60 mg, yield 51%) as a yellow solid. LC-MS: (ESI) m / z[M+H] + 436.1.

[0352] To a solution of compound 6 (50 mg, 0.115 mmol) in DCM (2.0 mL) was added prop-2-ynoic acid (12 mg, 0.172 mmol), pyridine (13 mg, 0.172 mmol) and POCl3 (35 mg, 0.230 mmol). The mixture was stirred at 0°C for 30 minutes. The mixture was concentrated in vacuo. The residue was purified by Prep HPLC (column: -WELCH Xtimate C18 21.2*250mm10um; mobile phase: ACN-H2O, 40-50% MeCN in 0.1% NH3 / water, 30 ml / min) to give compound 9265 (21.7 mg, 38% yield) as a white solid.

[0353] 1 H NMR (400MHz, DMSO-d6) δ11.01(s,1H),7.64(d,J=8.8Hz,2H),7.37(d,J=8.4Hz,2H),4.49-4.38(m,2H),3.34–3.33(m,2H),2 .63–2.57(m,3H),2.42(s,3H),1.65(s,3H),1.50–1.47(m,3H),0.82–0.76(m,2H),0.69–0.63(m,2H),LC-MS:(ESI)m / z[M+H] + 488.1.

[0354] Example 2-1

[0355]

[0356] Compound 1 (25.0 g, 0.15 mol) was stirred in Ac2O (140 mL) for 3.5 h at 140° C. The resulting mixture was concentrated under reduced pressure and triturated with Et2O to give Compound 2 (23.0 g, crude) as a brown solid.

[0357] At 0°C and N2 atmosphere, a THF solution of compound 3 (83.6mL, 83.60mmol, 1M) was added dropwise to a solution of PhMe / Et2O (200mL+100mL) of compound 2 (20.0g, 104.60mmol). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with 1M HCl and extracted with EA (100mL×2). The combined organic phase was concentrated under reduced pressure to obtain a residue, which was dissolved in ethanol (30mL) and concentrated hydrochloric acid (30mL) solution. The solution was heated to 80°C and stirred at 80°C for 3 hours. The reaction mixture was cooled to room temperature and concentrated in vacuo, then dispersed between DCM and 4N NaOH solution. The aqueous layer was re-extracted with DCM (100mL×2), and the combined organic portion was dried with anhydrous Na2SO4, filtered and concentrated. The crude product was purified by silica gel column chromatography to obtain compound 4 (6.0g, yield 17.78%) as a yellow oil.

[0358] Compound 4 (12.0 g, 39.20 mmol), compound 5 (16.8 g, 43.10 mmol) and NaHCO3 (6.6 g, 78.40 mmol) were dissolved in DCM (120 mL) and stirred at 0°C for 1 hour. The resulting mixture was extracted with DCM (100 mL×2), and the organic layer was collected, washed twice with saturated brine, and dried over Na2SO4. The organic layer was concentrated under reduced pressure. Compound 6 (24.0 g, crude product) was obtained as a yellow oil.

[0359] Stirring compound 6 (24.0 g, 36.40 mmol) and TEA (14.7 g, 145.60 mmol) were added to DCE (40 mL) and heated to reflux for 3 hours. The obtained mixture was concentrated to dryness, the crude product was dissolved in DCE (240 mL), then AcOH (23.7 mL) was added, and the reaction was stirred for another 2 hours at 80 ° C. The obtained mixture was extracted with DCM (100 mL × 3), the organic layer was collected, washed twice with saturated brine, and dried over Na2SO4. The organic layer was concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, DCM / MeOH=98 / 2) to obtain compound 7 (9.0 g, 20.50 mmol, yield 56.3%), which was a gray solid.

[0360] Compound 7 (6.0 g, 14.40 mmol), P4S10 (5.8 g, 25.90 mmol) and sodium carbonate (2.8 g, 25.90 mmol) were dissolved in DCE (60 mL) and stirred at 60 ° C for 5 hours. The resulting mixture was extracted with DCM (100 mL × 2), and the organic layer was collected, washed twice with saturated brine and NaHCO3 solution, and dried over Na2SO4. The organic layer was concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, DCM = 100%) to obtain a yellow solid compound 8 (3.0 g, 6.60 mmol, yield 45.8%).

[0361] To a solution of compound 8 (1.2 g, 2.80 mmol) in THF (12 mL) was added hydrazine hydrate (0.4 g, 8.40 mmol) and stirred at 0 ° C for 2.5 hours. TEA (0.9 g, 8.40 mmol) was added to the solution, the reaction mixture was cooled to 0 ° C, and then acetyl chloride (0.7 g, 8.40 mmol) was added and stirred at 0 ° C for another 30 minutes. The reaction mixture was diluted with water and DCM and separated. The aqueous layer was re-extracted with DCM (10 mL × 3), and the combined organic components were dried over anhydrous Na2SO4, filtered and concentrated in vacuo to give a yellow solid compound 9 (1.0 g, 1.90 mmol, 67.9% yield).

[0362] Compound 9 (1.0 g, 1.90 mmol), THF (10 mL) and AcOH (10 mL) were mixed and stirred at room temperature for 5 hours. The reaction mixture was diluted with water and DCM and separated. The aqueous layer was re-extracted with DCM (20 mL×3), and the combined organic fractions were dried over anhydrous Na2SO4. The organic layer was concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, DCM / MeOH=96 / 4) to obtain white solid compound 10 (700 g, 1.40 mmol, 66.7% yield).

[0363] Compound 10 (100 mg, 0.22 mmol), potassium iodide (109 mg, 0.66 mmol), Cu2O (3 mg, 0.02 mmol) and pyrrolidine-2-carboxylic acid (5 mg, 0.04 mmol) were mixed in EtOH (1.5 mL) and heated with stirring at 110 ° C under N2 for 24 hours. The mixture was extracted with ethyl acetate (10 mL × 3) and evaporated in vacuo. The residue was purified by column chromatography (DCM / MeOH=50 / 1) to obtain compound 11 (65 mg, 53% yield) as a yellow solid.

[0364] Compound 11 (30 mg, 0.06 mmol), prop-2-yn-1-amine (4 mg, 0.07 mmol), Pd(PPh3)4 (13 mg, 0.01 mmol), CuI (2 mg, 0.01 mmol) and triethylamine (18 mg, 0.17 mmol) were mixed in DMF (1 mL) and stirred at room temperature under N2 for 20 minutes. The mixture was diluted with MeOH, filtered and purified by prep-HPLC (chromatographic column: Phenomenexluna C18 250 mm×100 mm×10 um; mobile phase: [H2O(0.1% FA)-ACN]; B%: 15%~55%, 40 minutes) to give 9069 (5.8 mg, 23% yield) as a white solid.

[0365] LCMS: [M+H] + =444.1;

[0366] 1 H NMR: (400MHz, MeOD) δ7.744(d,J=9.0Hz,1H),7.535(d,J=8.4Hz,2H),7.483(d,J=8.4Hz,2H),7.386(dd,J=9.0,2.8Hz,1H),6.905(d,J=2.8Hz,1H ),4.636–4.581(m,1H),4.196(q,J=7.2Hz,2H),3.911(s,2H),3.827(s, 3H), 3.501 (dd, J=7.2, 4.4Hz, 2H), 2.631 (s, 3H), 1.287 (t, J=7.2Hz, 3H).

[0367] HPLC: 98.98% purity;

[0368] SFC: Rt = 3.932 min.

[0369] Example 2-2

[0370]

[0371] Compound 11 (400 mg, 0.796 mmol) and NaOH (10 mL, 2N) aqueous solution were dissolved in THF (10 mL) and stirred at room temperature for 20 hours. The pH of the solution was adjusted to about 5 with 2N HCl and extracted with DCM (20 mL x 3). The organic layer was dried and concentrated with Na2SO4 to give compound 12 (300 mg, 0.615 mmol, 90% yield).

[0372] To a solution of compound 12 (300 mg, 0.615 mmol) in t-BuOH (3 mL) was added Boc2O (781 mg, 3.584 mmol) and DMAP (18 mg, 0.123 mmol), and the mixture was stirred at 40 ° C for 3 hours. The reaction solution was diluted with water (20 mL), extracted with EA (20 mL x 3), and the obtained organic layer was dried over Na2SO4 and concentrated to give a crude product. The crude product was purified by FCC (DCM / MeOH=50:1) to give a yellow solid compound 13 (260 mg, 0.382 mmol, yield 53.3%).

[0373] Potassium carbonate (0.77 g, 5.579 mmol) and compound 15 (0.94 g, 7.836 mmol) were added to a DMF (10 mL) solution of compound 14 (1 g, 4.292 mmol). The mixture was stirred at room temperature for 4 hours. The solution was diluted with water (100 mL) and extracted with ethyl acetate (100 mL x 3). The organic layer was washed with a saturated sodium chloride aqueous solution (50 mL x 5), dried with Na2SO4, and then concentrated under reduced pressure. The residue was purified by FCC (eluent PE / EA=95 / 5) to obtain compound 16 (0.6 g, yield 46.5%) as a colorless oil.

[0374] Compound 16 (36 mg, 0.132 mmol), compound 13 (60 mg, 0.110 mmol), [t-Bu3PH] + [BF4] - To the product of 4-nitropropene (4-nitropropene) (64 mg, 0.220 mmol), Pd2(dba)3 (20 mg, 0.022 mmol) and Cy2NMe (43 mg, 0.220 mmol) were dissolved in NMP (1 mL), and the mixture was stirred for 90 minutes at 80 ° C under N2 under microwave irradiation. The mixture was then filtered, concentrated and diluted with water (10 mL). The resulting solution was extracted with EtOAc (10 ml × 3). The combined organic layers were dried over anhydrous Na2SO4 and concentrated. The residue was chromatographed on rapid silica gel (eluent DCM / MeOH=95 / 5) to give compound 17 (25 mg, 31.2% yield) as a white solid.

[0375] To a solution of compound 17 (25 mg, 0.036 mmol) in MeOH (1 mL) was added a solution of HCl in EA (2N, 1 mL), and the solution was stirred at room temperature for 3 hours. The solution was diluted with MeOH (2 mL) and dried with nitrogen. The crude product was dissolved in MeOH (2 mL) and purified by prep-HPLC (chromatographic column: Phenomenex luna C18 250 mm x 100 mm x 10 um; mobile phase: [H2O (0.1% FA)-ACN]; B%: 40 min) to afford 9077 (5.9 mg, 32.1% yield) as a white solid.

[0376] LCMS: [M+H] + =490.2;

[0377] 1 H NMR: (400MHz, MeOD) δ8.529 (s, 0.79H), 7.721 (d, J = 9.2Hz, 1H), 7.476 (q, J = 8.4 Hz,4H),7.380(dd,J=9.2,2.8Hz,1H),6.934(d,J=2.8Hz,1H),6.649(d,J=16.0 Hz,1H),6.439(dt,J=16.0,6.4Hz,1H),4.543(dd,J=8.4,6.0Hz,1H),3.821(s, 3H), 3.635 (d, J = 6.4Hz, 2H), 3.502–3.364 (m, 2H), 2.636 (s, 3H), 1.486 (s, 9H).

[0378] HPLC: 99.37% purity;

[0379] SFC: Rt = 9.747 min.

[0380] Example 2-3

[0381]

[0382] To a solution of 9077Int 12 (1.0 g, 0.002 mol) in DMF (10 mL) was added DIEA (0.78 g, 0.006 mol), HATU (1.14 g, 0.003 mmol) and NH4Cl (0.16 g, 0.003 mmol). The resulting mixture was stirred at room temperature for 2 hours. The crude product was directly purified by flash silica gel column (DCM / MeOH=5 / 1) to give yellow solid compound 1 (0.8 g, 80%).

[0383] Compound 1 (0.8 g, 0.002 mol) and Lawesson's reagent (2 mg, 0.004 mol) were mixed in THF (10 mL) and stirred at 40° C. for 3 hours. The solvent was removed under reduced pressure, and the crude product was directly purified by flash silica gel column (DCM / MeOH=8 / 1) to give compound 2 (0.6 g, 75%) as a yellow oil.

[0384] Compound 2 (500 mg, 0.993 mmol), compound 3 (336 mg, 1.987 mmol) and TsOH (34 mg, 0.199 mmol) were mixed in AcOH (10 mL) and heated at 50° C. for 2 hours. The crude product was directly purified by flash silica gel column (DCM / MeOH=10 / 1) to give compound 4 (400 mg, 75.6%) as a yellow oil.

[0385] Pd(PPh3)4 (88 mg, 0.076 mmol), copper(I) iodide (14 mg, 0.076 mmol), compound 5 (31 mg, 0.569 mmol) and TEA (115 mg, 1.138 mmol) were added to a solution of compound 4 (200 mg, 0.379 mmol) in DMF (5 mL). The reaction mixture was stirred at room temperature under N2 for 2 hours. The solution was directly filtered and purified by prep-HPLC (chromatographic column: Phenomenex luna C18 250 mm×100 mm×10 um; mobile phase: [H2O(0.1% FA)-ACN]; B%: 0%~540%, 40 minutes) to obtain a brown solid 9139 (25 mg, 9.91%).

[0386] LCMS: [M+H] + =455.2;

[0387] 1 H NMR: (400MHz, d4-MeOD) δ7.749-7.726(m,2H),7.586-7.494(m,5H),7.389(d,J=3 .6Hz,1H),6.876(s,1H),4.646(s,1H),4.085(m,4H),3.808(s,3H),2.646(s,3H).

[0388] HPLC: 95.13% purity.

[0389] Embodiment 2-4

[0390]

[0391] K2CO3 (10.9 g, 76.5 mmol) and compound 2 (15.4 g, 128 mmol) were added to a solution of compound 1 (15.0 g, 63.8 mmol) in DMF (100 mL). The reaction mixture was stirred at 20 ° C for 12 hours. The mixture was diluted with water (100 mL), extracted with EtOAc (150 mL×2), the organic layer was washed with saturated NaCl solution (200 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure, and the residue was purified by flash column chromatography (120 g silica gel column, PE / EtOAc and EtOAc from 0% to 10%) to obtain compound 3 (17.1 g, yield 88%) as a colorless oil. LC-MS: (ESI) m / z[m+Na]+296.4.

[0392] Pd(OAc)2(726mg, 4.36mmol) and TEA(6.61g, 65.4mmol) were added to a DMF(25mL) solution of compound 3(6.00g, 21.8mmol) and 1-bromo-2-fluoro-4-iodobenzene(6.56g, 21.78mmol). The reaction mixture was stirred for 5 hours at 90°C under N2 atmosphere. The mixture was diluted with DCM(100mL), washed with brine, dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography (120g silica gel column, PE / EtOAc and EtOAc from 0% to 20%) to obtain compound 4(3.6g, 33% yield), which was a yellow jelly.

[0393] Pd(PPh3)4 (103 mg, 0.0892 mmol) and potassium acetate (263 mg, 2.68 mmol) were added to a solution of compound 4 (400 mg, 0.892 mmol) and bipinacol borate (453 mg, 1.78 mmol) in dioxane (13 mL). The reaction mixture was stirred at 100 ° C for 4 hours to obtain a reaction solution containing compound 5. The reaction solution was used in the next step without further purification. LC-MS: (ESI) m / z [m+Na] + 516.4.

[0394] Pd(PPh3)4 (34.5 mg, 0.0298 mmol) and Na2CO3 (94.9 mg, 0.896 mmol) were added to a solution of compound 6 (100 mg, 0.298 mmol) and compound 5 (229 mg, 0.597 mmol) in dioxane (12 mL) and H2O (0.2 mL), and the reaction mixture was stirred at 100°C under N2 atmosphere for 5 hours. The mixture was concentrated under reduced pressure, and the residue was purified by flash column chromatography (25 g silica gel column, DCM / MeOH and MeOH from 0% to 5%) to give the product compound 7 (120 mg, yield 42%) as a yellow solid. LC-MS: (ESI) m / z [m+H] + 666.4.

[0395] LiOH (43.0 mg, 1.80 mmol) was added to a solution of compound 7 (120 mg, 0.180 mmol) in THF (5 mL) and H2O (2 mL). The reaction mixture was stirred at 25°C for 12 hours. EA (10 mL) was added to the mixture and extracted with water (40 mL). The aqueous layer was then acidified with HCl aqueous solution (1 M) until pH = 6-7. The mixture was washed with EtOAc (40 mL×3). The organic layer was dried over anhydrous Na2SO4. The mixture was purified by concentration under reduced pressure to give compound 8 (70 mg, yield 54%) as a yellow solid. LC-MS: (ESI) m / z[m+H] + 552.0.

[0396] To a solution of compound 8 (70 mg, 0.107 mmol) and Boc2O (117 mg, 0.536 mmol) in t-BuOH (3 mL) was added 4-DMAP (13.1 mg, 0.107 mmol). The reaction mixture was stirred at 25 ° C for 3 hours. The mixture was purified by concentration under reduced pressure. The mixture was purified by TLC (DCM / MeOH=20 / 1) to obtain compound 9 (45 mg, 53% yield) as a yellow solid.

[0397] A solution of compound 9 (30 mg, 0.0423 mmol) in FA (2 mL) was stirred at 25° C. for 2 hours. The mixture was purified by Prep HPLC (column: -Xbridge-C18 150×19 mm, 5 μm, mobile phase: ACN / H2O (0.1% FA), gradient: 25%-35%) to give compound 9149 (4.3 mg, yield 20%) as a white solid.

[0398] 1H NMR (400MHz, DMSO) δ7.81(d,J=8.8Hz,1H),7.45(t,J=8.0Hz,1H),7.40–7.33(m,2H),7.25(d,J=12.0Hz,1H),6.79(d,J=2.8Hz ,1H),6.61–6.43(m,2H),4.47–4.40(m,1H),3.76(s,3H),3.50(d,J=5.6Hz,3H),3.33–3.30(m,2H),2.53(s,3H),1.43(s,9H). 19 FNMR(376MHz,DMSO)δ-113.33.LC-MS:(ESI)m / z[M+H] + 508.2.

[0399] Embodiment 2-5

[0400]

[0401] At 100 ° C, compound 2 (109 mg, 0.603 mmol), Cs2CO3 (197 mg, 0.603 mmol), PD2(DBA)3 (18.4 mg, 0.0201 mmol) and SPhos (16.5 mg, 0.040)2 mmol) were added to a DMF (4 mL) solution of compound 1 (200 mg, 0.603 mmol) at 100 ° C. The reaction mixture was irradiated in a microwave reactor at 100 ° C for 1 hour. The reaction mixture was added to water (20 mL). The aqueous phase was extracted with ethyl acetate (50 milliliters). The combined organic phases were washed with an aqueous NaCl solution (15 mL x2), dried with Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluted with a petroleum ether solution of EtOAc from 0% to 50% to obtain compound 3 (110 mg, yield 44%) as a yellow solid. LC-MS: (ESI) m / z[M+H] + 589.2.

[0402] At 25 ° C, 1N HCl (1.0 mL) was added dropwise to a solution of compound 3 (110 mg, 0.184 mmol) in THF (5.0 mL) and stirred. The resulting mixture was stirred at 25 ° C for 30 minutes. The mixture was quenched with NaHCO3 solution (20 mL) and then extracted with ethyl acetate (20 mL x 2). The combined organic phases were dried over sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluted with a petroleum ether solution of THF from 0% to 80%, to give compound 4 (61.0 mg, yield 75%) as a yellow solid. LC-MS: (ESI) m / z[M+H] + 434.2.

[0403] To compound 4 (61.0mg, 0.141mmol) in DCM (4.0mL) solution, add propene-2-enoyl chloride (19.1mg, 0.211mmol) and TEA (42.7mg, 0.422mmol). The reaction mixture was stirred at 25°C for 10 minutes. The mixture was quenched with water (20mL), then extracted with DCM (20mL x 2). The combined organic phase was dried over sodium sulfate and filtered. The residue was purified by PrepHPLC (chromatographic column: -Xbridge-C18 150x 19mm, 5um mobile phase: ACN-H2O, 45-75%MeCN in 0.1%FA / water, 30mL / min), to obtain compound 9256 (31.6mg, 45% yield), which is a white solid.

[0404] 1 H NMR (400MHz, DMSO-d6) δ10.37(s,1H),7.79(d,J=8.8Hz,1H),7.71(d,J=8.8Hz,2H), 7.49(d,J=8.8Hz,2H),7.38(dd,J=8.8,2.8Hz,1H),6.90(d,J=2.8Hz,1H),6.50–6.40 (m,1H),6.28(dd,J=17.2,2.0Hz,1H),5.78(dd,J=10.0,2.0Hz,1H),4.44–4.33(m,1 H),3.80(s,3H),3.34–3.24(m,2H),2.54(s,3H),1.43(s,9H).LC-MS:(ESI)m / z[M+H] + 488.2.

[0405] Example 3-1

[0406]

[0407] Oxalyl chloride (19.2 g, 151 mmol) was added to a DCM (50 mL) solution of compound 1 (25 g, 100 mmol). The mixture was stirred at room temperature for 5 minutes, and then DMF (3 drops) was added. The resulting mixture was stirred at room temperature for 2 hours. The solvent was removed under reduced pressure, and the obtained residue was redispersed in compound 2 (150 mL). Anhydrous AlCl3 (20.2 g, 151.2 mmol) was then added at 0°C. The mixture was stirred at room temperature for 16 hours. The reaction was quenched with ice-cold water and extracted with CHCl3 (200 mL×3). The organic layer was dried and concentrated with Na2SO4. The residue was purified by FCC (PE / EA=20 / 1) to obtain white solid compound 3 (20 g, 41% yield).

[0408] Compound 4 (10 g, 42.70 mmol), compound 5 (8.2 g, 64.05 mmol), XPhos (2.04 g, 4.27 mmol), Pd(OAc)2 (0.48 g, 2.14 mmol) and TEA (12.96 g, 128.10 mmol) were mixed in dioxane (50 mL) and heated and stirred at 80 ° C under N2 atmosphere for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature and filtered. The filtrate was concentrated to obtain 20 g of brown oily compound 6.

[0409] Compound 6 (3 g, 0.011 mmol), compound 3 (4.56 g, 0.012 mmol), Pd (PPh 3 ) 4 (1.24 g, 0.001 mmol) and K 2 CO 3 (4.44 g, 0.032 mmol) were mixed in dioxane / H 2 O (5:1, 60 mL) and stirred at room temperature under N 2 for 12 hours. The reactant was diluted with EtOAc, filtered, extracted and concentrated. The crude product was purified by FCC (EA / PE=10%) to give yellow solid compound 7 (1.6 g, 32.7%).

[0410] At 0°C, LiOH·H2O (0.28 g, 0.012 mmol) was added to a solution of compound 7 (1.6 g, 0.004 mmol) in THF / MeOH / H2O (10 mL / 10 mL / 5 mL). The mixture solution was stirred at 0°C for 1 hour. The reaction was then quenched with water and extracted with EtOAc. The combined organics were washed with brine, dried over Na2SO4, filtered and concentrated. The solvent was removed under reduced pressure to obtain a yellow solid compound 8 (1.4 g, crude product).

[0411] A solution of compound 8 (700 mg, 1.881 mmol) and Dess Martin periodinane (1595 mg, 3.761 mmol) in DCM (10 mL) was stirred at room temperature for 1 hour. The solvent was removed in vacuo and the residue was purified by FCC (EA / PE=10%) to give compound 9 (600 mg, 77.57%) as a yellow oil.

[0412] Tetraethoxytitanium (739 mg, 3.242 mmol) and compound 10 (216 mg, 1.783 mmol) were added to a THF (6 mL) solution of compound 9 (600 mg, 1.621 mmol). The resulting mixture was stirred at room temperature for 16 hours. The mixture was then diluted with brine (10 m) and ethyl acetate (10 m). The resulting mixture was vigorously stirred at room temperature for 1.5 hours. The mixture was filtered and the organic layer was separated. The aqueous solution was extracted with EtOAc, and the combined organic layers were dried with Na2SO4, filtered and concentrated. The residue was purified by FCC (EA / PE=10%) to obtain yellow solid compound 11 (480 mg, 50%).

[0413] At 0 ° C, a solution of compound 12 (330 mg, 1.268 mmol) in THF (5 mL) was added to a solution of compound 11 (400 mg, 0.845 mmol) in THF (5 mL) within 10 minutes, and the resulting mixture was stirred at 0 ° C for 4 hours. The reaction was quenched with saturated NH4Cl and EA aqueous solution. The organic layer was separated and water was extracted with EA. The combined organic layer was washed with brine, dried over Na2SO4, filtered and concentrated. The residue was purified by FCC (usually gradient elution 5% MTBE: 95% hexane to 50% MTBE: 50% hexane) to give compound 13 (100 mg, 20%) as a white solid.

[0414] Compound 13 (100 mg, 0.169 mmol) and AlCl (34 mg, 0.254 mmol) were dissolved in EtOH (10 mL) and heated at 60 ° C under N for 30 minutes. The solution was then cooled to room temperature and quenched with NaHCO solution and extracted with EA. The combined organic layer was washed with brine, dried over NaSO, filtered and concentrated. The residue was purified by FCC (EA / PE=10%) to give yellow solid compound 14 (50 mg, yield 56.78%, purity 90%).

[0415] Compound 14 (50 mg, 0.107 mmol), compound 15 (12 mg, 0.214 mmol), XPhos-Pd-G3 (17 mg, 0.021 mmol) and Cs2CO3 (70 mg, 0.214 mmol) were mixed in ACN (3 mL) and heated with stirring at 60°C under N2 for 2 hours. The solvent was removed under reduced pressure, and the residue was extracted with EA (10 mL×3). The solvent was removed, and the residue was purified by prep-HPLC (chromatographic column: Phenomenex luna C18 250 mm×100 mm×10 um; mobile phase: [H2O (0.1% FA)-ACN]; B%: 10% to 55%, 40 minutes) to obtain a white solid 9098 (10.4 mg, 20.3% yield).

[0416] LCMS: [M+H] + =442.1;

[0417] 1 H NMR: (400MHz, MeOD) δ8.543(s,0.12H),7.810(d,J=7.6Hz,1H),7.712(d,J=4.0Hz,1H),7.436–7.372( m,4H),7.306(d,J=8.4Hz,2H),4.385(s,1H),3.648(s,2H),3.319(s,2H),2.564(s,3H),1.484(s,9H).

[0418] HPLC: 97.9% purity.

[0419] Example 3-2

[0420]

[0421] 9098Int 14 (50 mg, 0.107 mmol), compound 1 (44 mg, 0.161 mmol), Pd (OAc) 2 (12 mg, 0.054 mmol), P (o-tolyl) (33 mg, 0.107 mmol) and DIEA (60 mg, 0.110 mmol) were mixed in DMF (1 mL) and ACN (1 mL) and stirred for 3 hours at 75 ° C under microwave radiation under N 2. The mixture was then filtered, concentrated and diluted with water (10 mL). The resulting solution was extracted with EtOAc (10 mL × 3), and the combined organic layers were dried and concentrated with anhydrous Na 2 SO 4. The residue was purified by FCC (eluent PE / EtOAc = 90 / 10) to obtain compound 2 (30 mg, yield 42%) as a colorless oil.

[0422] A solution of compound 2 (30 mg, 0.045 mmol) in 1M HCl in MeOH / EA (3 mL / 3 mL) was stirred at room temperature for 4 hours. The solution was diluted with MeOH (2 mL) and dried under a stream of N2, and the crude product was dissolved in MeOH (2 mL) and purified by prep-HPLC (column: Phenomenex luna C18 250 mm×100 mm×10 um; mobile phase: [H2O (0.1% TFA)-ACN]; B%: 5% to 45%, 40 minutes) to give a yellow solid 9095 (14.4 mg, 70% yield).

[0423] LCMS: [M+H] + =460.1;

[0424] 1 H NMR: (400MHz, MeOD) δ7.828(d,J=7.6Hz,1H),7.729(dd,J=8.8,4.0Hz,1H),7.486(d,J=8.4Hz,2H),7.445–7.402(m,2H),7.369(t,J=7.6 Hz,2H),6.926(d,J=15.6Hz,1H),6.612–6.263(m,1H),4.412(s,1H),4.011(d,J=7.2Hz,2H),3.316(s,2H),2.572(s,3H),1.491(s,9H).

[0425] HPLC: 96.9% purity.

[0426] Example 4-1

[0427]

[0428] Compound 1 (500 mg, 0.912 mmol), compound 2 (207 mg, 1.367 mmol), Pd (PPh 3 ) 4 (211 mg, 0.182 mmol), CuI (35 mg, 0.182 mmol) and TEA (277 mg, 2.735 mmol) were dissolved in DMF (5 mL) and stirred at room temperature for 2 hours. The mixture was diluted with water (30 mL) and extracted with EA (30 mL x 3). The combined organic layers were dried over Na 2 SO 4 and concentrated. The crude product was purified by FCC (DCM / MeOH = 10: 1) to give brown solid compound 3 (400 mg, 0.629 mmol, 69.08% yield).

[0429] To a solution of compound 3 (400 mg, 0.699 mmol) in DCM (3 mL) was added TFA (3 mL), and the reaction mixture was stirred at room temperature for 2 hours. The solvent was then removed under reduced pressure to give a crude product, which was purified by FCC (DCM / MeOH=10:1) to give a brown solid compound 4 (280 mg, 0.489 mmol, 69.85% yield).

[0430] Compound 4 (280 mg, 0.543 mmol), compound 5 (133 mg, 0.652 mmol), HATU (310 mg, 0.815 mmol) and DIEA (211 mg, 1.629 mmol) were dissolved in DMF (5 mL) and stirred at room temperature for 2 hours. The mixture was then diluted with water (30 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were dried over Na2SO4 and concentrated. The crude product was purified by FCC (DCM / MeOH=10:1) to give yellow solid compound 6 (180 mg, 0.231 mmol, 42.49% yield).

[0431] Compound 6 (180 mg, 0.257 mmol) was dissolved in 1,4-dioxane (2 mL), and a solution of HCl in 1,4-dioxane (2 mL, 1 M) was added. The reaction mixture was stirred at room temperature for 2 hours. The solvent was removed under reduced pressure, and the crude product was purified by FCC (DCM / MeOH=10:1) to give a yellow solid compound 7 (150 mg, 0.224 mmol, 87.49% yield).

[0432] Compound 7 (150 mg, 0.249 mmol), 9154Int 6 (111 mg, 0.249 mmol), HATU (114 mg, 0.299 mmol) and DIEA (97 mg, 0.748 mmol) were dissolved in DMF (5 mL) and stirred at room temperature for 2 hours. The mixture was then diluted with water (25 mL) and extracted with EtOAc (25 mL x 3). The combined organic layers were dried and concentrated with Na2SO4. The crude product was purified by FCC (DCM / MeOH=10:1) to give compound 8 (120 mg, 0.111 mmol, 44.44% yield) as a white solid.

[0433] Compound 8 (100 mg, 0.097 mmol) and NaHCO (25 mg, 0.292 mmol) were dissolved in DMF (3 mL) and heated at 50° C. for 3 hours. The mixture was diluted with MeOH, filtered and purified by prep-HPLC (column: Phenomenexluna C18 250 mm x 100 mm x 10 um; mobile phase: [H2O (0.1% FA)-ACN]; B%: 30 minutes) to give a white solid 9157 (24.4 mg, 0.0261 mmol, yield 27.01%).

[0434] LCMS: [M+H] + =933.9;

[0435] 1 H NMR: (400MHz, d4-MeOD)δ7.427-7.325(m,8H),4.610(t,J=7.2Hz,1H),4.535(t,J=7.2Hz,1H),4.038(s,2H), 3.729(s,3H),3.729–3.430(m,12H),2.683(s,3H),2.612(s,3H),2.431(s,3H),2.009(s,3H),1.617(s,3H).

[0436] HPLC: 99.28% purity.

[0437] SFC: Rt = 2.920 min.

[0438] The following compounds were prepared with reference to the above examples:

[0439]

[0440]

[0441]

[0442]

[0443]

[0444]

[0445]

[0446]

[0447]

[0448]

[0449]

[0450]

[0451]

[0452]

[0453]

[0454]

[0455]

[0456]

[0457]

[0458]

[0459]

[0460]

[0461]

[0462]

[0463]

[0464]

[0465]

[0466]

[0467] Test Example 1 Cell Proliferation Assay

[0468] The cell lines used were 22RV1 or LNCaP, and the culture medium was RPMI-1640 plus 10% FBS and antibiotics. All incubations were performed at 37°C in a humidified carbon dioxide incubator.

[0469] On day 1, cells were digested with 0.25% trypsin (containing EDTA) and collected by centrifugation. Cells were seeded in a 96-well plate (white wall with transparent bottom) at 5000 cells / well and cultured overnight.

[0470] On the second day, the compound with a stock concentration of 10mM was diluted 8 times, and 2uL of each concentration was added to 98uL of fresh culture medium and mixed. Then, 5uL of the compound diluted in the culture medium at each concentration point was added to 100uL of overnight culture medium containing cells, and each concentration was repeated 2 times. Culture was continued for 72h.

[0471] After 72 hours of compound treatment, the assay plate was equilibrated to room temperature, 100uL CellTiterGlo (Promega) was added, mixed for 2 minutes, and allowed to stand at room temperature for 10 minutes before reading the luminescence on a SpectraMax M5 microplate reader to assess cell activity. The positive control was 1% Tween-20, and the negative control was 0.1% DMSO. IC values ​​were generated using a 4-parameter nonlinear regression fit in GraphPad Prism. 50 .

[0472]

[0473]

[0474]

[0475]

[0476]

[0477] Note: A<50nM,50nM <B<200nM,200nM<C<500nM,500nM<D

[0478] Test Example 2 BRD4 degradation assay

[0479] Conventional Western-blot method:

[0480] 1. Subculture and inoculate cells one day in advance (the most commonly used are 22RV1 and LnCap). Prepare the cell inoculation solution at a density of 0.67×10^6 / ml and inoculate into 12-well plates: 0.75ml / well or 24-well plates: 0.375ml / well. (The result is equivalent to 0.5×10^6 / well for 12-well plates; 0.25×10^6 / well for 24-well plates).

[0481] 2. Add the compound after 24-26 hours. Dissolve the compound in the culture medium at 4 times the concentration to be tested: 150ul (12-well plate) or 75ul (24-well plate). Add to the culture wells and mix thoroughly.

[0482] 3. Prepare cell homogenate with RIPA buffer (100ul / well for 12-well plate; 50ul / well for 24-well plate) according to the specified degradation experiment time. Vortex+ incubate on ice three times. Centrifuge at high speed 15800rpm (20000g) at 4℃ for 10min. Save the supernatant and determine the protein concentration by BCA method.

[0483] 4. Mix the sample, sample buffer and sample reducing agent in proportion, denature the protein sample at 75℃ for 10 minutes, and immediately cool on ice. Centrifuge for 5 minutes. Add 5μg of total protein to the gel sample well. Load the molecular weight markers into the wells on both sides. Run the gel at room temperature until the dye front reaches the lower opening.

[0484] 5. Transfer the proteins from the gel to the PVDF membrane using a mini blotting module at a constant voltage of 20 V for 60 min (refer to the manufacturer's manual for detailed steps).

[0485] 6. Cut and label the PVDF membrane. Block the membrane with 5% milk in TBST for 1 hour. Incubate with primary antibody in 1% bovine serum albumin at 4°C overnight. Incubate in 5% BSA / TBST with 1:10,000 anti-Rb-Ig G-HRP for 1.5 hours. Wash the membrane three times with TBST for 10 minutes each time between each step. Develop the color on ChemiDoc with chemiluminescent reagent. Acquire and analyze images. Use Chemidoc's own software for quantitative analysis. The blank control (DMSO) is 100%, and the degree of target degradation is expressed as a percentage of the control group. The grading method is: the target content is a percentage of the control: A<20%, 20% <B<50%,50%<C<80,80%<D<100%。

[0486] 7. Calculate dose-response curves and plot using GraphPad Prism.

[0487] Simple Western method:

[0488] 1-3. Same as the conventional Western-blot method above.

[0489] 4. Use the kit provided by ProteinSimple to prepare the sample and add the sample to the test plate according to the method in the concise user manual of the Wes fully automated protein expression analysis system.

[0490] 5. Perform detection and collect results on Wes or Jess fully automated protein expression analyzer.

[0491] 6. Use Compass SW software to edit the results and perform quantitative analysis. The quantitative and classification methods are the same as conventional Western-blot.

[0492]

[0493]

[0494] The above is an exemplary description of the implementation of the technical solution of the present invention. It should be understood that the protection scope of the present invention is not limited to the above implementation. Any modification, equivalent substitution, improvement, etc. made by those skilled in the art within the spirit and principle of the present invention should be included in the protection scope of the claims of this application.

Claims

1. A compound represented by formula (I) and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt: in, Represents a single bond or a double bond; Indicates that the ring in which it exists is aromatic; m is 1 or 2; Ring A is selected from unsubstituted or optionally substituted with one, two or more R a Substituted with the following groups: C 6-10 Aryl, 5-10 membered heteroaryl; each R a are the same or different and are independently selected from halogen, oxo (=O), C 1-12 Alkyl, C 1-12 Alkoxy; R1 is selected from unsubstituted or optionally substituted with one, two or more R b Substituted groups: 3-10 membered heterocyclic group, 5-10 membered heteroaryl group, -C(O)OR 11 、-C(O)N(R 12 )(R 13 ) or -OC(O)R 14 ; R 11 , R 12 , R 13 , R 14 The same or different, independently selected from H, OH, NH2, C 1-12 Alkyl, C 3-12 Cycloalkyl, 3-10 membered heterocyclyl, 5-10 membered heteroaryl, C 6-10 Aryl, 3-10 membered heterocyclic-C 1-12 Alkyl; each R b The same or different, independently selected from OH, NH2, C 1-12 Alkyl, C 2-12 Alkynyl; R2 is selected from C 1-12 Alkyl, C 1-12 Alkoxy, halogenated C 1-12 Alkyl, halogenated C 1-12 Alkoxy; R3 and R4 are the same or different and are independently selected from H, C 1-12 Alkyl, C 1-12 Alkoxy, halogenated C 1-12 Alkyl, halogenated C 1-12 Alkoxy; when middle When it is a double bond, R5 is selected from =NR 54 , when middle When it is a single bond, R5 is selected from H, halogen, unsubstituted or optionally substituted by one, two or more R c Substituted with the following groups: NH2, C 1-12 Alkyl, -C 1-12 Alkyl-N(R 51 )(R 52 )、-C(O)N(R 51 )(R 52 ),-C 1-12 Alkyl-OR 53 、-C(O)OR 53 , -C 1-12 Alkyl-NH-C(=NR 54 )(N(R 51 )(R 52 ))、-CH=N(R 54 )、-S(O)2R 55 、-C(O)R 55 ; R 51 , R 52 , R 53 , R 54 , R 55 The same or different, independently selected from H, OH, NH2, C 1-12 Alkyl, C 1-12 Alkoxy, C 1-12 Alkyl-C(O)-, C 2-12 Alkenyl-C(O)-, C 1-12 Alkyl-S(O)2-; each R c The same or different, independently selected from H, C 1-12 Alkyl-C(O)-, Boc, Boc-O-; X is selected from C or N; X1 is selected from S or C(R6); R6 is selected from H, C 1-12 Alkyl, C 1-12 Alkoxy; X2 is absent or is C(R7); R7 is selected from H, C 1-12 Alkyl, C 1-12 Alkoxy; X3 is selected from N or C(R8); R8 is selected from H, C 1-12 Alkyl, C 1-12 Alkoxy; L is absent or selected from halogen, NH, unsubstituted or optionally substituted with one, two or more R L Substituted with the following groups: 1-12 Alkyl, C 2-12 Alkenyl, C 2-12 Alkynyl, C 3-10 Cycloalkyl, 3-10 heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl; each R L are the same or different and are independently selected from H, halogen, oxo (=O), C 1-12 Alkyl, C 1-12 Alkoxy.

2. The compound according to claim 1 and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that: Ring A is selected from unsubstituted or optionally substituted with one, two or more R a Substituted with the following groups: phenyl or Preferably, ring A is selected from 3. The compound according to claim 1 or 2, and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that: R1 is selected from unsubstituted or optionally substituted with one, two or more R b Substituted with the following groups: C 1-6 Alkyl, 5-8 membered heterocyclic group, 5-6 membered heteroaryl (e.g., thiazolyl, oxazolyl, oxadiazolyl, triazolyl), -C(O)OR 11 、-C(O)N(R 12 )(R 13 ) or -OC(O)R 14 ; R 11 , R 12 , R 13 , R 14 The same or different, independently selected from H, OH, NH2, C 1-6 Alkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, 5-6 membered heteroaryl, C 6-10 Aryl, 3-8 membered heterocyclic group-C 1-6 Alkyl; each R b The same or different, independently selected from OH, NH2, C 1-6 Alkyl, C 2-6 Alkynyl or -OC(O)R 14 ; Preferably, R 11 , R 12 , R 13 , R 14 are the same or different and are independently selected from H, OH, NH2, methyl, ethyl, isopropyl, tert-butyl, hydroxyphenyl, propynyl, hydroxyethyl, methylcyclopropyl, Preferably, R1 is selected from Preferably, R2 is selected from C 1-6 Alkyl groups, such as methyl; Preferably, R3 and R4 are the same or different and are independently selected from H, C 1-6 Alkyl, C 1-6 Alkoxy, such as H, methyl, methoxy; Preferably, when middle When it is a double bond, R5 is selected from =NR 54 , when middle When it is a single bond, R5 is selected from H, halogen, unsubstituted or optionally substituted by one, two or more R c Substituted with the following groups: NH2, C 1-6 Alkyl, -C 1-6 Alkyl-N(R 51 )(R 52 )、-C(O)N(R 51 )(R 52 ),-C 1-6 Alkyl-OR 53 、-C(O)OR 53 , -C 1-6 Alkyl-NH-C(=NR 54 )(N(R 51 )(R 52 ))、-CH=N(R 54 )、-S(O)2R 55 、-C(O)R 55 ; R 51 , R 52 , R 53 , R 54 , R 55 The same or different, independently selected from H, OH, NH2, C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Alkyl-C(O)-, C 2-6 Alkenyl-C(O)-, C 1-6 Alkyl-S(O)2-; each R c The same or different, independently selected from H, C 1-6 Alkyl-C(O)-, Boc, Boc-O-; Preferably, when middle When it is a double bond, R5 is selected from when middle When it is a single bond, R5 is selected from H, Cl, NH2, methyl, 4. The compound according to any one of claims 1 to 3 and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that: X1 is selected from S or C(R6); R6 is selected from H, C 1-6 Alkyl, C 1-6 Alkoxy; X2 is absent or is C(R7); R7 is selected from H, C 1-6 Alkyl, C 1-6 Alkoxy; When X2 does not exist, middle is a single bond, X1 is S; when X2 is C(R7), middle is a double bond, X1 is C(R6); Preferably, X1 is selected from S or CH, X2 is absent or is CH; when X2 is absent, middle is a single bond, X1 is S; when X2 is CH, middle is a double bond, X1 is CH; Preferably, X3 is selected from N or CH; Preferably, the Selected from Preferably, the Selected from Preferably, the Selected from Preferably, L is absent or selected from halogen, NH, unsubstituted or optionally substituted by one, two or more R L Substituted with the following groups: 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, 3-8 heterocyclic group, C 6-10 Aryl, 5-6 membered heteroaryl; each R L are the same or different and are independently selected from H, halogen, oxo (=O), C 1-6 Alkyl, C 1-6 Alkoxy; Preferably, L is absent or selected from Cl, NH, unsubstituted or optionally substituted with one, two or more R L Substituted groups of the following: vinylene, ethynylene, 2,3-dihydroisoxazolylene, pyrazolylene, thiazolylene, furylene, azetidinylene, pyrrolylene, 2-azaspiro[3.3]heptanylene, thienylene, phenylene, cyclopropylene, cyclohexenylene, cyclopentenylene; Preferably, L is absent or selected from Cl, NH, 5. The compound according to any one of claims 1 to 4 and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that: Exemplary specific compounds of the compound represented by formula (I) are as follows: in, Represents a single bond or a double bond; Indicates that the ring in which it exists is aromatic; Ring A, R1, R2, R3, R4, R5, X1, X2, L, m have the definitions as described in any one of claims 1 to 4; Preferably, the compound represented by formula (I) is selected from the following structures: Wherein, ring A, R1, R2, R3, R4, R5, and L have the definitions as described in any one of claims 1 to 4.

6. The compound according to any one of claims 1 to 5 and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that: The compound represented by formula (I) is selected from the following compounds: Preferably, exemplary specific compounds of the compound represented by formula (I) are as follows: Preferably, exemplary specific compounds of the compound represented by formula (I) are as follows:

7. The compound represented by formula (III) and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof: in, Ring A, R1, R2, R3, R4, R5, L, m have the definitions as described in any one of claims 1 to 6, and Y is selected from -(CH2) p -[O-(CH2) q ] r , p is selected from 1, 2, 3, 4 or 5, q is selected from 1, 2, 3, 4 or 5, and r is selected from 0, 1, 2, 3, 4 or 5; Preferably, the compound represented by formula (III) has the following structure:

8. A pharmaceutical composition comprising the compound according to any one of claims 1 to 7 and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof.

9. Use of the compound according to any one of claims 1 to 7 and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof and a pharmaceutical composition comprising the same in the preparation of a medicament for preventing and / or treating a bromodomain-mediated disease or disorder; Preferably, the bromodomain protein is bromodomain protein 4 (BRD4); Preferably, the disease is cancer; Preferably, the cancer is selected from the group consisting of acoustic neuroma, acute leukemia, acute lymphocytic leukemia, acute myeloid leukemia, acute T-cell leukemia, basal cell carcinoma, bile duct cancer, bladder cancer, brain cancer, breast cancer, bronchial cancer, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic myeloid leukemia, colon cancer, colorectal cancer, craniopharyngioma, cystadenocarcinoma, diffuse Large B-cell lymphoma, dysplasia, embryonal carcinoma, endometrial carcinoma, endothelial sarcoma, ependymoma, epithelial carcinoma, erythroleukemia, esophageal carcinoma, estrogen receptor-positive breast cancer, essential thrombocythemia, Ewing's tumor, fibrosarcoma, follicular lymphoma, germ cell testicular cancer, glioma, glioblastoma, gliosarcoma, heavy chain disease, hemangioblastoma, hepatoma, hepatocellular carcinoma, hormone-insensitive prostate cancer, leiomyosarcoma, leukemia, liposarcoma, lung cancer, intralymphatic medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, multiple myeloma, myeloid leukemia, myeloma, myxosarcoma, neuroblastoma, NUT midline carcinoma (NMC), non-small cell lung cancer carcinoma, oligodendroglioma, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pinealoma, polycythemia vera, prostate cancer, rectal cancer, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, small cell lung cancer, solid tumors (carcinomas and sarcomas), small cell lung cancer, gastric cancer, squamous cell carcinoma, synovioma, sweat gland carcinoma, thyroid cancer, Waldenstrom's macroglobulinemia, testicular tumors, uterine cancer, and Wilms' tumor; Preferably, the disease is an autoimmune disease or inflammatory disease selected from the group consisting of Addison's disease, acute gout, ankylosing spondylitis, asthma, atherosclerosis, Behcet's disease, bullous dermatosis, chronic obstructive pulmonary disease (COPD), Crohn's disease, dermatitis, eczema, giant cell arteritis, glomerulonephritis, hepatitis, hypophysitis, inflammatory bowel disease, Kawasaki disease, lupus nephritis, multiple sclerosis, myocarditis, myositis, nephritis, organ transplant rejection, osteoarthritis, pancreatitis, pericarditis, polyarteritis nodosa, pneumonia, primary biliary cirrhosis, psoriasis, psoriatic arthritis, rheumatoid arthritis, scleritis, sclerosing cholangitis, sepsis, systemic lupus erythematosus, Takayasu's arteritis, toxic shock, thyroiditis, type I diabetes, ulcerative colitis, uveitis, vitiligo, vasculitis and Wegener's granulomatosis; Preferably, the disease or disorder is selected from the group consisting of: AIDS; chronic kidney disease, diabetic nephropathy, hypertensive nephropathy, HIV-related nephropathy, glomerulonephritis, lupus nephritis, IgA nephropathy, focal segmental glomerulosclerosis, membranous glomerulonephritis, minimal change disease, polycystic kidney disease and tubulointerstitial nephritis; acute kidney injury or disease or condition; obesity; dyslipidemia; hypercholesterolemia; Alzheimer's disease; metabolic syndrome; hepatic steatosis; type II diabetes; insulin resistance; and diabetic retinopathy.

10. A method for degrading a bromodomain-containing protein in a cell, comprising exposing the cell to an effective amount of the compound of any one of claims 1 to 7 and its racemate, stereoisomer, tautomer, isotope-labeled form, N-oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, wherein the compound achieves degradation of the bromodomain-containing protein; Preferably, the bromodomain-containing protein is bromodomain-containing protein 4 (BRD4).

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