WO2020088526A1 - DOUBLE PYRAZOLE COMPOUND AS TGF-βR1 KINASE INHIBITOR - Google Patents

DOUBLE PYRAZOLE COMPOUND AS TGF-βR1 KINASE INHIBITOR Download PDF

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WO2020088526A1
WO2020088526A1 PCT/CN2019/114347 CN2019114347W WO2020088526A1 WO 2020088526 A1 WO2020088526 A1 WO 2020088526A1 CN 2019114347 W CN2019114347 W CN 2019114347W WO 2020088526 A1 WO2020088526 A1 WO 2020088526A1
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isomer
compound
pharmaceutically acceptable
acceptable salt
compound according
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PCT/CN2019/114347
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French (fr)
Chinese (zh)
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付翔宇
丁照中
胡利红
陈曙辉
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南京明德新药研发有限公司
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Priority to CN201980067051.7A priority Critical patent/CN112839946B/en
Publication of WO2020088526A1 publication Critical patent/WO2020088526A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/72—Nitrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/32—One oxygen, sulfur or nitrogen atom
    • C07D239/42—One nitrogen atom
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D498/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D498/04—Ortho-condensed systems

Definitions

  • the invention relates to a class of bicyclic pyrazole compounds as TGF- ⁇ R1 inhibitors, and their application in the preparation of TGF- ⁇ R1 inhibitor drugs. Specifically, it relates to a compound represented by formula (I), a pharmaceutically acceptable salt thereof, or an isomer thereof.
  • Transforming growth factor ⁇ (Transforming growth factor- ⁇ , TGF- ⁇ ) is a multi-functional growth factor superfamily with a wide range of biological activities, involved in early embryonic development, cartilage and bone formation, synthesis of outer matrix, inflammation, Interstitial fibrosis, regulation of immune and endocrine functions, tumor formation and development.
  • TGF- ⁇ superfamily consists of a class of structural and functionally related polypeptide growth factors.
  • TGF- ⁇ is one of the important members of this family.
  • TGF- ⁇ mainly exists in three forms: TGF- ⁇ 1, TGF- ⁇ 2, and TGF- ⁇ 3, which are located on different chromosomes, of which TGF- ⁇ 1 accounts for the highest proportion (> 90%) in somatic cells. It has the strongest activity, the most functions, and the most widespread distribution.
  • TGF- ⁇ signaling molecule performs signal transduction through the transmembrane receptor complex.
  • TGF- ⁇ receptors are transmembrane proteins that exist on the cell surface and are divided into type I receptors (TGF- ⁇ RI), type II receptors (TGF- ⁇ R) II and type III receptors (TGF- ⁇ RIII), of which TGF - ⁇ R is also called activin-like receptor 5 (ALK5).
  • TGF- ⁇ RIII lacks intrinsic activity, which is mainly related to the storage of TGF- ⁇ .
  • TGF- ⁇ R I and TGF- ⁇ R II belong to the serine / threonine kinase family.
  • Type II receptors can bind to TGF- ⁇ ligands with high affinity and form heterologous receptor complexes with type I receptors.
  • a region rich in glycine and serine residues (GS domain) near the membrane of type I receptors is phosphorylated to initiate intracellular signaling cascade reactions.
  • TGF- ⁇ signal transduction and regulation molecule in the cell, which can directly transduce TGF- ⁇ signal from the cell membrane as in the nucleus.
  • activated TGF- ⁇ first binds to TGF- ⁇ R II on the cell membrane surface to form a heterodimer complex, and TGF- ⁇ R I recognizes and binds the binary complex.
  • TGF- ⁇ R II phosphorylates the serine / threonine of the GS domain of TGF- ⁇ R I cytoplasmic region, thereby activating TGF- ⁇ R I; activated TGF- ⁇ R I further phosphorylates R-Smads (Smad2 / Smad3) protein, The latter then combines with Co-Smad (Smad4) to form a heterotrimeric complex, which enters the nucleus and cooperates with other co-activator and co-inhibitor to regulate Target gene transcription. Changes in any part of the TGF- ⁇ / Smads signaling pathway will cause abnormalities in the signal transduction pathway.
  • TGF- ⁇ can directly affect the growth of tumors (extrinsic effects of TGF- ⁇ signaling), or by inducing epithelial-mesenchymal transition, blocking anti-tumor immune responses, and increasing tumor-related fibrosis And enhanced vascular regeneration indirectly affects tumor growth (intrinsic effect of TGF- ⁇ ).
  • TGF- ⁇ has a strong fibrosis induction effect, it is an activator of fibroblasts associated with tumors. These fibroblasts are the main source of collagen type I and other fibrotic factors. The induced products of fibroblasts and other fibrotic factors may continue to cultivate a microenvironment that will reduce the immune response, increase drug resistance and strengthen tumor angiogenesis.
  • TGF- ⁇ affects blood vessels Health regeneration.
  • TGF- ⁇ R type I-deficient mouse embryos show severe defects in vascular development, proving that TGF- ⁇ signaling channels are key regulators in the development of vascular endothelium and smooth muscle cells.
  • TGF- ⁇ is obviously associated with immune escape and has a greater impact on the anti-tumor immune response mediated by CD8 + T cells.
  • patients with high expression of TGF- ⁇ gene responded to PD-L1 monoclonal antibody and the simulated survival rate was low.
  • the basic research of TGF- ⁇ monoclonal antibody also proves that when it is used in conjunction with PD-L1 monoclonal antibody, more CD8 + T cells infiltrate and play a role, revealing the immune activation effect and mechanism of blocking TGF- ⁇ . Because of the immunomodulatory effect of TGF- ⁇ , single-agent small-molecule TGF- ⁇ R inhibitors or combined with PD- (L) 1 monoclonal antibody have great application prospects in the treatment of various solid tumors.
  • Lilly's patent application WO2002094833A1 reported that compound A (ie LY2157299 or Galunisertib) has TGF- ⁇ inhibitory activity. The compound can inhibit the invasion and metastasis of tumor cells, and at the same time inhibit the infiltration of tumor cells into blood vessels. There are currently multiple clinical trials in progress.
  • Another patent application WO2016057278A1 from Eli Lilly reported compound B (LY3200882), which is a newly developed TGF- ⁇ small molecule inhibitor. The first phase of clinical trials of this compound in combination with PD-L1 in the treatment of solid tumors is in progress.
  • the present invention provides a compound represented by formula (I), a pharmaceutically acceptable salt thereof, or an isomer thereof,
  • ring A is 5-6 membered heteroaryl, phenyl, C 5-6 cycloalkyl or 5-6 membered heterocycloalkyl;
  • R 1 , R 2 and R 3 are each independently H, F, Cl, Br, I, -CN, -OH, C 1-6 alkoxy, C 1-6 alkyl, C 2-4 alkenyl, C 2-4 alkynyl or C 3-6 cycloalkyl, wherein the C 1-6 alkoxy, C 1-6 alkyl, C 2-4 alkenyl, C 2-4 alkynyl and C 3- 6 cycloalkyl groups are optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, CN, -OH, -CH 3 , -OCH 3 and -NH 2 ;
  • n 1 or 2;
  • R 4 is 5-6 membered heteroaryl or phenyl, wherein the 5-6 membered heteroaryl and phenyl are optionally substituted with 1, 2 or 3 R a ;
  • T 1 is -O- or -CH 2- ;
  • T 2 is N or C (R 5 );
  • R 5 is H, F, Cl, Br, -CN, -OH, -NH 2 , -OCH 3 or -CH 3 ;
  • Each R d is independently F, Cl, Br, I, -OH, -CN, -NH 2, -OCH 3, -OCH 2 CH 3, -CH 3, -CF 3 or -CH 2 CH 3;
  • R b and R c are each independently H, -CH 3 , -CH 2 CH 3 , -CH 2 CH 2 CH 3 , -CH 2 (CH 3 ) 2 or cyclopropyl;
  • the 5-6 membered heterocycloalkyl group and 5-6 membered heteroaryl group respectively contain 1, 2, 3 or 4 heteroatoms or heteroatom groups independently selected from N, -O-, -S- and -NH- .
  • the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formula (I-A):
  • rings A, R 1 , R 2 , R 3 , R 4 , T 1 and m are as defined in the present invention.
  • the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formula (I-B):
  • rings A, R 1 , R 2 , R 3 , R 4 , R 5 , T 1 and m are as defined in the present invention.
  • R 4 is a 5-6 membered heteroaryl or phenyl, wherein said 5-6 membered heteroaryl and phenyl are optionally substituted with 1, 2, or 3 R a;
  • R 5 is H, F, Cl, Br, —CN, —OH, —OCH 3 , —NH 2 or —CH 3 , and other variables are as defined in the present invention.
  • the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formula (I-C):
  • rings A, R 1 , R 2 , R 3 , T 1 , m, R 6 and L are as defined in the present invention.
  • each of the above Ra is independently CN, -OCH 3 , Other variables are as defined in the present invention.
  • R 4 is pyrrolyl, pyrazolyl, pyridyl, pyrimidinyl, or phenyl.
  • the pyrrolyl, pyrazolyl, pyridyl, pyrimidinyl, and phenyl are optionally substituted by 1, 2 or 3 R a are substituted, other variables are as defined in the present invention.
  • R 4 is R a and the other variables are as defined in the present invention.
  • the above R 4 is Other variables are as defined in the present invention.
  • the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formulas (I-A1) to (I-A4):
  • Ring A, R 1, R 2, R 3, T 1, R a and m are as defined in the present invention.
  • the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formulas (I-A5) to (I-A12):
  • Ring A, R 1, R 2, R 3 and R a are as defined in the present invention.
  • the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formulas (I-B1) to (I-B4):
  • Ring A, R 1, R 2, R 3, R 5, T 1, m and R a are as defined in the present invention.
  • the above compound, its pharmaceutically acceptable salt, or its isomer has a structure represented by formulas (I-B5) to (I-B12):
  • Ring A, R 1, R 2, R 3, R 5 and R a are as defined in the present invention.
  • the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formula (I-C1) or (I-C2):
  • rings A, R 1 , R 2 , R 3 , R 6 and L are as defined in the present invention.
  • R 1 , R 2 and R 3 are each independently H, F, Cl, Br, I, -CN, -OH, -OCH 3 , -OCH 2 CH 3 , -CH 3 , -CH 2 CH 3 , vinyl, ethynyl, or cyclopropyl, wherein the -OCH 3 , -OCH 2 CH 3 , -CH 3 , -CH 2 CH 3 , vinyl, ethynyl, and cyclopropyl are is selected from substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, CN, -OH, -CH 3, -OCH 3 and -NH 2 to a substituent group, the other variables are as defined in the present invention.
  • R 1 , R 2 and R 3 are each independently H, F, Cl, Br, -CN, -OH, -OCH 3 , -CH 3 , -CH 2 CH 3 or- CF 3 , other variables are as defined in the present invention.
  • the above-mentioned ring A is thienyl, pyrrolyl, pyridyl, pyrimidinyl, phenyl, or tetrahydro-2H-pyranyl, and other variables are as defined in the present invention.
  • R 1 , R 2 and R 3 and other variables are as defined in the present invention.
  • the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formulas (I-A13) to (I-A16):
  • R 1 and R a are as defined in the present invention.
  • the above compound, its pharmaceutically acceptable salt, or its isomer has a structure represented by formulas (I-B13) to (I-B36):
  • R 1 and R a are defined in the present invention.
  • the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formulae (I-C3) to (I-C5):
  • R 1 , L and R 6 are as defined in the present invention.
  • the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formulas (I-C6) to (I-C9):
  • R 1 and R 6 are as defined in the present invention.
  • the above R 6 is H, —C ( ⁇ O) NH 2 , isopropyl, cyclopropyl, cyclohexyl, pyrazolyl, or pyridyl, wherein the isopropyl, cyclopropyl
  • the radical, cyclohexyl, pyrazolyl and pyridyl are optionally substituted with 1, 2 or 3 Rd , Rd and other variables are as defined in the present invention.
  • Other variables are as defined in the present invention.
  • Other variables are as defined in the present invention.
  • the above compound is selected from compounds of the formula, pharmaceutically acceptable salts or isomers thereof,
  • the present invention also provides the use of the above compound, its pharmaceutically acceptable salt or its isomer in the preparation of TGF- ⁇ R1 inhibitor drugs.
  • the invention also provides the application of the above compound, its pharmaceutically acceptable salt or its isomer in the preparation of a drug for treating solid cancer.
  • the compound of the present invention has high selectivity for TGF- ⁇ R1 and has significant kinase inhibitory activity. It showed obvious inhibition of the downstream signal of TGF- ⁇ in cells, and also had excellent pharmacokinetics, pharmacodynamic properties and in vivo pharmacodynamics.
  • pharmaceutically acceptable refers to those compounds, materials, compositions and / or dosage forms that are within the scope of reliable medical judgment and are suitable for use in contact with human and animal tissues Without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit / risk ratio.
  • pharmaceutically acceptable salt refers to a salt of a compound of the present invention, prepared from a compound having a specific substituent and a relatively non-toxic acid or base found in the present invention.
  • base addition salts can be obtained by contacting such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent.
  • Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts.
  • acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent.
  • Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, Bisulfate, hydroiodic acid, phosphorous acid, etc .; and organic acid salts, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, Fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; also includes salts of amino acids (such as arginine, etc.) , And salts of organic acids such as glucuronic acid. Certain compounds of the present invention contain basic and acidic functional groups and can be converted to any base or
  • the pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing acid radicals or bases by conventional chemical methods. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of both.
  • the compounds of the present invention may exist in specific geometric or stereoisomeric forms.
  • the present invention contemplates all such compounds, including cis and trans isomers, (-)-and (+)-enantiomers, (R)-and (S) -enantiomers, diastereomers Isomers, (D) -isomers, (L) -isomers, and their racemic mixtures and other mixtures, such as enantiomerically or diastereomerically enriched mixtures, all of which belong to this Within the scope of the invention. Additional asymmetric carbon atoms may be present in the substituents such as alkyl. All these isomers and mixtures thereof are included in the scope of the present invention.
  • enantiomer or “optical isomer” refers to stereoisomers in a mirror image relationship with each other.
  • cis-trans isomer or “geometric isomer” is caused by the fact that double bonds or single bonds of ring-forming carbon atoms cannot rotate freely.
  • diastereomers refers to stereoisomers in which molecules have two or more chiral centers and are in a non-mirror relationship.
  • the following formula (A) indicates that the compound exists as a single isomer of formula (A-1) or (A-2) or as two isomers of formula (A-1) and formula (A-2) Exists in the form of a mixture;
  • the following formula (B) indicates that the compound exists as a single isomer of formula (B-1) or formula (B-2) or as both formula (B-1) and formula (B-2) There is a mixture of isomers.
  • the following formula (C) indicates that the compound exists as a single isomer of formula (C-1) or formula (C-2) or as two isomers of formula (C-1) and formula (C-2) In the form of a mixture.
  • tautomer or “tautomeric form” means that at room temperature, isomers of different functional groups are in dynamic equilibrium and can quickly convert to each other. If tautomers are possible (as in solution), the chemical equilibrium of tautomers can be achieved.
  • proton tautomers also known as prototropic tautomers
  • proton tautomers include interconversion through proton migration, such as keto-enol isomerization and imine-ene Amine isomerization.
  • Valence tautomer (valence tautomer) includes some recombination of bond-forming electrons for mutual conversion.
  • keto-enol tautomerization is the interconversion between two tautomers of pentane-2,4-dione and 4-hydroxypent-3-en-2-one.
  • the terms “rich in one isomer”, “isomer enriched”, “rich in one enantiomer” or “enantiomerically enriched” refer to one of the isomers or pairs
  • the content of the enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or 96% or greater, or 97% or greater, or 98% or greater, or 99% or greater, or 99.5% or greater, or 99.6% or greater, or 99.7% or greater, or 99.8% or greater, or greater than or equal to 99.9%.
  • the term “isomer excess” or “enantiomeric excess” refers to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, then the excess of isomer or enantiomer (ee value) is 80% .
  • optically active (R)-and (S) -isomers and D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If an enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, where the resulting mixture of diastereomers is separated and the auxiliary groups are cleaved to provide pure The desired enantiomer.
  • a salt of a diastereomer is formed with an appropriate optically active acid or base, and then by conventional methods known in the art
  • the diastereomers are resolved and the pure enantiomers are recovered.
  • the separation of enantiomers and diastereomers is usually done by using chromatography, which uses a chiral stationary phase, and is optionally combined with chemical derivatization methods (for example, the formation of amino groups from amines) Formate).
  • the compound of the invention may contain unnatural proportions of atomic isotopes at one or more atoms constituting the compound.
  • compounds can be labeled with radioisotopes, such as tritium ( 3 H), iodine-125 ( 125 I) or C-14 ( 14 C).
  • the hydrogen can be replaced by heavy hydrogen to form a deuterated drug.
  • the bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have reduced toxic and side effects and increased drug stability. , Strengthen efficacy, prolong the biological half-life of drugs and other advantages.
  • the conversion of all isotopic compositions of the compounds of the present invention, whether radioactive or not, is included in the scope of the present invention.
  • substituted means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include heavy hydrogen and hydrogen variants, as long as the valence state of the particular atom is normal and the compound after substitution is stable of.
  • Oxygen substitution does not occur on aromatic groups.
  • optionally substituted means that it may or may not be substituted. Unless otherwise specified, the type and number of substituents may be arbitrary on the basis that they are chemically achievable.
  • any variable (such as R) appears more than once in the composition or structure of a compound, its definition in each case is independent.
  • R when any variable (such as R) appears more than once in the composition or structure of a compound, its definition in each case is independent.
  • the group can optionally be substituted with up to two Rs, and R in each case has independent options.
  • combinations of substituents and / or variants thereof are only allowed if such combinations will produce stable compounds.
  • linking group When the number of a linking group is 0, such as-(CRR) 0- , it means that the linking group is a single bond.
  • one of the variables When one of the variables is selected from a single bond, it means that the two groups to which it is connected are directly connected. For example, when L represents a single bond in A-L-Z, it means that the structure is actually A-Z.
  • substituents listed do not indicate through which atom they are connected to the substituted group, such substituents can be bonded through any of their atoms, for example, pyridyl as a substituent can be through any one of the pyridine rings The carbon atom is attached to the substituted group.
  • connection direction is arbitrary, for example,
  • the linking group L in the middle is -MW-, then -MW- can be formed by connecting ring A and ring B in the same direction as the reading order from left to right It can also be formed by connecting ring A and ring B in the opposite direction to the reading order from left to right
  • Combinations of the linking groups, substituents, and / or variants thereof are only allowed if such combinations will produce stable compounds.
  • any one or more sites of the group may be connected to other groups by chemical bonds.
  • the chemical bond connecting the site with other groups can be a straight solid bond Straight dotted key Or wavy lines Said.
  • the straight solid line bond in -OCH 3 indicates that it is connected to other groups through the oxygen atom in the group;
  • the straight dashed bond in means that the two ends of the nitrogen atom in the group are connected to other groups;
  • the wavy line in indicates that it is connected to other groups through the 1 and 2 carbon atoms in the phenyl group.
  • the number of atoms on a ring is usually defined as the number of members of the ring.
  • “5-7 membered ring” refers to a “ring” with 5-7 atoms arranged around it.
  • the term "5-6 membered ring” means cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, cycloalkynyl, heterocycloalkynyl, consisting of 5 to 6 ring atoms, Aryl or heteroaryl.
  • the ring includes a single ring, and also includes a double ring system such as a spiro ring, a bicyclic ring, and a bridge ring. Unless otherwise specified, the ring optionally contains 1, 2 or 3 heteroatoms independently selected from O, S and N.
  • the 5-6 member ring includes 5 member, 6 member ring, and the like.
  • 5-6 membered ring includes, for example, phenyl, pyridyl, piperidinyl, and the like; on the other hand, the term “5-6 membered heterocycloalkyl” includes piperidinyl and the like, but does not include phenyl.
  • ring also includes ring systems containing at least one ring, where each "ring” independently conforms to the above definition.
  • C 1-6 alkyl is used to indicate a linear or branched saturated hydrocarbon group composed of 1 to 6 carbon atoms.
  • the C 1-6 alkyl group includes C 1-5 , C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-4 , C 6 and C 5 alkyl groups; etc .; Is monovalent (such as methyl), divalent (such as methylene) or multivalent (such as methine).
  • C 1-6 alkyl examples include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl , S-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl and so on.
  • C 1-4 alkyl is used to indicate a linear or branched saturated hydrocarbon group composed of 1 to 4 carbon atoms.
  • the C 1-4 alkyl group includes C 1-2 , C 1-3 and C 2-3 alkyl groups, etc .; it may be monovalent (such as methyl), divalent (such as methylene) or multivalent ( Such as methine).
  • Examples of C 1-4 alkyl include but are not limited to methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl , S-butyl and t-butyl) and so on.
  • C 2-4 alkenyl is used to denote a linear or branched hydrocarbon group consisting of at least one carbon-carbon double bond consisting of 2 to 4 carbon atoms, carbon-carbon double The bond can be located anywhere on the group.
  • the C 2-4 alkenyl group includes C 2-3 , C 4 , C 3 and C 2 alkenyl groups; the C 2-4 alkenyl group may be monovalent, divalent or multivalent. Examples of C 2-4 alkenyl include, but are not limited to, vinyl, propenyl, butenyl, butadienyl, and the like.
  • C 2-4 alkynyl is used to denote a linear or branched hydrocarbon group consisting of 2 to 4 carbon atoms containing at least one carbon-carbon triple bond, carbon-carbon triple The bond can be located anywhere on the group.
  • the C 2-4 alkynyl group includes C 2-3 , C 4 , C 3 and C 2 alkynyl groups. It can be monovalent, bivalent or multivalent. Examples of C 2-4 alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, and the like.
  • C 1-6 alkoxy refers to those alkyl groups containing 1 to 6 carbon atoms connected to the rest of the molecule through one oxygen atom.
  • the C 1-6 alkoxy group includes C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-4 , C 6 , C 5 , C 4 and C 3 alkoxy groups, etc. .
  • C 1-6 alkoxy examples include but are not limited to methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy Oxy, s-butoxy and t-butoxy), pentyloxy (including n-pentyloxy, isopentyloxy and neopentyloxy), hexyloxy, etc.
  • C 1-4 alkoxy refers to those alkyl groups containing 1 to 4 carbon atoms connected to the rest of the molecule through one oxygen atom.
  • the C 1-4 alkoxy group includes C 1-3 , C 1-2 , C 2-4 , C 4 and C 3 alkoxy groups and the like.
  • Examples of C 1-6 alkoxy include but are not limited to methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy Oxy, s-butoxy and t-butoxy), pentyloxy (including n-pentyloxy, isopentyloxy and neopentyloxy), hexyloxy, etc.
  • halogen or halogen itself or as part of another substituent means a fluorine, chlorine, bromine or iodine atom.
  • C 3-6 cycloalkyl means a saturated cyclic hydrocarbon group consisting of 3 to 6 carbon atoms, which is a monocyclic and bicyclic ring system, the C 3-6 cycloalkyl Including C 3-5 , C 4-5 and C 5-6 cycloalkyl, etc .; it can be monovalent, divalent or multivalent.
  • Examples of C 3-6 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.
  • C 5-6 cycloalkyl means a saturated cyclic hydrocarbon group consisting of 5 to 6 carbon atoms, which is a monocyclic and bicyclic ring system, the C 5-6 cycloalkyl Including C 5 and C 6 cycloalkyl and the like; it can be monovalent, divalent or polyvalent.
  • Examples of C 5-6 cycloalkyl include, but are not limited to, cyclopentyl, cyclohexyl, and the like.
  • the term "5-6 membered heterocycloalkyl" by itself or in combination with other terms means a saturated cyclic group consisting of 5 to 6 ring atoms with 1, 2, 3 or 4 ring atoms Are heteroatoms independently selected from O, S, and N, and the rest are carbon atoms, wherein nitrogen atoms are optionally quaternized, and nitrogen and sulfur heteroatoms may be optionally oxidized (ie, NO and S (O) p , p Is 1 or 2). It includes single-ring and double-ring systems, wherein the double-ring system includes spiro ring, parallel ring and bridge ring.
  • the hetero atom may occupy the connection position of the heterocyclic alkyl group to the rest of the molecule.
  • the 5-6 membered heterocycloalkyl group includes 5-membered and 6-membered heterocycloalkyl groups.
  • 5-6 membered heterocycloalkyl examples include, but are not limited to, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothienyl (including tetrahydrothien-2-yl and tetrahydrothien-3-yl, etc.) , Tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl, and 3-piperidinyl, etc.), piperazinyl (including 1 -Piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazole Alkyl, 1,2-oxazinyl, 1,2-thiazinyl, hexa
  • C 6-12 aryl ring and “C 6-12 aryl group” can be used interchangeably.
  • the term “C 6-12 aryl ring” or “C 6-12 aryl group” means from 6 to A cyclic hydrocarbon group consisting of 12 carbon atoms with a conjugated ⁇ -electron system. It can be a single ring, a fused bicyclic ring, or a fused tricyclic ring system, where each ring is aromatic. It may be monovalent, divalent or multivalent, and C 6-12 aryl groups include C 6-10 , C 6-9 , C 6-8 , C 12 , C 10 and C 6 aryl groups and the like. Examples of C 6-12 aryl groups include, but are not limited to, phenyl, naphthyl (including 1-naphthyl, 2-naphthyl, etc.).
  • C 6-10 aryl ring and “C 6-10 aryl group” can be used interchangeably.
  • the term “C 6-10 aryl ring” or “C 6-10 aryl group” means from 6 to A cyclic hydrocarbon group consisting of 10 carbon atoms with a conjugated ⁇ -electron system. It can be a single ring, a fused bicyclic ring or a fused tricyclic ring system, where each ring is aromatic. It may be monovalent, divalent or multivalent, and C 6-10 aryl groups include C 6-9 , C 9 , C 10 and C 6 aryl groups.
  • Example C 6- 10 aryl group include, but are not limited to, phenyl, naphthyl (including 1-naphthyl and 2-naphthyl and the like).
  • the terms “5-6 membered heteroaryl ring” and “5-6 membered heteroaryl group” are used interchangeably, and the term “5-6 membered heteroaryl group” means composed of 5 to 6 ring atoms In a monocyclic group with a conjugated ⁇ electron system, 1, 2, 3, or 4 ring atoms are heteroatoms independently selected from O, S, and N, and the rest are carbon atoms. Where nitrogen atoms are optionally quaternized, nitrogen and sulfur heteroatoms can be optionally oxidized (ie NO and S (O) p , p is 1 or 2). The 5-6 membered heteroaryl group can be attached to the rest of the molecule through a heteroatom or carbon atom.
  • the 5-6 membered heteroaryl group includes 5-membered and 6-membered heteroaryl groups.
  • Examples of the 5-6 membered heteroaryl group include but are not limited to pyrrolyl (including N-pyrrolyl, 2-pyrrolyl, and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyryl Oxazolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl, and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl, and 5- Oxazolyl, etc.), triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl and 4H-1, 2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isox
  • C n-n + m or C n -C n + m includes any specific case of n to n + m carbons, for example, C 1-12 includes C 1 , C 2 , C 3 , C 4 , C 5 , C 6 , C 7 , C 8 , C 9 , C 10 , C 11 , and C 12 , and also includes any range from n to n + m, for example, C 1-12 includes C 1- 3 , C 1-6 , C 1-9 , C 3-6 , C 3-9 , C 3-12 , C 6-9 , C 6-12 , and C 9-12, etc .; similarly, n yuan to n + m member means that the number of atoms in the ring is n to n + m, for example, 3-12 member ring includes 3 member ring, 4 member ring, 5 member ring, 6 member ring, 7 member ring, 8 member ring, 9 member ring , 10-membered
  • leaving group refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (eg, an affinity substitution reaction).
  • representative leaving groups include triflate; chlorine, bromine, and iodine; sulfonate groups such as mesylate, tosylate, p-bromobenzenesulfonate, and p-toluenesulfonate Ester, etc .; acyloxy, such as acetoxy, trifluoroacetoxy, etc.
  • protecting group includes but is not limited to "amino protecting group", “hydroxy protecting group” or “mercapto protecting group”.
  • amino protecting group refers to a protecting group suitable for preventing side reactions at the amino nitrogen position.
  • Representative amino protecting groups include, but are not limited to: formyl; acyl, such as alkanoyl (such as acetyl, trichloroacetyl, or trifluoroacetyl); alkoxycarbonyl, such as tert-butoxycarbonyl (Boc) ; Arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), trityl (Tr), 1,1-di -(4'-methoxyphenyl) methyl; silyl, such as trimethylsilyl (TMS) and tert-butyld
  • hydroxyl protecting group refers to a protecting group suitable for preventing side reactions of hydroxyl groups.
  • Representative hydroxy protecting groups include, but are not limited to: alkyl groups, such as methyl, ethyl, and tert-butyl; acyl groups, such as alkanoyl groups (such as acetyl); arylmethyl groups, such as benzyl (Bn), p-methyl Oxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl, such as trimethylsilyl (TMS) and tert-butyl Dimethylsilyl (TBS) and so on.
  • alkyl groups such as methyl, ethyl, and tert-butyl
  • acyl groups such as alkanoyl groups (such as acetyl)
  • arylmethyl groups such as benzyl (Bn), p-methyl Oxybenzyl (
  • the compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by the combination with other chemical synthesis methods, and well known to those skilled in the art Equivalently, preferred embodiments include but are not limited to the embodiments of the present invention.
  • the solvent used in the present invention is commercially available.
  • CDCl 3 stands for deuterated chloroform
  • DMSO stands for dimethyl sulfoxide
  • Boc stands for tert-butoxycarbonyl
  • DPBS Dulbecco's Phosphate Buffered Saline.
  • the compounds of the present invention are based on conventional naming principles or use in the art
  • the software is named, and the commercially available compounds adopt the supplier catalog name.
  • Step A Dissolve compound 1-1 (200 g, 1.02 mol, 1.0 equiv) in 1 liter of dichloromethane, add pyridine (84.64 g, 1.07 mol, 1.05 equiv), and control the reaction temperature at 30-40 degrees Celsius
  • 4-chlorobutyryl chloride 150.88 g, 1.07 mol, 1.05 equiv was added dropwise. After the dropwise addition, the mixture was stirred at 25 degrees Celsius for 12 hours. 1 liter of water was added to the reaction solution at 25 degrees Celsius, the organic phase was separated after stirring for 30 minutes, the organic phase was dried over anhydrous sodium sulfate, and concentrated to obtain compound 1-2.
  • Step B Compound 1-2 (319.60 g, 1.06 mol, 1.0 equiv) was dissolved in 1.8 liters of tetrahydrofuran, and potassium tert-butoxide (125.19 g, 1.12 mol, 1.05 equiv) was added in portions at 25 ° C. Stir for 8 hours at 25 degrees Celsius. At 25 degrees Celsius, 1.8 liters of water was added to the reaction solution, and the organic phase was separated after stirring for 30 minutes. The aqueous phase was extracted with ethyl acetate (1.2 liters). The organic phases were combined, washed with saturated brine (1.2 liters), and dried over anhydrous sodium sulfate. Concentrate to give compound 1-3.
  • Step C Compound 25 (267.08 g, 1.01 mol, 1.0 equiv) was added to 1.5 liters of toluene at 25 degrees Celsius, stirred, and water (18.20 g, 1.01 mol, 1.0 equiv) was added. P-toluenesulfonic acid monohydrate (176.83 g, 929.60 mmol, 0.92 equiv) was added in portions and the feed temperature was controlled between 40-45 degrees Celsius. After the addition is complete, stir at 40-45 degrees Celsius for 3 hours. Then cool to 15 degrees Celsius and stir for 1 hour. The reaction solution was filtered, and the filter cake was rinsed with toluene (600 ml) and dried to obtain compound 1-4.
  • P-toluenesulfonic acid monohydrate 176.83 g, 929.60 mmol, 0.92 equiv
  • Step D Ethyl acetate (349.71 grams, 3.97 moles, 3.0 equivalents) was added to 1 liter of toluene, and sodium ethoxide (180.07 grams, 2.65 moles, 2.0 equivalents) was added in portions at 20-30 degrees Celsius. After the addition is complete, stir at 20-30 ° C for 1 hour. Compound 1-5 (200 grams, 1.32 moles, 1.0 equivalent) was added in three batches, and after the addition was completed, it was stirred at 100 degrees Celsius for 12 hours. The reaction solution was cooled to 25 degrees Celsius, the pH was adjusted to 6 with glacial acetic acid, 1 liter of water was added, and the organic phase was separated after stirring for 30 minutes. The aqueous phase was extracted with toluene (600 mL), and the organic phases were combined and concentrated to give compound 1-6.
  • Step E Compound 1-6 (137 g, 661.12 moles, 1.0 equivalent) was added to 350 ml of pyridine, and then compound 1-4 (216.04 g, 793.34 mmol, 1.2 equivalents) was added. Stir for 12 hours at 40-45 degrees Celsius. The reaction liquid was cooled to 25 ° C, 700 ml of water was added, and extracted with toluene (560 ml ⁇ 2). The organic phases were combined, washed with saturated brine (560 ml), dried over anhydrous sodium sulfate, and concentrated to obtain compound 1-7.
  • Step F Compound 1-7 (148.50 g, 513.26 mmol, 1.0 equiv) was dissolved in 900 ml of toluene, and sodium ethoxide (69.85 g, 1.03 mol, 2.0 equiv) was added in portions. After the addition, the mixture was stirred at 100-110 degrees Celsius for 12 hours. The reaction solution was cooled to 25 degrees Celsius, 600 ml of water was added, and stirred for 15 minutes. The aqueous phase was separated, and the aqueous phase was washed with ethyl acetate (300 ml ⁇ 3), and then the pH value of the aqueous phase was adjusted to 6 with concentrated hydrochloric acid and filtered. The filter cake was slurried with 150 ml of isopropanol, filtered and dried to give compound 1-8. MS (ESI) m / z: 244.0 [M + H + ].
  • Step G Compound 1-8 (77.22 g, 317.44 mmol, 1.0 equiv) was dissolved in 800 ml of N, N-dimethylformamide, and N-bromosuccinimide (59.32 g) was added in portions , 333.31 mmol, 1.05 equiv), stirred at 25 degrees Celsius for 8 hours. N-bromosuccinimide (59.32 g, 333.31 mmol, 1.05 equiv) was added, and the mixture was stirred at 50 degrees Celsius for 12 hours. The reaction solution was cooled to 0-10 degrees Celsius, 2 liters of water was added, stirred at 0-10 degrees Celsius for 30 minutes, and filtered. The filter cake was slurried with water (500 mL), filtered, and dried to give compound 1-9. MS (ESI) m / z: 277.8, 279.8 [M + H + ].
  • Step H Compound 1-9 (20 g, 71.90 mmol, 1.0 equiv), triisopropyl borate (35.84 g, 190.55 mmol, 2.65 equiv) were added to 200 mL of anhydrous tetrahydrofuran, and cooled to -70 ⁇ -60 degrees Celsius. N-Butyllithium (2.5 moles per liter of n-heptane solution, 68.74 ml, 2.39 equiv) was added dropwise at -70 to -60 degrees Celsius. After the addition, the mixture was stirred at -70 to -60 degrees Celsius for 2 hours.
  • Step I Compound 1-10 (26.62 g, 109.52 mmol, 1.0 equiv) and hydrogen peroxide (37.25 g, 328.55 mmol, 30% concentration, 3.0 equiv) were added to 250 ml of tetrahydrofuran, stirred, and cooled to 0-5 ° C . A 2 mol per liter aqueous sodium hydroxide solution (219.03 mL, 4.0 equiv) was added dropwise at 0-5 degrees Celsius. After the addition, the mixture was stirred at 25 degrees Celsius for 12 hours.
  • Step J To a flask containing 100 ml of anhydrous dichloromethane, compound 1-12 (5 g, 25.38 mmol, 1.0 equiv), di-tert-butyl dicarbonate (6.65 g, 30.45) were added in sequence with constant stirring. Mmol, 1.2 equiv), 4-dimethylaminopyridine (310.03 mg, 2.54 mmol, 0.1 equiv) and triethylamine (7.70 g, 76.13 mmol, 3.0 equiv). The reaction solution was stirred at 25 degrees Celsius for 3 hours. 150 ml of water was added to the reaction solution, and extracted with dichloromethane (100 ml ⁇ 2). The organic phase was washed with 100 ml of saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain compound 1-13.
  • Step K Dissolve compound 1-13 (2 g, 6.73 mmol, 1 equiv) in 20 mL of 1,4-dioxane, and add double pinacol borate (2.05 g, 8.08 mmol, 1.2 equivalents), [1,1'-bis (diphenylphosphino)] palladium dichloride (492.49 mg, 673.07 micromolar, 0.1 equivalent) and potassium acetate (1.32 g, 13.46 mmol, 2 equivalents). The reaction system was replaced with nitrogen three times, and then heated to 100 degrees Celsius under nitrogen and stirred for 8 hours.
  • Step L Dissolve compound 1-14 (3.26 g, 9.47 mmol, 1 equiv) in a mixed solvent of 20 ml of acetone and 10 ml of water, and add sodium periodate (6.28 g, 29.36 mmol) in sequence with continuous stirring , 1.63 mL, 3.1 equiv) and ammonium acetate (2.19 g, 28.41 mmol, 3 equiv). Stir for 12 hours at 25 degrees Celsius. 30 ml of water was added to the reaction solution, and ethyl acetate was extracted (30 ml ⁇ 3). The organic phases were combined, washed with saturated brine (30 mL), and concentrated.
  • Step M Dissolve compounds 1-15 (200 mg, 763.16 ⁇ mol, 1 equiv) and 1-11 (246.41 mg, 1.14 mmol, 1.5 equiv) in 5 ml of dichloromethane, and add copper acetate (207.92 mg, 1.14 mmol, 1.5 equiv), triethylamine (231.67 mg, 2.29 mmol, 318.67 ⁇ l, 3 equiv) and Molecular sieve (500 mg), the system was replaced with oxygen three times. React for 12 hours in an oxygen atmosphere at 25 degrees Celsius. 20 ml of water was added to the reaction solution, and extracted with dichloromethane (30 ml ⁇ 3). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain compound 1-16.
  • Step N Compound 1-16 (200 mg, 463.51 micromolar, 1 equivalent) was added to 4 moles per liter of hydrochloric acid methanol solution (5 mL, 43.15 equivalent), and stirred at 25 degrees Celsius for 8 hours.
  • the reaction solution was poured into 20 ml of water, and the pH was adjusted to 8 to 9 with 4 mol per liter of sodium hydroxide aqueous solution.
  • Methanol was distilled off under reduced pressure, and extracted with ethyl acetate (30 mL ⁇ 3). The organic phases were combined, and the organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to prepare high performance liquid phase separation to obtain compound 1.
  • Step A Compound 2-1 (10 g, 46.29 mmol, 1 equiv) was dissolved in tetrahydrofuran (100 mL), and this solution was added dropwise to methyl magnesium bromide (3 mol per liter of tetrahydrofuran under the protection of nitrogen) The solution, 33.95 ml, 2.2 equivalents), keep the temperature at 0-5 degrees Celsius. After the dropwise addition, the temperature was raised to 25 degrees Celsius and stirred for 1 hour. The pH of the reaction solution was adjusted to 7 with 1 mol per liter of hydrochloric acid.
  • Step B Compound 2-2 (10.33 g, 31.99 mmol, 1 equiv) was dissolved in ammonia water (76.53 g, 611.36 mmol, 84.10 mL, 28%, 19.11 equiv), and copper powder (1.22 g, 19.19 mmol) was added , 0.6 equivalent). After stirring at 25 degrees Celsius in the air for 1 hour, it was stirred at 100 degrees Celsius under nitrogen for 12 hours. The reaction solution was filtered, the filtrate was lyophilized, and purified by column chromatography to obtain compound 2-3. MS (ESI) m / z: 153.3 [M + H + ].
  • Step C Dissolve compound 1-11 (5.3 g, 21.80 mmol, 1 equiv) and compound 2-4 (6.86 g, 43.60 mmol, 2 equiv) in dichloromethane (250 mL), followed by stirring Add anhydrous copper acetate (5.94 g, 32.70 mmol, 1.5 equiv), triethylamine (6.62 g, 65.40 mmol, 9.10 mL, 3 equiv), Molecular sieve (5g). Stir for 12 hours at 35 degrees Celsius in an oxygen atmosphere.
  • Step D Compound 2-5 (1.5 g, 4.59 mmol, 1 equiv) was dissolved in 1,4-dioxane (30 mL), and then compound 2-3 (1.05 g, 6.89 mmol, 1.5 equivalents), bis (dibenzylideneacetone) palladium (527.89 mg, 918.05 micromolar, 0.2 equivalents), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (531.20 mg , 918.05 micromolar, 0.2 equivalent), sodium phenolate (2.13 g, 18.36 mmol, 4 equivalent). After replacing the nitrogen three times, it was heated to 100 degrees Celsius and stirred for 12 hours.
  • Step A At 5 degrees Celsius, in batches in a solution of 3-1 (5 g, 32.34 mmol, 4.17 mL, 1 equivalent) of diethyl formate (39.00 g, 330.14 mmol, 40 mL, 10.21 equivalent) Sodium hydrogen (2.68 g, 66.95 mmol, purity: 60%, 2.07 equiv) was added, the temperature was slowly raised to 85 degrees Celsius, and after reacting at this temperature for 2 hours, 100 ml of water was added and extracted with ethyl acetate (100 ml ⁇ 2).
  • Step B To a solution of compound 3-2 (5.3 g, 23.38 mmol, 1 equiv) in pyridine (15 mL) was added 1-4 (7.75 g, 28.06 mmol, 1.2 equiv. Tosylate), after reacting at 40 degrees Celsius for 16 hours, 50 ml of water and ethyl acetate (100 ml ⁇ 3) were added. The organic phases were combined, washed with saturated brine (100 mL ⁇ 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain compound 3-3. MS (ESI) m / z: 309.1 [M + H + ].
  • Step C To a solution of compound 3-3 (4.3 g, 13.93 mmol, 1 equiv) in toluene (25 mL) was added sodium ethoxide (1.90 g, 27.85 mmol, 2 equiv) in batches at 100-110 ° C After 16 hours of reaction, 150 ml of ethyl acetate and 100 ml of water were added. The aqueous phase was washed twice with 25 ml of ethyl acetate each time, and then the pH of the aqueous phase was adjusted to 4 with 36% hydrochloric acid. ML ⁇ 2) extraction. The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 3-4.
  • Step D To a solution of compound 3-4 (1.27 g, 4.83 mmol, 1 equiv) in N, N-dimethylformamide (50 mL) was added portionwise N-bromosuccinimide (860.48 mg , 4.83 mmol, 1 equiv), after reacting at 40 degrees Celsius for 12 hours, 100 ml of water was added, and ethyl acetate (100 ml ⁇ 2) was extracted. The organic phases were combined, washed with saturated brine (50 mL ⁇ 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain compound 3-5. MS (ESI) m / z: 298.9 [M + H + ].
  • Step E A solution of 3-5 (763 mg, 2.56 mmol, 1 equiv) and isopropoxy pinacol borate (1.26 g, 6.79 mmol, 1.39 mL, 2.65 equiv) in tetrahydrofuran under a nitrogen atmosphere ( 20 ml) Cooled to -70 degrees Celsius, then added n-butyllithium (2.5 moles per liter, 2.45 ml, 2.39 equiv) dropwise, slowly warmed to 20 degrees Celsius, and reacted under nitrogen protection and 20 degrees Celsius for 12 hours, then added 50 The reaction was quenched with mL of saturated ammonium chloride solution, diluted with 50 mL of ethyl acetate, and extracted with ethyl acetate (100 mL ⁇ 2). The organic phases were combined, washed with saturated brine (100 ml ⁇ 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 3-6.
  • Step F To a solution of compound 3-6 (1.65 g, 4.79 mmol, 1 equiv) in tetrahydrofuran (30 mL) was added dropwise hydrogen peroxide (2.17 g, 19.15 mmol, 1.84 mL) at 0 ° C. Purity: 30%, 4 equiv.), Then add 5 ml of an aqueous solution of sodium hydroxide (383.01 mg, 9.58 mmol, 2 equiv.). After reacting at 25 degrees Celsius for 12 hours, adjust the pH to 6 with 1 mol of hydrochloric acid solution. , Add 20 ml of ethyl acetate and extract with ethyl acetate (50 ml x 2).
  • Step G To a solution of compound 3-7 (1.09 g, 4.64 mmol, 1 equiv) and 2-4 (1.46 g, 9.29 mmol, 2 equiv) in dichloromethane (70 mL) was added copper acetate (1.27 g , 6.97 mmol, 1.5 eq), triethylamine (1.41 g, 13.93 mmol, 1.94 mL, 3 eq) and Molecular sieve (5 g), after reacting at 25 degrees Celsius for 12 hours, 50 ml of water and 50 ml of dichloromethane were added, and extracted with dichloromethane (50 ml ⁇ 2).
  • Step H To a solution of compound 3-8 (200 mg, 577.68 micromolar, 1 equivalent) and 2-3 (131.88 mg, 866.53 micromolar, 1.5 equivalent) in 1,4-dioxane (10 mL) was added 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (66.85 mg, 115.54 ⁇ mol, 0.2 equivalent), bis (dibenzylideneacetone) palladium (66.43 mg, 115.54 ⁇ mol , 0.2 equiv.) And cesium carbonate (752.88 mg, 2.31 mmol, 4 equiv.), After reacting under nitrogen atmosphere and at 100 degrees Celsius for 12 hours, add 10 ml of water and 30 ml of ethyl acetate, ethyl acetate (25 ml ⁇ 2 )extraction.
  • 1,4-dioxane 10 mL
  • Step A Compound 4-1 (5 g, 26.58 mmol, 3.97 mL, 1 equiv) was added to diethyl carbonate (34.13 g, 288.87 mmol, 35 mL, 10.87 equiv), cooled to 0 ° C, then Sodium hydrogen (2.13 g, 53.16 mmol, 60%, 2 equivalents) was added in portions at 0 degrees Celsius. The mixed system was heated to 85 degrees Celsius and stirred for 2 hours. The reaction solution was quenched with hydrochloric acid (2 mol per liter), and the pH was adjusted to 2 with hydrochloric acid, then water (50 mL) was added, and extracted with ethyl acetate (30 mL ⁇ 3). The combined organic phases were saturated brine (30 ML), washed with anhydrous sodium sulfate, filtered, and concentrated to give compound 4-2. MS (ESI) m / z: 261.0 [M + H + ].
  • Step B To a solution of compound 4-2 (7.81 g, 30.01 mmol, 1 equiv) in pyridine (15 mL) was added 1-4 (9.81 g, 36.02 mmol, 1.2 equiv, p-toluenesulfonate), and The mixture was heated to 40 degrees Celsius and stirred for 12 hours. Water (50 mL) was added to the reaction solution, and extracted with ethyl acetate (50 mL ⁇ 4). The combined organic phase was washed with saturated brine (50 mL ⁇ 2), dried over anhydrous sodium sulfate, filtered and concentrated to give Compound 4-3. MS (ESI) m / z: 343.1 [M + H + ].
  • Step C To a solution of compound 4-3 (1 g, 2.92 mmol, 1 equiv) in toluene (15 mL) was added sodium ethoxide (596.39 mg, 8.76 mmol, 3 equiv), and the mixture was heated to 100 to 110 React for 12 hours between degrees Celsius.
  • the reaction solution was poured into water (60 ml), and the pH was adjusted to between 6 and 7 with hydrochloric acid (2 moles per liter).
  • the aqueous phase was extracted with ethyl acetate (60 mL ⁇ 2), and the combined organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 4-4.
  • Step D To a solution of compound 4-4 (0.78 g, 2.63 mmol, 1 equiv) in N, N-dimethylformamide (12 mL) was added N-bromosuccinimide (445.19 mg, 2.50 Mmol, 0.95 equiv), the mixture was heated to 50 degrees Celsius and stirred for 4 hours. The reaction solution was poured into water (60 mL), the aqueous phase was extracted with ethyl acetate (60 mL ⁇ 2), and the combined organic phase was washed with saturated brine (60 mL ⁇ 2), dried over anhydrous sodium sulfate, and filtered. Concentrate and purify by column chromatography to obtain compound 4-5.
  • Step E At -78 degrees Celsius, apply compound 4-5 (360 mg, 1.09 mmol, 1 equiv) and isopropoxy pinacol borate (405.60 mg, 2.18 mmol, 444.74 ⁇ l, 2 equiv) To the tetrahydrofuran solution (10 ml) was added n-butyllithium (2.5 moles per liter, 872.00 microliters, 2 equivalents), and the reaction solution was naturally heated to 25 degrees Celsius and stirred for 12 hours.
  • Step F To a solution of compound 4-6 (450 mg, 1.52 mmol, 1 equivalent) in tetrahydrofuran (10 mL) was added hydrogen peroxide (689.37 mg, 6.08 mmol, 584.21 ⁇ L, 30%, 4 equivalents), Cool to between 0 and 5 degrees Celsius, then add sodium hydroxide (4 moles per liter, 1.52 milliliters, 4 equivalents), and stir the mixture at 25 degrees Celsius for 12 hours.
  • hydrogen peroxide 689.37 mg, 6.08 mmol, 584.21 ⁇ L, 30%, 4 equivalents
  • Step G To a solution of compound 4-7 (240 mg, 894.74 ⁇ mol, 1 equiv) and compound 2-4 (211.20 mg, 1.34 mmol, 1.5 equiv) in methylene chloride (10 mL) was added copper acetate (243.77 Mg, 1.34 mmol, 1.5 equiv), triethylamine (271.62 mg, 2.68 mmol, 373.61 ⁇ l, 3 equiv) and Molecular sieve (1 g). The system was replaced with oxygen several times, and then the mixture was reacted at 25 degrees Celsius for 12 hours. The reaction solution was filtered, the filtrate was concentrated, and purified by thin layer chromatography to obtain compound 4-8.
  • Step H To a solution of compound 4-8 (130 mg, 342.32 micromolar, 1 equivalent) and compound 2-3 (104.20 mg, 684.64 micromolar, 2 equivalents) in 1,4-dioxane (6 mL) Add bis (dibenzylideneacetone) palladium (19.68 mg, 34.23 ⁇ mol, 0.1 equivalent), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (19.81 mg, 34.23 ⁇ m) Molar, 0.1 equivalent) and cesium carbonate (223.07 mg, 684.64 micromolar, 2 equivalent). The system was replaced with nitrogen and then heated to 100 degrees Celsius for 12 hours.
  • Step A Dissolve compound 5-1 (3.13 g, 20.71 mmol, 1 equiv) and ethyl acetate (5.47 g, 62.14 mmol, 6.08 mL, 3 equiv) in tetrahydrofuran (20 mL) and cool to 0 ° C Then, sodium hydrogen (1.66 g, 41.43 mmol, 60%, 2 equivalents) was added in portions at 0 ° C. The mixed system was heated to 70 degrees Celsius and stirred for 4 hours.
  • Step B To a solution of compound 5-2 (5.52 g, 26.64 mmol, 1 equivalent) in pyridine (15 mL) was added 1-4 (8.70 g, 31.97 mmol, 1.2 equivalent, p-toluenesulfonate), and The mixture was heated to 40 degrees Celsius and stirred for 12 hours. Water (50 mL) was added to the reaction solution, and extracted with ethyl acetate (50 mL ⁇ 4). The combined organic phase was washed with saturated brine (50 mL ⁇ 2), dried over anhydrous sodium sulfate, filtered, and concentrated to give Compound 5-3. MS (ESI) m / z: 290.1 [M + H + ].
  • Step C To a solution of compound 5-3 (5.6 g, 19.36 mmol, 1 equiv) in toluene (50 mL) was added sodium ethoxide (3.95 g, 58.07 mmol, 3 equiv), and the mixture was heated to 110 ° C for reaction 12 hours. Then water (3.00 g, 166.53 mmol, 3 mL, 8.60 equiv) was added, and the reaction was continued at 100 ° C for 12 hours. The reaction solution was poured into water (50 ml), and its pH was adjusted to 4 with hydrochloric acid (2 mol per liter).
  • Step D To a solution of compound 5-4 (3.97 g, 16.32 mmol, 1 equivalent) in N, N-dimethylformamide (50 mL) was added N-bromosuccinimide (3.49 g, 19.58 Mmol, 1.2 equivalents), the mixture was heated to 50 degrees Celsius and stirred for 12 hours. The reaction solution was poured into water (200 mL), the aqueous phase was extracted with ethyl acetate (100 mL ⁇ 3), and the combined organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. Column chromatography purification gave compound 5-5. MS (ESI) m / z: 277.9, 279.9 [M + H + ].
  • Step E At -78 to -65 degrees Celsius, apply compound 5-5 (2.82 g, 10.14 mmol, 1 equivalent) and isopropoxy pinacol borate (3.77 g, 20.28 mmol, 4.14 mL, 2 equivalents) )
  • tetrahydrofuran solution (30 ml) was added n-butyllithium (2.5 moles per liter, 8.11 ml, 2 equivalents), and the reaction solution was naturally heated to 25 ° C and stirred for 12 hours.
  • Step F To a solution of compound 5-6 (1 g, 3.07 mmol, 1 equiv) in tetrahydrofuran (15 mL) was added hydrogen peroxide (1.39 g, 12.30 mmol, 1.18 mL, 30%, 4 equiv), and the system was cooled Between 0 and 5 degrees Celsius, additional sodium hydroxide (4 moles per liter, 3.07 milliliters, 4 equivalents) was added and the mixture was stirred at 25 degrees Celsius for 12 hours.
  • Step G To a solution of compound 5-7 (300 mg, 1.39 mmol, 1 equiv) and compound 2-4 (328.98 mg, 2.09 mmol, 1.5 equiv) in dichloromethane (15 mL) was added copper acetate (379.72 Mg, 2.09 mmol, 1.5 equiv), triethylamine (423.09 mg, 4.18 mmol, 581.97 ⁇ l, 3 equiv) and Molecular sieve (1 g). The system was replaced with oxygen several times, and then the mixture was reacted at 25 degrees Celsius for 12 hours.
  • reaction solution was poured into water (60 mL), the aqueous phase was extracted with ethyl acetate (60 mL ⁇ 2), and the combined organic phase was washed with brine (60 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and thin Layer chromatography purification gave compounds 5-8.
  • Step H To a solution of compound 5-8 (200 mg, 612.03 micromolar, 1 equivalent) and compound 2-3 (93.15 mg, 612.03 micromolar, 1 equivalent) in 1,4-dioxane (8 mL) Add bis (dibenzylideneacetone) palladium (35.19 mg, 61.20 ⁇ mol, 0.1 equivalent), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (35.41 mg, 61.20 ⁇ m) Molar, 0.1 equivalent) and cesium carbonate (398.82 mg, 1.22 mmol, 2 equivalent). The system was replaced with nitrogen, and then heated to 90 degrees Celsius for 3 hours. The reaction solution was concentrated and purified by high performance liquid chromatography (first formic acid system, then basic system) to obtain compound 5. MS (ESI) m / z: 443.4 [M + H + ];
  • Step A Ethyl acetate (17.6 g, 199.77 mmol, 3.0 equiv) was added to toluene (50 mL), and sodium ethoxide (9.06 g, 133.18 mmol, 2.0 equiv) was added at 20 degrees Celsius. After the addition, the mixture was stirred at 20 degrees Celsius for 1 hour. Compound 6-1 (10 g, 66.59 mmol, 1.0 equiv) was added in three batches, and after the addition was completed, it was stirred at 100 degrees Celsius for 12 hours.
  • reaction solution was cooled to 25 degrees Celsius, the pH value was adjusted to 6 with glacial acetic acid, water (50 ml) was added, extracted with toluene (30 ml ⁇ 2), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain compound 6-2.
  • Step B Compound 6-2 (10.05 g, 48.73 mmol, 1.0 equiv) was added to pyridine (25 mL), and then compound 1-4 (16.10 g, 58.48 mmol, 1.2 equiv) was added. Stir at 40 degrees Celsius for 12 hours. The reaction liquid was cooled to 20 degrees Celsius, water (50 ml) was added, and extracted with toluene (40 ml ⁇ 2). The organic phases were combined, washed with water (50 mL), dried over anhydrous sodium sulfate, and concentrated to give compound 6-3.
  • Step C Compound 6-3 (12.54 g, 43.49 mmol, 1.0 equiv) was dissolved in toluene (80 mL), and sodium ethoxide (8.88 g, 130.47 mmol, 3.0 equiv) was added in portions with constant stirring. After the addition, the mixture was stirred at 100-110 degrees Celsius for 12 hours. The reaction solution was cooled to 20 degrees Celsius, and the pH was adjusted to 6 with 1 mole of hydrochloric acid. 50 ml of water was added, extracted with ethyl acetate (50 ml ⁇ 3), the organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, and concentrated to give compound 6-4. MS (ESI) m / z: 271.2 [M + H + ].
  • Step D Sodium hydroxide (16.69 g, 417.27 mmol, 10 equiv) was dissolved in water (25 mL), and the resulting solution was added to compound 6-4 (11.28 g, 41.73 mmol, 1.0 equiv) in methanol (50 Ml) solution. Stir for 12 hours at 40 degrees Celsius. Methanol was removed, water (25 ml) was added, and washed with ethyl acetate (25 ml ⁇ 3). The pH of the aqueous phase was adjusted to 4 with concentrated hydrochloric acid and filtered. The filter cake was washed with water (25 ml ⁇ 3) and dried to obtain compound 6-5.
  • Step E Compound 6-5 (10.01 g, 41.32 mmol, 1.0 equivalent) was dissolved in N, N-dimethylformamide (120 mL), and bromosuccinimide (7.35 g, 41.32 mmol, 1.0 equivalent) and sodium hydroxide (1.98 g, 49.58 mmol, 1.2 equivalent). Stir for 12 hours at 40 degrees Celsius. The reaction solution was cooled to 20-25 degrees Celsius, water (300 mL) was added, stirred for 10 minutes, filtered, and the filter cake was washed with water (20 mL ⁇ 2) and dried to obtain compound 6-6. MS (ESI) m / z: 277.2 [M + H + ].
  • Step F Compound 6-6 (1.4 g, 5.05 mmol, 1.0 equiv), isopropoxy pinacol borate (2.49 g, 13.39 mmol, 2.65 equiv) was added to anhydrous tetrahydrofuran (30 mL), Replace nitrogen three times, stir, and cool to -70 ⁇ -60 degrees Celsius.
  • N-Butyllithium 2.5 moles per liter of n-heptane solution, 4.83 ml, 2.39 equiv was added dropwise at -70 to -60 degrees Celsius. After the addition, the mixture was stirred at -70 to -60 degrees Celsius for 1 hour. It was then stirred at 25 degrees Celsius for 12 hours.
  • reaction solution was added dropwise to 60 ml of saturated ammonium chloride solution, extracted with ethyl acetate (40 ml ⁇ 3), the organic phases were combined, washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, and concentrated to obtain compound 6 7.
  • Step G Add compound 6-7 (2.08 g, 6.42 mmol, 1.0 equiv) and hydrogen peroxide (2.91 g, 25.66 mmol, 30% concentration, 4 equiv) to tetrahydrofuran (20 mL), stir, and cool to 0 ⁇ 5 degrees Celsius. A 4 mol per liter aqueous solution of sodium hydroxide (6.42 ml, 4.0 equivalents) was added dropwise at 0-5 degrees Celsius. After the addition, the mixture was stirred at 25 degrees Celsius for 12 hours.
  • Step H Dissolve compound 6-8 (336 mg, 1.57 mmol, 1 equiv), compound 2-4 (494.12 mg, 3.14 mmol, 2 equiv) in dichloromethane (20 mL), and add copper acetate in sequence (427.75 mg, 2.36 mmol, 1.5 equiv), triethylamine (476.60 mg, 4.71 mmol, 655.58 ⁇ l, 3 equiv) and Molecular sieve (3.5 g). The oxygen atmosphere was stirred for 12 hours at 25 degrees Celsius.
  • Step I Dissolve compound 6-9 (200 mg, 613.89 micromolar, 1 equivalent) and compound 2-3 (186.86 mg, 1.23 mmol, 2 equivalents) in 1,4-dioxane (20 mL) , followeded by the addition of bis (dibenzylideneacetone) palladium (70.60 mg, 112.78 micromolar, 0.2 equivalent), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (71.04 mg, 122.78 micromolar, 0.2 equivalent), cesium carbonate (800.07 mg, 2.46 mmol, 4 equivalent). The nitrogen was replaced three times, and the temperature was raised to 100 degrees Celsius and stirred for 12 hours under the protection of nitrogen.
  • Step A Add lithium hexamethyldisilazide (1 mol per liter, 728.32 ml, 3 equivalents) to a three-necked bottle, cool the reaction solution to -78 degrees Celsius, and then add ethyl acetate (64.17 g, 728.32 dropwise) Mmol, 71.30 mL, 3 equivalents). The reaction solution was stirred at -78 degrees Celsius for half an hour. At -78 degrees Celsius, 7-1 (35 g, 242.77 mmol, 32.41 mL, 1 equivalent) was dissolved in 400 mL of tetrahydrofuran and added dropwise to the reaction solution. The reaction solution was stirred at -78 degrees Celsius for 1 hour.
  • reaction was quenched with 150 ml of saturated ammonium chloride solution at -70 degrees Celsius, diluted with 400 ml of water and extracted with ethyl acetate (500 ml ⁇ 2). The organic phases were combined, washed with saturated brine (400 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 7-2.
  • Step B Dissolve 7-2 (14.5 g, 72.42 mmol, 1 equiv) in pyridine (140 mL) and add 1-4 (31.55 g, 115.87 mmol, 1.6 equiv, p-toluenesulfonate). The mixture was replaced with nitrogen three times, and stirred at 40 degrees Celsius for 12 hours. Dilute with 300 ml of water and extract with ethyl acetate (300 ml x 2). The organic phases were combined, washed with saturated brine (100 ml ⁇ 2), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to obtain compound 7-3. MS (ESI) m / z: 283.1 [M + H + ].
  • Step C Dissolve compound 7-3 (6.3 g, 22.31 mmol, 1 equiv) in toluene (80 mL), add sodium hydride (2.68 g, 66.94 mmol, 60% purity, 3 equiv) under a nitrogen atmosphere The mixture was heated to 110 degrees Celsius and reacted for 12 hours. 250 ml of ethyl acetate and 250 ml of water were added to the mixture, and the organic phase was washed with saturated brine (100 ml), dried over anhydrous sodium sulfate, filtered, and concentrated.
  • Step D Dissolve 7-4 (7 g, 26.48 mmol, 1 eq) in water (20 mL) and methanol (40 mL), and add sodium hydroxide (3.18 g, 79.45 mmol, 3 eq) under a nitrogen atmosphere ), Stirring at 60 degrees Celsius for 5 hours. The solution was adjusted to pH 6 with 4 moles per liter of hydrochloric acid, filtered, and the filter cake was dried to obtain compound 7-5. MS (ESI) m / z: 237.2 [M + H + ].
  • Step E Dissolve 7-5 (5 g, 21.16 mmol, 1 equivalent) in dimethylformamide (20 mL), and add bromosuccinimide (4.52 g, 25.40 mmol, under nitrogen atmosphere). 1.2 equivalent). The mixture was stirred at 25 degrees Celsius for 12 hours. Dilute with water (100 ml) and extract with ethyl acetate (100 ml x 2). The organic phases were combined, washed with saturated brine (50 ml ⁇ 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain compound 7-6. MS (ESI) m / z: 271,273 [M + H + ].
  • Step F Compound 7-6 (4.26 g, 15.71 mmol, 1 equiv) was dissolved in tetrahydrofuran (40 mL). Under nitrogen atmosphere, n-butyllithium (2.5 mol per liter, 18.85 mL, 3 equivalents). The mixed solution was stirred at -70 degrees Celsius for 1 hour, maintaining temperature to add dropwise isopropoxy pinacol borate (7.60 g, 40.85 mmol, 8.33 mL, 2.6 equiv). Slowly warm up to 0 degrees Celsius and stir for 1 hour.
  • reaction solution was poured into a saturated aqueous ammonium chloride solution (100 ml) at 0 degrees Celsius, extracted with ethyl acetate (100 ml ⁇ 2), the organic phases were combined, washed with saturated brine (50 ml ⁇ 2), and dried over anhydrous sodium sulfate , Filtered, and concentrated to give compound 7-7.
  • Step G Dissolve 7-7 (5.2 g, 16.34 mmol, 1 equiv) in tetrahydrofuran (50 mL), then add hydrogen peroxide (4.63 g, 40.85 mmol, 3.92 mL, 30% purity, 2.5 equiv) .
  • the reaction solution was cooled to 0-5 degrees Celsius, and an aqueous solution of sodium hydroxide (4 moles per liter, 10.21 ml, 2.5 equivalents) was added dropwise.
  • the reaction solution was stirred at 25 degrees Celsius for 12 hours.
  • Sodium sulfite (1 g) was added for quenching, 4 mol / L hydrochloric acid was used to adjust the pH to 8, and the mixture was stirred at 20 degrees Celsius for 1 hour.
  • Step H Compounds 7-8 (800 mg, 3.84 mmol, 1 equivalent) and 7-9 (757.92 mg, 5.76 mmol, 1.5 equivalent) were dissolved in N, N-dimethylformamide (8 mL) Under a nitrogen atmosphere, potassium carbonate (1.59 g, 11.52 mmol, 3 equivalents) was added. The mixed solution was reacted at 120 degrees Celsius for 12 hours. Diluted with 60 ml of water, extracted with ethyl acetate (80 ml ⁇ 2), the organic phase was washed with 60 ml of saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain compound 7-10. MS (ESI) m / z: 320.1 [M + H + ].
  • Step I Compound 7-10 (1.05 g, 3.28 mmol, 1 equivalent), 7-11 (426.68 mg, 3.61 mmol, 96.31 ⁇ L, 1.1 equivalent), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (189.99 mg, 328.35 ⁇ mol, 0.1 equivalent), cesium carbonate (3.21 g, 9.85 mmol, 3 equivalent) and palladium acetate (73.72 mg, 328.35 ⁇ mol, 0.1 equivalent) Dissolved in dioxane (10 ml). The reaction was carried out at 100 degrees Celsius for 3 hours under a nitrogen atmosphere.
  • Step J Dissolve sodium hydroxide (179.35 mg, 4.48 mmol, 1 equivalent) in water (1.8 mL), add ethanol (18 mL) at 25 ° C, 7-12 (1.80 g, 4.48 mmol, 1 equivalent) And dimethyl sulfoxide (525.46 mg, 6.73 mmol, 525.46 ⁇ l, 1.5 equivalents). Hydrogen peroxide (762.44 mg, 6.73 mmol, 646.14 ⁇ L, 30% purity, 1.5 equivalents) was diluted with water (0.8 mL), and the reaction solution was added dropwise. Stir at 25 degrees Celsius for 2 hours. Dilute with 100 ml of water and extract with ethyl acetate (150 ml ⁇ 2).
  • Step A Compound 8-1 (3 g, 26.53 mmol, 1 equiv), compound 8-2 (5.74 g, 79.59 mmol, 7.07 mL, 3 equiv), cesium carbonate (17.29 g, 53.06 mmol, 2 Equivalent) was dissolved in N, N-dimethylformamide (40 mL), and the resulting mixture was replaced with nitrogen three times and stirred at 100 degrees Celsius for 5 hours. Water (50 mL) was added to quench the reaction and extracted with ethyl acetate (50 mL ⁇ 2).
  • Step B Dissolve compound 8-3 (3 g, 16.20 mmol, 1 equiv) in methanol (30 mL), add wet palladium on carbon (500 mg, 10%), replace with hydrogen, under a hydrogen atmosphere at 25 ° C React for 12 hours. Filter and concentrate to give compound 8-4.
  • Step C Compound 1-11 (0.3 g, 1.39 mmol, 1 equivalent) was dissolved in N, N-dimethylformamide (20 mL), and 7-9 (219.99 mg, 1.67 mmol) was added to the solution , 1.2 equivalents), potassium carbonate (577.86 mg, 4.18 mmol, 3 equivalents), and the resulting mixture was stirred at 80 degrees Celsius for 12 hours. Water (50 mL) was added to quench the reaction. Extracted with ethyl acetate (20 ml ⁇ 2), the organic phase was washed with saturated sodium chloride solution (30 ml ⁇ 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column separation to obtain compound 2-5.
  • Step D Compound 2-5 (100 mg, 0.306 mmol, 1 equivalent) and compound 8-4 (49.87 mg, 0.321 mmol, 1.05 equivalent), bis (dibenzylideneacetone) palladium (17.60 mg, 30.6 Micromolar, 0.1 equivalent), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (35.41 mg, 61.20 micromolar, 0.2 equivalent) and cesium carbonate (199.41 mg, 612.03 micromolar) , 2 equivalents) dissolved in 1,4-dioxane (15 ml). The system was replaced with nitrogen, then heated to 100 degrees Celsius, and reacted for 12 hours under a nitrogen atmosphere.
  • Step A To a solution of compound 2-5 (200 mg, 612.03 micromolar, 1 equivalent) and compound 9-1 (120.27 mg, 795.64 micromolar, 1.3 equivalent) in 1,4-dioxane (8 mL) Add bis (dibenzylideneacetone) palladium (35.19 mg, 61.20 ⁇ mol, 0.1 equivalent), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (35.41 mg, 61.20 ⁇ m) Molar, 0.1 equivalent) and cesium carbonate (398.82 mg, 1.22 mmol, 2 equivalent). The system was replaced with nitrogen and then heated to 100 degrees Celsius for 12 hours.
  • Step B At -78 degrees Celsius, ammonia gas was bubbled into ethylene glycol (15 ml) and kept aerated for 5 minutes.
  • Compound 9-2 (390 mg, 883.39 micromolar, 1 equivalent) was dissolved in the above solution. Then, the mixed solution was poured into a Teflon pot, heated to 80 degrees Celsius and stirred for 12 hours. The reaction solution was poured into water (60 ml) and stirred for 3 minutes. The aqueous phase was extracted with ethyl acetate (60 mL ⁇ 2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated, and purified by preparative high performance liquid chromatography (formic acid system) to obtain the formate salt of compound 9.
  • Step A To a solution of compound 2-5 (100 mg, 306.02 micromolar, 1 equivalent) and compound 10-1 (60.66 mg, 367.22 micromolar, 1.2 equivalent) in 1,4-dioxane (3 mL) Add bis (dibenzylideneacetone) palladium (17.6 mg, 30.6 ⁇ mol, 0.1 equivalent), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (17.71 mg, 30.60 ⁇ m) Molar, 0.1 equivalent) and cesium carbonate (199.41 mg, 612.03 micromolar, 2 equivalent). The system was replaced with nitrogen, heated to 65 degrees Celsius for 4 hours, and then heated to 100 degrees Celsius for 12 hours.
  • Step B At -78 degrees Celsius, ammonia gas was bubbled into 10 ml of ethylene glycol for 15 minutes, and then compound 10-2 (147.54 mg, 323.91 micromolar, 1.2 equivalents) was added, and the solution was added to the stuffy tank After reacting at 100 degrees Celsius for 12 hours, add 20 ml of water and 50 ml of ethyl acetate at 25 degrees Celsius, extract twice with ethyl acetate, 50 ml each time, combine organic phases, wash with saturated brine (25 ml ⁇ 2) , Dried over anhydrous sodium sulfate, filtered, concentrated, purified by preparative high performance liquid chromatography (basic) to obtain compound 10.
  • compound 10-2 147.54 mg, 323.91 micromolar, 1.2 equivalents
  • Step A To a solution of compound 2-5 (200 mL, 612.03 micromolar, 1 equivalent) and compound 15-1 (121.06 mg, 795.64 micromolar, 1.3 equivalent) in 1,4-dioxane (8 mL) Add bis (dibenzylideneacetone) palladium (35.19 mg, 61.20 ⁇ mol, 0.1 equivalent), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (35.41 mg, 61.20 ⁇ m) Molar, 0.1 equivalent) and cesium carbonate (398.82 mg, 1.22 mmol, 2 equivalent). The system was replaced with nitrogen, and then heated to 90 degrees Celsius for 12 hours.
  • Step B To a solution of compound 15-2 (200 mg, 466.81 micromolar, 1 equivalent) in dichloromethane (3 mL) was added oxalyl chloride (177.75 mg, 1.40 mmol, 122.59 microliter, 3 equivalent), reaction 1 After hours, the mixture was added to stirred ammonia water (5.60 g, 47.94 mmol, 6.15 mL, 30%, 102.69 equiv), and stirred at 25 degrees Celsius for 15 minutes.
  • Step A To a solution of compound 1-11 (300 mg, 1.39 mmol, 1 equiv) in N, N-dimethylformamide (10 mL) was added potassium carbonate (385.24 mg, 2.79 mmol, 2 equiv) and Compound 16-1 (249.16 mg, 1.67 mmol, 1.2 equiv) was reacted at 80 degrees Celsius for 12 hours, 40 ml of water was added, filtered, and the filter cake was dried to obtain compound 16-2.
  • Step B To a solution of compound 16-2 (350 mg, 1.07 mmol, 1 equivalent) and 7-11 (138.76 mg, 1.17 mmol, 1.1 equivalent) in 1,4-dioxane (10 mL) was added Cesium carbonate (1.04 g, 3.20 mmol, 3 equivalents), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (123.57 mg, 213.57 micromolar, 0.2 equivalents) and tri ( Dibenzylideneacetone) dipalladium (97.78 mg, 106.78 micromolar, 0.1 equiv), after reacting under nitrogen atmosphere and at 100 degrees Celsius for 12 hours, 40 ml of ethyl acetate was added, filtered, and the filtrate was saturated with sodium chloride solution Ml ⁇ 3) washed, the organic phase was added to 20 ml of 1 mol per liter of hydrochloric acid, the aqueous phase obtained by separation was washed with
  • Step C To a solution of compound 16-3 (350 mg, 854.82 micromolar, 1 equivalent) in ethanol (5 ml) and water (5 ml) was added dimethyl sulfoxide (133.58 mg, 1.71 mmol, 133.58 ⁇ l , 2 equiv.) And sodium hydroxide (68.38 mg, 1.71 mmol, 2 equiv.), Then add hydrogen peroxide (166.15 mg, 1.71 mmol, 140.81 ⁇ l, 35%, 2 equiv.), And react at 25 degrees Celsius for 12 hours.
  • Step A To compound 1 (3 g, 9.05 mmol, 1 equiv), triethylamine (1.10 g, 10.86 mmol, 1.51 mL, 1.2 equiv) and 4-dimethylaminopyridine (55.30 mg, 452.67 ⁇ mol, To a solution of 0.05 equivalent) in dichloromethane (20 mL), benzenesulfonyl chloride (1.92 g, 10.86 mmol, 1.39 mL, 1.2 equivalent) was added. React for 12 hours at 25 degrees Celsius. Water (100 ml) was added, and dichloromethane (300 ml ⁇ 3) was extracted.
  • Step B Dissolve compound 17-1 (100 mg, 212.08 micromolar, 1 equivalent) in tetrahydrofuran (5 mL), and add lithium diisopropylamide (2 mol per liter, 159.06) dropwise at -78 ° C under a nitrogen atmosphere Microliter, 318.11 micromolar, 1.5 equivalents). The reaction was maintained at -78 degrees Celsius for 1 hour, at this temperature a solution of iodine (80.74 mg, 318.11 micromolar, 1.5 equivalents) in tetrahydrofuran (5 mL) was added, and the reaction was continued for 2 hours. Water (30 ml) was added, and ethyl acetate (50 ml ⁇ 3) was extracted.
  • Step C Compound 17-2 (150 mg, 251.08 micromolar, 1 equivalent), 1-methylpyrazole-4-boronic acid (33.20 mg, 263.63 micromolar, 1.05 equivalent), potassium carbonate (104.10 mg, 753.23 micromolar) Molar, 3 equivalents) and [1,1'-bis (diphenylphosphino) ferrocene] palladium dichloride (18.37 mg, 25.11 micromolar, 0.1 equivalent) dissolved in 1,4-dioxane (5 ML) and water (1 mL) under nitrogen atmosphere at 80 degrees Celsius for 12 hours. The reaction solution was filtered, the filtrate was diluted with water (30 mL), and extracted with ethyl acetate (50 mL ⁇ 3).
  • Step D To a solution of compound 17-3 (120 mg, 217.54 ⁇ mol, 1 equiv) in methanol (5 mL), add an aqueous solution of sodium hydroxide (2 mol per liter, 543.85 ⁇ l, 1.09 mmol, 5 equiv) . React for 4 hours at 60 degrees Celsius. Water (50 ml) was added, and ethyl acetate (50 ml ⁇ 3) was extracted. The organic phases were combined, washed with saturated brine (50 mL ⁇ 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by preparative high-performance liquid chromatography (formic acid conditions) to obtain the formate salt of compound 17.
  • TGF- ⁇ R1, TGF- ⁇ R1 (T204D), LCK, p38 ⁇ Kinase Enzyme System (kinase system) were purchased from Promega. Envision Multi-Mark Analyzer (PerkinElmer).
  • the compound to be tested was diluted 5 times to the eighth concentration with an exhaust gun, that is, from 50 micromoles per liter to 0.65 nanomoles per liter, and the final concentration of dimethyl sulfoxide was 5%.
  • a double-multiwell experiment was set up. Add 1 ⁇ l inhibitor concentration gradient to the microplate, 2 ⁇ l TGF- ⁇ R1 enzyme (30 ng), 2 ⁇ l substrate and ATP mixture (50 ⁇ mol / L ATP, 0.2 ⁇ g / L) TGF- ⁇ R1 peptide), at this time the final concentration gradient of the compound is diluted from 10 micromoles per liter to 0.13 nanomoles per liter.
  • the reaction system was placed at 30 degrees Celsius for 120 minutes.
  • TGF- ⁇ R1 T204D
  • the compound to be tested was diluted 5 times to the eighth concentration with an exhaust gun, that is, from 50 micromoles per liter to 0.65 nanomoles per liter, and the final concentration of dimethyl sulfoxide was 5%.
  • a double-multiwell experiment was set up. Add 1 ⁇ l inhibitor concentration gradient to the microplate, 2 ⁇ l TGF- ⁇ R1 (T204D) enzyme (15 ng), 2 ⁇ l substrate and ATP mixture (50 ⁇ mol per liter ATP, 0.2 ⁇ g Per microliter of TGF- ⁇ R1 peptide), then the final concentration gradient of the compound is 10 micromoles per liter diluted to 0.13 nanomoles per liter.
  • the reaction system was placed at 30 degrees Celsius for 120 minutes.
  • the compound to be tested was diluted 5 times to the eighth concentration with an exhaust gun, that is, from 500 micromoles per liter to 6.5 nanomoles per liter, and the final concentration of dimethyl sulfoxide was 5%.
  • a double-multiwell experiment was set up. Add 1 ⁇ l inhibitor concentration gradient to the microplate, 2 ⁇ l LCK enzyme (1.55 ng), 2 ⁇ l substrate and ATP mixture (30 ⁇ mol / L ATP, 0.4 ⁇ g / L Poly E 4 Y 1 ), at this time the final concentration gradient of the compound is diluted from 100 micromoles per liter to 1.3 nanomoles per liter.
  • the reaction system was placed at 30 degrees Celsius and reacted for 60 minutes.
  • the compound to be tested was diluted 5 times to the eighth concentration with an exhaust gun, that is, from 50 micromoles per liter to 0.65 nanomoles per liter, and the final concentration of dimethyl sulfoxide was 5%.
  • a double-multiwell experiment was set up. Add 1 ⁇ l inhibitor concentration gradient to the microplate, 2 ⁇ l p38 ⁇ enzyme (4 ng), 2 ⁇ l substrate and ATP mixture (150 ⁇ mol / L ATP, 0.2 ⁇ g / L p38 peptide) ), At this time the final concentration gradient of the compound is diluted from 10 micromoles per liter to 0.13 nanomoles per liter.
  • the reaction system was placed at 30 degrees Celsius and reacted for 60 minutes.
  • Table 1 provides enzymatic inhibitory activities of the compounds of the examples of the present invention on TGF- ⁇ R1, TGF- ⁇ R1 (T204D), LCK, and p38 ⁇ .
  • the compound of the present invention has excellent in vitro inhibitory activity against TGF- ⁇ R1, TGF- ⁇ R1 (T204D), and weak in vitro inhibitory activity against LCK and p38 ⁇ , exhibiting excellent in vitro activity and selectivity.
  • This experiment integrated a SMAD-responsive element controlled luciferase gene into HEK293 cells. This cell line was verified by the stimulation response of human TGF ⁇ 1 and the treatment of TGF ⁇ / SMAD signaling pathway inhibitors.
  • each well contains approximately 35,000 seed cells and 100 microliters of gene-free growth medium. And incubate at 37 degrees Celsius.
  • Each concentration point should include three sets of experiments. a. Add 10 microliters of human-derived TGF ⁇ 1 analysis medium solution to the medium (the final TGF ⁇ 1 concentration is 20 ng / ml). b. Add 10 ⁇ l of blank culture medium to the unstimulated group. c. Add 100 ⁇ l of blank medium to the cell-free group.
  • Measurement using a luciferase analysis system add 100 ⁇ l of ONE-Step TM luciferase reagent to each well at room temperature and leave at room temperature for 15 to 30 minutes. Use a photometer to measure fluorescence.
  • the compound of the present invention has excellent pSmad inhibitory activity. It is proved that the compound of the present invention can play a role in inhibiting the TGF- ⁇ / SMAD signaling pathway.
  • the main purpose of this study is to study the anti-tumor efficacy of the tested compounds on the CT26 mouse allograft model.
  • Cell culture Mouse colon cancer CT-26 cells (Typical Culture Depository of the Chinese Academy of Sciences Cell Bank) in vitro monolayer culture, culture conditions are RPMI-1640 medium (manufacturer: Gibco) plus 10% fetal bovine serum, 37 Cultivation in 5% carbon dioxide incubator. Twice a week, try digestion with EDTA (Ethylene Diamine Tetraacetic Acid). When the cell saturation is 80% -90%, and the quantity reaches the requirement, collect the cells, count and inoculate.
  • RPMI-1640 medium manufactured by Gibco
  • fetal bovine serum 37 Cultivation in 5% carbon dioxide incubator. Twice a week, try digestion with EDTA (Ethylene Diamine Tetraacetic Acid).
  • EDTA Ethylene Diamine Tetraacetic Acid
  • Tumor inoculation 0.1 ml of a DPBS cell suspension containing 3 ⁇ 10 5 CT26 cells was subcutaneously inoculated into the right groin of each mouse, and the administration started on the day of inoculation.
  • the experimental index is to investigate whether the tumor growth is inhibited, delayed or cured.
  • the diameter of the tumor was measured with vernier calipers twice a week.
  • the compound of the invention has obvious antitumor effect in vivo in mouse colon cancer CT-26 cell BALB / c mouse subcutaneous allograft model. At the same dose (50 mg per kg, twice a day), it showed a significantly better tumor suppressive effect than compound A.

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Abstract

Disclosed are a double pyrazole compound as TGF-βR1 kinase inhibitor and the uses thereof in preparing a drug for inhibiting a TGF-βR1 kinase. Specifically disclosed are said compound represented by the formula (I), a pharmaceutically acceptable salt and tautomer thereof.

Description

作为TGF-βR1激酶抑制剂的双环吡唑类化合物Bicyclic pyrazole compounds as TGF-βR1 kinase inhibitors
相关申请的交叉引用Cross-reference of related applications
本申请主张如下优先权：CN201811288439.3，申请日2018年10月31日；CN201910288923.4，申请日2019年04月11日。This application claims the following priority: CN201811288439.3, application date October 31, 2018; CN201910288923.4, application date April 11, 2019.
技术领域Technical field
本发明涉及一类作为TGF-βR1抑制剂的双环吡唑类化合物，以及它们在制备TGF-βR1抑制剂药物中的应用。具体涉及式(I)所示化合物、其药学上可接受的盐或其异构体。The invention relates to a class of bicyclic pyrazole compounds as TGF-βR1 inhibitors, and their application in the preparation of TGF-βR1 inhibitor drugs. Specifically, it relates to a compound represented by formula (I), a pharmaceutically acceptable salt thereof, or an isomer thereof.
背景技术Background technique
转化生长因子β(Transforming growth factor-β，TGF-β)是一个多功能生长因子超家族，具有广泛的生物学活性，参与早期胚胎发育，软骨和骨的形成，包外基质的合成，炎症，间质纤维化，免疫和内分泌功能的调节，肿瘤的形成和发展。Transforming growth factor β (Transforming growth factor-β, TGF-β) is a multi-functional growth factor superfamily with a wide range of biological activities, involved in early embryonic development, cartilage and bone formation, synthesis of outer matrix, inflammation, Interstitial fibrosis, regulation of immune and endocrine functions, tumor formation and development.
TGF-β超家族由一类结构和功能相关的多肽生长因子组成，TGF-β是该家族的重要成员之一。在哺乳动物中TGF-β主要以TGF-β1、TGF-β2和TGF-β3三种形式存在，它们位于不同的染色体上，其中TGF-β1在体细胞中所占比例最高(>90％)，它活性最强、功能最多，分布也最广泛。The TGF-β superfamily consists of a class of structural and functionally related polypeptide growth factors. TGF-β is one of the important members of this family. In mammals, TGF-β mainly exists in three forms: TGF-β1, TGF-β2, and TGF-β3, which are located on different chromosomes, of which TGF-β1 accounts for the highest proportion (> 90%) in somatic cells. It has the strongest activity, the most functions, and the most widespread distribution.
TGF-β信号分子通过跨膜的受体复合物进行信号转导。TGF-β受体是存在于细胞表面的跨膜蛋白，分为Ⅰ型受体(TGF-βRI)、Ⅱ型受体(TGF-βR Ⅱ)和Ⅲ型受体(TGF-βRⅢ)，其中TGF-βR Ⅰ又被称作活化素样受体5(activin receptor-like kinase 5，ALK5)。TGF-βRⅢ缺乏内在活性，主要与TGF-β的储存有关。TGF-βR Ⅰ和TGF-βR Ⅱ属于丝氨酸/苏氨酸激酶家族，Ⅱ型受体能以较高的亲和力与TGF-β配体结合，并与Ⅰ型受体形成异源受体复合物，将Ⅰ型受体近膜的一段富含甘氨酸、丝氨酸残基的区域(GS结构域)磷酸化，启动细胞内信号联级反应。The TGF-β signaling molecule performs signal transduction through the transmembrane receptor complex. TGF-β receptors are transmembrane proteins that exist on the cell surface and are divided into type I receptors (TGF-βRI), type II receptors (TGF-βR) II and type III receptors (TGF-βRⅢ), of which TGF -βR is also called activin-like receptor 5 (ALK5). TGF-βRⅢ lacks intrinsic activity, which is mainly related to the storage of TGF-β. TGF-βR I and TGF-βR II belong to the serine / threonine kinase family. Type II receptors can bind to TGF-β ligands with high affinity and form heterologous receptor complexes with type I receptors. A region rich in glycine and serine residues (GS domain) near the membrane of type I receptors is phosphorylated to initiate intracellular signaling cascade reactions.
Smads是细胞内重要的TGF-β信号转导和调节分子，可以将TGF-β信号直接由细胞膜转导如细胞核内，TGF-β/Smads信号通路在肿瘤的发生和发展中起到重要的作用。在TGF-β/Smads信号转导中，活化的TGF-β首先与细胞膜表面的TGF-βR Ⅱ结合，形成异源二聚体复合物，TGF-βR Ⅰ识别并结合该二元复合物。Smads is an important TGF-β signal transduction and regulation molecule in the cell, which can directly transduce TGF-β signal from the cell membrane as in the nucleus. . In TGF-β / Smads signal transduction, activated TGF-β first binds to TGF-βR II on the cell membrane surface to form a heterodimer complex, and TGF-βR I recognizes and binds the binary complex.
TGF-βR Ⅱ将TGF-βR Ⅰ胞浆区GS结构域的丝氨酸/苏氨酸磷酸化，从而激活TGF-βR Ⅰ；活化的TGF-βR Ⅰ进一步磷酸化R-Smads(Smad2/Smad3)蛋白，后者再与Co-Smad(Smad4)结合成为异三聚体复合物，这一复合物进入细胞核内，与其他辅助活化因子(co-activator)和辅助抑制因子(co-inhibitor)协同作用，调节靶基因的转录。在TGF-β/Smads信号通路中任何一个环节发生改变，都会导致信号转导通路的异常。TGF-βR Ⅱ phosphorylates the serine / threonine of the GS domain of TGF-βR Ⅰ cytoplasmic region, thereby activating TGF-βR Ⅰ; activated TGF-βR Ⅰ further phosphorylates R-Smads (Smad2 / Smad3) protein, The latter then combines with Co-Smad (Smad4) to form a heterotrimeric complex, which enters the nucleus and cooperates with other co-activator and co-inhibitor to regulate Target gene transcription. Changes in any part of the TGF-β / Smads signaling pathway will cause abnormalities in the signal transduction pathway.
目前的研究表明，在肿瘤细胞中，TGF-β能直接影响肿瘤的生长(TGF-β信号的非固有影响)，或者通过诱导上皮间质转化、阻断抗肿瘤免疫应答、增加肿瘤相关纤维化和强化血管再生间接地影响肿瘤生长(TGF-β的固有影响)。同时，TGF-β具有很强的纤维化诱导作用，它是与肿瘤相关的成纤维细胞的激活剂。这些成纤维细胞是胶原Ⅰ型和其他纤维化因子的主要来源。成纤维细胞和其他纤维化因子的诱导产物可能继续培育出一个微环境，这个环境会减少免疫应答，增加抗药性和强化肿瘤血管生成另外，在个体发育和肿瘤生长过程中，TGF-β影响血管生再生。例如，TGF-βR Ⅰ型缺陷的小鼠胚胎显示出了严重的血管发育缺陷，证明TGF-β信号通道是血管内皮及平滑肌细胞发育中的关键调节器。Current research shows that in tumor cells, TGF-β can directly affect the growth of tumors (extrinsic effects of TGF-β signaling), or by inducing epithelial-mesenchymal transition, blocking anti-tumor immune responses, and increasing tumor-related fibrosis And enhanced vascular regeneration indirectly affects tumor growth (intrinsic effect of TGF-β). At the same time, TGF-β has a strong fibrosis induction effect, it is an activator of fibroblasts associated with tumors. These fibroblasts are the main source of collagen type I and other fibrotic factors. The induced products of fibroblasts and other fibrotic factors may continue to cultivate a microenvironment that will reduce the immune response, increase drug resistance and strengthen tumor angiogenesis. In addition, during individual development and tumor growth, TGF-β affects blood vessels Health regeneration. For example, TGF-βR type I-deficient mouse embryos show severe defects in vascular development, proving that TGF-β signaling channels are key regulators in the development of vascular endothelium and smooth muscle cells.
近期的研究报道同时指出，TGF-β明显与免疫逃逸相关，对CD8+T细胞介导的抗肿瘤免疫反应影响较大。在针对转移型泌尿上皮癌的临床试验中，TGF-β基因高表达的患者对PD-L1单抗响应及模拟生存率低。TGF-β单抗的基础研究也证明，当其与PD-L1单抗协同使用时，更多CD8+T细胞浸润并发挥作用，揭示了阻断TGF-β对免疫的激活作用及其机理。由于TGF-β的免疫调节作用，小分子TGF-βR Ⅰ抑制剂单药或与PD-(L)1单抗联用在多种实体瘤治疗上具有极大的应用前景。Recent research reports also pointed out that TGF-β is obviously associated with immune escape and has a greater impact on the anti-tumor immune response mediated by CD8 + T cells. In clinical trials for metastatic urothelial carcinoma, patients with high expression of TGF-β gene responded to PD-L1 monoclonal antibody and the simulated survival rate was low. The basic research of TGF-β monoclonal antibody also proves that when it is used in conjunction with PD-L1 monoclonal antibody, more CD8 + T cells infiltrate and play a role, revealing the immune activation effect and mechanism of blocking TGF-β. Because of the immunomodulatory effect of TGF-β, single-agent small-molecule TGF-βR inhibitors or combined with PD- (L) 1 monoclonal antibody have great application prospects in the treatment of various solid tumors.
礼来公司的专利申请WO2002094833A1报道了化合物A(即LY2157299或者Galunisertib)具有TGF-β抑制活性。该化合物能抑制肿瘤细胞侵袭和转移，同时抑制肿瘤细胞向血管內渗。目前有多个临床实验在进行。礼来公司的另一篇专利申请WO2016057278A1报道了化合物B(即LY3200882)，是该公司新研发的TGF-β小分子抑制剂。该化合物与PD-L1联用治疗实体瘤的临床一期实验正在进行中。Lilly's patent application WO2002094833A1 reported that compound A (ie LY2157299 or Galunisertib) has TGF-β inhibitory activity. The compound can inhibit the invasion and metastasis of tumor cells, and at the same time inhibit the infiltration of tumor cells into blood vessels. There are currently multiple clinical trials in progress. Another patent application WO2016057278A1 from Eli Lilly reported compound B (LY3200882), which is a newly developed TGF-β small molecule inhibitor. The first phase of clinical trials of this compound in combination with PD-L1 in the treatment of solid tumors is in progress.
Figure PCTCN2019114347-appb-000001
Figure PCTCN2019114347-appb-000001
发明内容Summary of the invention
一方面，本发明提供了式(I)所示化合物、其药学上可接受的盐或其异构体，In one aspect, the present invention provides a compound represented by formula (I), a pharmaceutically acceptable salt thereof, or an isomer thereof,
Figure PCTCN2019114347-appb-000002
Figure PCTCN2019114347-appb-000002
其中，环A为5-6元杂芳基、苯基、C 5-6环烷基或5-6元杂环烷基； Among them, ring A is 5-6 membered heteroaryl, phenyl, C 5-6 cycloalkyl or 5-6 membered heterocycloalkyl;
R 1、R 2和R 3各自独立地为H、F、Cl、Br、I、-CN、-OH、C 1-6烷氧基、C 1-6烷基、C 2-4烯基、C 2-4炔基或C 3-6环烷基，其中所述C 1-6烷氧基、C 1-6烷基、C 2-4烯基、C 2-4炔基和C 3-6环烷基任选被1、2或3个独立选自F、Cl、Br、CN、-OH、-CH 3、-OCH 3和-NH 2的取代基所取代； R 1 , R 2 and R 3 are each independently H, F, Cl, Br, I, -CN, -OH, C 1-6 alkoxy, C 1-6 alkyl, C 2-4 alkenyl, C 2-4 alkynyl or C 3-6 cycloalkyl, wherein the C 1-6 alkoxy, C 1-6 alkyl, C 2-4 alkenyl, C 2-4 alkynyl and C 3- 6 cycloalkyl groups are optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, CN, -OH, -CH 3 , -OCH 3 and -NH 2 ;
m为1或2；m is 1 or 2;
R 4为5-6元杂芳基或苯基，其中所述5-6元杂芳基和苯基任选被1、2或3个R a所取代； R 4 is 5-6 membered heteroaryl or phenyl, wherein the 5-6 membered heteroaryl and phenyl are optionally substituted with 1, 2 or 3 R a ;
T 1为-O-或-CH 2-； T 1 is -O- or -CH 2- ;
T 2为N或C(R 5)； T 2 is N or C (R 5 );
R 5为H、F、Cl、Br、-CN、-OH、-NH 2、-OCH 3或-CH 3； R 5 is H, F, Cl, Br, -CN, -OH, -NH 2 , -OCH 3 or -CH 3 ;
各R a独立地为-CN、-C(＝O)NR bR c、C 1-6烷氧基或任选被1、2或3个独立选自F、Cl、Br、I、-OH、-OCH 3、-CN或和NH 2的取代基所取代的C 1-6烷基； Each R a is independently -CN, -C (= O) NR b R c , C 1-6 alkoxy or optionally 1, 2, or 3 independently selected from F, Cl, Br, I, -OH , -OCH 3 , -CN or C 1-6 alkyl substituted with a substituent of NH 2 ;
或R 4和R 5与其连接的吡啶基和
Figure PCTCN2019114347-appb-000003
一起形成式(B)所示结构： Or the pyridyl group to which R 4 and R 5 are linked
Figure PCTCN2019114347-appb-000003
Together form the structure shown in formula (B):
Figure PCTCN2019114347-appb-000004
Figure PCTCN2019114347-appb-000004
L为单键、
Figure PCTCN2019114347-appb-000005
L is a single key,
Figure PCTCN2019114347-appb-000005
R 6独立地为H、-CN、-C(＝O)NR bR c、C 1-4烷基、C 3-6环烷基或5-6元杂芳基，其中所述C 1-4烷基、C 3-6环烷基和5-6元杂芳基任选被1、2或3个R d所取代； R 6 is independently H, -CN, -C (= O) NR b R c , C 1-4 alkyl, C 3-6 cycloalkyl, or 5-6 membered heteroaryl, wherein the C 1- 4 alkyl, C 3-6 cycloalkyl and 5-6 membered heteroaryl are optionally substituted with 1, 2 or 3 Rd ;
各R d独立地为F、Cl、Br、I、-OH、-CN、-NH 2、-OCH 3、-OCH 2CH 3、-CH 3、-CF 3或-CH 2CH 3； Each R d is independently F, Cl, Br, I, -OH, -CN, -NH 2, -OCH 3, -OCH 2 CH 3, -CH 3, -CF 3 or -CH 2 CH 3;
R b和R c各自独立地为H、-CH 3、-CH 2CH 3、-CH 2CH 2CH 3、-CH 2(CH 3) 2或环丙基； R b and R c are each independently H, -CH 3 , -CH 2 CH 3 , -CH 2 CH 2 CH 3 , -CH 2 (CH 3 ) 2 or cyclopropyl;
所述5-6元杂环烷基和5-6元杂芳基分别包含1、2、3或4个独立选自N、-O-、-S-和-NH-的杂原子或杂原子团。The 5-6 membered heterocycloalkyl group and 5-6 membered heteroaryl group respectively contain 1, 2, 3 or 4 heteroatoms or heteroatom groups independently selected from N, -O-, -S- and -NH- .
在本发明的一些方案中，上述化合物、其药学上可接受的盐或其异构体，其具有式(I-A)所示结构：In some embodiments of the present invention, the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formula (I-A):
Figure PCTCN2019114347-appb-000006
Figure PCTCN2019114347-appb-000006
其中，环A、R 1、R 2、R 3、R 4、T 1和m如本发明所定义。 Among them, rings A, R 1 , R 2 , R 3 , R 4 , T 1 and m are as defined in the present invention.
在本发明的一些方案中，上述化合物、其药学上可接受的盐或其异构体，其具有式(I-B)所示结构：In some embodiments of the present invention, the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formula (I-B):
Figure PCTCN2019114347-appb-000007
Figure PCTCN2019114347-appb-000007
其中，环A、R 1、R 2、R 3、R 4、R 5、T 1和m如本发明所定义。 Among them, rings A, R 1 , R 2 , R 3 , R 4 , R 5 , T 1 and m are as defined in the present invention.
在本发明的一些方案中，上述R 4为5-6元杂芳基或苯基，其中所述5-6元杂芳基和苯基任选被1、2或3个R a所取代；R 5为H、F、Cl、Br、-CN、-OH、-OCH 3、-NH 2或-CH 3，其他变量如本发明所定义。 In some aspects of the present invention, the above R 4 is a 5-6 membered heteroaryl or phenyl, wherein said 5-6 membered heteroaryl and phenyl are optionally substituted with 1, 2, or 3 R a; R 5 is H, F, Cl, Br, —CN, —OH, —OCH 3 , —NH 2 or —CH 3 , and other variables are as defined in the present invention.
在本发明的一些方案中，上述化合物、其药学上可接受的盐或其异构体，其具有式(I-C)所示结构：In some embodiments of the present invention, the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formula (I-C):
Figure PCTCN2019114347-appb-000008
Figure PCTCN2019114347-appb-000008
其中，环A、R 1、R 2、R 3、T 1、m、R 6和L如本发明所定义。 Among them, rings A, R 1 , R 2 , R 3 , T 1 , m, R 6 and L are as defined in the present invention.
在本发明的一些方案中，上述各R a独立地为CN、-OCH 3、
Figure PCTCN2019114347-appb-000009
Figure PCTCN2019114347-appb-000010
其他变量如本发明所定义。 In some aspects of the present invention, each of the above Ra is independently CN, -OCH 3 ,
Figure PCTCN2019114347-appb-000009
Figure PCTCN2019114347-appb-000010
Other variables are as defined in the present invention.
在本发明的一些方案中，上述R 4为吡咯基、吡唑基、吡啶基、嘧啶基或苯基，所述吡咯基、吡唑基、吡啶基、嘧啶基和苯基任选被1、2或3个R a所取代，其他变量如本发明所定义。 In some embodiments of the present invention, the above R 4 is pyrrolyl, pyrazolyl, pyridyl, pyrimidinyl, or phenyl. The pyrrolyl, pyrazolyl, pyridyl, pyrimidinyl, and phenyl are optionally substituted by 1, 2 or 3 R a are substituted, other variables are as defined in the present invention.
在本发明的一些方案中，上述R 4为
Figure PCTCN2019114347-appb-000011
Figure PCTCN2019114347-appb-000012
R a及其他变量如本发明所定义。 In some aspects of the invention, the above R 4 is
Figure PCTCN2019114347-appb-000011
Figure PCTCN2019114347-appb-000012
R a and the other variables are as defined in the present invention.
在本发明的一些方案中，上述R 4为
Figure PCTCN2019114347-appb-000013
Figure PCTCN2019114347-appb-000014
其他变量如本发明所定义。 In some aspects of the invention, the above R 4 is
Figure PCTCN2019114347-appb-000013
Figure PCTCN2019114347-appb-000014
Other variables are as defined in the present invention.
在本发明的一些方案中，上述R 4为
Figure PCTCN2019114347-appb-000015
Figure PCTCN2019114347-appb-000016
其他变量如本发明所定义。在本发明的一些方案中，上述化合物、其药学上可接受的盐或其异构体，其具有式(I-A1)～(I-A4)所示结构： In some aspects of the invention, the above R 4 is
Figure PCTCN2019114347-appb-000015
Figure PCTCN2019114347-appb-000016
Other variables are as defined in the present invention. In some embodiments of the present invention, the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formulas (I-A1) to (I-A4):
Figure PCTCN2019114347-appb-000017
Figure PCTCN2019114347-appb-000017
其中，环A、R 1、R 2、R 3、T 1、R a和m如本发明所定义。 Wherein Ring A, R 1, R 2, R 3, T 1, R a and m are as defined in the present invention.
在本发明的一些方案中，上述化合物、其药学上可接受的盐或其异构体，其具有式(I-A5)～(I-A12)所示结构：In some embodiments of the present invention, the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formulas (I-A5) to (I-A12):
Figure PCTCN2019114347-appb-000018
Figure PCTCN2019114347-appb-000018
Figure PCTCN2019114347-appb-000019
Figure PCTCN2019114347-appb-000019
其中，环A、R 1、R 2、R 3和R a如本发明所定义。 Wherein Ring A, R 1, R 2, R 3 and R a are as defined in the present invention.
在本发明的一些方案中，上述化合物、其药学上可接受的盐或其异构体，其具有式(I-B1)～(I-B4)所示结构：In some embodiments of the present invention, the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formulas (I-B1) to (I-B4):
Figure PCTCN2019114347-appb-000020
Figure PCTCN2019114347-appb-000020
其中，环A、R 1、R 2、R 3、R 5、T 1、m和R a如本发明所定义。 Wherein Ring A, R 1, R 2, R 3, R 5, T 1, m and R a are as defined in the present invention.
在本发明的一些方案中，上述化合物、其药学上可接受的盐或其异构体，其具有式(I-B5)～(I-B12)所示结构：In some embodiments of the present invention, the above compound, its pharmaceutically acceptable salt, or its isomer has a structure represented by formulas (I-B5) to (I-B12):
Figure PCTCN2019114347-appb-000021
Figure PCTCN2019114347-appb-000021
Figure PCTCN2019114347-appb-000022
Figure PCTCN2019114347-appb-000022
其中，环A、R 1、R 2、R 3、R 5和R a如本发明所定义。 Wherein Ring A, R 1, R 2, R 3, R 5 and R a are as defined in the present invention.
在本发明的一些方案中，上述化合物、其药学上可接受的盐或其异构体，其具有式(I-C1)或(I-C2)所示结构：In some embodiments of the present invention, the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formula (I-C1) or (I-C2):
Figure PCTCN2019114347-appb-000023
Figure PCTCN2019114347-appb-000023
其中，环A、R 1、R 2、R 3、R 6和L如本发明所定义。 Among them, rings A, R 1 , R 2 , R 3 , R 6 and L are as defined in the present invention.
在本发明的一些方案中，上述R 1、R 2和R 3各自独立地为H、F、Cl、Br、I、-CN、-OH、-OCH 3、-OCH 2CH 3、-CH 3、-CH 2CH 3、乙烯基、乙炔基或环丙基，其中所述-OCH 3、-OCH 2CH 3、-CH 3、-CH 2CH 3、乙烯基、乙炔基和环丙基任选被1、2或3个独立选自F、Cl、Br、CN、-OH、-CH 3、-OCH 3和-NH 2的取代基所取代，其他变量如本发明所定义。 In some aspects of the present invention, the above R 1 , R 2 and R 3 are each independently H, F, Cl, Br, I, -CN, -OH, -OCH 3 , -OCH 2 CH 3 , -CH 3 , -CH 2 CH 3 , vinyl, ethynyl, or cyclopropyl, wherein the -OCH 3 , -OCH 2 CH 3 , -CH 3 , -CH 2 CH 3 , vinyl, ethynyl, and cyclopropyl are is selected from substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, CN, -OH, -CH 3, -OCH 3 and -NH 2 to a substituent group, the other variables are as defined in the present invention.
在本发明的一些方案中，上述R 1、R 2和R 3各自独立地为H、F、Cl、Br、-CN、-OH、-OCH 3、-CH 3、-CH 2CH 3或-CF 3，其他变量如本发明所定义。 In some aspects of the present invention, the above R 1 , R 2 and R 3 are each independently H, F, Cl, Br, -CN, -OH, -OCH 3 , -CH 3 , -CH 2 CH 3 or- CF 3 , other variables are as defined in the present invention.
在本发明的一些方案中，上述环A为噻吩基、吡咯基、吡啶基、嘧啶基、苯基或四氢-2H-吡喃基，其他变量如本发明所定义。In some embodiments of the present invention, the above-mentioned ring A is thienyl, pyrrolyl, pyridyl, pyrimidinyl, phenyl, or tetrahydro-2H-pyranyl, and other variables are as defined in the present invention.
在本发明的一些方案中，上述
Figure PCTCN2019114347-appb-000024
为
Figure PCTCN2019114347-appb-000025
R 1、R 2和R 3及其他变量如本发明所定义。 In some aspects of the invention, the above
Figure PCTCN2019114347-appb-000024
for
Figure PCTCN2019114347-appb-000025
R 1 , R 2 and R 3 and other variables are as defined in the present invention.
在本发明的一些方案中，上述
Figure PCTCN2019114347-appb-000026
为
Figure PCTCN2019114347-appb-000027
Figure PCTCN2019114347-appb-000028
R 1及其他变量如本发明所定义。 In some aspects of the invention, the above
Figure PCTCN2019114347-appb-000026
for
Figure PCTCN2019114347-appb-000027
Figure PCTCN2019114347-appb-000028
R 1 and other variables are as defined in the present invention.
在本发明的一些方案中，上述
Figure PCTCN2019114347-appb-000029
为
Figure PCTCN2019114347-appb-000030
Figure PCTCN2019114347-appb-000031
其他变量如本发明所定义。 In some aspects of the invention, the above
Figure PCTCN2019114347-appb-000029
for
Figure PCTCN2019114347-appb-000030
Figure PCTCN2019114347-appb-000031
Other variables are as defined in the present invention.
在本发明的一些方案中，上述
Figure PCTCN2019114347-appb-000032
为
Figure PCTCN2019114347-appb-000033
Figure PCTCN2019114347-appb-000034
其他变量如本发明所定义。 In some aspects of the invention, the above
Figure PCTCN2019114347-appb-000032
for
Figure PCTCN2019114347-appb-000033
Figure PCTCN2019114347-appb-000034
Other variables are as defined in the present invention.
在本发明的一些方案中，上述化合物、其药学上可接受的盐或其异构体，其具有式(I-A13)～(I-A16)所示结构：In some embodiments of the present invention, the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formulas (I-A13) to (I-A16):
Figure PCTCN2019114347-appb-000035
Figure PCTCN2019114347-appb-000035
其中，R 1和R a如本发明所定义。 Among them, R 1 and R a are as defined in the present invention.
在本发明的一些方案中，上述化合物、其药学上可接受的盐或其异构体，其具有式(I-B13)～(I-B36)所示结构：In some embodiments of the present invention, the above compound, its pharmaceutically acceptable salt, or its isomer has a structure represented by formulas (I-B13) to (I-B36):
Figure PCTCN2019114347-appb-000036
Figure PCTCN2019114347-appb-000036
Figure PCTCN2019114347-appb-000037
Figure PCTCN2019114347-appb-000037
其中，R 1和R a本发明所定义。 Among them, R 1 and R a are defined in the present invention.
在本发明的一些方案中，上述化合物、其药学上可接受的盐或其异构体，具有式(I-C3)～(I-C5)所示结构：In some embodiments of the present invention, the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formulae (I-C3) to (I-C5):
Figure PCTCN2019114347-appb-000038
Figure PCTCN2019114347-appb-000038
其中，R 1、L和R 6如本发明所定义。 Among them, R 1 , L and R 6 are as defined in the present invention.
在本发明的一些方案中，上述化合物、其药学上可接受的盐或其异构体，其具有式(I-C6)～(I-C9)所示结构：In some embodiments of the present invention, the above compound, its pharmaceutically acceptable salt, or its isomer has the structure represented by formulas (I-C6) to (I-C9):
Figure PCTCN2019114347-appb-000039
Figure PCTCN2019114347-appb-000039
其中，R 1和R 6如本发明所定义。 Among them, R 1 and R 6 are as defined in the present invention.
在本发明的一些方案中，上述R 6为H、-C(＝O)NH 2、异丙基、环丙基、环己基、吡唑基或吡啶基，其中所述异丙基、环丙基、环己基、吡唑基和吡啶基任选被1、2或3个R d所取代，R d及其他变量如本发明所定义。 In some embodiments of the present invention, the above R 6 is H, —C (═O) NH 2 , isopropyl, cyclopropyl, cyclohexyl, pyrazolyl, or pyridyl, wherein the isopropyl, cyclopropyl The radical, cyclohexyl, pyrazolyl and pyridyl are optionally substituted with 1, 2 or 3 Rd , Rd and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 6独立地为H、-CN、-C(＝O)NH 2、异丙基、环丙基、环己基、吡唑基或吡啶基，其中所述异丙基、环丙基、环己基、吡唑基和吡啶基任选被1、2或3个R d所取代，R d及其他变量如本发明所定义。 In some embodiments of the present invention, the above R 6 is independently H, -CN, -C (= O) NH 2 , isopropyl, cyclopropyl, cyclohexyl, pyrazolyl, or pyridyl, wherein the iso Propyl, cyclopropyl, cyclohexyl, pyrazolyl and pyridyl are optionally substituted with 1, 2 or 3 Rd , Rd and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 6为H、-C(＝O)NH 2、
Figure PCTCN2019114347-appb-000040
Figure PCTCN2019114347-appb-000041
R d及其他变量如本发明所定义。 In some aspects of the present invention, the above R 6 is H, -C (= O) NH 2 ,
Figure PCTCN2019114347-appb-000040
Figure PCTCN2019114347-appb-000041
Rd and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 6独立地为H、-CN、-C(＝O)NH 2、
Figure PCTCN2019114347-appb-000042
Figure PCTCN2019114347-appb-000043
R d及其他变量如本发明所定义。 In some aspects of the invention, the above R 6 is independently H, -CN, -C (= O) NH 2 ,
Figure PCTCN2019114347-appb-000042
Figure PCTCN2019114347-appb-000043
Rd and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 6为H、-C(＝O)NH 2、
Figure PCTCN2019114347-appb-000044
Figure PCTCN2019114347-appb-000045
其他变量如本发明所定义。 In some aspects of the present invention, the above R 6 is H, -C (= O) NH 2 ,
Figure PCTCN2019114347-appb-000044
Figure PCTCN2019114347-appb-000045
Other variables are as defined in the present invention.
在本发明的一些方案中，上述R 6独立地为H、-CN、-C(＝O)NH 2、
Figure PCTCN2019114347-appb-000046
Figure PCTCN2019114347-appb-000047
其他变量如本发明所定义。 In some aspects of the invention, the above R 6 is independently H, -CN, -C (= O) NH 2 ,
Figure PCTCN2019114347-appb-000046
Figure PCTCN2019114347-appb-000047
Other variables are as defined in the present invention.
本发明还有一些方案是由上述变量任意组合而来。There are still some solutions of the present invention that are derived from any combination of the above variables.
在本发明的一些方案中，上述化合物选自下式化合物，其药学上可接受的盐或其异构体，In some embodiments of the present invention, the above compound is selected from compounds of the formula, pharmaceutically acceptable salts or isomers thereof,
Figure PCTCN2019114347-appb-000048
Figure PCTCN2019114347-appb-000048
Figure PCTCN2019114347-appb-000049
Figure PCTCN2019114347-appb-000049
Figure PCTCN2019114347-appb-000050
Figure PCTCN2019114347-appb-000050
另一方面，本发明还提供了上述化合物、其药学上可接受的盐或其异构体在制备TGF-βR1抑制剂药物中的应用。On the other hand, the present invention also provides the use of the above compound, its pharmaceutically acceptable salt or its isomer in the preparation of TGF-βR1 inhibitor drugs.
本发明还提供了上述化合物、其药学上可接受的盐或其异构体在制备治疗实体癌药物中的应用。The invention also provides the application of the above compound, its pharmaceutically acceptable salt or its isomer in the preparation of a drug for treating solid cancer.
技术效果Technical effect
本发明化合物对TGF-βR1具有很高的选择性，且具有显著的激酶抑制活性。在细胞中展示出了对TGF-β下游信号的明显抑制，同时也具有优良的药代动力学、药效动力学性质和体内药效。The compound of the present invention has high selectivity for TGF-βR1 and has significant kinase inhibitory activity. It showed obvious inhibition of the downstream signal of TGF-β in cells, and also had excellent pharmacokinetics, pharmacodynamic properties and in vivo pharmacodynamics.
定义和说明Definition and description
除非另有说明，本文所用的下列术语和短语旨在具有下列含义。一个特定的术语或短语在没有特别定义的情况下不应该被认为是不确定的或不清楚的，而应该按照普通的含义去理解。当本文中出现商品名时，意在指代其对应的商品或其活性成分。Unless otherwise stated, the following terms and phrases used herein are intended to have the following meanings. A specific term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary meaning. When a trade name appears in this article, it is intended to refer to its corresponding trade product or its active ingredient.
这里所采用的术语“药学上可接受的”，是针对那些化合物、材料、组合物和/或剂型而言，它们在可靠的医学判断的范围之内，适用于与人类和动物的组织接触使用，而没有过多的毒性、刺激性、过敏性反应或其它问题或并发症，与合理的利益/风险比相称。The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms that are within the scope of reliable medical judgment and are suitable for use in contact with human and animal tissues Without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit / risk ratio.
术语“药学上可接受的盐”是指本发明化合物的盐，由本发明发现的具有特定取代基的化合物与相对无毒的酸或碱制备。当本发明的化合物中含有相对酸性的功能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的碱与这类化合物接触的方式获得碱加成盐。药学上可接受的碱加成盐包括钠、钾、钙、铵、有机胺或镁盐或类似的盐。当本发明的化合物中含有相对碱性的官能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的酸与这类化合物接触的方式获得酸加成盐。药学上可接受的酸加成盐的实例包括无机酸盐，所述无机酸包括例如盐酸、氢溴酸、硝酸、碳酸，碳酸氢根，磷酸、磷酸一氢根、磷酸二氢根、硫酸、硫酸氢根、氢碘酸、亚磷酸等；以及有机酸盐，所述有机酸包括如乙酸、丙酸、异丁酸、马来酸、丙二酸、苯甲酸、琥珀酸、辛二酸、反丁烯二酸、乳酸、扁桃酸、邻苯二甲酸、苯磺酸、对甲苯磺酸、柠檬酸、酒石酸和甲磺酸等类似的酸；还包括氨基酸(如精氨酸等)的盐，以及如葡糖醛酸等有机酸的盐。本发明的某些特定的化合物含有碱性和酸性的官能团，从而可以被转换成任一碱或酸加成盐。The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention, prepared from a compound having a specific substituent and a relatively non-toxic acid or base found in the present invention. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, Bisulfate, hydroiodic acid, phosphorous acid, etc .; and organic acid salts, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, Fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; also includes salts of amino acids (such as arginine, etc.) , And salts of organic acids such as glucuronic acid. Certain compounds of the present invention contain basic and acidic functional groups and can be converted to any base or acid addition salt.
本发明的药学上可接受的盐可由含有酸根或碱基的母体化合物通过常规化学方法合成。一般情况下，这样的盐的制备方法是：在水或有机溶剂或两者的混合物中，经由游离酸或碱形式的这些化合物与化学计量的适当的碱或酸反应来制备。The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing acid radicals or bases by conventional chemical methods. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of both.
本发明的化合物可以存在特定的几何或立体异构体形式。本发明设想所有的这类化合物，包括顺式和反式异构体、(-)-和(+)-对映体、(R)-和(S)-对映体、非对映异构体、(D)-异构体、(L)-异构体，及其外消旋混合物和其他混合物，例如对映异构体或非对映体富集的混合物，所有这些混合物都属于本发明的范围 之内。烷基等取代基中可存在另外的不对称碳原子。所有这些异构体以及它们的混合物，均包括在本发明的范围之内。The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)-and (+)-enantiomers, (R)-and (S) -enantiomers, diastereomers Isomers, (D) -isomers, (L) -isomers, and their racemic mixtures and other mixtures, such as enantiomerically or diastereomerically enriched mixtures, all of which belong to this Within the scope of the invention. Additional asymmetric carbon atoms may be present in the substituents such as alkyl. All these isomers and mixtures thereof are included in the scope of the present invention.
除非另有说明，术语“对映异构体”或者“旋光异构体”是指互为镜像关系的立体异构体。Unless otherwise stated, the term "enantiomer" or "optical isomer" refers to stereoisomers in a mirror image relationship with each other.
除非另有说明，术语“顺反异构体”或者“几何异构体”系由因双键或者成环碳原子单键不能自由旋转而引起。Unless otherwise stated, the term "cis-trans isomer" or "geometric isomer" is caused by the fact that double bonds or single bonds of ring-forming carbon atoms cannot rotate freely.
除非另有说明，术语“非对映异构体”是指分子具有两个或多个手性中心，并且分子间为非镜像的关系的立体异构体。Unless otherwise stated, the term "diastereomers" refers to stereoisomers in which molecules have two or more chiral centers and are in a non-mirror relationship.
除非另有说明，“(+)”表示右旋，“(-)”表示左旋，“(±)”表示外消旋。Unless otherwise stated, "(+)" means right-handed, "(-)" means left-handed, and "(±)" means racemic.
除非另有说明，用楔形实线键
Figure PCTCN2019114347-appb-000051
和楔形虚线键
Figure PCTCN2019114347-appb-000052
表示一个立体中心的绝对构型，用波浪线
Figure PCTCN2019114347-appb-000053
表示楔形实线键
Figure PCTCN2019114347-appb-000054
或楔形虚线键
Figure PCTCN2019114347-appb-000055
Unless otherwise stated, use a solid wedge key
Figure PCTCN2019114347-appb-000051
And wedge-shaped dotted keys
Figure PCTCN2019114347-appb-000052
Represents the absolute configuration of a three-dimensional center, with wavy lines
Figure PCTCN2019114347-appb-000053
Represents a solid wedge key
Figure PCTCN2019114347-appb-000054
Or wedge-shaped dotted key
Figure PCTCN2019114347-appb-000055
除非另有说明，当化合物中存在双键结构，如碳碳双键、碳氮双键和氮氮双键，且双键上的各个原子均连接有两个不同的取代基时(包含氮原子的双键中，氮原子上的一对孤对电子视为其连接的一个取代基)，如果该化合物中双键上的原子与其取代基之间用波浪线
Figure PCTCN2019114347-appb-000056
连接，则表示该化合物的(Z)型异构体、(E)型异构体或两种异构体的混合物。例如下式(A)表示该化合物以式(A-1)或式(A-2)的单一异构体形式存在或以式(A-1)和式(A-2)两种异构体的混合物形式存在；下式(B)表示该化合物以式(B-1)或式(B-2)的单一异构体形式存在或以式(B-1)和式(B-2)两种异构体的混合物形式存在。下式(C)表示该化合物以式(C-1)或式(C-2)的单一异构体形式存在或以式(C-1)和式(C-2)两种异构体的混合物形式存在。 Unless otherwise stated, when a double bond structure exists in a compound, such as a carbon-carbon double bond, a carbon-nitrogen double bond, and a nitrogen-nitrogen double bond, and each atom on the double bond is connected to two different substituents (including nitrogen atoms In the double bond of, a lone pair of electrons on the nitrogen atom is regarded as a substituent to which it is connected), if the compound on the double bond in the compound is wavy
Figure PCTCN2019114347-appb-000056
Linked means the (Z) isomer, (E) isomer or a mixture of two isomers of the compound. For example, the following formula (A) indicates that the compound exists as a single isomer of formula (A-1) or (A-2) or as two isomers of formula (A-1) and formula (A-2) Exists in the form of a mixture; the following formula (B) indicates that the compound exists as a single isomer of formula (B-1) or formula (B-2) or as both formula (B-1) and formula (B-2) There is a mixture of isomers. The following formula (C) indicates that the compound exists as a single isomer of formula (C-1) or formula (C-2) or as two isomers of formula (C-1) and formula (C-2) In the form of a mixture.
Figure PCTCN2019114347-appb-000057
Figure PCTCN2019114347-appb-000057
本发明的化合物可以存在特定的。除非另有说明，术语“互变异构体”或“互变异构体形式”是指在室温下，不同官能团异构体处于动态平衡，并能很快的相互转化。若互变异构体是可能的(如在溶液中)，则可以达到互变异构体的化学平衡。例如，质子互变异构体(proton tautomer)(也称质子转移互变异构体(prototropic tautomer))包括通过质子迁移来进行的互相转化，如酮-烯醇异构化和亚胺-烯胺异构化。价键异构体(valence tautomer)包括一些成键电子的重组来进行的相互转化。其中酮-烯醇互变异构化的具体实例是戊烷-2,4-二酮与4-羟基戊-3-烯-2-酮两个互变异构体之间的互变。The compounds of the present invention may be present in specific. Unless otherwise stated, the term "tautomer" or "tautomeric form" means that at room temperature, isomers of different functional groups are in dynamic equilibrium and can quickly convert to each other. If tautomers are possible (as in solution), the chemical equilibrium of tautomers can be achieved. For example, proton tautomers (also known as prototropic tautomers) include interconversion through proton migration, such as keto-enol isomerization and imine-ene Amine isomerization. Valence tautomer (valence tautomer) includes some recombination of bond-forming electrons for mutual conversion. A specific example of keto-enol tautomerization is the interconversion between two tautomers of pentane-2,4-dione and 4-hydroxypent-3-en-2-one.
除非另有说明，术语“富含一种异构体”、“异构体富集”、“富含一种对映体”或者“对映体富集”指其中一种异构体或对映体的含量小于100％，并且，该异构体或对映体的含量大于等于60％，或者大于等于70％， 或者大于等于80％，或者大于等于90％，或者大于等于95％，或者大于等于96％，或者大于等于97％，或者大于等于98％，或者大于等于99％，或者大于等于99.5％，或者大于等于99.6％，或者大于等于99.7％，或者大于等于99.8％，或者大于等于99.9％。Unless otherwise stated, the terms "rich in one isomer", "isomer enriched", "rich in one enantiomer" or "enantiomerically enriched" refer to one of the isomers or pairs The content of the enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or 96% or greater, or 97% or greater, or 98% or greater, or 99% or greater, or 99.5% or greater, or 99.6% or greater, or 99.7% or greater, or 99.8% or greater, or greater than or equal to 99.9%.
除非另有说明，术语“异构体过量”或“对映体过量”指两种异构体或两种对映体相对百分数之间的差值。例如，其中一种异构体或对映体的含量为90％，另一种异构体或对映体的含量为10％，则异构体或对映体过量(ee值)为80％。Unless otherwise stated, the term "isomer excess" or "enantiomeric excess" refers to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, then the excess of isomer or enantiomer (ee value) is 80% .
可以通过的手性合成或手性试剂或者其他常规技术制备光学活性的(R)-和(S)-异构体以及D和L异构体。如果想得到本发明某化合物的一种对映体，可以通过不对称合成或者具有手性助剂的衍生作用来制备，其中将所得非对映体混合物分离，并且辅助基团裂开以提供纯的所需对映异构体。或者，当分子中含有碱性官能团(如氨基)或酸性官能团(如羧基)时，与适当的光学活性的酸或碱形成非对映异构体的盐，然后通过本领域所公知的常规方法进行非对映异构体拆分，然后回收得到纯的对映体。此外，对映异构体和非对映异构体的分离通常是通过使用色谱法完成的，所述色谱法采用手性固定相，并任选地与化学衍生法相结合(例如由胺生成氨基甲酸盐)。The optically active (R)-and (S) -isomers and D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If an enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, where the resulting mixture of diastereomers is separated and the auxiliary groups are cleaved to provide pure The desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a salt of a diastereomer is formed with an appropriate optically active acid or base, and then by conventional methods known in the art The diastereomers are resolved and the pure enantiomers are recovered. In addition, the separation of enantiomers and diastereomers is usually done by using chromatography, which uses a chiral stationary phase, and is optionally combined with chemical derivatization methods (for example, the formation of amino groups from amines) Formate).
发明的化合物可以在一个或多个构成该化合物的原子上包含非天然比例的原子同位素。例如，可用放射性同位素标记化合物，比如氚( 3H)，碘-125( 125I)或C-14( 14C)。又例如，可用重氢取代氢形成氘代药物，氘与碳构成的键比普通氢与碳构成的键更坚固，相比于未氘化药物，氘代药物有降低毒副作用、增加药物稳定性、增强疗效、延长药物生物半衰期等优势。本发明的化合物的所有同位素组成的变换，无论放射性与否，都包括在本发明的范围之内。 The compound of the invention may contain unnatural proportions of atomic isotopes at one or more atoms constituting the compound. For example, compounds can be labeled with radioisotopes, such as tritium ( 3 H), iodine-125 ( 125 I) or C-14 ( 14 C). For another example, the hydrogen can be replaced by heavy hydrogen to form a deuterated drug. The bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have reduced toxic and side effects and increased drug stability. , Strengthen efficacy, prolong the biological half-life of drugs and other advantages. The conversion of all isotopic compositions of the compounds of the present invention, whether radioactive or not, is included in the scope of the present invention.
“任选”或“任选地”指的是随后描述的事件或状况可能但不是必需出现的，并且该描述包括其中所述事件或状况发生的情况以及所述事件或状况不发生的情况。"Optional" or "optionally" means that the subsequently described events or conditions may, but need not necessarily occur, and that the description includes situations in which the events or conditions occur and situations in which the events or conditions do not occur.
术语“被取代的”是指特定原子上的任意一个或多个氢原子被取代基取代，可以包括重氢和氢的变体，只要特定原子的价态是正常的并且取代后的化合物是稳定的。当取代基为氧(即＝O)时，意味着两个氢原子被取代。氧取代不会发生在芳香基上。术语“任选被取代的”是指可以被取代，也可以不被取代，除非另有规定，取代基的种类和数目在化学上可以实现的基础上可以是任意的。The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include heavy hydrogen and hydrogen variants, as long as the valence state of the particular atom is normal and the compound after substitution is stable of. When the substituent is oxygen (ie = O), it means that two hydrogen atoms are substituted. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may or may not be substituted. Unless otherwise specified, the type and number of substituents may be arbitrary on the basis that they are chemically achievable.
当任何变量(例如R)在化合物的组成或结构中出现一次以上时，其在每一种情况下的定义都是独立的。因此，例如，如果一个基团被0-2个R所取代，则所述基团可以任选地至多被两个R所取代，并且每种情况下的R都有独立的选项。此外，取代基和/或其变体的组合只有在这样的组合会产生稳定的化合物的情况下才是被允许的。When any variable (such as R) appears more than once in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group can optionally be substituted with up to two Rs, and R in each case has independent options. In addition, combinations of substituents and / or variants thereof are only allowed if such combinations will produce stable compounds.
当一个连接基团的数量为0时，比如-(CRR) 0-，表示该连接基团为单键。 When the number of a linking group is 0, such as-(CRR) 0- , it means that the linking group is a single bond.
当其中一个变量选自单键时，表示其连接的两个基团直接相连，比如A-L-Z中L代表单键时表示该结构实际上是A-Z。When one of the variables is selected from a single bond, it means that the two groups to which it is connected are directly connected. For example, when L represents a single bond in A-L-Z, it means that the structure is actually A-Z.
一个取代基为空缺时，表示该取代基是不存在的，比如A-X中X为空缺时表示该结构实际上是A。当所列举的取代基中没有指明其通过哪一个原子连接到被取代的基团上时，这种取代基可以通过其任何原子相键合，例如，吡啶基作为取代基可以通过吡啶环上任意一个碳原子连接到被取代的基团上。When a substituent is vacant, it means that the substituent does not exist. For example, when X is vacant in A-X, it means that the structure is actually A. When the substituents listed do not indicate through which atom they are connected to the substituted group, such substituents can be bonded through any of their atoms, for example, pyridyl as a substituent can be through any one of the pyridine rings The carbon atom is attached to the substituted group.
当所列举的连接基团没有指明其连接方向，其连接方向是任意的，例如，
Figure PCTCN2019114347-appb-000058
中连接基团L为-M-W-，此时-M-W-既可以按与从左往右的读取顺序相同的方向连接环A和环B构成
Figure PCTCN2019114347-appb-000059
也可以按照与从左往右的读取顺序相反的方向连接环A和环B构成
Figure PCTCN2019114347-appb-000060
所述连接基团、取代基和/或其变体的组合只有在这样的组合会产生稳定的化合物的情况下才是被允许的。 When the listed linking group does not indicate the connection direction, the connection direction is arbitrary, for example,
Figure PCTCN2019114347-appb-000058
The linking group L in the middle is -MW-, then -MW- can be formed by connecting ring A and ring B in the same direction as the reading order from left to right
Figure PCTCN2019114347-appb-000059
It can also be formed by connecting ring A and ring B in the opposite direction to the reading order from left to right
Figure PCTCN2019114347-appb-000060
Combinations of the linking groups, substituents, and / or variants thereof are only allowed if such combinations will produce stable compounds.
除非另有规定，当某一基团具有一个或多个可连接位点时，该基团的任意一个或多个位点可以通过化学键与其他基团相连。所述位点与其他基团连接的化学键可以用直形实线键
Figure PCTCN2019114347-appb-000061
直形虚线键
Figure PCTCN2019114347-appb-000062
或波浪线
Figure PCTCN2019114347-appb-000063
表示。例如-OCH 3中的直形实线键表示通过该基团中的氧原子与其他基团相连；
Figure PCTCN2019114347-appb-000064
中的直形虚线键表示通过该基团中的氮原子的两端与其他基团相连；
Figure PCTCN2019114347-appb-000065
中的波浪线表示通过该苯基基团中的1位和2位碳原子与其他基团相连。 Unless otherwise specified, when a group has one or more connectable sites, any one or more sites of the group may be connected to other groups by chemical bonds. The chemical bond connecting the site with other groups can be a straight solid bond
Figure PCTCN2019114347-appb-000061
Straight dotted key
Figure PCTCN2019114347-appb-000062
Or wavy lines
Figure PCTCN2019114347-appb-000063
Said. For example, the straight solid line bond in -OCH 3 indicates that it is connected to other groups through the oxygen atom in the group;
Figure PCTCN2019114347-appb-000064
The straight dashed bond in means that the two ends of the nitrogen atom in the group are connected to other groups;
Figure PCTCN2019114347-appb-000065
The wavy line in indicates that it is connected to other groups through the 1 and 2 carbon atoms in the phenyl group.
除非另有规定，环上原子的数目通常被定义为环的元数，例如，“5-7元环”是指环绕排列5-7个原子的“环”。Unless otherwise specified, the number of atoms on a ring is usually defined as the number of members of the ring. For example, "5-7 membered ring" refers to a "ring" with 5-7 atoms arranged around it.
除非另有规定，术语“5-6元环”表示由5至6个环原子组成的环烷基、杂环烷基、环烯基、杂环烯基、环炔基、杂环炔基、芳基或杂芳基。所述的环包括单环，也包括螺环、并环和桥环等双环体系。除非另有规定，该环任选地包含1、2或3个独立选自O、S和N的杂原子。所述5-6元环包括5元、6元环等。“5-6元环”包括例如苯基、吡啶基和哌啶基等；另一方面，术语“5-6元杂环烷基”包括哌啶基等，但不包括苯基。术语“环”还包括含有至少一个环的环系，其中的每一个“环”均独立地符合上述定义。Unless otherwise specified, the term "5-6 membered ring" means cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, cycloalkynyl, heterocycloalkynyl, consisting of 5 to 6 ring atoms, Aryl or heteroaryl. The ring includes a single ring, and also includes a double ring system such as a spiro ring, a bicyclic ring, and a bridge ring. Unless otherwise specified, the ring optionally contains 1, 2 or 3 heteroatoms independently selected from O, S and N. The 5-6 member ring includes 5 member, 6 member ring, and the like. "5-6 membered ring" includes, for example, phenyl, pyridyl, piperidinyl, and the like; on the other hand, the term "5-6 membered heterocycloalkyl" includes piperidinyl and the like, but does not include phenyl. The term "ring" also includes ring systems containing at least one ring, where each "ring" independently conforms to the above definition.
除非另有规定，术语“C 1-6烷基”用于表示直链或支链的由1至6个碳原子组成的饱和碳氢基团。所述C 1-6烷基包括C 1-5、C 1-4、C 1-3、C 1-2、C 2-6、C 2-4、C 6和C 5烷基等；其可以是一价(如甲基)、二价(如亚甲基)或者多价(如次甲基)。C 1-6烷基的实例包括但不限于甲基(Me)、乙基(Et)、丙基(包括n-丙基和异丙基)、丁基(包括n-丁基，异丁基，s-丁基和t-丁基)、戊基(包括n-戊基，异戊基和新戊基)、己基等。 Unless otherwise specified, the term "C 1-6 alkyl" is used to indicate a linear or branched saturated hydrocarbon group composed of 1 to 6 carbon atoms. The C 1-6 alkyl group includes C 1-5 , C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-4 , C 6 and C 5 alkyl groups; etc .; Is monovalent (such as methyl), divalent (such as methylene) or multivalent (such as methine). Examples of C 1-6 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl , S-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl and so on.
除非另有规定，术语“C 1-4烷基”用于表示直链或支链的由1至4个碳原子组成的饱和碳氢基团。所述C 1-4烷基包括C 1-2、C 1-3和C 2-3烷基等；其可以是一价(如甲基)、二价(如亚甲基)或者多价(如次甲基)。C 1-4烷基的实例包括但不限于甲基(Me)、乙基(Et)、丙基(包括n-丙基和异丙基)、丁基(包括n-丁基，异丁基，s-丁基和t-丁基)等。 Unless otherwise specified, the term "C 1-4 alkyl" is used to indicate a linear or branched saturated hydrocarbon group composed of 1 to 4 carbon atoms. The C 1-4 alkyl group includes C 1-2 , C 1-3 and C 2-3 alkyl groups, etc .; it may be monovalent (such as methyl), divalent (such as methylene) or multivalent ( Such as methine). Examples of C 1-4 alkyl include but are not limited to methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl , S-butyl and t-butyl) and so on.
除非另有规定，术语“C 2-4烯基”用于表示直链或支链的包含至少一个碳-碳双键的由2至4个碳原子组成的碳氢基团，碳-碳双键可以位于该基团的任何位置上。所述C 2-4烯基包括C 2-3、C 4、C 3和C 2烯基等；所述C 2-4烯基可以是一价、二价或者多价。C 2-4烯基的实例包括但不限于乙烯基、丙烯基、丁烯基、丁间二烯基等。 Unless otherwise specified, the term "C 2-4 alkenyl" is used to denote a linear or branched hydrocarbon group consisting of at least one carbon-carbon double bond consisting of 2 to 4 carbon atoms, carbon-carbon double The bond can be located anywhere on the group. The C 2-4 alkenyl group includes C 2-3 , C 4 , C 3 and C 2 alkenyl groups; the C 2-4 alkenyl group may be monovalent, divalent or multivalent. Examples of C 2-4 alkenyl include, but are not limited to, vinyl, propenyl, butenyl, butadienyl, and the like.
除非另有规定，术语“C 2-4炔基”用于表示直链或支链的包含至少一个碳-碳三键的由2至4个碳原子组 成的碳氢基团，碳-碳三键可以位于该基团的任何位置上。所述C 2-4炔基包括C 2-3、C 4、C 3和C 2炔基等。其可以是一价、二价或者多价。C 2-4炔基的实例包括但不限于乙炔基、丙炔基、丁炔基等。 Unless otherwise specified, the term "C 2-4 alkynyl" is used to denote a linear or branched hydrocarbon group consisting of 2 to 4 carbon atoms containing at least one carbon-carbon triple bond, carbon-carbon triple The bond can be located anywhere on the group. The C 2-4 alkynyl group includes C 2-3 , C 4 , C 3 and C 2 alkynyl groups. It can be monovalent, bivalent or multivalent. Examples of C 2-4 alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, and the like.
除非另有规定，术语“C 1-6烷氧基”表示通过一个氧原子连接到分子的其余部分的那些包含1至6个碳原子的烷基基团。所述C 1-6烷氧基包括C 1-4、C 1-3、C 1-2、C 2-6、C 2-4、C 6、C 5、C 4和C 3烷氧基等。C 1-6烷氧基的实例包括但不限于甲氧基、乙氧基、丙氧基(包括正丙氧基和异丙氧基)、丁氧基(包括n-丁氧基、异丁氧基、s-丁氧基和t-丁氧基)、戊氧基(包括n-戊氧基、异戊氧基和新戊氧基)、己氧基等。 Unless otherwise specified, the term "C 1-6 alkoxy" refers to those alkyl groups containing 1 to 6 carbon atoms connected to the rest of the molecule through one oxygen atom. The C 1-6 alkoxy group includes C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-4 , C 6 , C 5 , C 4 and C 3 alkoxy groups, etc. . Examples of C 1-6 alkoxy include but are not limited to methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy Oxy, s-butoxy and t-butoxy), pentyloxy (including n-pentyloxy, isopentyloxy and neopentyloxy), hexyloxy, etc.
除非另有规定，术语“C 1-4烷氧基”表示通过一个氧原子连接到分子的其余部分的那些包含1至4个碳原子的烷基基团。所述C 1-4烷氧基包括C 1-3、C 1-2、C 2-4、C 4和C 3烷氧基等。C 1-6烷氧基的实例包括但不限于甲氧基、乙氧基、丙氧基(包括正丙氧基和异丙氧基)、丁氧基(包括n-丁氧基、异丁氧基、s-丁氧基和t-丁氧基)、戊氧基(包括n-戊氧基、异戊氧基和新戊氧基)、己氧基等。 Unless otherwise specified, the term "C 1-4 alkoxy" refers to those alkyl groups containing 1 to 4 carbon atoms connected to the rest of the molecule through one oxygen atom. The C 1-4 alkoxy group includes C 1-3 , C 1-2 , C 2-4 , C 4 and C 3 alkoxy groups and the like. Examples of C 1-6 alkoxy include but are not limited to methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy Oxy, s-butoxy and t-butoxy), pentyloxy (including n-pentyloxy, isopentyloxy and neopentyloxy), hexyloxy, etc.
除非另有规定，术语“卤代素”或“卤素”本身或作为另一取代基的一部分表示氟、氯、溴或碘原子。Unless otherwise specified, the term "halogen" or "halogen" itself or as part of another substituent means a fluorine, chlorine, bromine or iodine atom.
除非另有规定，术语“C 3-6环烷基”表示由3至6个碳原子组成的饱和环状碳氢基团，其为单环和双环体系，所述C 3-6环烷基包括C 3-5、C 4-5和C 5-6环烷基等；其可以是一价、二价或者多价。C 3-6环烷基的实例包括，但不限于，环丙基、环丁基、环戊基、环己基等。 Unless otherwise specified, the term "C 3-6 cycloalkyl" means a saturated cyclic hydrocarbon group consisting of 3 to 6 carbon atoms, which is a monocyclic and bicyclic ring system, the C 3-6 cycloalkyl Including C 3-5 , C 4-5 and C 5-6 cycloalkyl, etc .; it can be monovalent, divalent or multivalent. Examples of C 3-6 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.
除非另有规定，术语“C 5-6环烷基”表示由5至6个碳原子组成的饱和环状碳氢基团，其为单环和双环体系，所述C 5-6环烷基包括C 5和C 6环烷基等；其可以是一价、二价或者多价。C 5-6环烷基的实例包括，但不限于，环戊基、环己基等。 Unless otherwise specified, the term "C 5-6 cycloalkyl" means a saturated cyclic hydrocarbon group consisting of 5 to 6 carbon atoms, which is a monocyclic and bicyclic ring system, the C 5-6 cycloalkyl Including C 5 and C 6 cycloalkyl and the like; it can be monovalent, divalent or polyvalent. Examples of C 5-6 cycloalkyl include, but are not limited to, cyclopentyl, cyclohexyl, and the like.
除非另有规定，术语“5-6元杂环烷基”本身或者与其他术语联合分别表示由5至6个环原子组成的饱和环状基团，其1、2、3或4个环原子为独立选自O、S和N的杂原子，其余为碳原子，其中氮原子任选地被季铵化，氮和硫杂原子可任选被氧化(即NO和S(O) p，p是1或2)。其包括单环和双环体系，其中双环体系包括螺环、并环和桥环。此外，就该“5-6元杂环烷基”而言，杂原子可以占据杂环烷基与分子其余部分的连接位置。所述5-6元杂环烷基包括5元和6元杂环烷基。5-6元杂环烷基的实例包括但不限于吡咯烷基、吡唑烷基、咪唑烷基、四氢噻吩基(包括四氢噻吩-2-基和四氢噻吩-3-基等)、四氢呋喃基(包括四氢呋喃-2-基等)、四氢吡喃基、哌啶基(包括1-哌啶基、2-哌啶基和3-哌啶基等)、哌嗪基(包括1-哌嗪基和2-哌嗪基等)、吗啉基(包括3-吗啉基和4-吗啉基等)、二噁烷基、二噻烷基、异噁唑烷基、异噻唑烷基、1,2-噁嗪基、1,2-噻嗪基、六氢哒嗪基、高哌嗪基或高哌啶基等。 Unless otherwise specified, the term "5-6 membered heterocycloalkyl" by itself or in combination with other terms means a saturated cyclic group consisting of 5 to 6 ring atoms with 1, 2, 3 or 4 ring atoms Are heteroatoms independently selected from O, S, and N, and the rest are carbon atoms, wherein nitrogen atoms are optionally quaternized, and nitrogen and sulfur heteroatoms may be optionally oxidized (ie, NO and S (O) p , p Is 1 or 2). It includes single-ring and double-ring systems, wherein the double-ring system includes spiro ring, parallel ring and bridge ring. In addition, as far as the "5-6 membered heterocycloalkyl group" is concerned, the hetero atom may occupy the connection position of the heterocyclic alkyl group to the rest of the molecule. The 5-6 membered heterocycloalkyl group includes 5-membered and 6-membered heterocycloalkyl groups. Examples of 5-6 membered heterocycloalkyl include, but are not limited to, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothienyl (including tetrahydrothien-2-yl and tetrahydrothien-3-yl, etc.) , Tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl, and 3-piperidinyl, etc.), piperazinyl (including 1 -Piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazole Alkyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, homopiperazinyl or homopiperidinyl, etc.
除非另有规定，术语“C 6-12芳环”和“C 6-12芳基”可以互换使用，术语“C 6-12芳环”或“C 6-12芳基”表示由6至12个碳原子组成的具有共轭π电子体系的环状碳氢基团，它可以是单环、稠合双环或稠合三环体系，其中各个环均为芳香性的。其可以是一价、二价或者多价，C 6-12芳基包括C 6-10、C 6-9、C 6-8、C 12、C 10和C 6芳基等。C 6-12芳基的实例包括但不限于苯基、萘基(包括1-萘基和2-萘基等)。 Unless otherwise specified, the terms "C 6-12 aryl ring" and "C 6-12 aryl group" can be used interchangeably. The term "C 6-12 aryl ring" or "C 6-12 aryl group" means from 6 to A cyclic hydrocarbon group consisting of 12 carbon atoms with a conjugated π-electron system. It can be a single ring, a fused bicyclic ring, or a fused tricyclic ring system, where each ring is aromatic. It may be monovalent, divalent or multivalent, and C 6-12 aryl groups include C 6-10 , C 6-9 , C 6-8 , C 12 , C 10 and C 6 aryl groups and the like. Examples of C 6-12 aryl groups include, but are not limited to, phenyl, naphthyl (including 1-naphthyl, 2-naphthyl, etc.).
除非另有规定，术语“C 6-10芳环”和“C 6-10芳基”可以互换使用，术语“C 6-10芳环”或“C 6-10芳基”表示由6至10个碳原子组成的具有共轭π电子体系的环状碳氢基团，它可以是单环、稠合双环或稠合三环体系，其中各个环均为芳香性的。其可以是一价、二价或者多价，C 6-10芳基包括C 6-9、C 9、C 10和C 6芳基等。C 6- 10芳基的实例包括但不限于苯基、萘基(包括1-萘基和2-萘基等)。 Unless otherwise specified, the terms "C 6-10 aryl ring" and "C 6-10 aryl group" can be used interchangeably. The term "C 6-10 aryl ring" or "C 6-10 aryl group" means from 6 to A cyclic hydrocarbon group consisting of 10 carbon atoms with a conjugated π-electron system. It can be a single ring, a fused bicyclic ring or a fused tricyclic ring system, where each ring is aromatic. It may be monovalent, divalent or multivalent, and C 6-10 aryl groups include C 6-9 , C 9 , C 10 and C 6 aryl groups. Example C 6- 10 aryl group include, but are not limited to, phenyl, naphthyl (including 1-naphthyl and 2-naphthyl and the like).
除非另有规定，术语“5-6元杂芳环”和“5-6元杂芳基”可以互换使用，术语“5-6元杂芳基”表示由5至6 个环原子组成的具有共轭π电子体系的单环基团，其1、2、3或4个环原子为独立选自O、S和N的杂原子，其余为碳原子。其中氮原子任选地被季铵化，氮和硫杂原子可任选被氧化(即NO和S(O) p，p是1或2)。5-6元杂芳基可通过杂原子或碳原子连接到分子的其余部分。所述5-6元杂芳基包括5元和6元杂芳基。所述5-6元杂芳基的实例包括但不限于吡咯基(包括N-吡咯基、2-吡咯基和3-吡咯基等)、吡唑基(包括2-吡唑基和3-吡唑基等)、咪唑基(包括N-咪唑基、2-咪唑基、4-咪唑基和5-咪唑基等)、噁唑基(包括2-噁唑基、4-噁唑基和5-噁唑基等)、三唑基(1H-1,2,3-三唑基、2H-1,2,3-三唑基、1H-1,2,4-三唑基和4H-1,2,4-三唑基等)、四唑基、异噁唑基(3-异噁唑基、4-异噁唑基和5-异噁唑基等)、噻唑基(包括2-噻唑基、4-噻唑基和5-噻唑基等)、呋喃基(包括2-呋喃基和3-呋喃基等)、噻吩基(包括2-噻吩基和3-噻吩基等)、吡啶基(包括2-吡啶基、3-吡啶基和4-吡啶基等)、吡嗪基或嘧啶基(包括2-嘧啶基和4-嘧啶基等)。 Unless otherwise specified, the terms "5-6 membered heteroaryl ring" and "5-6 membered heteroaryl group" are used interchangeably, and the term "5-6 membered heteroaryl group" means composed of 5 to 6 ring atoms In a monocyclic group with a conjugated π electron system, 1, 2, 3, or 4 ring atoms are heteroatoms independently selected from O, S, and N, and the rest are carbon atoms. Where nitrogen atoms are optionally quaternized, nitrogen and sulfur heteroatoms can be optionally oxidized (ie NO and S (O) p , p is 1 or 2). The 5-6 membered heteroaryl group can be attached to the rest of the molecule through a heteroatom or carbon atom. The 5-6 membered heteroaryl group includes 5-membered and 6-membered heteroaryl groups. Examples of the 5-6 membered heteroaryl group include but are not limited to pyrrolyl (including N-pyrrolyl, 2-pyrrolyl, and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyryl Oxazolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl, and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl, and 5- Oxazolyl, etc.), triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl and 4H-1, 2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl , 4-thiazolyl and 5-thiazolyl, etc.), furyl (including 2-furanyl and 3-furanyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.), pyridyl (including 2 -Pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl or pyrimidinyl (including 2-pyrimidyl and 4-pyrimidyl, etc.).
除非另有规定，C n-n+m或C n-C n+m包括n至n+m个碳的任何一种具体情况，例如C 1-12包括C 1、C 2、C 3、C 4、C 5、C 6、C 7、C 8、C 9、C 10、C 11、和C 12，也包括n至n+m中的任何一个范围，例如C 1-12包括C 1- 3、C 1-6、C 1-9、C 3-6、C 3-9、C 3-12、C 6-9、C 6-12、和C 9-12等；同理，n元至n+m元表示环上原子数为n至n+m个，例如3-12元环包括3元环、4元环、5元环、6元环、7元环、8元环、9元环、10元环、11元环、和12元环，也包括n至n+m中的任何一个范围，例如3-12元环包括3-6元环、3-9元环、5-6元环、5-7元环、6-7元环、6-8元环、和6-10元环等。 Unless otherwise specified, C n-n + m or C n -C n + m includes any specific case of n to n + m carbons, for example, C 1-12 includes C 1 , C 2 , C 3 , C 4 , C 5 , C 6 , C 7 , C 8 , C 9 , C 10 , C 11 , and C 12 , and also includes any range from n to n + m, for example, C 1-12 includes C 1- 3 , C 1-6 , C 1-9 , C 3-6 , C 3-9 , C 3-12 , C 6-9 , C 6-12 , and C 9-12, etc .; similarly, n yuan to n + m member means that the number of atoms in the ring is n to n + m, for example, 3-12 member ring includes 3 member ring, 4 member ring, 5 member ring, 6 member ring, 7 member ring, 8 member ring, 9 member ring , 10-membered ring, 11-membered ring, and 12-membered ring, also including any range from n to n + m, for example, 3-12 membered ring includes 3-6 membered ring, 3-9 membered ring, 5-6 membered ring Rings, 5-7 member rings, 6-7 member rings, 6-8 member rings, 6-10 member rings, etc.
术语“离去基团”是指可以被另一种官能团或原子通过取代反应(例如亲和取代反应)所取代的官能团或原子。例如，代表性的离去基团包括三氟甲磺酸酯；氯、溴、碘；磺酸酯基，如甲磺酸酯、甲苯磺酸酯、对溴苯磺酸酯、对甲苯磺酸酯等；酰氧基，如乙酰氧基、三氟乙酰氧基等等。The term "leaving group" refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (eg, an affinity substitution reaction). For example, representative leaving groups include triflate; chlorine, bromine, and iodine; sulfonate groups such as mesylate, tosylate, p-bromobenzenesulfonate, and p-toluenesulfonate Ester, etc .; acyloxy, such as acetoxy, trifluoroacetoxy, etc.
术语“保护基”包括但不限于“氨基保护基”、“羟基保护基”或“巯基保护基”。术语“氨基保护基”是指适合用于阻止氨基氮位上副反应的保护基团。代表性的氨基保护基包括但不限于：甲酰基；酰基，例如链烷酰基(如乙酰基、三氯乙酰基或三氟乙酰基)；烷氧基羰基，如叔丁氧基羰基(Boc)；芳基甲氧羰基，如苄氧羰基(Cbz)和9-芴甲氧羰基(Fmoc)；芳基甲基，如苄基(Bn)、三苯甲基(Tr)、1,1-二-(4'-甲氧基苯基)甲基；甲硅烷基，如三甲基甲硅烷基(TMS)和叔丁基二甲基甲硅烷基(TBS)等等。术语“羟基保护基”是指适合用于阻止羟基副反应的保护基。代表性羟基保护基包括但不限于：烷基，如甲基、乙基和叔丁基；酰基，例如链烷酰基(如乙酰基)；芳基甲基，如苄基(Bn)，对甲氧基苄基(PMB)、9-芴基甲基(Fm)和二苯基甲基(二苯甲基，DPM)；甲硅烷基，如三甲基甲硅烷基(TMS)和叔丁基二甲基甲硅烷基(TBS)等等。The term "protecting group" includes but is not limited to "amino protecting group", "hydroxy protecting group" or "mercapto protecting group". The term "amino protecting group" refers to a protecting group suitable for preventing side reactions at the amino nitrogen position. Representative amino protecting groups include, but are not limited to: formyl; acyl, such as alkanoyl (such as acetyl, trichloroacetyl, or trifluoroacetyl); alkoxycarbonyl, such as tert-butoxycarbonyl (Boc) ; Arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), trityl (Tr), 1,1-di -(4'-methoxyphenyl) methyl; silyl, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), etc. The term "hydroxyl protecting group" refers to a protecting group suitable for preventing side reactions of hydroxyl groups. Representative hydroxy protecting groups include, but are not limited to: alkyl groups, such as methyl, ethyl, and tert-butyl; acyl groups, such as alkanoyl groups (such as acetyl); arylmethyl groups, such as benzyl (Bn), p-methyl Oxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl, such as trimethylsilyl (TMS) and tert-butyl Dimethylsilyl (TBS) and so on.
本发明的化合物可以通过本领域技术人员所熟知的多种合成方法来制备，包括下面列举的具体实施方式、其与其他化学合成方法的结合所形成的实施方式以及本领域技术上人员所熟知的等同替换方式，优选的实施方式包括但不限于本发明的实施例。The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by the combination with other chemical synthesis methods, and well known to those skilled in the art Equivalently, preferred embodiments include but are not limited to the embodiments of the present invention.
本发明所使用的溶剂可经市售获得。The solvent used in the present invention is commercially available.
本发明采用下述缩略词：CDCl 3代表氘代氯仿；DMSO代表二甲基亚砜；Boc代表叔丁氧羰基；DPBS代表杜氏磷酸盐缓冲液(Dulbecco's Phosphate Buffered Saline)。 The present invention uses the following abbreviations: CDCl 3 stands for deuterated chloroform; DMSO stands for dimethyl sulfoxide; Boc stands for tert-butoxycarbonyl; DPBS stands for Dulbecco's Phosphate Buffered Saline.
本发明化合物依据本领域常规命名原则或者使用
Figure PCTCN2019114347-appb-000066
软件命名，市售化合物采用供应商目录名称。 The compounds of the present invention are based on conventional naming principles or use in the art
Figure PCTCN2019114347-appb-000066
The software is named, and the commercially available compounds adopt the supplier catalog name.
具体实施方式detailed description
下面通过实施例对本发明进行详细描述，但并不意味着对本发明任何不利限制。本文已经详细地描述了本发明，其中也公开了其具体实施例方式，对本领域的技术人员而言，在不脱离本发明精神和范围的情况下针对本发明具体实施方式进行各种变化和改进将是显而易见的。The following describes the present invention in detail through examples, but it does not mean any adverse limitation of the present invention. The present invention has been described in detail herein, and the specific embodiments thereof are also disclosed. For those skilled in the art, various changes and improvements are made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention Will be obvious.
实施例1：化合物1Example 1: Compound 1
Figure PCTCN2019114347-appb-000067
Figure PCTCN2019114347-appb-000067
Figure PCTCN2019114347-appb-000068
Figure PCTCN2019114347-appb-000068
步骤A:将化合物1-1(200克，1.02摩尔，1.0当量)溶于1升二氯甲烷中，加入吡啶(84.64克，1.07摩尔，1.05当量)，控制反应温度在30～40摄氏度的条件下，将4-氯丁酰氯(150.88克，1.07摩尔，1.05当量)逐滴加入。滴加完毕后在25摄氏度下搅拌12小时。25摄氏度下向反应液中加入1升水，搅拌30分钟后分离有机相，有机相用无水硫酸钠干燥，浓缩得到化合物1-2。Step A: Dissolve compound 1-1 (200 g, 1.02 mol, 1.0 equiv) in 1 liter of dichloromethane, add pyridine (84.64 g, 1.07 mol, 1.05 equiv), and control the reaction temperature at 30-40 degrees Celsius Next, 4-chlorobutyryl chloride (150.88 g, 1.07 mol, 1.05 equiv) was added dropwise. After the dropwise addition, the mixture was stirred at 25 degrees Celsius for 12 hours. 1 liter of water was added to the reaction solution at 25 degrees Celsius, the organic phase was separated after stirring for 30 minutes, the organic phase was dried over anhydrous sodium sulfate, and concentrated to obtain compound 1-2.
步骤B：将化合物1-2(319.60克，1.06摩尔，1.0当量)溶于1.8升四氢呋喃中，在25摄氏度下分批加入叔丁醇钾(125.19克，1.12摩尔，1.05当量)。25摄氏度下搅拌8小时。25摄氏度下向反应液中加入1.8升水，搅拌30分钟后分离有机相，水相用乙酸乙酯萃取(1.2升)，合并有机相，用饱和食盐水洗(1.2升)，无水硫酸钠干燥，浓缩得化合物1-3。Step B: Compound 1-2 (319.60 g, 1.06 mol, 1.0 equiv) was dissolved in 1.8 liters of tetrahydrofuran, and potassium tert-butoxide (125.19 g, 1.12 mol, 1.05 equiv) was added in portions at 25 ° C. Stir for 8 hours at 25 degrees Celsius. At 25 degrees Celsius, 1.8 liters of water was added to the reaction solution, and the organic phase was separated after stirring for 30 minutes. The aqueous phase was extracted with ethyl acetate (1.2 liters). The organic phases were combined, washed with saturated brine (1.2 liters), and dried over anhydrous sodium sulfate. Concentrate to give compound 1-3.
步骤C：25摄氏度下将化合物1-3(267.08克，1.01摩尔，1.0当量)加入1.5升甲苯中，搅拌，加入水(18.20克，1.01摩尔，1.0当量)。分批加入一水合对甲苯磺酸(176.83克，929.60毫摩尔，0.92当量)并控制加料温度在40-45摄氏度之间。加完后在40-45摄氏度下搅拌3小时。然后冷却至15摄氏度，搅拌1小时。将反应液过滤，滤饼用甲苯淋洗(600毫升)，干燥，得到化合物1-4。Step C: Compound 25 (267.08 g, 1.01 mol, 1.0 equiv) was added to 1.5 liters of toluene at 25 degrees Celsius, stirred, and water (18.20 g, 1.01 mol, 1.0 equiv) was added. P-toluenesulfonic acid monohydrate (176.83 g, 929.60 mmol, 0.92 equiv) was added in portions and the feed temperature was controlled between 40-45 degrees Celsius. After the addition is complete, stir at 40-45 degrees Celsius for 3 hours. Then cool to 15 degrees Celsius and stir for 1 hour. The reaction solution was filtered, and the filter cake was rinsed with toluene (600 ml) and dried to obtain compound 1-4.
Figure PCTCN2019114347-appb-000069
Figure PCTCN2019114347-appb-000069
步骤D：将乙酸乙酯(349.71克，3.97摩尔，3.0当量)加入1升甲苯中，在20-30摄氏度下分批加入乙醇钠(180.07克，2.65摩尔，2.0当量)。加完后在20-30摄氏度下搅拌1小时。将化合物1-5(200克， 1.32摩尔，1.0当量)分三批加入，加完后在100摄氏度下搅拌12小时。将反应液冷却至25摄氏度，用冰醋酸调节pH值至6，加入1升水，搅拌30分钟后分离有机相。水相用甲苯萃取(600毫升)，合并有机相，浓缩得到化合物1-6。Step D: Ethyl acetate (349.71 grams, 3.97 moles, 3.0 equivalents) was added to 1 liter of toluene, and sodium ethoxide (180.07 grams, 2.65 moles, 2.0 equivalents) was added in portions at 20-30 degrees Celsius. After the addition is complete, stir at 20-30 ° C for 1 hour. Compound 1-5 (200 grams, 1.32 moles, 1.0 equivalent) was added in three batches, and after the addition was completed, it was stirred at 100 degrees Celsius for 12 hours. The reaction solution was cooled to 25 degrees Celsius, the pH was adjusted to 6 with glacial acetic acid, 1 liter of water was added, and the organic phase was separated after stirring for 30 minutes. The aqueous phase was extracted with toluene (600 mL), and the organic phases were combined and concentrated to give compound 1-6.
步骤E:将化合物1-6(137克，661.12摩尔，1.0当量)加入350毫升吡啶中，然后加入化合物1-4(216.04克，793.34毫摩尔，1.2当量)。在40-45摄氏度下搅拌12小时。将反应液冷却至25摄氏度，加入700毫升水，用甲苯萃取(560毫升×2)。合并有机相，用饱和食盐水洗(560毫升)，无水硫酸钠干燥，浓缩得到化合物1-7。Step E: Compound 1-6 (137 g, 661.12 moles, 1.0 equivalent) was added to 350 ml of pyridine, and then compound 1-4 (216.04 g, 793.34 mmol, 1.2 equivalents) was added. Stir for 12 hours at 40-45 degrees Celsius. The reaction liquid was cooled to 25 ° C, 700 ml of water was added, and extracted with toluene (560 ml × 2). The organic phases were combined, washed with saturated brine (560 ml), dried over anhydrous sodium sulfate, and concentrated to obtain compound 1-7.
步骤F：将化合物1-7(148.50克，513.26毫摩尔，1.0当量)溶于900毫升甲苯中，分批加入乙醇钠(69.85克，1.03摩尔，2.0当量)。加完后在100-110摄氏度下搅拌12小时。将反应液冷却至25摄氏度，加入600毫升水，搅拌15分钟。分离水相，水相用乙酸乙酯洗(300毫升×3)，然后将水相的pH值用浓盐酸调至6，过滤。滤饼用150毫升异丙醇打浆，过滤，干燥，得到化合物1-8。MS(ESI)m/z:244.0[M+H +]。 Step F: Compound 1-7 (148.50 g, 513.26 mmol, 1.0 equiv) was dissolved in 900 ml of toluene, and sodium ethoxide (69.85 g, 1.03 mol, 2.0 equiv) was added in portions. After the addition, the mixture was stirred at 100-110 degrees Celsius for 12 hours. The reaction solution was cooled to 25 degrees Celsius, 600 ml of water was added, and stirred for 15 minutes. The aqueous phase was separated, and the aqueous phase was washed with ethyl acetate (300 ml × 3), and then the pH value of the aqueous phase was adjusted to 6 with concentrated hydrochloric acid and filtered. The filter cake was slurried with 150 ml of isopropanol, filtered and dried to give compound 1-8. MS (ESI) m / z: 244.0 [M + H + ].
步骤G:将化合物1-8(77.22克，317.44毫摩尔，1.0当量)，溶于800毫升N,N-二甲基甲酰胺中，分批加入N-溴代丁二酰亚胺(59.32克，333.31毫摩尔，1.05当量)，25摄氏度下搅拌8小时。再加入N-溴代丁二酰亚胺(59.32克，333.31毫摩尔，1.05当量)，50摄氏度下搅拌12小时。将反应液冷却至0-10摄氏度，加入2升水，在0-10摄氏度下搅拌30分钟，过滤。滤饼用水(500毫升)打浆，过滤，干燥，得到化合物1-9。MS(ESI)m/z:277.8，279.8[M+H +]。 Step G: Compound 1-8 (77.22 g, 317.44 mmol, 1.0 equiv) was dissolved in 800 ml of N, N-dimethylformamide, and N-bromosuccinimide (59.32 g) was added in portions , 333.31 mmol, 1.05 equiv), stirred at 25 degrees Celsius for 8 hours. N-bromosuccinimide (59.32 g, 333.31 mmol, 1.05 equiv) was added, and the mixture was stirred at 50 degrees Celsius for 12 hours. The reaction solution was cooled to 0-10 degrees Celsius, 2 liters of water was added, stirred at 0-10 degrees Celsius for 30 minutes, and filtered. The filter cake was slurried with water (500 mL), filtered, and dried to give compound 1-9. MS (ESI) m / z: 277.8, 279.8 [M + H + ].
步骤H：将化合物1-9(20克，71.90毫摩尔，1.0当量)，硼酸三异丙酯(35.84克，190.55毫摩尔，2.65当量)加入200毫升无水四氢呋喃中，在氮气气氛下冷却至-70～-60摄氏度。在-70～-60摄氏度下逐滴滴加正丁基锂(2.5摩尔每升正庚烷溶液，68.74毫升，2.39当量)。加完后在-70～-60摄氏度下搅拌2小时。将反应液倒入0-5摄氏度的500毫升饱和氯化铵溶液中，过滤，滤饼用水洗(100毫升×2)，干燥得到化合物1-10。MS(ESI)m/z:244.2[M+H +]。 Step H: Compound 1-9 (20 g, 71.90 mmol, 1.0 equiv), triisopropyl borate (35.84 g, 190.55 mmol, 2.65 equiv) were added to 200 mL of anhydrous tetrahydrofuran, and cooled to -70 ～ -60 degrees Celsius. N-Butyllithium (2.5 moles per liter of n-heptane solution, 68.74 ml, 2.39 equiv) was added dropwise at -70 to -60 degrees Celsius. After the addition, the mixture was stirred at -70 to -60 degrees Celsius for 2 hours. The reaction solution was poured into 500 ml of saturated ammonium chloride solution at 0-5 degrees Celsius, filtered, and the filter cake was washed with water (100 ml × 2) and dried to obtain compound 1-10. MS (ESI) m / z: 244.2 [M + H + ].
步骤I:将化合物1-10(26.62克，109.52毫摩尔，1.0当量)和双氧水(37.25克，328.55毫摩尔，浓度30％，3.0当量)加入250毫升四氢呋喃中，搅拌，冷却至0-5摄氏度。在0-5摄氏度下逐滴加入2摩尔每升的氢氧化钠水溶液(219.03毫升，4.0当量)。加完后在25摄氏度下搅拌12小时。向反应液中加入250毫升水，用浓盐酸调节pH值至7，乙酸乙酯萃取(200毫升×3)，饱和亚硫酸钠溶液洗(100毫升×2)，无水硫酸钠干燥，浓缩，柱层析纯化得到化合物1-11。MS(ESI)m/z:215.9[M+H +]。 Step I: Compound 1-10 (26.62 g, 109.52 mmol, 1.0 equiv) and hydrogen peroxide (37.25 g, 328.55 mmol, 30% concentration, 3.0 equiv) were added to 250 ml of tetrahydrofuran, stirred, and cooled to 0-5 ° C . A 2 mol per liter aqueous sodium hydroxide solution (219.03 mL, 4.0 equiv) was added dropwise at 0-5 degrees Celsius. After the addition, the mixture was stirred at 25 degrees Celsius for 12 hours. Add 250 ml of water to the reaction solution, adjust pH to 7 with concentrated hydrochloric acid, extract with ethyl acetate (200 ml × 3), wash with saturated sodium sulfite solution (100 ml × 2), dry over anhydrous sodium sulfate, concentrate, and column layer Analysis and purification gave compound 1-11. MS (ESI) m / z: 215.9 [M + H + ].
Figure PCTCN2019114347-appb-000070
Figure PCTCN2019114347-appb-000070
步骤J：向装有100毫升无水二氯甲烷的烧瓶中，在不断搅拌下依次加入化合物1-12(5克，25.38毫摩尔，1.0当量)，二碳酸二叔丁酯(6.65克，30.45毫摩尔，1.2当量)，4-二甲基氨基吡啶(310.03毫克，2.54毫摩尔，0.1当量)和三乙胺(7.70克，76.13毫摩尔，3.0当量)。反应液在25摄氏度下搅拌3小时。向反应液中加入150毫升水，二氯甲烷萃取(100毫升×2)，有机相用100毫升饱和食盐水洗涤，无水硫酸钠干燥，过滤，浓缩，柱层析纯化得到化合物1-13。Step J: To a flask containing 100 ml of anhydrous dichloromethane, compound 1-12 (5 g, 25.38 mmol, 1.0 equiv), di-tert-butyl dicarbonate (6.65 g, 30.45) were added in sequence with constant stirring. Mmol, 1.2 equiv), 4-dimethylaminopyridine (310.03 mg, 2.54 mmol, 0.1 equiv) and triethylamine (7.70 g, 76.13 mmol, 3.0 equiv). The reaction solution was stirred at 25 degrees Celsius for 3 hours. 150 ml of water was added to the reaction solution, and extracted with dichloromethane (100 ml × 2). The organic phase was washed with 100 ml of saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain compound 1-13.
步骤K：将化合物1-13(2克，6.73毫摩尔，1当量)溶于20毫升1,4-二氧六环中，加入双联频哪醇硼酸酯(2.05克，8.08毫摩尔，1.2当量)，[1,1’-双(二苯基膦基)]二氯化钯(492.49毫克，673.07微摩尔，0.1当量)和乙酸钾(1.32克，13.46毫摩尔，2当量)。反应体系用氮气置换三次，然后在氮气保护下升温至100摄氏度搅拌8小时。将反应液冷却至25摄氏度，加入50毫升水，乙酸乙酯萃取(40毫升×3)。合并有机相，50毫升饱和食盐水洗涤，无水硫酸钠干燥，过滤，浓缩得到化合物1-14。MS(ESI)m/z:345.1[M+H +]。 Step K: Dissolve compound 1-13 (2 g, 6.73 mmol, 1 equiv) in 20 mL of 1,4-dioxane, and add double pinacol borate (2.05 g, 8.08 mmol, 1.2 equivalents), [1,1'-bis (diphenylphosphino)] palladium dichloride (492.49 mg, 673.07 micromolar, 0.1 equivalent) and potassium acetate (1.32 g, 13.46 mmol, 2 equivalents). The reaction system was replaced with nitrogen three times, and then heated to 100 degrees Celsius under nitrogen and stirred for 8 hours. The reaction solution was cooled to 25 degrees Celsius, 50 ml of water was added, and extracted with ethyl acetate (40 ml × 3). The organic phases were combined, washed with 50 ml of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 1-14. MS (ESI) m / z: 345.1 [M + H + ].
步骤L:将化合物1-14(3.26克，9.47毫摩尔，1当量)溶于20毫升丙酮和10毫升水的混合溶剂中，在不断搅拌下依次加入高碘酸钠(6.28克，29.36毫摩尔，1.63毫升，3.1当量)和乙酸铵(2.19克，28.41毫摩尔，3当量)。25摄氏度下搅拌12小时。向反应液中加30毫升水，乙酸乙酯萃取(30毫升×3)。合并有机相，饱和食盐水洗涤(30毫升)，浓缩。粗产物溶于30毫升1摩尔每升的NaOH溶液，乙酸乙酯洗涤(30毫升×3)。用2摩尔每升的盐酸将水相的pH调到6。过滤，滤饼用水洗涤(5毫升×2)，干燥得到化合物1-15。MS(ESI)m/z:263.2[M+H +]。 Step L: Dissolve compound 1-14 (3.26 g, 9.47 mmol, 1 equiv) in a mixed solvent of 20 ml of acetone and 10 ml of water, and add sodium periodate (6.28 g, 29.36 mmol) in sequence with continuous stirring , 1.63 mL, 3.1 equiv) and ammonium acetate (2.19 g, 28.41 mmol, 3 equiv). Stir for 12 hours at 25 degrees Celsius. 30 ml of water was added to the reaction solution, and ethyl acetate was extracted (30 ml × 3). The organic phases were combined, washed with saturated brine (30 mL), and concentrated. The crude product was dissolved in 30 ml of 1 mol NaOH solution and washed with ethyl acetate (30 ml × 3). The pH of the aqueous phase was adjusted to 6 with 2 molar hydrochloric acid. Filter and wash the filter cake with water (5 mL x 2) and dry to give compound 1-15. MS (ESI) m / z: 263.2 [M + H + ].
步骤M:将化合物1-15(200毫克，763.16微摩尔，1当量)和1-11(246.41毫克，1.14毫摩尔，1.5当量)溶解在5毫升二氯甲烷中，加入醋酸铜(207.92毫克，1.14毫摩尔，1.5当量)，三乙胺(231.67毫克，2.29毫摩尔，318.67微升，3当量)和
Figure PCTCN2019114347-appb-000071
分子筛(500毫克)，体系用氧气置换三次。于25摄氏度下氧气氛围中反应12小时。向反应液中加入20毫升水，二氯甲烷萃取(30毫升×3)。合并有机相，饱和食盐水洗涤(30毫升)，无水硫酸钠干燥，过滤，浓缩，柱层析纯化得到化合物1-16。 Step M: Dissolve compounds 1-15 (200 mg, 763.16 μmol, 1 equiv) and 1-11 (246.41 mg, 1.14 mmol, 1.5 equiv) in 5 ml of dichloromethane, and add copper acetate (207.92 mg, 1.14 mmol, 1.5 equiv), triethylamine (231.67 mg, 2.29 mmol, 318.67 μl, 3 equiv) and
Figure PCTCN2019114347-appb-000071
Molecular sieve (500 mg), the system was replaced with oxygen three times. React for 12 hours in an oxygen atmosphere at 25 degrees Celsius. 20 ml of water was added to the reaction solution, and extracted with dichloromethane (30 ml × 3). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain compound 1-16.
步骤N:将化合物1-16(200毫克，463.51微摩尔，1当量)加入到4摩尔每升盐酸甲醇溶液(5毫升，43.15当量)中，25摄氏度下搅拌8小时。将反应液倒入20毫升水中，用4摩尔每升氢氧化钠水溶液将pH调至8到9。减压蒸馏除去甲醇，乙酸乙酯萃取(30毫升×3)。合并有机相，有机相用饱和食盐水洗涤(30毫升)，无水硫酸钠干燥，过滤，浓缩，制备高效液相分离得到化合物1。MS(ESI)m/z:332.1[M+H +]； 1H NMR(400MHz,DMSO-d 6):δ11.59(s,1H),7.96(d,J＝5.6Hz,1H),7.58-7.51(m,2H),7.32(t,J＝2.8Hz,1H),6.97(d,J＝7.2Hz,1H),6.41-6.39(m,2H),4.20(t,J＝7.4Hz,2H),2.82(t,J＝7.2Hz,2H),2.62-2.55(m,2H),2.05(s,3H)。 Step N: Compound 1-16 (200 mg, 463.51 micromolar, 1 equivalent) was added to 4 moles per liter of hydrochloric acid methanol solution (5 mL, 43.15 equivalent), and stirred at 25 degrees Celsius for 8 hours. The reaction solution was poured into 20 ml of water, and the pH was adjusted to 8 to 9 with 4 mol per liter of sodium hydroxide aqueous solution. Methanol was distilled off under reduced pressure, and extracted with ethyl acetate (30 mL × 3). The organic phases were combined, and the organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to prepare high performance liquid phase separation to obtain compound 1. MS (ESI) m / z: 332.1 [M + H + ]; 1 H NMR (400 MHz, DMSO-d 6 ): δ 11.59 (s, 1H), 7.96 (d, J = 5.6 Hz, 1H), 7.58 -7.51 (m, 2H), 7.32 (t, J = 2.8Hz, 1H), 6.97 (d, J = 7.2Hz, 1H), 6.41-6.39 (m, 2H), 4.20 (t, J = 7.4Hz, 2H), 2.82 (t, J = 7.2 Hz, 2H), 2.62-2.55 (m, 2H), 2.05 (s, 3H).
实施例2：化合物2Example 2: Compound 2
Figure PCTCN2019114347-appb-000072
Figure PCTCN2019114347-appb-000072
Figure PCTCN2019114347-appb-000073
Figure PCTCN2019114347-appb-000073
步骤A：将化合物2-1(10克，46.29毫摩尔，1当量)溶于四氢呋喃(100毫升)中，将该溶液在氮气保护下逐滴加入甲基溴化镁(3摩尔每升的四氢呋喃溶液，33.95毫升，2.2当量)中，保持温度为0～5摄氏度。滴加完毕后升温至25摄氏度，搅拌1小时。用1摩尔每升的盐酸将反应液的pH调到7。加入水(200毫升)稀释，乙酸乙酯(200毫升×3)萃取，合并有机相，水(200毫升)洗涤，无水硫酸钠干燥，过滤，浓缩得到化合物2-2。Step A: Compound 2-1 (10 g, 46.29 mmol, 1 equiv) was dissolved in tetrahydrofuran (100 mL), and this solution was added dropwise to methyl magnesium bromide (3 mol per liter of tetrahydrofuran under the protection of nitrogen) The solution, 33.95 ml, 2.2 equivalents), keep the temperature at 0-5 degrees Celsius. After the dropwise addition, the temperature was raised to 25 degrees Celsius and stirred for 1 hour. The pH of the reaction solution was adjusted to 7 with 1 mol per liter of hydrochloric acid. Diluted with water (200 mL), extracted with ethyl acetate (200 mL × 3), combined organic phases, washed with water (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 2-2.
步骤B：将化合物2-2(10.33克，31.99毫摩尔，1当量)溶于氨水(76.53克，611.36毫摩尔，84.10毫升，28％，19.11当量)，加入铜粉(1.22克，19.19毫摩尔，0.6当量)。25摄氏度于空气中搅拌1小时后在氮气保护下于100摄氏度搅拌12小时。将反应液过滤，滤液冻干，柱层析纯化得到化合物2-3。MS(ESI)m/z:153.3[M+H +]。 Step B: Compound 2-2 (10.33 g, 31.99 mmol, 1 equiv) was dissolved in ammonia water (76.53 g, 611.36 mmol, 84.10 mL, 28%, 19.11 equiv), and copper powder (1.22 g, 19.19 mmol) was added , 0.6 equivalent). After stirring at 25 degrees Celsius in the air for 1 hour, it was stirred at 100 degrees Celsius under nitrogen for 12 hours. The reaction solution was filtered, the filtrate was lyophilized, and purified by column chromatography to obtain compound 2-3. MS (ESI) m / z: 153.3 [M + H + ].
Figure PCTCN2019114347-appb-000074
Figure PCTCN2019114347-appb-000074
步骤C：将化合物1-11(5.3克，21.80毫摩尔，1当量)和化合物2-4(6.86克，43.60毫摩尔，2当量)溶于二氯甲烷(250毫升)中，在搅拌下依次加入无水乙酸铜(5.94克，32.70毫摩尔，1.5当量)，三乙胺(6.62克，65.40毫摩尔，9.10毫升，3当量)，
Figure PCTCN2019114347-appb-000075
分子筛(5克)。35摄氏度下氧气氛围中搅拌12小时。将体系温度降到20摄氏度到25摄氏度之间，缓慢加入水(300毫升)稀释，乙酸乙酯(200毫升×3)萃取，合并有机相，饱和食盐水(200毫升)洗涤，无水硫酸钠干燥，浓缩，柱层析纯化得到化合物2-5。MS(ESI)m/z:327.1[M+H +]。 Step C: Dissolve compound 1-11 (5.3 g, 21.80 mmol, 1 equiv) and compound 2-4 (6.86 g, 43.60 mmol, 2 equiv) in dichloromethane (250 mL), followed by stirring Add anhydrous copper acetate (5.94 g, 32.70 mmol, 1.5 equiv), triethylamine (6.62 g, 65.40 mmol, 9.10 mL, 3 equiv),
Figure PCTCN2019114347-appb-000075
Molecular sieve (5g). Stir for 12 hours at 35 degrees Celsius in an oxygen atmosphere. Reduce the temperature of the system to between 20 ° C and 25 ° C, slowly add water (300 mL) to dilute, extract with ethyl acetate (200 mL × 3), combine organic phases, wash with saturated brine (200 mL), anhydrous sodium sulfate Dry, concentrate, and purify by column chromatography to obtain compound 2-5. MS (ESI) m / z: 327.1 [M + H + ].
步骤D：将化合物2-5(1.5克，4.59毫摩尔，1当量)溶于1,4-二氧六环(30毫升)中，再依次加入化合物2-3(1.05克，6.89毫摩尔，1.5当量)，二(二亚苄基丙酮)钯(527.89毫克，918.05微摩尔，0.2当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(531.20毫克，918.05微摩尔，0.2当量)，苯酚钠(2.13克，18.36毫摩尔，4当量)。置换氮气3次后加热至100摄氏度搅拌12小时。将反应液过滤，滤液中加入水(50毫升)，用乙酸乙酯(50毫升×3)萃取，合并有机相，饱和食盐水(50毫升)洗涤，无水硫酸钠干燥，浓缩，高效液相色谱纯化得到化合物2。MS(ESI)m/z:443.2[M+H +]； Step D: Compound 2-5 (1.5 g, 4.59 mmol, 1 equiv) was dissolved in 1,4-dioxane (30 mL), and then compound 2-3 (1.05 g, 6.89 mmol, 1.5 equivalents), bis (dibenzylideneacetone) palladium (527.89 mg, 918.05 micromolar, 0.2 equivalents), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (531.20 mg , 918.05 micromolar, 0.2 equivalent), sodium phenolate (2.13 g, 18.36 mmol, 4 equivalent). After replacing the nitrogen three times, it was heated to 100 degrees Celsius and stirred for 12 hours. The reaction solution was filtered, water (50 mL) was added to the filtrate, and extracted with ethyl acetate (50 mL × 3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated. Chromatographic purification gave compound 2. MS (ESI) m / z: 443.2 [M + H + ];
1H NMR(400MHz,DMSO-d 6)δ＝9.35(s,1H),8.17(d,J＝5.6Hz,1H),8.10(d,J＝5.6Hz,1H),7.68(s,1H),7.65-7.61(m,2H),7.56(d,J＝8.0Hz,1H),7.04(d,J＝7.2Hz,1H),6.59(dd,J＝2.0,5.6Hz,1H),6.39(d,J＝2.0Hz,1H),5.08(s,1H),4.20(t,J＝7.2Hz,2H),2.86(t,J＝7.6Hz,2H),2.63-2.56(m,2H),2.22(s,3H),1.39(s,6H)。 1 H NMR (400 MHz, DMSO-d 6 ) δ = 9.35 (s, 1H), 8.17 (d, J = 5.6 Hz, 1H), 8.10 (d, J = 5.6 Hz, 1H), 7.68 (s, 1H) , 7.65-7.61 (m, 2H), 7.56 (d, J = 8.0 Hz, 1H), 7.04 (d, J = 7.2 Hz, 1H), 6.59 (dd, J = 2.0, 5.6 Hz, 1H), 6.39 ( d, J = 2.0 Hz, 1H), 5.08 (s, 1H), 4.20 (t, J = 7.2 Hz, 2H), 2.86 (t, J = 7.6 Hz, 2H), 2.63-2.56 (m, 2H), 2.22 (s, 3H), 1.39 (s, 6H).
实施例3：化合物3Example 3: Compound 3
Figure PCTCN2019114347-appb-000076
Figure PCTCN2019114347-appb-000076
步骤A：在5摄氏度下，在3-1(5克，32.34毫摩尔，4.17毫升，1当量)的甲酸二乙酯(39.00克，330.14毫摩尔，40毫升，10.21当量)的溶液中分批加入钠氢(2.68克，66.95毫摩尔，纯度：60％，2.07当量)，缓慢升温到85摄氏度并在此温度反应2小时后，加入100毫升水，乙酸乙酯(100毫升×2)萃取。合并有机相，饱和食盐水(100毫升×2)洗，无水硫酸钠干燥，过滤，浓缩，柱层析纯化得到化合物3-2。MS(ESI)m/z:227.1[M+H +]。 Step A: At 5 degrees Celsius, in batches in a solution of 3-1 (5 g, 32.34 mmol, 4.17 mL, 1 equivalent) of diethyl formate (39.00 g, 330.14 mmol, 40 mL, 10.21 equivalent) Sodium hydrogen (2.68 g, 66.95 mmol, purity: 60%, 2.07 equiv) was added, the temperature was slowly raised to 85 degrees Celsius, and after reacting at this temperature for 2 hours, 100 ml of water was added and extracted with ethyl acetate (100 ml × 2). The organic phases were combined, washed with saturated brine (100 ml × 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain compound 3-2. MS (ESI) m / z: 227.1 [M + H + ].
步骤B：在25摄氏度下，向化合物3-2(5.3克，23.38毫摩尔，1当量)的吡啶(15毫升)溶液中分批加入1-4(7.75克，28.06毫摩尔，1.2当量，对甲苯磺酸盐)，在40摄氏度下反应16小时后，加入50毫升水，乙酸乙酯(100毫升×3)。合并有机相，饱和食盐水(100毫升×2)洗，无水硫酸钠干燥，过滤，浓缩，柱层析纯化得到化合物3-3。MS(ESI)m/z:309.1[M+H +]。 Step B: To a solution of compound 3-2 (5.3 g, 23.38 mmol, 1 equiv) in pyridine (15 mL) was added 1-4 (7.75 g, 28.06 mmol, 1.2 equiv. Tosylate), after reacting at 40 degrees Celsius for 16 hours, 50 ml of water and ethyl acetate (100 ml × 3) were added. The organic phases were combined, washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain compound 3-3. MS (ESI) m / z: 309.1 [M + H + ].
步骤C：向化合物3-3(4.3克，13.93毫摩尔，1当量)的甲苯(25毫升)溶液中分批加入乙醇钠(1.90克，27.85毫摩尔，2当量)，在100～110摄氏度下反应16小时后，加入150毫升乙酸乙酯和100毫升水，水相用乙酸乙酯洗涤两次，每次25毫升，然后用36％的盐酸将水相pH调节到4，乙酸乙酯(50毫升×2)萃取。合并有机相，饱和食盐水(100毫升)洗，无水硫酸钠干燥，过滤，浓缩得到化合物3-4。Step C: To a solution of compound 3-3 (4.3 g, 13.93 mmol, 1 equiv) in toluene (25 mL) was added sodium ethoxide (1.90 g, 27.85 mmol, 2 equiv) in batches at 100-110 ° C After 16 hours of reaction, 150 ml of ethyl acetate and 100 ml of water were added. The aqueous phase was washed twice with 25 ml of ethyl acetate each time, and then the pH of the aqueous phase was adjusted to 4 with 36% hydrochloric acid. ML × 2) extraction. The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 3-4.
MS(ESI)m/z:263.1[M+H +]。 MS (ESI) m / z: 263.1 [M + H + ].
步骤D：向化合物3-4(1.27克，4.83毫摩尔，1当量)的N,N-二甲基甲酰胺(50毫升)溶液中分批 加入N-溴代丁二酰亚胺(860.48毫克，4.83毫摩尔，1当量)，在40摄氏度下反应12小时后，加入100毫升水，乙酸乙酯(100毫升×2)萃取。合并有机相，饱和食盐水(50毫升×2)洗，无水硫酸钠干燥，过滤，浓缩，柱层析纯化得到化合物3-5。MS(ESI)m/z:298.9[M+H +]。 Step D: To a solution of compound 3-4 (1.27 g, 4.83 mmol, 1 equiv) in N, N-dimethylformamide (50 mL) was added portionwise N-bromosuccinimide (860.48 mg , 4.83 mmol, 1 equiv), after reacting at 40 degrees Celsius for 12 hours, 100 ml of water was added, and ethyl acetate (100 ml × 2) was extracted. The organic phases were combined, washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain compound 3-5. MS (ESI) m / z: 298.9 [M + H + ].
步骤E：在氮气保护下将3-5(763毫克，2.56毫摩尔，1当量)和异丙氧基频哪醇硼酯(1.26克，6.79毫摩尔，1.39毫升，2.65当量)的四氢呋喃溶液(20毫升)冷却到-70摄氏度，然后逐滴加入正丁基锂(2.5摩尔每升，2.45毫升，2.39当量)，缓慢升温到20摄氏度，在氮气保护和20摄氏度下反应12小时后，加入50毫升饱和氯化铵溶液淬灭反应，加入50毫升乙酸乙酯稀释，用乙酸乙酯(100毫升×2)萃取。合并有机相，饱和食盐水(100毫升×2)洗，无水硫酸钠干燥，过滤，浓缩得到化合物3-6。Step E: A solution of 3-5 (763 mg, 2.56 mmol, 1 equiv) and isopropoxy pinacol borate (1.26 g, 6.79 mmol, 1.39 mL, 2.65 equiv) in tetrahydrofuran under a nitrogen atmosphere ( 20 ml) Cooled to -70 degrees Celsius, then added n-butyllithium (2.5 moles per liter, 2.45 ml, 2.39 equiv) dropwise, slowly warmed to 20 degrees Celsius, and reacted under nitrogen protection and 20 degrees Celsius for 12 hours, then added 50 The reaction was quenched with mL of saturated ammonium chloride solution, diluted with 50 mL of ethyl acetate, and extracted with ethyl acetate (100 mL × 2). The organic phases were combined, washed with saturated brine (100 ml × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 3-6.
MS(ESI)m/z:345.1[M+H +]。 MS (ESI) m / z: 345.1 [M + H + ].
步骤F：在0摄氏度下，向化合物3-6(1.65克，4.79毫摩尔，1当量)的四氢呋喃(30毫升)溶液中逐滴的加入双氧水(2.17克，19.15毫摩尔，1.84毫升，纯度：30％，4当量)，然后加入氢氧化钠(383.01毫克，9.58毫摩尔，2当量)的5毫升水溶液，在25摄氏度下反应12小时后，用1摩尔每升的盐酸溶液将pH调节到6，加入20毫升乙酸乙酯，用乙酸乙酯(50毫升×2)萃取。合并有机相，饱和食盐水(50毫升×2)洗，无水硫酸钠干燥，过滤，浓缩得到化合物3-7。MS(ESI)m/z:235.1[M+H +]。 Step F: To a solution of compound 3-6 (1.65 g, 4.79 mmol, 1 equiv) in tetrahydrofuran (30 mL) was added dropwise hydrogen peroxide (2.17 g, 19.15 mmol, 1.84 mL) at 0 ° C. Purity: 30%, 4 equiv.), Then add 5 ml of an aqueous solution of sodium hydroxide (383.01 mg, 9.58 mmol, 2 equiv.). After reacting at 25 degrees Celsius for 12 hours, adjust the pH to 6 with 1 mol of hydrochloric acid solution. , Add 20 ml of ethyl acetate and extract with ethyl acetate (50 ml x 2). The organic phases were combined, washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 3-7. MS (ESI) m / z: 235.1 [M + H + ].
步骤G：向化合物3-7(1.09克，4.64毫摩尔，1当量)和2-4(1.46克，9.29毫摩尔，2当量)的二氯甲烷(70毫升)溶液中加入醋酸铜(1.27克，6.97毫摩尔，1.5当量)，三乙胺(1.41克，13.93毫摩尔，1.94毫升，3当量)和
Figure PCTCN2019114347-appb-000077
分子筛(5克)，25摄氏度下反应12小时后，加入50毫升水和50毫升二氯甲烷，用二氯甲烷(50毫升×2)萃取。合并有机相，饱和食盐水(50毫升×2)洗，无水硫酸钠干燥，过滤，浓缩，柱层析纯化得到化合物3-8。MS(ESI)m/z:346.0[M+H +]。 Step G: To a solution of compound 3-7 (1.09 g, 4.64 mmol, 1 equiv) and 2-4 (1.46 g, 9.29 mmol, 2 equiv) in dichloromethane (70 mL) was added copper acetate (1.27 g , 6.97 mmol, 1.5 eq), triethylamine (1.41 g, 13.93 mmol, 1.94 mL, 3 eq) and
Figure PCTCN2019114347-appb-000077
Molecular sieve (5 g), after reacting at 25 degrees Celsius for 12 hours, 50 ml of water and 50 ml of dichloromethane were added, and extracted with dichloromethane (50 ml × 2). The organic phases were combined, washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain compound 3-8. MS (ESI) m / z: 346.0 [M + H + ].
步骤H：向化合物3-8(200毫克，577.68微摩尔，1当量)和2-3(131.88毫克，866.53微摩尔，1.5当量)的1,4-二氧六环(10毫升)溶液中加入4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(66.85毫克，115.54微摩尔，0.2当量)，二(二亚苄基丙酮)钯(66.43毫克，115.54微摩尔，0.2当量)和碳酸铯(752.88毫克，2.31毫摩尔，4当量)，在氮气保护和100摄氏度下反应12小时后，加入10毫升水和30毫升乙酸乙酯，乙酸乙酯(25毫升×2)萃取。合并有机相，饱和食盐水(25毫升×2)洗，无水硫酸钠干燥，过滤，浓缩，制备薄层色谱纯化后再用制备高效液相色谱(甲酸体系)纯化得到化合物3。MS(ESI)m/z:462.3[M+H +]； Step H: To a solution of compound 3-8 (200 mg, 577.68 micromolar, 1 equivalent) and 2-3 (131.88 mg, 866.53 micromolar, 1.5 equivalent) in 1,4-dioxane (10 mL) was added 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (66.85 mg, 115.54 μmol, 0.2 equivalent), bis (dibenzylideneacetone) palladium (66.43 mg, 115.54 μmol , 0.2 equiv.) And cesium carbonate (752.88 mg, 2.31 mmol, 4 equiv.), After reacting under nitrogen atmosphere and at 100 degrees Celsius for 12 hours, add 10 ml of water and 30 ml of ethyl acetate, ethyl acetate (25 ml × 2 )extraction. The organic phases were combined, washed with saturated brine (25 ml × 2), dried over anhydrous sodium sulfate, filtered, concentrated, purified by preparative thin layer chromatography and purified by preparative high performance liquid chromatography (formic acid system) to obtain compound 3. MS (ESI) m / z: 462.3 [M + H + ];
1H NMR(400MHz,CDCl 3):δ9.42(br s,1H),8.52(br s,1H),8.18(br d,J＝5.50Hz,2H),7.85-7.56(m,4H),7.32-7.17(m,3H),6.75-6.63(m,2H),4.22(br t,J＝6.66Hz,2H),2.84(br d,J＝6.85Hz,2H),2.65(br d,J＝6.72Hz,2H),1.53(br s,6H)。 1 H NMR (400 MHz, CDCl 3 ): δ 9.42 (br s, 1H), 8.52 (br s, 1H), 8.18 (br d, J = 5.50 Hz, 2H), 7.85-7.56 (m, 4H), 7.32-7.17 (m, 3H), 6.75-6.63 (m, 2H), 4.22 (br t, J = 6.66 Hz, 2H), 2.84 (br d, J = 6.85 Hz, 2H), 2.65 (br d, J = 6.72 Hz, 2H), 1.53 (br s, 6H).
实施例4：化合物4的甲酸盐Example 4: Formate of compound 4
Figure PCTCN2019114347-appb-000078
Figure PCTCN2019114347-appb-000078
Figure PCTCN2019114347-appb-000079
Figure PCTCN2019114347-appb-000079
步骤A：将化合物4-1(5克，26.58毫摩尔，3.97毫升，1当量)加入到碳酸二乙酯(34.13克，288.87毫摩尔，35毫升，10.87当量)中，冷却到0摄氏度，再在0摄氏度分批加入钠氢(2.13克，53.16毫摩尔，60％，2当量)。将该混合体系加热到85摄氏度搅拌2小时。反应液用盐酸(2摩尔每升)淬灭，并用盐酸调节pH到2，再加入水(50毫升)，用乙酸乙酯(30毫升×3)萃取，合并的有机相用饱和食盐水(30毫升)洗涤，无水硫酸钠干燥，过滤，浓缩，得到化合物4-2。MS(ESI)m/z:261.0[M+H +]。 Step A: Compound 4-1 (5 g, 26.58 mmol, 3.97 mL, 1 equiv) was added to diethyl carbonate (34.13 g, 288.87 mmol, 35 mL, 10.87 equiv), cooled to 0 ° C, then Sodium hydrogen (2.13 g, 53.16 mmol, 60%, 2 equivalents) was added in portions at 0 degrees Celsius. The mixed system was heated to 85 degrees Celsius and stirred for 2 hours. The reaction solution was quenched with hydrochloric acid (2 mol per liter), and the pH was adjusted to 2 with hydrochloric acid, then water (50 mL) was added, and extracted with ethyl acetate (30 mL × 3). The combined organic phases were saturated brine (30 ML), washed with anhydrous sodium sulfate, filtered, and concentrated to give compound 4-2. MS (ESI) m / z: 261.0 [M + H + ].
步骤B：向化合物4-2(7.81克，30.01毫摩尔，1当量)的吡啶(15毫升)溶液中加入1-4(9.81克，36.02毫摩尔，1.2当量，对甲苯磺酸盐)，将该混合物加热到40摄氏度搅拌12小时。向反应液中加入水(50毫升)，并用乙酸乙酯(50毫升×4)萃取，合并的有机相用饱和食盐水(50毫升×2)洗涤，无水硫酸钠干燥，过滤，浓缩，得到化合物4-3。MS(ESI)m/z:343.1[M+H +]。 Step B: To a solution of compound 4-2 (7.81 g, 30.01 mmol, 1 equiv) in pyridine (15 mL) was added 1-4 (9.81 g, 36.02 mmol, 1.2 equiv, p-toluenesulfonate), and The mixture was heated to 40 degrees Celsius and stirred for 12 hours. Water (50 mL) was added to the reaction solution, and extracted with ethyl acetate (50 mL × 4). The combined organic phase was washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered and concentrated to give Compound 4-3. MS (ESI) m / z: 343.1 [M + H + ].
步骤C：向化合物4-3(1克，2.92毫摩尔，1当量)的甲苯(15毫升)溶液中加入乙醇钠(596.39毫克，8.76毫摩尔，3当量)，将该混合物加热到100到110摄氏度之间反应12小时。将反应液倒入水(60毫升)中，用盐酸(2摩尔每升)调节pH到6到7之间。水相用乙酸乙酯(60毫升×2)萃取，合并的有机相用饱和食盐水(100毫升)洗涤，无水硫酸钠干燥，过滤，浓缩，得到化合物4-4。Step C: To a solution of compound 4-3 (1 g, 2.92 mmol, 1 equiv) in toluene (15 mL) was added sodium ethoxide (596.39 mg, 8.76 mmol, 3 equiv), and the mixture was heated to 100 to 110 React for 12 hours between degrees Celsius. The reaction solution was poured into water (60 ml), and the pH was adjusted to between 6 and 7 with hydrochloric acid (2 moles per liter). The aqueous phase was extracted with ethyl acetate (60 mL × 2), and the combined organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 4-4.
MS(ESI)m/z:297.0[M+H +]。 MS (ESI) m / z: 297.0 [M + H + ].
步骤D：向化合物4-4(0.78克，2.63毫摩尔，1当量)的N,N-二甲基甲酰胺(12毫升)溶液中加入N-溴代丁二酰亚胺(445.19毫克，2.50毫摩尔，0.95当量)，将该混合物加热到50摄氏度搅拌4小时。将反应液倒入水(60毫升)中，水相用乙酸乙酯(60毫升×2)萃取，合并的有机相用饱和食盐水(60毫升×2)洗涤，无水硫酸钠干燥，过滤，浓缩，柱层析纯化得到化合物4-5。Step D: To a solution of compound 4-4 (0.78 g, 2.63 mmol, 1 equiv) in N, N-dimethylformamide (12 mL) was added N-bromosuccinimide (445.19 mg, 2.50 Mmol, 0.95 equiv), the mixture was heated to 50 degrees Celsius and stirred for 4 hours. The reaction solution was poured into water (60 mL), the aqueous phase was extracted with ethyl acetate (60 mL × 2), and the combined organic phase was washed with saturated brine (60 mL × 2), dried over anhydrous sodium sulfate, and filtered. Concentrate and purify by column chromatography to obtain compound 4-5.
步骤E：在-78摄氏度，向化合物4-5(360毫克，1.09毫摩尔，1当量)和异丙氧基频哪醇硼酯(405.60毫克，2.18毫摩尔，444.74微升，2当量)的四氢呋喃溶液(10毫升)中加入正丁基锂(2.5摩尔每升，872.00微升，2当量)，反应液自然升温到25摄氏度并搅拌12小时。将饱和氯化铵水溶液(60毫升)倒 入反应液中，水相用乙酸乙酯(60毫升×2)萃取，合并的有机相用饱和用食盐水(60毫升)洗涤，无水硫酸钠干燥，过滤，浓缩，得到化合物4-6。MS(ESI)m/z:297.1[M+H +]。 Step E: At -78 degrees Celsius, apply compound 4-5 (360 mg, 1.09 mmol, 1 equiv) and isopropoxy pinacol borate (405.60 mg, 2.18 mmol, 444.74 μl, 2 equiv) To the tetrahydrofuran solution (10 ml) was added n-butyllithium (2.5 moles per liter, 872.00 microliters, 2 equivalents), and the reaction solution was naturally heated to 25 degrees Celsius and stirred for 12 hours. Saturated aqueous ammonium chloride solution (60 mL) was poured into the reaction solution, the aqueous phase was extracted with ethyl acetate (60 mL × 2), and the combined organic phase was washed with saturated brine (60 mL) and dried over anhydrous sodium sulfate. , Filtered, and concentrated to give compound 4-6. MS (ESI) m / z: 297.1 [M + H + ].
步骤F：向化合物4-6(450毫克，1.52毫摩尔，1当量)的四氢呋喃(10毫升)溶液中加入双氧水(689.37毫克，6.08毫摩尔，584.21微升，30％，4当量)，将体系冷却到0到5摄氏度之间，再加入氢氧化钠(4摩尔每升，1.52毫升，4当量)，将该混合物在25摄氏度搅拌12小时。将反应液倒入水(60毫升)中，水相用乙酸乙酯(80毫升)萃取，合并的有机相用饱和食盐水(80毫升)洗涤，无水硫酸钠干燥，过滤，浓缩，薄层色谱纯化得到化合物4-7。MS(ESI)m/z:269.0[M+H +]。 Step F: To a solution of compound 4-6 (450 mg, 1.52 mmol, 1 equivalent) in tetrahydrofuran (10 mL) was added hydrogen peroxide (689.37 mg, 6.08 mmol, 584.21 μL, 30%, 4 equivalents), Cool to between 0 and 5 degrees Celsius, then add sodium hydroxide (4 moles per liter, 1.52 milliliters, 4 equivalents), and stir the mixture at 25 degrees Celsius for 12 hours. The reaction solution was poured into water (60 mL), the aqueous phase was extracted with ethyl acetate (80 mL), and the combined organic phase was washed with saturated brine (80 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and a thin layer Chromatographic purification gave compound 4-7. MS (ESI) m / z: 269.0 [M + H + ].
步骤G：向化合物4-7(240毫克，894.74微摩尔，1当量)和化合物2-4(211.20毫克，1.34毫摩尔，1.5当量)的二氯甲烷(10毫升)溶液中加入醋酸铜(243.77毫克，1.34毫摩尔，1.5当量)，三乙胺(271.62毫克，2.68毫摩尔，373.61微升，3当量)和
Figure PCTCN2019114347-appb-000080
分子筛(1克)。该体系用氧气置换若干次，然后该混合物在25摄氏度反应12小时。将反应液过滤，滤液浓缩，薄层色谱纯化得到化合物4-8。 Step G: To a solution of compound 4-7 (240 mg, 894.74 μmol, 1 equiv) and compound 2-4 (211.20 mg, 1.34 mmol, 1.5 equiv) in methylene chloride (10 mL) was added copper acetate (243.77 Mg, 1.34 mmol, 1.5 equiv), triethylamine (271.62 mg, 2.68 mmol, 373.61 μl, 3 equiv) and
Figure PCTCN2019114347-appb-000080
Molecular sieve (1 g). The system was replaced with oxygen several times, and then the mixture was reacted at 25 degrees Celsius for 12 hours. The reaction solution was filtered, the filtrate was concentrated, and purified by thin layer chromatography to obtain compound 4-8.
MS(ESI)m/z:380.0,382.0[M+H +]。 MS (ESI) m / z: 380.0, 382.0 [M + H + ].
步骤H：向化合物4-8(130毫克，342.32微摩尔，1当量)和化合物2-3(104.20毫克，684.64微摩尔，2当量)的1,4-二氧六环(6毫升)溶液中加入双(二亚苄基丙酮)钯(19.68毫克，34.23微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(19.81毫克，34.23微摩尔，0.1当量)和碳酸铯(223.07毫克，684.64微摩尔，2当量)。该体系用氮气置换，然后加热到100摄氏度反应12小时。将反应液浓缩，柱色谱纯化后再使用制备高效液相色谱(甲酸体系)纯化得到化合物4的甲酸盐。MS(ESI)m/z:496.3[M+H +]； 1H NMR(400MHz,DMSO-d 6):δ9.48(s,1H),8.24-8.13(m,3H),8.02-7.95(m,2H),7.72-7.59(m,4H),6.68(dd,J＝5.6Hz,2.4Hz,1H),6.48(d,J＝2.4Hz,1H),4.23(t,J＝7.2Hz,2H),2.84(t,J＝7.2Hz,2H),2.64–2.57(m,2H),1.40(s,6H)。 Step H: To a solution of compound 4-8 (130 mg, 342.32 micromolar, 1 equivalent) and compound 2-3 (104.20 mg, 684.64 micromolar, 2 equivalents) in 1,4-dioxane (6 mL) Add bis (dibenzylideneacetone) palladium (19.68 mg, 34.23 μmol, 0.1 equivalent), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (19.81 mg, 34.23 μm) Molar, 0.1 equivalent) and cesium carbonate (223.07 mg, 684.64 micromolar, 2 equivalent). The system was replaced with nitrogen and then heated to 100 degrees Celsius for 12 hours. The reaction solution was concentrated, purified by column chromatography and then purified by preparative high performance liquid chromatography (formic acid system) to obtain the formate salt of compound 4. MS (ESI) m / z: 496.3 [M + H + ]; 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.48 (s, 1H), 8.24-8.13 (m, 3H), 8.02-7.95 ( m, 2H), 7.72-7.59 (m, 4H), 6.68 (dd, J = 5.6 Hz, 2.4 Hz, 1H), 6.48 (d, J = 2.4 Hz, 1H), 4.23 (t, J = 7.2 Hz, 2H), 2.84 (t, J = 7.2 Hz, 2H), 2.64-2.57 (m, 2H), 1.40 (s, 6H).
实施例5：化合物5Example 5: Compound 5
Figure PCTCN2019114347-appb-000081
Figure PCTCN2019114347-appb-000081
Figure PCTCN2019114347-appb-000082
Figure PCTCN2019114347-appb-000082
步骤A：将化合物5-1(3.13克，20.71毫摩尔，1当量)和乙酸乙酯(5.47克，62.14毫摩尔，6.08毫升，3当量)溶于四氢呋喃(20毫升)中，冷却到0摄氏度，再在0摄氏度分批加入钠氢(1.66克，41.43毫摩尔，60％，2当量)。将该混合体系加热到70摄氏度搅拌4小时。反应液用醋酸和水的混合溶液(醋酸：水＝1:1)淬灭，并调节pH到7，分离出有机相，水相用乙酸乙酯(30毫升×3)萃取，合并的有机相用饱和食盐水(20毫升)洗涤，无水硫酸钠干燥，过滤，浓缩，得到化合物5-2。Step A: Dissolve compound 5-1 (3.13 g, 20.71 mmol, 1 equiv) and ethyl acetate (5.47 g, 62.14 mmol, 6.08 mL, 3 equiv) in tetrahydrofuran (20 mL) and cool to 0 ° C Then, sodium hydrogen (1.66 g, 41.43 mmol, 60%, 2 equivalents) was added in portions at 0 ° C. The mixed system was heated to 70 degrees Celsius and stirred for 4 hours. The reaction solution was quenched with a mixed solution of acetic acid and water (acetic acid: water = 1: 1), and the pH was adjusted to 7, the organic phase was separated, the aqueous phase was extracted with ethyl acetate (30 ml × 3), and the combined organic phases It was washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 5-2.
MS(ESI)m/z:208.1[M+H +]。 MS (ESI) m / z: 208.1 [M + H + ].
步骤B：向化合物5-2(5.52克，26.64毫摩尔，1当量)的吡啶(15毫升)溶液中加入1-4(8.70克，31.97毫摩尔，1.2当量，对甲苯磺酸盐)，将该混合物加热到40摄氏度搅拌12小时。向反应液中加入水(50毫升)，并用乙酸乙酯(50毫升×4)萃取，合并的有机相用饱和食盐水(50毫升×2)洗涤，无水硫酸钠干燥，过滤，浓缩，得到化合物5-3。MS(ESI)m/z:290.1[M+H +]。 Step B: To a solution of compound 5-2 (5.52 g, 26.64 mmol, 1 equivalent) in pyridine (15 mL) was added 1-4 (8.70 g, 31.97 mmol, 1.2 equivalent, p-toluenesulfonate), and The mixture was heated to 40 degrees Celsius and stirred for 12 hours. Water (50 mL) was added to the reaction solution, and extracted with ethyl acetate (50 mL × 4). The combined organic phase was washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to give Compound 5-3. MS (ESI) m / z: 290.1 [M + H + ].
步骤C：向化合物5-3(5.6克，19.36毫摩尔，1当量)的甲苯(50毫升)溶液中加入乙醇钠(3.95克，58.07毫摩尔，3当量)，将该混合物加热到110摄氏度反应12小时。然后加入水(3.00克，166.53毫摩尔，3毫升，8.60当量)，继续在100摄氏度反应12小时。将反应液倒入水(50毫升)中，用盐酸(2摩尔每升)调节其pH到4。分离出有机相，水相用异丙醇/二氯甲烷(1/10,50毫升×4)萃取，合并的有机相用饱和食盐水(50毫升)洗涤，无水硫酸钠干燥，过滤，浓缩，得到化合物5-4。MS(ESI)m/z:244.0[M+H +]。 Step C: To a solution of compound 5-3 (5.6 g, 19.36 mmol, 1 equiv) in toluene (50 mL) was added sodium ethoxide (3.95 g, 58.07 mmol, 3 equiv), and the mixture was heated to 110 ° C for reaction 12 hours. Then water (3.00 g, 166.53 mmol, 3 mL, 8.60 equiv) was added, and the reaction was continued at 100 ° C for 12 hours. The reaction solution was poured into water (50 ml), and its pH was adjusted to 4 with hydrochloric acid (2 mol per liter). The organic phase was separated, the aqueous phase was extracted with isopropanol / dichloromethane (1/10, 50 ml × 4), and the combined organic phase was washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, filtered, and concentrated To give compound 5-4. MS (ESI) m / z: 244.0 [M + H + ].
步骤D：向化合物5-4(3.97克，16.32毫摩尔，1当量)的N,N-二甲基甲酰胺(50毫升)溶液中加入N-溴代丁二酰亚胺(3.49克，19.58毫摩尔，1.2当量)，将该混合物加热到50摄氏度搅拌12小时。将反应液倒入水(200毫升)中，水相用乙酸乙酯(100毫升×3)萃取，合并的有机相用饱和食盐水(50毫升)洗涤，无水硫酸钠干燥，过滤，浓缩，柱层析纯化得到化合物5-5。MS(ESI)m/z:277.9,279.9[M+H +]。 Step D: To a solution of compound 5-4 (3.97 g, 16.32 mmol, 1 equivalent) in N, N-dimethylformamide (50 mL) was added N-bromosuccinimide (3.49 g, 19.58 Mmol, 1.2 equivalents), the mixture was heated to 50 degrees Celsius and stirred for 12 hours. The reaction solution was poured into water (200 mL), the aqueous phase was extracted with ethyl acetate (100 mL × 3), and the combined organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. Column chromatography purification gave compound 5-5. MS (ESI) m / z: 277.9, 279.9 [M + H + ].
步骤E：在-78～-65摄氏度，向化合物5-5(2.82克，10.14毫摩尔，1当量)和异丙氧基频哪醇硼酯(3.77克，20.28毫摩尔，4.14毫升，2当量)的四氢呋喃溶液(30毫升)中加入正丁基锂(2.5摩尔每升， 8.11毫升，2当量)，反应液自然升温到25摄氏度并搅拌12小时。将饱和氯化铵水溶液(50毫升)倒入反应液中，水相用乙酸乙酯(50毫升×3)萃取，合并的有机相用饱和食盐水(50毫升)洗涤，无水硫酸钠干燥，过滤，浓缩，得到化合物5-6。MS(ESI)m/z:325.8[M+H +]。 Step E: At -78 to -65 degrees Celsius, apply compound 5-5 (2.82 g, 10.14 mmol, 1 equivalent) and isopropoxy pinacol borate (3.77 g, 20.28 mmol, 4.14 mL, 2 equivalents) ) In tetrahydrofuran solution (30 ml) was added n-butyllithium (2.5 moles per liter, 8.11 ml, 2 equivalents), and the reaction solution was naturally heated to 25 ° C and stirred for 12 hours. Saturated aqueous ammonium chloride solution (50 mL) was poured into the reaction solution, the aqueous phase was extracted with ethyl acetate (50 mL × 3), the combined organic phase was washed with saturated brine (50 mL), and dried over anhydrous sodium sulfate, Filtration and concentration provided compound 5-6. MS (ESI) m / z: 325.8 [M + H + ].
步骤F：向化合物5-6(1克，3.07毫摩尔，1当量)的四氢呋喃(15毫升)溶液中加入双氧水(1.39克，12.30毫摩尔，1.18毫升，30％，4当量)，将体系冷却到0到5摄氏度之间，再加入氢氧化钠(4摩尔每升，3.07毫升，4当量)，将该混合物在25摄氏度搅拌12小时。向反应液中加入饱和亚硫酸氢钠(5毫升)，将混合液倒入水(60毫升)中，水相用乙酸乙酯(60毫升×2)萃取，合并有机相，无水硫酸钠干燥，过滤，浓缩，柱层析纯化得到化合物5-7。MS(ESI)m/z:216.1[M+H +]。 Step F: To a solution of compound 5-6 (1 g, 3.07 mmol, 1 equiv) in tetrahydrofuran (15 mL) was added hydrogen peroxide (1.39 g, 12.30 mmol, 1.18 mL, 30%, 4 equiv), and the system was cooled Between 0 and 5 degrees Celsius, additional sodium hydroxide (4 moles per liter, 3.07 milliliters, 4 equivalents) was added and the mixture was stirred at 25 degrees Celsius for 12 hours. Saturated sodium bisulfite (5 mL) was added to the reaction solution, the mixture was poured into water (60 mL), the aqueous phase was extracted with ethyl acetate (60 mL × 2), the organic phases were combined, and dried over anhydrous sodium sulfate , Filtered, concentrated, and purified by column chromatography to obtain compound 5-7. MS (ESI) m / z: 216.1 [M + H + ].
步骤G：向化合物5-7(300毫克，1.39毫摩尔，1当量)和化合物2-4(328.98毫克，2.09毫摩尔，1.5当量)的二氯甲烷(15毫升)溶液中加入醋酸铜(379.72毫克，2.09毫摩尔，1.5当量)，三乙胺(423.09毫克，4.18毫摩尔，581.97微升，3当量)和
Figure PCTCN2019114347-appb-000083
分子筛(1克)。该体系用氧气置换若干次，然后该混合物在25摄氏度反应12小时。将反应液倒入水(60毫升)中，水相用乙酸乙酯(60毫升×2)萃取，合并的有机相用食盐水(60毫升)洗涤，无水硫酸钠干燥，过滤，浓缩，薄层色谱纯化得到化合物5-8。 Step G: To a solution of compound 5-7 (300 mg, 1.39 mmol, 1 equiv) and compound 2-4 (328.98 mg, 2.09 mmol, 1.5 equiv) in dichloromethane (15 mL) was added copper acetate (379.72 Mg, 2.09 mmol, 1.5 equiv), triethylamine (423.09 mg, 4.18 mmol, 581.97 μl, 3 equiv) and
Figure PCTCN2019114347-appb-000083
Molecular sieve (1 g). The system was replaced with oxygen several times, and then the mixture was reacted at 25 degrees Celsius for 12 hours. The reaction solution was poured into water (60 mL), the aqueous phase was extracted with ethyl acetate (60 mL × 2), and the combined organic phase was washed with brine (60 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and thin Layer chromatography purification gave compounds 5-8.
MS(ESI)m/z:326.9,328.9[M+H +]。 MS (ESI) m / z: 326.9, 328.9 [M + H + ].
步骤H：向化合物5-8(200毫克，612.03微摩尔，1当量)和化合物2-3(93.15毫克，612.03微摩尔，1当量)的1,4-二氧六环(8毫升)溶液中加入双(二亚苄基丙酮)钯(35.19毫克，61.20微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(35.41毫克，61.20微摩尔，0.1当量)和碳酸铯(398.82毫克，1.22毫摩尔，2当量)。该体系用氮气置换，然后加热到90摄氏度反应3小时。将反应液浓缩，制备高效液相色谱(先甲酸体系，再碱性体系)提纯得到化合物5。MS(ESI)m/z:443.4[M+H +]； Step H: To a solution of compound 5-8 (200 mg, 612.03 micromolar, 1 equivalent) and compound 2-3 (93.15 mg, 612.03 micromolar, 1 equivalent) in 1,4-dioxane (8 mL) Add bis (dibenzylideneacetone) palladium (35.19 mg, 61.20 μmol, 0.1 equivalent), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (35.41 mg, 61.20 μm) Molar, 0.1 equivalent) and cesium carbonate (398.82 mg, 1.22 mmol, 2 equivalent). The system was replaced with nitrogen, and then heated to 90 degrees Celsius for 3 hours. The reaction solution was concentrated and purified by high performance liquid chromatography (first formic acid system, then basic system) to obtain compound 5. MS (ESI) m / z: 443.4 [M + H + ];
1H NMR(400MHz,DMSO-d 6)δ＝9.43(s,1H),8.64(d,J＝2.0Hz,1H),8.30(d,J＝1.6Hz,1H),8.19(d,J＝5.6Hz,1H),8.16(d,J＝6.0Hz,1H),7.86(s,1H),7.71-7.61(m,2H),6.67(dd,J＝2.0,6.0Hz,1H),6.45(d,J＝2.0Hz,1H),5.09(br s,1H),4.21(t,J＝7.2Hz,2H),2.88-2.77(m,2H),2.60(m,2H),2.28(s,3H),1.39(s,6H)。 1 H NMR (400 MHz, DMSO-d 6 ) δ = 9.43 (s, 1H), 8.64 (d, J = 2.0 Hz, 1 H), 8.30 (d, J = 1.6 Hz, 1 H), 8.19 (d, J = 5.6Hz, 1H), 8.16 (d, J = 6.0Hz, 1H), 7.86 (s, 1H), 7.71-7.61 (m, 2H), 6.67 (dd, J = 2.0, 6.0Hz, 1H), 6.45 ( d, J = 2.0 Hz, 1H), 5.09 (br s, 1H), 4.21 (t, J = 7.2 Hz, 2H), 2.88-2.77 (m, 2H), 2.60 (m, 2H), 2.28 (s, 3H), 1.39 (s, 6H).
实施例6：化合物6Example 6: Compound 6
Figure PCTCN2019114347-appb-000084
Figure PCTCN2019114347-appb-000084
Figure PCTCN2019114347-appb-000085
Figure PCTCN2019114347-appb-000085
步骤A：将乙酸乙酯(17.6克，199.77毫摩尔，3.0当量)加入甲苯(50毫升)中，在20摄氏度下加入乙醇钠(9.06克，133.18毫摩尔，2.0当量)。加完后在20摄氏度下搅拌1小时。将化合物6-1(10克，66.59毫摩尔，1.0当量)分三批加入，加完后在100摄氏度下搅拌12小时。将反应液冷却至25摄氏度，用冰醋酸调节pH值至6，加入水(50毫升)，甲苯萃取(30毫升×2)，合并有机相，无水硫酸钠干燥，浓缩得到化合物6-2。Step A: Ethyl acetate (17.6 g, 199.77 mmol, 3.0 equiv) was added to toluene (50 mL), and sodium ethoxide (9.06 g, 133.18 mmol, 2.0 equiv) was added at 20 degrees Celsius. After the addition, the mixture was stirred at 20 degrees Celsius for 1 hour. Compound 6-1 (10 g, 66.59 mmol, 1.0 equiv) was added in three batches, and after the addition was completed, it was stirred at 100 degrees Celsius for 12 hours. The reaction solution was cooled to 25 degrees Celsius, the pH value was adjusted to 6 with glacial acetic acid, water (50 ml) was added, extracted with toluene (30 ml × 2), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain compound 6-2.
步骤B：将化合物6-2(10.05克，48.73毫摩尔，1.0当量)加入吡啶(25毫升)中，然后加入化合物1-4(16.10克，58.48毫摩尔，1.2当量)。在40摄氏度下搅拌12小时。将反应液冷却至20摄氏度，加入水(50毫升)，用甲苯萃取(40毫升×2)。合并有机相，水(50毫升)洗，无水硫酸钠干燥，浓缩得到化合物6-3。Step B: Compound 6-2 (10.05 g, 48.73 mmol, 1.0 equiv) was added to pyridine (25 mL), and then compound 1-4 (16.10 g, 58.48 mmol, 1.2 equiv) was added. Stir at 40 degrees Celsius for 12 hours. The reaction liquid was cooled to 20 degrees Celsius, water (50 ml) was added, and extracted with toluene (40 ml × 2). The organic phases were combined, washed with water (50 mL), dried over anhydrous sodium sulfate, and concentrated to give compound 6-3.
步骤C：将化合物6-3(12.54克，43.49毫摩尔，1.0当量)溶于甲苯(80毫升)中，在不断搅拌下分批加入乙醇钠(8.88克，130.47毫摩尔，3.0当量)。加完后在100-110摄氏度下搅拌12小时。将反应液冷却至20摄氏度，用1摩尔每升的盐酸调节pH至6。加入50毫升水，乙酸乙酯(50毫升×3)萃取，合并有机相，饱和食盐水洗(50毫升)，无水硫酸钠干燥，浓缩得到化合物6-4。MS(ESI)m/z:271.2[M+H +]。 Step C: Compound 6-3 (12.54 g, 43.49 mmol, 1.0 equiv) was dissolved in toluene (80 mL), and sodium ethoxide (8.88 g, 130.47 mmol, 3.0 equiv) was added in portions with constant stirring. After the addition, the mixture was stirred at 100-110 degrees Celsius for 12 hours. The reaction solution was cooled to 20 degrees Celsius, and the pH was adjusted to 6 with 1 mole of hydrochloric acid. 50 ml of water was added, extracted with ethyl acetate (50 ml × 3), the organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, and concentrated to give compound 6-4. MS (ESI) m / z: 271.2 [M + H + ].
步骤D：将氢氧化钠(16.69克，417.27毫摩尔，10当量)溶于水(25毫升)中，所得溶液加入到化合物6-4(11.28克，41.73毫摩尔，1.0当量)的甲醇(50毫升)溶液中。40摄氏度下搅拌12小时。除去甲醇，加入水(25毫升)，用乙酸乙酯(25毫升×3)洗涤。将水相的pH用浓盐酸调到4，过滤。滤饼用水(25毫升×3)洗，干燥，得到化合物6-5。MS(ESI)m/z:243.1[M+H +]。 Step D: Sodium hydroxide (16.69 g, 417.27 mmol, 10 equiv) was dissolved in water (25 mL), and the resulting solution was added to compound 6-4 (11.28 g, 41.73 mmol, 1.0 equiv) in methanol (50 Ml) solution. Stir for 12 hours at 40 degrees Celsius. Methanol was removed, water (25 ml) was added, and washed with ethyl acetate (25 ml × 3). The pH of the aqueous phase was adjusted to 4 with concentrated hydrochloric acid and filtered. The filter cake was washed with water (25 ml × 3) and dried to obtain compound 6-5. MS (ESI) m / z: 243.1 [M + H + ].
步骤E:将化合物6-5(10.01克，41.32毫摩尔，1.0当量)溶于N,N-二甲基甲酰胺(120毫升)中，分批加入溴代丁二酰亚胺(7.35克，41.32毫摩尔，1.0当量)和氢氧化钠(1.98克，49.58毫摩尔，1.2当量)。40摄氏度下搅拌12小时。将反应液冷却至20-25摄氏度，加入水(300毫升)，搅拌10分钟，过滤，滤饼用水(20毫升×2)洗，干燥，得到化合物6-6。MS(ESI)m/z:277.2[M+H +]。 Step E: Compound 6-5 (10.01 g, 41.32 mmol, 1.0 equivalent) was dissolved in N, N-dimethylformamide (120 mL), and bromosuccinimide (7.35 g, 41.32 mmol, 1.0 equivalent) and sodium hydroxide (1.98 g, 49.58 mmol, 1.2 equivalent). Stir for 12 hours at 40 degrees Celsius. The reaction solution was cooled to 20-25 degrees Celsius, water (300 mL) was added, stirred for 10 minutes, filtered, and the filter cake was washed with water (20 mL × 2) and dried to obtain compound 6-6. MS (ESI) m / z: 277.2 [M + H + ].
步骤F：将化合物6-6(1.4克，5.05毫摩尔，1.0当量)，异丙氧基频哪醇硼酯(2.49克，13.39毫摩尔，2.65当量)加入无水四氢呋喃(30毫升)中，置换氮气三次，搅拌，冷却至-70～-60摄氏度。在-70～ -60摄氏度下逐滴滴加正丁基锂(2.5摩尔每升正庚烷溶液，4.83毫升，2.39当量)。加完后在-70～-60摄氏度下搅拌1小时。然后在25摄氏度下搅拌12小时。将反应液逐滴加入60毫升饱和氯化铵溶液中，乙酸乙酯(40毫升×3)萃取，合并有机相，饱和食盐水(30毫升)洗，无水硫酸钠干燥，浓缩得到化合物6-7。Step F: Compound 6-6 (1.4 g, 5.05 mmol, 1.0 equiv), isopropoxy pinacol borate (2.49 g, 13.39 mmol, 2.65 equiv) was added to anhydrous tetrahydrofuran (30 mL), Replace nitrogen three times, stir, and cool to -70 ~ -60 degrees Celsius. N-Butyllithium (2.5 moles per liter of n-heptane solution, 4.83 ml, 2.39 equiv) was added dropwise at -70 to -60 degrees Celsius. After the addition, the mixture was stirred at -70 to -60 degrees Celsius for 1 hour. It was then stirred at 25 degrees Celsius for 12 hours. The reaction solution was added dropwise to 60 ml of saturated ammonium chloride solution, extracted with ethyl acetate (40 ml × 3), the organic phases were combined, washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, and concentrated to obtain compound 6 7.
步骤G:将化合物6-7(2.08克，6.42毫摩尔，1.0当量)和双氧水(2.91克，25.66毫摩尔，浓度30％，4当量)加入四氢呋喃(20毫升)中，搅拌，冷却至0～5摄氏度。在0～5摄氏度下逐滴加入4摩尔每升的氢氧化钠水溶液(6.42毫升，4.0当量)。加完后在25摄氏度下搅拌12小时。向反应液中加入饱和亚硫酸氢钠水溶液(20毫升)，用2摩尔每升的盐酸调节pH值至7，乙酸乙酯萃取(30毫升×2)，饱和食盐水洗(30毫升)，无水硫酸钠干燥，过滤，浓缩，柱层析纯化得到化合物6-8。MS(ESI)m/z:215.1[M+H +]。 Step G: Add compound 6-7 (2.08 g, 6.42 mmol, 1.0 equiv) and hydrogen peroxide (2.91 g, 25.66 mmol, 30% concentration, 4 equiv) to tetrahydrofuran (20 mL), stir, and cool to 0 ～ 5 degrees Celsius. A 4 mol per liter aqueous solution of sodium hydroxide (6.42 ml, 4.0 equivalents) was added dropwise at 0-5 degrees Celsius. After the addition, the mixture was stirred at 25 degrees Celsius for 12 hours. Saturated aqueous sodium bisulfite solution (20 mL) was added to the reaction solution, and the pH was adjusted to 7 with 2 mol per liter of hydrochloric acid, extracted with ethyl acetate (30 mL × 2), washed with saturated brine (30 mL), anhydrous Dry over sodium sulfate, filter, concentrate, and purify by column chromatography to obtain compound 6-8. MS (ESI) m / z: 215.1 [M + H + ].
步骤H：将化合物6-8(336毫克，1.57毫摩尔，1当量)，化合物2-4(494.12毫克，3.14毫摩尔，2当量)溶于二氯甲烷(20毫升)中，依次加入醋酸铜(427.75毫克，2.36毫摩尔，1.5当量)，三乙胺(476.60毫克，4.71毫摩尔，655.58微升，3当量)和
Figure PCTCN2019114347-appb-000086
分子筛(3.5克)。在25摄氏度下氧气氛围搅拌12小时。过滤，滤液加水(30毫升)稀释，乙酸乙酯萃取(30毫升×3)，合并有机相，饱和食盐水洗(40毫升)，无水硫酸钠干燥，浓缩，柱层析纯化得到化合物6-9。MS(ESI)m/z:326.0[M+H +]。 Step H: Dissolve compound 6-8 (336 mg, 1.57 mmol, 1 equiv), compound 2-4 (494.12 mg, 3.14 mmol, 2 equiv) in dichloromethane (20 mL), and add copper acetate in sequence (427.75 mg, 2.36 mmol, 1.5 equiv), triethylamine (476.60 mg, 4.71 mmol, 655.58 μl, 3 equiv) and
Figure PCTCN2019114347-appb-000086
Molecular sieve (3.5 g). The oxygen atmosphere was stirred for 12 hours at 25 degrees Celsius. Filtration, the filtrate was diluted with water (30 mL), extracted with ethyl acetate (30 mL × 3), the organic phases were combined, washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain compound 6-9 . MS (ESI) m / z: 326.0 [M + H + ].
步骤I：将化合物6-9(200毫克，613.89微摩尔，1当量)，化合物2-3(186.86毫克，1.23毫摩尔，2当量)溶于1,4-二氧六环(20毫升)中，依次加入二(二亚苄基丙酮)钯(70.60毫克，112.78微摩尔，0.2当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(71.04毫克，122.78微摩尔，0.2当量)，碳酸铯(800.07毫克，2.46毫摩尔，4当量)。置换氮气三次，在氮气保护下升温至100摄氏度搅拌12小时。冷却至25摄氏度，加入水(50毫升)淬灭，乙酸乙酯萃取(50毫升×2)，合并有机相，饱和食盐水洗(40毫升)，无水硫酸钠干燥，浓缩得到粗产品，粗产品经制备高效液相色谱分离，冻干，得到化合物6。Step I: Dissolve compound 6-9 (200 mg, 613.89 micromolar, 1 equivalent) and compound 2-3 (186.86 mg, 1.23 mmol, 2 equivalents) in 1,4-dioxane (20 mL) , Followed by the addition of bis (dibenzylideneacetone) palladium (70.60 mg, 112.78 micromolar, 0.2 equivalent), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (71.04 mg, 122.78 micromolar, 0.2 equivalent), cesium carbonate (800.07 mg, 2.46 mmol, 4 equivalent). The nitrogen was replaced three times, and the temperature was raised to 100 degrees Celsius and stirred for 12 hours under the protection of nitrogen. Cool to 25 degrees Celsius, quench with water (50 mL), extract with ethyl acetate (50 mL × 2), combine organic phases, wash with saturated brine (40 mL), dry over anhydrous sodium sulfate, and concentrate to obtain crude product. After separation by preparative high performance liquid chromatography and lyophilization, compound 6 was obtained.
MS(ESI)m/z:442.2[M+H +]。 MS (ESI) m / z: 442.2 [M + H + ].
1H NMR(400MHz,DMSO-d 6)δ9.50(s,1H),8.21–8.15(m,3H),7.71–7.67(m,2H),7.55(s,1H),7.47(d,J＝8.0Hz,1H),7.21(t,J＝7.8Hz,1H),7.06(d,J＝7.6Hz,1H),6.67(dd,J＝2.4,5.6Hz,1H),6.46(d,J＝2.0Hz,1H),4.17(t,J＝7.0Hz,2H),2.80(t,J＝7.2Hz,2H),2.61–2.54(m,2H),2.26(s,3H),1.40(s,6H)。 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.50 (s, 1H), 8.21–8.15 (m, 3H), 7.71–7.67 (m, 2H), 7.55 (s, 1H), 7.47 (d, J = 8.0Hz, 1H), 7.21 (t, J = 7.8Hz, 1H), 7.06 (d, J = 7.6Hz, 1H), 6.67 (dd, J = 2.4, 5.6Hz, 1H), 6.46 (d, J = 2.0 Hz, 1H), 4.17 (t, J = 7.0 Hz, 2H), 2.80 (t, J = 7.2 Hz, 2H), 2.61–2.54 (m, 2H), 2.26 (s, 3H), 1.40 (s , 6H).
实施例7：化合物7Example 7: Compound 7
Figure PCTCN2019114347-appb-000087
Figure PCTCN2019114347-appb-000087
Figure PCTCN2019114347-appb-000088
Figure PCTCN2019114347-appb-000088
步骤A：将六甲基二硅基氨基锂(1摩尔每升，728.32毫升，3当量)加入三口瓶中，将反应液冷却到-78摄氏度，之后逐滴加入乙酸乙酯(64.17克，728.32毫摩尔，71.30毫升，3当量)。将反应液在-78摄氏度搅拌半个小时。在-78摄氏度，将7-1(35克，242.77毫摩尔，32.41毫升，1当量)溶解于400毫升四氢呋喃中并逐滴加入反应液中。将反应液在-78摄氏度搅拌1小时。在-70摄氏度使用150毫升饱和氯化铵溶液淬灭反应，加入400毫升水稀释并用乙酸乙酯(500毫升×2)萃取。合并有机相，饱和食盐水(400毫升)洗，无水硫酸钠干燥，过滤，浓缩得到化合物7-2。Step A: Add lithium hexamethyldisilazide (1 mol per liter, 728.32 ml, 3 equivalents) to a three-necked bottle, cool the reaction solution to -78 degrees Celsius, and then add ethyl acetate (64.17 g, 728.32 dropwise) Mmol, 71.30 mL, 3 equivalents). The reaction solution was stirred at -78 degrees Celsius for half an hour. At -78 degrees Celsius, 7-1 (35 g, 242.77 mmol, 32.41 mL, 1 equivalent) was dissolved in 400 mL of tetrahydrofuran and added dropwise to the reaction solution. The reaction solution was stirred at -78 degrees Celsius for 1 hour. The reaction was quenched with 150 ml of saturated ammonium chloride solution at -70 degrees Celsius, diluted with 400 ml of water and extracted with ethyl acetate (500 ml × 2). The organic phases were combined, washed with saturated brine (400 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 7-2.
步骤B：将7-2(14.5克，72.42毫摩尔，1当量)溶解于吡啶(140毫升)中，加入1-4(31.55克，115.87毫摩尔，1.6当量，对甲苯磺酸盐)。将混合液使用氮气置换三次，40摄氏度搅拌12小时。加300毫升水稀释，乙酸乙酯(300毫升×2)萃取。合并有机相，饱和食盐水(100毫升×2)洗，无水硫酸钠干燥，过滤并在真空中浓缩得到化合物7-3。MS(ESI)m/z:283.1[M+H +]。 Step B: Dissolve 7-2 (14.5 g, 72.42 mmol, 1 equiv) in pyridine (140 mL) and add 1-4 (31.55 g, 115.87 mmol, 1.6 equiv, p-toluenesulfonate). The mixture was replaced with nitrogen three times, and stirred at 40 degrees Celsius for 12 hours. Dilute with 300 ml of water and extract with ethyl acetate (300 ml x 2). The organic phases were combined, washed with saturated brine (100 ml × 2), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to obtain compound 7-3. MS (ESI) m / z: 283.1 [M + H + ].
步骤C：将化合物7-3(6.3克，22.31毫摩尔，1当量)溶解于甲苯(80毫升)中，加入氢化钠(2.68克，66.94毫摩尔，60％纯度，3当量)，在氮气氛围将混合液加热到110摄氏度反应12小时。向混合液中加入250毫升乙酸乙酯和250毫升水，有机相用饱和食盐水(100毫升)洗，无水硫酸钠干燥，过滤，浓缩。水相用4摩尔每升盐酸调pH至4，乙酸乙酯(100毫升×2)萃取，合并有机相，饱和食盐水(80毫升)洗，无水硫酸钠干燥，过滤，浓缩。将两部分产品合并即得7-4。MS(ESI)m/z:265.1[M+H +]。 Step C: Dissolve compound 7-3 (6.3 g, 22.31 mmol, 1 equiv) in toluene (80 mL), add sodium hydride (2.68 g, 66.94 mmol, 60% purity, 3 equiv) under a nitrogen atmosphere The mixture was heated to 110 degrees Celsius and reacted for 12 hours. 250 ml of ethyl acetate and 250 ml of water were added to the mixture, and the organic phase was washed with saturated brine (100 ml), dried over anhydrous sodium sulfate, filtered, and concentrated. The aqueous phase was adjusted to pH 4 with 4 moles per liter of hydrochloric acid, extracted with ethyl acetate (100 mL × 2), the organic phases were combined, washed with saturated brine (80 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. 7-4 is obtained by combining the two parts. MS (ESI) m / z: 265.1 [M + H + ].
步骤D：将7-4(7克，26.48毫摩尔，1当量)溶于水(20毫升)和甲醇(40毫升)中，氮气氛围下加入氢氧化钠(3.18克，79.45毫摩尔，3当量)，60摄氏度搅拌5小时。溶液用4摩尔每升盐酸调pH至6，过滤，滤饼干燥得到化合物7-5。MS(ESI)m/z:237.2[M+H +]。 Step D: Dissolve 7-4 (7 g, 26.48 mmol, 1 eq) in water (20 mL) and methanol (40 mL), and add sodium hydroxide (3.18 g, 79.45 mmol, 3 eq) under a nitrogen atmosphere ), Stirring at 60 degrees Celsius for 5 hours. The solution was adjusted to pH 6 with 4 moles per liter of hydrochloric acid, filtered, and the filter cake was dried to obtain compound 7-5. MS (ESI) m / z: 237.2 [M + H + ].
步骤E：将7-5(5克，21.16毫摩尔，1当量)溶解于二甲基甲酰胺(20毫升)中，在氮气氛围加入溴代丁二酰亚胺(4.52克，25.40毫摩尔，1.2当量)。将混合液在25摄氏度搅拌12小时。加水(100毫升)稀释，乙酸乙酯(100毫升×2)萃取。合并有机相，饱和食盐水(50毫升×2)洗，无水硫酸钠干燥，过滤，浓缩，柱层析纯化得到化合物7-6。MS(ESI)m/z:271,273[M+H +]。 Step E: Dissolve 7-5 (5 g, 21.16 mmol, 1 equivalent) in dimethylformamide (20 mL), and add bromosuccinimide (4.52 g, 25.40 mmol, under nitrogen atmosphere). 1.2 equivalent). The mixture was stirred at 25 degrees Celsius for 12 hours. Dilute with water (100 ml) and extract with ethyl acetate (100 ml x 2). The organic phases were combined, washed with saturated brine (50 ml × 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain compound 7-6. MS (ESI) m / z: 271,273 [M + H + ].
步骤F：将化合物7-6(4.26克，15.71毫摩尔，1当量)溶解于四氢呋喃(40毫升)，氮气气氛下，- 70摄氏度逐滴添加正丁基锂(2.5摩尔每升，18.85毫升，3当量)。将混合液在-70摄氏度搅拌1小时，保持温度逐滴加入异丙氧基频哪醇硼酯(7.60克，40.85毫摩尔，8.33毫升，2.6当量)。缓缓升温到0摄氏度并搅拌1小时。将反应液倾入0摄氏度的饱和氯化铵水溶液(100毫升)中，乙酸乙酯(100毫升×2)萃取，合并有机相，饱和食盐水(50毫升×2)洗，无水硫酸钠干燥，过滤，浓缩得到化合物7-7。Step F: Compound 7-6 (4.26 g, 15.71 mmol, 1 equiv) was dissolved in tetrahydrofuran (40 mL). Under nitrogen atmosphere, n-butyllithium (2.5 mol per liter, 18.85 mL, 3 equivalents). The mixed solution was stirred at -70 degrees Celsius for 1 hour, maintaining temperature to add dropwise isopropoxy pinacol borate (7.60 g, 40.85 mmol, 8.33 mL, 2.6 equiv). Slowly warm up to 0 degrees Celsius and stir for 1 hour. The reaction solution was poured into a saturated aqueous ammonium chloride solution (100 ml) at 0 degrees Celsius, extracted with ethyl acetate (100 ml × 2), the organic phases were combined, washed with saturated brine (50 ml × 2), and dried over anhydrous sodium sulfate , Filtered, and concentrated to give compound 7-7.
MS(ESI)m/z:319.1[M+H +]。 MS (ESI) m / z: 319.1 [M + H + ].
步骤G：将7-7(5.2克，16.34毫摩尔，1当量)溶解于四氢呋喃(50毫升)中，之后添加过氧化氢(4.63克，40.85毫摩尔，3.92毫升，30％纯度，2.5当量)。将反应液冷却到0-5摄氏度，逐滴加入氢氧化钠(4摩尔每升，10.21毫升，2.5当量)水溶液。将反应液在25摄氏度搅拌12小时。加入亚硫酸钠(1克)淬灭，4摩尔每升盐酸调pH至8，20摄氏度搅拌1小时。加200毫升水稀释，乙酸乙酯(300毫升×2)萃取，有机相用饱和食盐水(200毫升)洗，无水硫酸钠干燥，过滤，浓缩得到化合物7-8。Step G: Dissolve 7-7 (5.2 g, 16.34 mmol, 1 equiv) in tetrahydrofuran (50 mL), then add hydrogen peroxide (4.63 g, 40.85 mmol, 3.92 mL, 30% purity, 2.5 equiv) . The reaction solution was cooled to 0-5 degrees Celsius, and an aqueous solution of sodium hydroxide (4 moles per liter, 10.21 ml, 2.5 equivalents) was added dropwise. The reaction solution was stirred at 25 degrees Celsius for 12 hours. Sodium sulfite (1 g) was added for quenching, 4 mol / L hydrochloric acid was used to adjust the pH to 8, and the mixture was stirred at 20 degrees Celsius for 1 hour. It was diluted with 200 ml of water, extracted with ethyl acetate (300 ml × 2), and the organic phase was washed with saturated brine (200 ml), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 7-8.
MS(ESI)m/z:209.1[M+H +]。 MS (ESI) m / z: 209.1 [M + H + ].
步骤H：将化合物7-8(800毫克，3.84毫摩尔，1当量)和7-9(757.92毫克，5.76毫摩尔，1.5当量)溶解于N,N-二甲基甲酰胺(8毫升)中，在氮气氛围下加入碳酸钾(1.59克，11.52毫摩尔，3当量)。混合液在120摄氏度反应12小时。加入60毫升水稀释，乙酸乙酯(80毫升×2)萃取，有机相用60毫升饱和食盐水洗，无水硫酸钠干燥，过滤，浓缩，柱层析纯化得到化合物7-10。MS(ESI)m/z:320.1[M+H +]。 Step H: Compounds 7-8 (800 mg, 3.84 mmol, 1 equivalent) and 7-9 (757.92 mg, 5.76 mmol, 1.5 equivalent) were dissolved in N, N-dimethylformamide (8 mL) Under a nitrogen atmosphere, potassium carbonate (1.59 g, 11.52 mmol, 3 equivalents) was added. The mixed solution was reacted at 120 degrees Celsius for 12 hours. Diluted with 60 ml of water, extracted with ethyl acetate (80 ml × 2), the organic phase was washed with 60 ml of saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain compound 7-10. MS (ESI) m / z: 320.1 [M + H + ].
步骤I：将化合物7-10(1.05克，3.28毫摩尔，1当量)，7-11(426.68毫克，3.61毫摩尔，96.31微升，1.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(189.99毫克，328.35微摩尔，0.1当量)，碳酸铯(3.21克，9.85毫摩尔，3当量)和醋酸钯(73.72毫克，328.35微摩，0.1当量)溶解于二氧六环(10毫升)中。氮气气氛下于100摄氏度反应3小时。加入40毫升水稀释，乙酸乙酯(70毫升×2)萃取，有机相用60毫升饱和食盐水洗，无水硫酸钠干燥，过滤，浓缩。加入70毫升乙酸乙酯溶解并加入1克巯基硅胶，25摄氏度搅拌1小时。过滤并浓缩得到化合物7-12。MS(ESI)m/z:402.1[M+H +]。 Step I: Compound 7-10 (1.05 g, 3.28 mmol, 1 equivalent), 7-11 (426.68 mg, 3.61 mmol, 96.31 μL, 1.1 equivalent), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (189.99 mg, 328.35 μmol, 0.1 equivalent), cesium carbonate (3.21 g, 9.85 mmol, 3 equivalent) and palladium acetate (73.72 mg, 328.35 μmol, 0.1 equivalent) Dissolved in dioxane (10 ml). The reaction was carried out at 100 degrees Celsius for 3 hours under a nitrogen atmosphere. It was diluted with 40 ml of water, and extracted with ethyl acetate (70 ml × 2). The organic phase was washed with 60 ml of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. Add 70 ml of ethyl acetate to dissolve and add 1 g of mercapto silica gel, and stir at 25 ° C for 1 hour. Filtration and concentration gave compound 7-12. MS (ESI) m / z: 402.1 [M + H + ].
步骤J：将氢氧化钠(179.35毫克，4.48毫摩尔，1当量)溶解于水(1.8毫升)中，25摄氏度加入乙醇(18毫升)，7-12(1.80克，4.48毫摩尔，1当量)和二甲基亚砜(525.46毫克，6.73毫摩尔，525.46微升，1.5当量)。将双氧水(762.44毫克，6.73毫摩尔，646.14微升，30％纯度，1.5当量)使用水(0.8毫升)稀释，逐滴加入反应液。25摄氏度搅拌2小时。加入100毫升水稀释，乙酸乙酯(150毫升×2)萃取，有机相用100毫升饱和食盐水洗，无水硫酸钠干燥，过滤，浓缩，制备高效液相色谱(甲酸条件)提纯得到化合物7。MS(ESI)m/z:420.1[M+H +]； Step J: Dissolve sodium hydroxide (179.35 mg, 4.48 mmol, 1 equivalent) in water (1.8 mL), add ethanol (18 mL) at 25 ° C, 7-12 (1.80 g, 4.48 mmol, 1 equivalent) And dimethyl sulfoxide (525.46 mg, 6.73 mmol, 525.46 μl, 1.5 equivalents). Hydrogen peroxide (762.44 mg, 6.73 mmol, 646.14 μL, 30% purity, 1.5 equivalents) was diluted with water (0.8 mL), and the reaction solution was added dropwise. Stir at 25 degrees Celsius for 2 hours. Dilute with 100 ml of water and extract with ethyl acetate (150 ml × 2). The organic phase was washed with 100 ml of saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by preparative high performance liquid chromatography (formic acid conditions) to obtain compound 7. MS (ESI) m / z: 420.1 [M + H + ];
1H NMR(400MHz,DMSO-d 6):δ9.11(s,1H),8.08-7.98(m,2H),7.91-7.77(m,2H),7.39-7.21(m,3H),6.47(dd,J＝2.0,6.0Hz,1H),6.29(d,J＝2.0Hz,1H),4.05(br t,J＝7.2Hz,2H),3.88-3.77(m,2H),3.29(br s,4H),2.78-2.64(m,3H),1.73-1.59(m,4H)。 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.11 (s, 1H), 8.08-7.98 (m, 2H), 7.91-7.77 (m, 2H), 7.39-7.21 (m, 3H), 6.47 ( dd, J = 2.0, 6.0 Hz, 1H), 6.29 (d, J = 2.0 Hz, 1H), 4.05 (br t, J = 7.2 Hz, 2H), 3.88-3.77 (m, 2H), 3.29 (br s , 4H), 2.78-2.64 (m, 3H), 1.73-1.59 (m, 4H).
实施例8:化合物8Example 8: Compound 8
Figure PCTCN2019114347-appb-000089
Figure PCTCN2019114347-appb-000089
步骤A：将化合物8-1(3克，26.53毫摩尔，1当量)，化合物8-2(5.74克，79.59毫摩尔，7.07毫升，3当量)，碳酸铯(17.29克，53.06毫摩尔，2当量)溶于N,N-二甲基甲酰胺中(40毫升)中，所得混合物用氮气置换三次并在100摄氏度下搅拌5小时。加入水(50毫升)淬灭反应并用乙酸乙酯(50毫升×2)萃取。合并有机相，饱和氯化钠溶液(50毫升×2)洗涤，无水硫酸钠干燥，过滤，浓缩得到化合物8-3。MS(ESI)m/z:186.2[M+H +]。 Step A: Compound 8-1 (3 g, 26.53 mmol, 1 equiv), compound 8-2 (5.74 g, 79.59 mmol, 7.07 mL, 3 equiv), cesium carbonate (17.29 g, 53.06 mmol, 2 Equivalent) was dissolved in N, N-dimethylformamide (40 mL), and the resulting mixture was replaced with nitrogen three times and stirred at 100 degrees Celsius for 5 hours. Water (50 mL) was added to quench the reaction and extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed with saturated sodium chloride solution (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 8-3. MS (ESI) m / z: 186.2 [M + H + ].
步骤B：将化合物8-3(3克，16.20毫摩尔，1当量)溶于甲醇(30毫升)中，加入湿钯碳(500毫克，10％)，用氢气置换，在25摄氏度氢气气氛下反应12小时。过滤，浓缩得到化合物8-4。Step B: Dissolve compound 8-3 (3 g, 16.20 mmol, 1 equiv) in methanol (30 mL), add wet palladium on carbon (500 mg, 10%), replace with hydrogen, under a hydrogen atmosphere at 25 ° C React for 12 hours. Filter and concentrate to give compound 8-4.
Figure PCTCN2019114347-appb-000090
Figure PCTCN2019114347-appb-000090
步骤C：将化合物1-11(0.3克，1.39毫摩尔，1当量)溶于N,N-二甲基甲酰胺(20毫升)中，往溶液中加入7-9(219.99毫克，1.67毫摩尔，1.2当量)，碳酸钾(577.86毫克，4.18毫摩尔，3当量)，所得混合物在80摄氏度下搅拌12小时。加入水(50毫升)淬灭反应。乙酸乙酯(20毫升×2)萃取，有机相用饱和氯化钠溶液(30毫升×2)洗涤，无水硫酸钠干燥，过滤，浓缩，柱分离纯化得到化合物2-5。Step C: Compound 1-11 (0.3 g, 1.39 mmol, 1 equivalent) was dissolved in N, N-dimethylformamide (20 mL), and 7-9 (219.99 mg, 1.67 mmol) was added to the solution , 1.2 equivalents), potassium carbonate (577.86 mg, 4.18 mmol, 3 equivalents), and the resulting mixture was stirred at 80 degrees Celsius for 12 hours. Water (50 mL) was added to quench the reaction. Extracted with ethyl acetate (20 ml × 2), the organic phase was washed with saturated sodium chloride solution (30 ml × 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column separation to obtain compound 2-5.
MS(ESI)m/z:327.1[M+H +]。 MS (ESI) m / z: 327.1 [M + H + ].
步骤D：将化合物2-5(100毫克，0.306毫摩尔，1当量)和化合物8-4(49.87毫克，0.321毫摩尔，1.05当量)，二(二亚苄基丙酮)钯(17.60毫克，30.6微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(35.41毫克，61.20微摩尔，0.2当量)和碳酸铯(199.41毫克，612.03微摩尔，2当量)溶于1,4-二氧六环(15毫升)中。该体系用氮气置换，然后加热到100摄氏度，在氮气氛围下反应12小时。加入水(50毫升)淬灭反应，乙酸乙酯(30毫升×2)萃取。有机相用饱和氯化钠溶液(30毫升×2)洗涤，无水硫酸钠干燥，过滤，浓缩，制备级高效液相色谱(盐酸条件)纯化得到化合物8。MS(ESI)m/z:446.4[M+H +]； Step D: Compound 2-5 (100 mg, 0.306 mmol, 1 equivalent) and compound 8-4 (49.87 mg, 0.321 mmol, 1.05 equivalent), bis (dibenzylideneacetone) palladium (17.60 mg, 30.6 Micromolar, 0.1 equivalent), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (35.41 mg, 61.20 micromolar, 0.2 equivalent) and cesium carbonate (199.41 mg, 612.03 micromolar) , 2 equivalents) dissolved in 1,4-dioxane (15 ml). The system was replaced with nitrogen, then heated to 100 degrees Celsius, and reacted for 12 hours under a nitrogen atmosphere. Water (50 mL) was added to quench the reaction, and ethyl acetate (30 mL × 2) was extracted. The organic phase was washed with a saturated sodium chloride solution (30 ml × 2), dried over anhydrous sodium sulfate, filtered, and concentrated, and purified by preparative high-performance liquid chromatography (hydrochloric acid condition) to obtain compound 8. MS (ESI) m / z: 446.4 [M + H + ];
1H NMR(400MHz,DMSO-d 6):δ10.49(s,1H),8.16–8.14(m,1H),7.97–7.95(m,1H),7.86(s,1H),7.81(d,J＝7.6Hz,1H),7.53(d,J＝7.2Hz,1H),7.48(s,1H),6.86–6.83(m,1H),6.72(s,1H),4.30–4.19(m,2H),4.01(s,1H),2.96–2.93(m,2H),2.67–2.62(m,2H),2.33(s,3H),1.08(s,6H)。 1 H NMR (400 MHz, DMSO-d 6 ): δ 10.49 (s, 1H), 8.16–8.14 (m, 1H), 7.97–7.95 (m, 1H), 7.86 (s, 1H), 7.81 (d, J = 7.6Hz, 1H), 7.53 (d, J = 7.2Hz, 1H), 7.48 (s, 1H), 6.86-6.83 (m, 1H), 6.72 (s, 1H), 4.30-4.19 (m, 2H ), 4.01 (s, 1H), 2.96–2.93 (m, 2H), 2.67–2.62 (m, 2H), 2.33 (s, 3H), 1.08 (s, 6H).
实施例9：化合物9的甲酸盐Example 9: Formate salt of compound 9
Figure PCTCN2019114347-appb-000091
Figure PCTCN2019114347-appb-000091
步骤A：向化合物2-5(200毫克，612.03微摩尔，1当量)和化合物9-1(120.27毫克，795.64微摩尔，1.3当量)的1,4-二氧六环(8毫升)溶液中加入双(二亚苄基丙酮)钯(35.19毫克，61.20微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(35.41毫克，61.20微摩尔，0.1当量)和碳酸铯(398.82毫克，1.22毫摩尔，2当量)。该体系用氮气置换，然后加热到100摄氏度反应12小时。将反应液倒入水(80毫升)中并搅拌3分钟。水相用乙酸乙酯(80毫升×2)萃取，合并有机相，用饱和食盐水(80毫升)洗涤，无水硫酸钠干燥，过滤，浓缩得到化合物9-2。MS(ESI)m/z:442.3[M+H +]。 Step A: To a solution of compound 2-5 (200 mg, 612.03 micromolar, 1 equivalent) and compound 9-1 (120.27 mg, 795.64 micromolar, 1.3 equivalent) in 1,4-dioxane (8 mL) Add bis (dibenzylideneacetone) palladium (35.19 mg, 61.20 μmol, 0.1 equivalent), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (35.41 mg, 61.20 μm) Molar, 0.1 equivalent) and cesium carbonate (398.82 mg, 1.22 mmol, 2 equivalent). The system was replaced with nitrogen and then heated to 100 degrees Celsius for 12 hours. The reaction solution was poured into water (80 ml) and stirred for 3 minutes. The aqueous phase was extracted with ethyl acetate (80 mL × 2), and the organic phases were combined, washed with saturated brine (80 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 9-2. MS (ESI) m / z: 442.3 [M + H + ].
步骤B：在-78摄氏度，将氨气通入乙二醇(15毫升)中，保持通气5分钟。将化合物9-2(390毫克，883.39微摩尔，1当量)溶于上述溶液中。再将该混合液倒入聚四氟乙烯闷罐中，加热到80摄氏度并搅拌12小时。将反应液倒入水(60毫升)中并搅拌3分钟。水相用乙酸乙酯(60毫升×2)萃取，合并有机相，无水硫酸钠干燥，过滤，浓缩，制备高效液相色谱(甲酸体系)提纯得到化合物9的甲酸盐。Step B: At -78 degrees Celsius, ammonia gas was bubbled into ethylene glycol (15 ml) and kept aerated for 5 minutes. Compound 9-2 (390 mg, 883.39 micromolar, 1 equivalent) was dissolved in the above solution. Then, the mixed solution was poured into a Teflon pot, heated to 80 degrees Celsius and stirred for 12 hours. The reaction solution was poured into water (60 ml) and stirred for 3 minutes. The aqueous phase was extracted with ethyl acetate (60 mL × 2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated, and purified by preparative high performance liquid chromatography (formic acid system) to obtain the formate salt of compound 9.
MS(ESI)m/z:427.3[M+H +]； MS (ESI) m / z: 427.3 [M + H + ];
1H NMR(400MHz,DMSO-d 6):δ9.01(s,1H),8.24(s,1H),8.05-7.98(m,2H),7.88-7.75(m,2H),7.63(t,J＝8.0Hz,1H),7.54(d,J＝8.0Hz,1H),7.34-7.21(m,3H),7.05(d,J＝7.2Hz,1H),6.49(dd,J＝6.0Hz,2.0Hz,1H),6.29(s,1H),4.19(t,J＝7.2Hz,2H),2.86(t,J＝7.2Hz,2H),2.68-2.54(m,2H),2.25(s,3H)。 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.01 (s, 1H), 8.24 (s, 1H), 8.05-7.98 (m, 2H), 7.88-7.75 (m, 2H), 7.63 (t, J = 8.0Hz, 1H), 7.54 (d, J = 8.0Hz, 1H), 7.34-7.21 (m, 3H), 7.05 (d, J = 7.2Hz, 1H), 6.49 (dd, J = 6.0Hz, 2.0Hz, 1H), 6.29 (s, 1H), 4.19 (t, J = 7.2Hz, 2H), 2.86 (t, J = 7.2Hz, 2H), 2.68-2.54 (m, 2H), 2.25 (s, 3H).
实施例10：化合物10Example 10: Compound 10
Figure PCTCN2019114347-appb-000092
Figure PCTCN2019114347-appb-000092
Figure PCTCN2019114347-appb-000093
Figure PCTCN2019114347-appb-000093
步骤A：向化合物2-5(100毫克，306.02微摩尔，1当量)和化合物10-1(60.66毫克，367.22微摩尔，1.2当量)的1,4-二氧六环(3毫升)溶液中加入双(二亚苄基丙酮)钯(17.6毫克，30.6微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(17.71毫克，30.60微摩尔，0.1当量)和碳酸铯(199.41毫克，612.03微摩尔，2当量)。该体系用氮气置换，加热到65摄氏度反应4小时，然后加热到100摄氏度反应12小时。加入水(20毫升)稀释，乙酸乙酯(20毫升×3)萃取，合并有机相，用2摩尔每升盐酸(10毫升×4)萃取，合并水相，用4摩尔每升的氢氧化钠水溶液调节pH至7，再用乙酸乙酯(20毫升×3)萃取。合并有机相，用饱和食盐水(20毫升)洗涤，无水硫酸钠干燥，过滤，浓缩得到化合物10-2。Step A: To a solution of compound 2-5 (100 mg, 306.02 micromolar, 1 equivalent) and compound 10-1 (60.66 mg, 367.22 micromolar, 1.2 equivalent) in 1,4-dioxane (3 mL) Add bis (dibenzylideneacetone) palladium (17.6 mg, 30.6 μmol, 0.1 equivalent), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (17.71 mg, 30.60 μm) Molar, 0.1 equivalent) and cesium carbonate (199.41 mg, 612.03 micromolar, 2 equivalent). The system was replaced with nitrogen, heated to 65 degrees Celsius for 4 hours, and then heated to 100 degrees Celsius for 12 hours. Diluted with water (20 ml), extracted with ethyl acetate (20 ml x 3), combined organic phases, extracted with 2 mol per liter hydrochloric acid (10 ml x 4), combined aqueous phase, combined with 4 mol per liter sodium hydroxide The aqueous solution was adjusted to pH 7, and extracted with ethyl acetate (20 ml × 3). The organic phases were combined, washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 10-2.
MS(ESI)m/z:456.1[M+H +]。 MS (ESI) m / z: 456.1 [M + H + ].
步骤B：在-78摄氏度下，向10毫升乙二醇中鼓泡氨气15分钟，然后加入化合物10-2(147.54毫克，323.91微摩尔，1.2当量)，将此溶液加入闷罐中，在100摄氏度下反应12小时后，在25摄氏度下加入20毫升水和50毫升乙酸乙酯，用乙酸乙酯萃取两次，每次50毫升，合并有机相，饱和食盐水(25毫升×2)洗，无水硫酸钠干燥，过滤，浓缩，制备级高效液相色谱(碱性)纯化得到化合物10。Step B: At -78 degrees Celsius, ammonia gas was bubbled into 10 ml of ethylene glycol for 15 minutes, and then compound 10-2 (147.54 mg, 323.91 micromolar, 1.2 equivalents) was added, and the solution was added to the stuffy tank After reacting at 100 degrees Celsius for 12 hours, add 20 ml of water and 50 ml of ethyl acetate at 25 degrees Celsius, extract twice with ethyl acetate, 50 ml each time, combine organic phases, wash with saturated brine (25 ml × 2) , Dried over anhydrous sodium sulfate, filtered, concentrated, purified by preparative high performance liquid chromatography (basic) to obtain compound 10.
MS(ESI)m/z:427.2[M+H +]； MS (ESI) m / z: 427.2 [M + H + ];
1H NMR(400MHz,DMSO-d 6)δ＝9.01(s,1H),8.02(d,J＝6.0Hz,1H),7.73(t,J＝1.6Hz,1H),7.65-7.61(m,1H),7.55-7.52(m,2H),7.24(t,J＝8.0Hz,1H),7.04(d,J＝7.2Hz,1H),6.82(d,J＝7.6Hz,1H),6.50(dd,J＝2.4,6.0Hz,1H),6.27(d,J＝2.0Hz,1H),4.19(t,J＝7.2Hz,2H),3.00-2.84(m,8H),2.63-2.55(m,2H),2.24(s,3H)。 1 H NMR (400 MHz, DMSO-d 6 ) δ = 9.01 (s, 1H), 8.02 (d, J = 6.0 Hz, 1H), 7.73 (t, J = 1.6 Hz, 1H), 7.65-7.61 (m, 1H), 7.55-7.52 (m, 2H), 7.24 (t, J = 8.0Hz, 1H), 7.04 (d, J = 7.2Hz, 1H), 6.82 (d, J = 7.6Hz, 1H), 6.50 ( dd, J = 2.4, 6.0 Hz, 1H), 6.27 (d, J = 2.0 Hz, 1H), 4.19 (t, J = 7.2 Hz, 2H), 3.00-2.84 (m, 8H), 2.63-2.55 (m , 2H), 2.24 (s, 3H).
实施例11：化合物11Example 11: Compound 11
Figure PCTCN2019114347-appb-000094
Figure PCTCN2019114347-appb-000094
将化合物2-5(200毫克，612.03微毫摩尔，1当量)溶于溶于1,4-二氧六环(5毫升)中，依次加入化合物11-1(150.75毫克，918.05微摩尔，1.5当量)，二(二亚苄基丙酮)钯(35.19毫克，61.20微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(70.83毫克，122.41微摩尔，0.2当量)，碳酸铯(598.24 毫克，1.84毫摩尔，3当量)。置换氮气三次，然后在氮气保护下于100摄氏度搅拌2小时。冷却至25摄氏度，加入1克硅胶粉，搅拌15分钟后硅藻土垫层过滤。滤液用1摩尔每升的盐酸调节pH至4，加入水(20毫升)稀释，乙酸乙酯(20毫升×3)洗涤。水相用1摩尔每升的氢氧化钠水溶液调节pH至9，乙酸乙酯(20毫升×3)萃取，合并有机相，饱和食盐水(20毫升)洗，无水硫酸钠干燥，浓缩，制备高效液相色谱提纯到化合物11。MS(ESI)m/z:455.3[M+H +]； Compound 2-5 (200 mg, 612.03 μmole, 1 equivalent) was dissolved in 1,4-dioxane (5 mL), and compound 11-1 (150.75 mg, 918.05 μmol, 1.5 Equivalent), bis (dibenzylideneacetone) palladium (35.19 mg, 61.20 μmol, 0.1 equivalent), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (70.83 mg, 122.41 micromolar, 0.2 equivalent), cesium carbonate (598.24 mg, 1.84 mmol, 3 equivalent). Nitrogen was replaced three times, and then stirred at 100 degrees Celsius for 2 hours under nitrogen protection. Cooled to 25 degrees Celsius, added 1 gram of silica gel powder, stirred for 15 minutes and filtered through a pad of diatomaceous earth. The filtrate was adjusted to pH 4 with 1 molar hydrochloric acid, diluted with water (20 mL), and washed with ethyl acetate (20 mL × 3). The aqueous phase was adjusted to pH 9 with 1 mol per liter of sodium hydroxide aqueous solution, extracted with ethyl acetate (20 mL × 3), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated to prepare HPLC purified to compound 11. MS (ESI) m / z: 455.3 [M + H + ];
1H NMR(400MHz,DMSO-d 6)δ＝9.01(s,1H),8.02(d,J＝6.0Hz,1H),7.73(t,J＝1.6Hz,1H),7.65-7.61(m,1H),7.55-7.52(m,2H),7.24(t,J＝8.0Hz,1H),7.04(d,J＝7.2Hz,1H),6.82(d,J＝7.6Hz,1H),6.50(dd,J＝2.4,6.0Hz,1H),6.27(d,J＝2.0Hz,1H),4.19(t,J＝7.2Hz,2H),3.00-2.84(m,8H),2.63-2.55(m,2H),2.24(s,3H)。 1 H NMR (400 MHz, DMSO-d 6 ) δ = 9.01 (s, 1H), 8.02 (d, J = 6.0 Hz, 1H), 7.73 (t, J = 1.6 Hz, 1H), 7.65-7.61 (m, 1H), 7.55-7.52 (m, 2H), 7.24 (t, J = 8.0Hz, 1H), 7.04 (d, J = 7.2Hz, 1H), 6.82 (d, J = 7.6Hz, 1H), 6.50 ( dd, J = 2.4, 6.0 Hz, 1H), 6.27 (d, J = 2.0 Hz, 1H), 4.19 (t, J = 7.2 Hz, 2H), 3.00-2.84 (m, 8H), 2.63-2.55 (m , 2H), 2.24 (s, 3H).
实施例12：化合物12Example 12: Compound 12
Figure PCTCN2019114347-appb-000095
Figure PCTCN2019114347-appb-000095
将化合物2-5(200毫克，612.03微毫摩尔，1当量)溶于溶于1,4-二氧六环(5毫升)中，依次加入化合物12-1(113.06毫克，918.05微摩尔，1.5当量)，二(二亚苄基丙酮)钯(35.19毫克，61.20微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(70.83毫克，122.41微摩尔，0.2当量)，碳酸铯(598.24毫克，1.84毫摩尔，3当量)。置换氮气三次，然后在氮气保护下于100摄氏度搅拌12小时。冷却至25摄氏度，加入1克硅胶粉，搅拌15分钟后用硅藻土垫层过滤。滤液用1摩尔每升的盐酸调节pH至4，加入水(20毫升)稀释，乙酸乙酯(20毫升×3)洗涤。水相用1摩尔每升的氢氧化钠水溶液调节pH至9，乙酸乙酯(20毫升×3)萃取，合并有机相，饱和食盐水(20毫升)洗涤，无水硫酸钠干燥，浓缩，制备高效液相色谱提纯得到化合物12。MS(ESI)m/z:414.4[M+H +]； Compound 2-5 (200 mg, 612.03 μmole, 1 equivalent) was dissolved in 1,4-dioxane (5 mL), and compound 12-1 (113.06 mg, 918.05 μmol, 1.5 Equivalent), bis (dibenzylideneacetone) palladium (35.19 mg, 61.20 μmol, 0.1 equivalent), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (70.83 mg, 122.41 micromolar, 0.2 equivalent), cesium carbonate (598.24 mg, 1.84 mmol, 3 equivalent). Nitrogen was replaced three times, and then stirred at 100 degrees Celsius for 12 hours under nitrogen protection. Cool to 25 degrees Celsius, add 1 gram of silica gel powder, stir for 15 minutes and filter through a pad of celite. The filtrate was adjusted to pH 4 with 1 molar hydrochloric acid, diluted with water (20 mL), and washed with ethyl acetate (20 mL × 3). The aqueous phase was adjusted to pH 9 with 1 mol per liter of sodium hydroxide aqueous solution, extracted with ethyl acetate (20 mL × 3), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated to prepare High performance liquid chromatography purification to obtain compound 12. MS (ESI) m / z: 414.4 [M + H + ];
1H NMR(400MHz,DMSO-d 6)δ＝8.85(s,1H),8.00(d,J＝5.6Hz,1H),7.63(t,J＝7.6Hz,1H),7.54(d,J＝7.6Hz,1H),7.33-7.32(m,1H),7.11-7.02(m,3H),6.47(dd,J＝2.0,6.0Hz,1H),6.44-6.39(m,1H),6.26(d,J＝2.0Hz,1H),4.19(t,J＝7.2Hz,2H),3.68(s,3H),2.85(d,J＝7.2Hz,2H),2.63-2.55(m,2H),2.25(s,3H)。 1 H NMR (400 MHz, DMSO-d 6 ) δ = 8.85 (s, 1H), 8.00 (d, J = 5.6 Hz, 1H), 7.63 (t, J = 7.6 Hz, 1H), 7.54 (d, J = 7.6Hz, 1H), 7.33-7.32 (m, 1H), 7.11-7.02 (m, 3H), 6.47 (dd, J = 2.0, 6.0Hz, 1H), 6.44-6.39 (m, 1H), 6.26 (d , J = 2.0 Hz, 1H), 4.19 (t, J = 7.2 Hz, 2H), 3.68 (s, 3H), 2.85 (d, J = 7.2 Hz, 2H), 2.63-2.55 (m, 2H), 2.25 (s, 3H).
实施例13：化合物13Example 13: Compound 13
Figure PCTCN2019114347-appb-000096
Figure PCTCN2019114347-appb-000096
Figure PCTCN2019114347-appb-000097
Figure PCTCN2019114347-appb-000097
将化合物2-5(4.7克，14.38毫摩尔，1当量)、化合物7-11(1.87克，15.82毫摩尔，1.1当量)、醋酸钯(322.90毫克，1.44毫摩尔，0.1当量)、4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(832.21毫克，1.44毫摩尔，0.1当量)和碳酸铯(14.06克，43.15毫摩尔，3当量)加入到1,4二氧六环(50毫升)中。在100摄氏度和氮气保护下搅拌10小时。将体系温度降至室温，加入100毫升水稀释，乙酸乙酯(100毫升×3)萃取。合并有机相，饱和食盐水(100毫升×3)洗涤，无水硫酸钠干燥，浓缩，制备高效液相色谱(碱性体系)纯化得到化合物13。MS(ESI)m/z:409.1[M+H +]； Compound 2-5 (4.7 g, 14.38 mmol, 1 equiv), compound 7-11 (1.87 g, 15.82 mmol, 1.1 equiv), palladium acetate (322.90 mg, 1.44 mmol, 0.1 equiv), 4,5 -Bis (diphenylphosphine) -9,9-dimethylxanthene (832.21 mg, 1.44 mmol, 0.1 equiv) and cesium carbonate (14.06 g, 43.15 mmol, 3 equiv) were added to 1,4 di Oxygen ring (50 ml). Stir at 100 degrees Celsius under nitrogen for 10 hours. The temperature of the system was reduced to room temperature, diluted with 100 ml of water, and extracted with ethyl acetate (100 ml × 3). The organic phases were combined, washed with saturated brine (100 ml × 3), dried over anhydrous sodium sulfate, concentrated, and purified by preparative high performance liquid chromatography (basic system) to obtain compound 13. MS (ESI) m / z: 409.1 [M + H + ];
1H NMR(400MHz,DMSO-d 6)δ＝9.28(s,1H),8.30(s,1H),8.09(d,J＝6.0Hz,1H),7.73(dd,J＝1.2,8.4Hz,1H),7.67-7.60(m,1H),7.59-7.53(m,1H),7.41(s,1H),7.26(d,J＝7.6Hz,1H),7.05(d,J＝7.6Hz,1H),6.59(dd,J＝2.2,5.9Hz,1H),6.29(d,J＝2.2Hz,1H),4.25-4.16(m,2H),2.91-2.83(m,2H),2.65-2.58(m,2H),2.23(s,3H)。 1 H NMR (400 MHz, DMSO-d 6 ) δ = 9.28 (s, 1H), 8.30 (s, 1H), 8.09 (d, J = 6.0 Hz, 1H), 7.73 (dd, J = 1.2, 8.4 Hz, 1H), 7.67-7.60 (m, 1H), 7.59-7.53 (m, 1H), 7.41 (s, 1H), 7.26 (d, J = 7.6Hz, 1H), 7.05 (d, J = 7.6Hz, 1H) ), 6.59 (dd, J = 2.2, 5.9 Hz, 1H), 6.29 (d, J = 2.2 Hz, 1H), 4.25-4.16 (m, 2H), 2.91-2.83 (m, 2H), 2.65-2.58 ( m, 2H), 2.23 (s, 3H).
实施例14：化合物14Example 14: Compound 14
Figure PCTCN2019114347-appb-000098
Figure PCTCN2019114347-appb-000098
将化合物2-5(200毫克，612.03微毫摩尔，1当量)溶于溶于1,4-二氧六环(5毫升)中，依次加入化合物14-1(150.75毫克，918.05微摩尔，1.5当量)，二(二亚苄基丙酮)钯(35.19毫克，61.20微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(70.83毫克，122.41微摩尔，0.2当量)，碳酸铯(598.24毫克，1.84毫摩尔，3当量)。置换氮气三次，然后在氮气保护下于100摄氏度搅拌12小时。冷却至25摄氏度，加入1克硅胶粉，搅拌15分钟后硅藻土垫层过滤。滤液用1摩尔每升的盐酸调节pH至4，加入水(20毫升)稀释，乙酸乙酯(20毫升×3)洗涤。水相用1摩尔每升的氢氧化钠水溶液调节pH至9，乙酸乙酯(20毫升×3)萃取，合并有机相，用饱和食盐水(20毫升)洗涤，无水硫酸钠干燥，浓缩，制备高效液相色谱提纯得到化合物14。MS(ESI)m/z:455.3[M+H +]； Compound 2-5 (200 mg, 612.03 μmole, 1 equivalent) was dissolved in 1,4-dioxane (5 mL), and compound 14-1 (150.75 mg, 918.05 μmol, 1.5 Equivalent), bis (dibenzylideneacetone) palladium (35.19 mg, 61.20 μmol, 0.1 equivalent), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (70.83 mg, 122.41 micromolar, 0.2 equivalent), cesium carbonate (598.24 mg, 1.84 mmol, 3 equivalent). Nitrogen was replaced three times, and then stirred at 100 degrees Celsius for 12 hours under nitrogen protection. Cooled to 25 degrees Celsius, added 1 gram of silica gel powder, stirred for 15 minutes and filtered through a pad of diatomaceous earth. The filtrate was adjusted to pH 4 with 1 molar hydrochloric acid, diluted with water (20 mL), and washed with ethyl acetate (20 mL × 3). The aqueous phase was adjusted to pH 9 with 1 mol of sodium hydroxide aqueous solution, extracted with ethyl acetate (20 ml × 3), the organic phases were combined, washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, and concentrated Preparative high performance liquid chromatography was purified to obtain compound 14. MS (ESI) m / z: 455.3 [M + H + ];
1H NMR(400MHz,DMSO-d 6)δ＝9.10(s,1H),8.03(d,J＝6.0Hz,1H),7.65-7.61(m,3H),7.5(d,J＝8.0Hz,1H),7.28(d,J＝8.8Hz,2H),7.04(d,J＝7.6Hz,1H),6.52(dd,J＝2.4,6.0Hz,1H),6.29(d,J＝2.4Hz,1H),4.19(t,J＝7.2Hz,2H),2.94(s,6H),2.86(t,J＝7.2Hz,2H),2.63-2.56(m,2H),2.23(s,3H)。 1 H NMR (400 MHz, DMSO-d 6 ) δ = 9.10 (s, 1H), 8.03 (d, J = 6.0 Hz, 1H), 7.65-7.61 (m, 3H), 7.5 (d, J = 8.0 Hz, 1H), 7.28 (d, J = 8.8 Hz, 2H), 7.04 (d, J = 7.6 Hz, 1H), 6.52 (dd, J = 2.4, 6.0 Hz, 1H), 6.29 (d, J = 2.4 Hz, 1H), 4.19 (t, J = 7.2 Hz, 2H), 2.94 (s, 6H), 2.86 (t, J = 7.2 Hz, 2H), 2.63-2.56 (m, 2H), 2.23 (s, 3H).
实施例15：化合物15Example 15: Compound 15
Figure PCTCN2019114347-appb-000099
Figure PCTCN2019114347-appb-000099
步骤A：向化合物2-5(200毫升，612.03微摩尔，1当量)和化合物15-1(121.06毫克,795.64微摩尔,1.3当量)的1,4-二氧六环(8毫升)溶液中加入双(二亚苄基丙酮)钯(35.19毫克,61.20微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(35.41毫克，61.20微摩尔，0.1当量)和碳酸铯(398.82毫克，1.22毫摩尔，2当量)。该体系用氮气置换，然后加热到90摄氏度反应12小时。将反应液倒入水(60毫升)中，水相用乙酸乙酯(80毫升)萃取，再将水相浓缩得到化合物15-2。MS(ESI)m/z:429.1[M+H +]。 Step A: To a solution of compound 2-5 (200 mL, 612.03 micromolar, 1 equivalent) and compound 15-1 (121.06 mg, 795.64 micromolar, 1.3 equivalent) in 1,4-dioxane (8 mL) Add bis (dibenzylideneacetone) palladium (35.19 mg, 61.20 μmol, 0.1 equivalent), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (35.41 mg, 61.20 μm) Molar, 0.1 equivalent) and cesium carbonate (398.82 mg, 1.22 mmol, 2 equivalent). The system was replaced with nitrogen, and then heated to 90 degrees Celsius for 12 hours. The reaction solution was poured into water (60 mL), the aqueous phase was extracted with ethyl acetate (80 mL), and the aqueous phase was concentrated to obtain compound 15-2. MS (ESI) m / z: 429.1 [M + H + ].
步骤B：向化合物15-2(200毫克,466.81微摩尔,1当量)的二氯甲烷(3毫升)溶液中加入草酰氯(177.75毫克,1.40毫摩尔,122.59微升,3当量)，反应1小时后，将该混合物加入到搅拌的氨水(5.60克,47.94毫摩尔,6.15毫升,30％,102.69当量)中，并在25摄氏度搅拌15分钟。将反应液倒入冰水(60毫升)中，用乙酸乙酯(60毫升×2)萃取，合并有机相，饱和食盐水(60毫升)洗，无水硫酸钠干燥，过滤，浓缩，制备高效液相色谱(中性)分离得到化合物15。MS(ESI)m/z:428.4[M+H +]； Step B: To a solution of compound 15-2 (200 mg, 466.81 micromolar, 1 equivalent) in dichloromethane (3 mL) was added oxalyl chloride (177.75 mg, 1.40 mmol, 122.59 microliter, 3 equivalent), reaction 1 After hours, the mixture was added to stirred ammonia water (5.60 g, 47.94 mmol, 6.15 mL, 30%, 102.69 equiv), and stirred at 25 degrees Celsius for 15 minutes. The reaction solution was poured into ice water (60 mL), extracted with ethyl acetate (60 mL × 2), the organic phases were combined, washed with saturated brine (60 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and prepared efficiently Liquid chromatographic (neutral) separation gave compound 15. MS (ESI) m / z: 428.4 [M + H + ];
1H NMR(400MHz,DMSO-d 6)δ＝9.59(s,1H),8.29(d,J＝5.6Hz,1H),8.23(s,1H),8.14(d,J＝5.6Hz,1H),7.98(d,J＝2.8Hz,1H),7.81(d,J＝5.6Hz,1H),7.65-7.60(m,1H),7.58-7.48(m,2H),7.04(d,J＝7.2Hz,1H),6.65(d,J＝6.0Hz,1H),6.38(s,1H),4.20(t,J＝7.2Hz,2H),2.87(t,J＝7.2Hz,2H),2.60(t,J＝7.2Hz,2H),2.21(s,3H)。 1 H NMR (400 MHz, DMSO-d 6 ) δ = 9.59 (s, 1H), 8.29 (d, J = 5.6 Hz, 1H), 8.23 (s, 1H), 8.14 (d, J = 5.6 Hz, 1H) , 7.98 (d, J = 2.8 Hz, 1H), 7.81 (d, J = 5.6 Hz, 1H), 7.65-7.60 (m, 1H), 7.58-7.48 (m, 2H), 7.04 (d, J = 7.2 Hz, 1H), 6.65 (d, J = 6.0 Hz, 1H), 6.38 (s, 1H), 4.20 (t, J = 7.2 Hz, 2H), 2.87 (t, J = 7.2 Hz, 2H), 2.60 ( t, J = 7.2 Hz, 2H), 2.21 (s, 3H).
实施例16：化合物16Example 16: Compound 16
Figure PCTCN2019114347-appb-000100
Figure PCTCN2019114347-appb-000100
Figure PCTCN2019114347-appb-000101
Figure PCTCN2019114347-appb-000101
步骤A:向化合物1-11(300毫克，1.39毫摩尔，1当量)的N,N-二甲基甲酰胺(10毫升)溶液中加入碳酸钾(385.24毫克，2.79毫摩尔，2当量)和化合物16-1(249.16毫克，1.67毫摩尔，1.2当量)，在80摄氏度下反应12小时后，加入40毫升水，过滤，滤饼干燥得到化合物16-2。MS(ESI)m/z:328.1[M+H +]。 Step A: To a solution of compound 1-11 (300 mg, 1.39 mmol, 1 equiv) in N, N-dimethylformamide (10 mL) was added potassium carbonate (385.24 mg, 2.79 mmol, 2 equiv) and Compound 16-1 (249.16 mg, 1.67 mmol, 1.2 equiv) was reacted at 80 degrees Celsius for 12 hours, 40 ml of water was added, filtered, and the filter cake was dried to obtain compound 16-2. MS (ESI) m / z: 328.1 [M + H + ].
步骤B:向化合物16-2(350毫克，1.07毫摩尔，1当量)和7-11(138.76毫克，1.17毫摩尔，1.1当量)的1,4-二氧六环(10毫升)溶液中加入碳酸铯(1.04克，3.20毫摩尔，3当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(123.57毫克，213.57微摩尔，0.2当量)和三(二亚苄基丙酮)二钯(97.78毫克，106.78微摩尔，0.1当量)，在氮气保护和100摄氏度下反应12小时后，加入40毫升乙酸乙酯，过滤，滤液用饱和氯化钠溶液(40毫升×3)洗涤，将有机相加入到20毫升1摩尔每升盐酸中，分液得到的水相用乙酸乙酯(40毫升)洗，然后用饱和碳酸钠溶液将水相pH调节到9，用40毫升乙酸乙酯萃取一次，有机相用食盐水(40毫升×2)洗涤，无水硫酸钠干燥，浓缩得到化合物16-3。MS(ESI)m/z:410.2[M+H +]。 Step B: To a solution of compound 16-2 (350 mg, 1.07 mmol, 1 equivalent) and 7-11 (138.76 mg, 1.17 mmol, 1.1 equivalent) in 1,4-dioxane (10 mL) was added Cesium carbonate (1.04 g, 3.20 mmol, 3 equivalents), 4,5-bis (diphenylphosphine) -9,9-dimethylxanthene (123.57 mg, 213.57 micromolar, 0.2 equivalents) and tri ( Dibenzylideneacetone) dipalladium (97.78 mg, 106.78 micromolar, 0.1 equiv), after reacting under nitrogen atmosphere and at 100 degrees Celsius for 12 hours, 40 ml of ethyl acetate was added, filtered, and the filtrate was saturated with sodium chloride solution Ml × 3) washed, the organic phase was added to 20 ml of 1 mol per liter of hydrochloric acid, the aqueous phase obtained by separation was washed with ethyl acetate (40 ml), and then the pH of the aqueous phase was adjusted to 9 with saturated sodium carbonate solution. It was extracted once with 40 ml of ethyl acetate, and the organic phase was washed with brine (40 ml × 2), dried over anhydrous sodium sulfate, and concentrated to give compound 16-3. MS (ESI) m / z: 410.2 [M + H + ].
步骤C:向化合物16-3(350毫克，854.82微摩尔，1当量)的乙醇(5毫升)和水(5毫升)溶液中加入二甲基亚砜(133.58毫克，1.71毫摩尔，133.58微升，2当量)和氢氧化钠(68.38毫克，1.71毫摩尔，2当量)，然后加入双氧水(166.15毫克，1.71毫摩尔，140.81微升，35％，2当量)，在25摄氏度下反应12小时后，加入乙酸乙酯(20毫升)，有机相用饱和氯化钠溶液(20毫升×3)洗涤，无水硫酸钠干燥，过滤，浓缩，制备级高效液相色谱(甲酸体系)纯化得到化合物16。MS(ESI)m/z:428.2[M+H +]； Step C: To a solution of compound 16-3 (350 mg, 854.82 micromolar, 1 equivalent) in ethanol (5 ml) and water (5 ml) was added dimethyl sulfoxide (133.58 mg, 1.71 mmol, 133.58 μl , 2 equiv.) And sodium hydroxide (68.38 mg, 1.71 mmol, 2 equiv.), Then add hydrogen peroxide (166.15 mg, 1.71 mmol, 140.81 μl, 35%, 2 equiv.), And react at 25 degrees Celsius for 12 hours. ， Add ethyl acetate (20 ml), the organic phase was washed with saturated sodium chloride solution (20 ml × 3), dried over anhydrous sodium sulfate, filtered, concentrated, purified by preparative high performance liquid chromatography (formic acid system) to obtain compound 16 . MS (ESI) m / z: 428.2 [M + H + ];
1H NMR(400MHz,DMSO-d 6):δ9.56(s,1H),8.32(d,J＝5.6Hz,1H),7.99(s,1H),7.80(s,1H),7.72(d,J＝8.0Hz,1H),7.63-7.52(m,2H),7.36(d,J＝7.6Hz,1H),7.28(s,1H),7.20(t,J＝8.0Hz,1H),,7.02(d,J＝7.2Hz,1H),6.48(d,J＝5.6Hz,1H),4.20(t,J＝7.2Hz,2H),2.87(br t,J＝7.2Hz,2H),2.61-2.52(m,2H),2.21(s,3H)。 1 H NMR (400 MHz, DMSO-d 6 ): δ 9.56 (s, 1H), 8.32 (d, J = 5.6 Hz, 1H), 7.99 (s, 1H), 7.80 (s, 1H), 7.72 (d , J = 8.0Hz, 1H), 7.63-7.52 (m, 2H), 7.36 (d, J = 7.6Hz, 1H), 7.28 (s, 1H), 7.20 (t, J = 8.0Hz, 1H) ,, 7.02 (d, J = 7.2 Hz, 1H), 6.48 (d, J = 5.6 Hz, 1H), 4.20 (t, J = 7.2 Hz, 2H), 2.87 (br t, J = 7.2 Hz, 2H), 2.61 -2.52 (m, 2H), 2.21 (s, 3H).
实施例17：化合物17的甲酸盐Example 17: Formate salt of compound 17
Figure PCTCN2019114347-appb-000102
Figure PCTCN2019114347-appb-000102
Figure PCTCN2019114347-appb-000103
Figure PCTCN2019114347-appb-000103
步骤A：向化合物1(3克，9.05毫摩尔，1当量)，三乙胺(1.10克，10.86毫摩尔，1.51毫升，1.2当量)和4-二甲氨基吡啶(55.30毫克，452.67微摩尔，0.05当量)的二氯甲烷(20毫升)溶液中，加入苯磺酰氯(1.92克，10.86毫摩尔，1.39毫升，1.2当量)。在25摄氏度下反应12小时。加入水(100毫升)，二氯甲烷(300毫升×3)萃取。合并有机相，饱和食盐水(300毫升×2)洗，无水硫酸钠干燥，过滤，浓缩，柱分离纯化得到化合物17-1。MS(ESI)m/z:472.2[M+H +]。 Step A: To compound 1 (3 g, 9.05 mmol, 1 equiv), triethylamine (1.10 g, 10.86 mmol, 1.51 mL, 1.2 equiv) and 4-dimethylaminopyridine (55.30 mg, 452.67 μmol, To a solution of 0.05 equivalent) in dichloromethane (20 mL), benzenesulfonyl chloride (1.92 g, 10.86 mmol, 1.39 mL, 1.2 equivalent) was added. React for 12 hours at 25 degrees Celsius. Water (100 ml) was added, and dichloromethane (300 ml × 3) was extracted. The organic phases were combined, washed with saturated brine (300 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column separation to obtain compound 17-1. MS (ESI) m / z: 472.2 [M + H + ].
步骤B：将化合物17-1(100毫克，212.08微摩尔，1当量)溶解于四氢呋喃(5毫升)，氮气气氛下，-78摄氏度逐滴添加二异丙基氨基锂(2摩尔每升，159.06微升，318.11微摩尔，1.5当量)。保持-78摄氏度反应1小时，在此温度下加入碘(80.74毫克，318.11微摩尔，1.5当量)的四氢呋喃(5毫升)溶液，并继续反应2小时。加入水(30毫升)，乙酸乙酯(50毫升×3)萃取。合并有机相，饱和食盐水(50毫升×2)洗，无水硫酸钠干燥，过滤，浓缩得到化合物17-2。MS(ESI)m/z:598.1[M+H +]。 Step B: Dissolve compound 17-1 (100 mg, 212.08 micromolar, 1 equivalent) in tetrahydrofuran (5 mL), and add lithium diisopropylamide (2 mol per liter, 159.06) dropwise at -78 ° C under a nitrogen atmosphere Microliter, 318.11 micromolar, 1.5 equivalents). The reaction was maintained at -78 degrees Celsius for 1 hour, at this temperature a solution of iodine (80.74 mg, 318.11 micromolar, 1.5 equivalents) in tetrahydrofuran (5 mL) was added, and the reaction was continued for 2 hours. Water (30 ml) was added, and ethyl acetate (50 ml × 3) was extracted. The organic phases were combined, washed with saturated brine (50 ml × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 17-2. MS (ESI) m / z: 598.1 [M + H + ].
步骤C：将化合物17-2(150毫克，251.08微摩尔，1当量)，1-甲基吡唑-4-硼酸(33.20毫克，263.63微摩尔，1.05当量)，碳酸钾(104.10毫克，753.23微摩尔，3当量)和[1,1'-双(二苯基膦)二茂铁]二氯化钯(18.37毫克，25.11微摩尔，0.1当量)溶于1,4-二氧六环(5毫升)和水(1毫升)中，氮气气氛下于80摄氏度反应12小时。反应液过滤，滤液加水(30毫升)稀释，乙酸乙酯(50毫升×3)萃取。合并有机相，饱和食盐水(50毫升×2)洗，无水硫酸钠干燥，过滤，浓缩得到化合物17-3。MS(ESI)m/z:552.3[M+H +]。 Step C: Compound 17-2 (150 mg, 251.08 micromolar, 1 equivalent), 1-methylpyrazole-4-boronic acid (33.20 mg, 263.63 micromolar, 1.05 equivalent), potassium carbonate (104.10 mg, 753.23 micromolar) Molar, 3 equivalents) and [1,1'-bis (diphenylphosphino) ferrocene] palladium dichloride (18.37 mg, 25.11 micromolar, 0.1 equivalent) dissolved in 1,4-dioxane (5 ML) and water (1 mL) under nitrogen atmosphere at 80 degrees Celsius for 12 hours. The reaction solution was filtered, the filtrate was diluted with water (30 mL), and extracted with ethyl acetate (50 mL × 3). The organic phases were combined, washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 17-3. MS (ESI) m / z: 552.3 [M + H + ].
步骤D：向化合物17-3(120毫克，217.54微摩尔，1当量)的甲醇(5毫升)溶液中，加入氢氧化钠水溶液(2摩尔每升，543.85微升，1.09毫摩尔，5当量)。60摄氏度反应4小时。加入水(50毫升)，乙酸乙酯(50毫升×3)萃取。合并有机相，饱和食盐水(50毫升×2)洗，无水硫酸钠干燥，过滤，浓缩，制备级高效液相色谱(甲酸条件)纯化得到化合物17的甲酸盐。MS(ESI)m/z:412.2[M+H +]； Step D: To a solution of compound 17-3 (120 mg, 217.54 μmol, 1 equiv) in methanol (5 mL), add an aqueous solution of sodium hydroxide (2 mol per liter, 543.85 μl, 1.09 mmol, 5 equiv) . React for 4 hours at 60 degrees Celsius. Water (50 ml) was added, and ethyl acetate (50 ml × 3) was extracted. The organic phases were combined, washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by preparative high-performance liquid chromatography (formic acid conditions) to obtain the formate salt of compound 17. MS (ESI) m / z: 412.2 [M + H + ];
1H NMR(300MHz,DMSO-d 6)δppm 2.08(s,3H),2.56-2.64(m,2H),2.79-2.90(m,2H),3.89(s,3H),4.22(br t,J＝7.04Hz,2H),6.36(d,J＝5.49Hz,1H),6.62(s,1H),6.99(d,J＝7.23Hz,1H),7.50-7.55(m,1H),7.57(br d,J＝7.32Hz,1H),7.89(d,J＝5.49Hz,1H),7.97(s,1H),8.16(s,1H),8.25(br s,1H),11.90(br s,1H)。 1 H NMR (300 MHz, DMSO-d 6 ) δ ppm 2.08 (s, 3H), 2.56-2.64 (m, 2H), 2.79-2.90 (m, 2H), 3.89 (s, 3H), 4.22 (br t, J = 7.04 Hz, 2H), 6.36 (d, J = 5.49 Hz, 1H), 6.62 (s, 1H), 6.99 (d, J = 7.23 Hz, 1H), 7.50-7.55 (m, 1H), 7.57 (br d, J = 7.32 Hz, 1H), 7.89 (d, J = 5.49 Hz, 1H), 7.97 (s, 1H), 8.16 (s, 1H), 8.25 (br s, 1H), 11.90 (br s, 1H ).
活性测试Activity test
1.体外酶抑制活性实验1. In vitro enzyme inhibitory activity test
实验材料：Experimental Materials:
TGF-βR1、TGF-βR1(T204D)、LCK、p38αKinase Enzyme System(激酶系统)购自Promega。Envision多标记分析仪(PerkinElmer)。TGF-βR1, TGF-βR1 (T204D), LCK, p38αKinase Enzyme System (kinase system) were purchased from Promega. Envision Multi-Mark Analyzer (PerkinElmer).
实验方法：experimental method:
使用试剂盒里的缓冲溶液稀释酶，底物，ATP(腺苷酸三磷酸)和抑制剂。Use the buffer solution in the kit to dilute the enzyme, substrate, ATP (adenosine triphosphate) and inhibitor.
TGF-βR1：TGF-βR1:
将待测化合物用排枪进行5倍稀释至第8个浓度，即从50微摩尔每升稀释至0.65纳摩尔每升，二甲亚砜终浓度为5％，设置双复孔实验。向微孔板中加入1微升抑制剂各浓度梯度，2微升TGF-βR1酶(30纳克)，2微升底物和ATP的混合物(50微摩尔每升ATP，0.2微克每微升TGF-βR1肽)，此时化合物终浓度梯度为10微摩尔每升稀释至0.13纳摩尔每升。反应体系置于30摄氏度反应120分钟。反应结束后，每孔加入5微升ADP-Glo试剂，30摄氏度继续反应40分钟，结束反应后每孔加入10微升的激酶检测试剂，30摄氏度反应30分钟后采用PerkinElmer Envision多标记分析仪读数化学发光，积分时间0.5秒。The compound to be tested was diluted 5 times to the eighth concentration with an exhaust gun, that is, from 50 micromoles per liter to 0.65 nanomoles per liter, and the final concentration of dimethyl sulfoxide was 5%. A double-multiwell experiment was set up. Add 1 μl inhibitor concentration gradient to the microplate, 2 μl TGF-βR1 enzyme (30 ng), 2 μl substrate and ATP mixture (50 μmol / L ATP, 0.2 μg / L) TGF-βR1 peptide), at this time the final concentration gradient of the compound is diluted from 10 micromoles per liter to 0.13 nanomoles per liter. The reaction system was placed at 30 degrees Celsius for 120 minutes. After the reaction, add 5 microliters of ADP-Glo reagent to each well and continue the reaction at 30 degrees Celsius for 40 minutes. After the reaction, add 10 microliters of kinase detection reagent to each well. After 30 minutes at 30 degrees Celsius, use the PerkinElmer Envision multi-label analyzer to read Chemiluminescence, integration time 0.5 seconds.
TGF-βR1(T204D)：TGF-βR1 (T204D):
将待测化合物用排枪进行5倍稀释至第8个浓度，即从50微摩尔每升稀释至0.65纳摩尔每升，二甲亚砜终浓度为5％，设置双复孔实验。向微孔板中加入1微升抑制剂各浓度梯度，2微升TGF-βR1(T204D)酶(15纳克)，2微升底物和ATP的混合物(50微摩尔每升ATP，0.2微克每微升TGF-βR1肽)，此时化合物终浓度梯度为10微摩尔每升稀释至0.13纳摩尔每升。反应体系置于30摄氏度反应120分钟。反应结束后，每孔加入5微升ADP-Glo试剂，30摄氏度继续反应40分钟，结束反应后每孔加入10微升的激酶检测试剂，30摄氏度反应30分钟后采用PerkinElmer Envision多标记分析仪读数化学发光，积分时间0.5秒。The compound to be tested was diluted 5 times to the eighth concentration with an exhaust gun, that is, from 50 micromoles per liter to 0.65 nanomoles per liter, and the final concentration of dimethyl sulfoxide was 5%. A double-multiwell experiment was set up. Add 1 μl inhibitor concentration gradient to the microplate, 2 μl TGF-βR1 (T204D) enzyme (15 ng), 2 μl substrate and ATP mixture (50 μmol per liter ATP, 0.2 μg Per microliter of TGF-βR1 peptide), then the final concentration gradient of the compound is 10 micromoles per liter diluted to 0.13 nanomoles per liter. The reaction system was placed at 30 degrees Celsius for 120 minutes. After the reaction, add 5 microliters of ADP-Glo reagent to each well and continue the reaction at 30 degrees Celsius for 40 minutes. After the reaction, add 10 microliters of kinase detection reagent to each well. After 30 minutes at 30 degrees Celsius, use the PerkinElmer Envision multi-label analyzer to read Chemiluminescence, integration time 0.5 seconds.
LCK：LCK:
将待测化合物用排枪进行5倍稀释至第8个浓度，即从500微摩尔每升稀释至6.5纳摩尔每升，二甲亚砜终浓度为5％，设置双复孔实验。向微孔板中加入1微升抑制剂各浓度梯度，2微升LCK酶(1.55纳克)，2微升底物和ATP的混合物(30微摩尔每升ATP，0.4微克每微升Poly E 4Y 1)，此时化合物终浓度梯度为100微摩尔每升稀释至1.3纳摩尔每升。反应体系置于30摄氏度反应60分钟。反应结束后，每孔加入5微升ADP-Glo试剂，30摄氏度继续反应40分钟，结束反应后每孔加入10微升的激酶检测试剂，30摄氏度反应30分钟后采用PerkinElmer Envision多标记分析仪读数化学发光，积分时间0.5秒。 The compound to be tested was diluted 5 times to the eighth concentration with an exhaust gun, that is, from 500 micromoles per liter to 6.5 nanomoles per liter, and the final concentration of dimethyl sulfoxide was 5%. A double-multiwell experiment was set up. Add 1 μl inhibitor concentration gradient to the microplate, 2 μl LCK enzyme (1.55 ng), 2 μl substrate and ATP mixture (30 μmol / L ATP, 0.4 μg / L Poly E 4 Y 1 ), at this time the final concentration gradient of the compound is diluted from 100 micromoles per liter to 1.3 nanomoles per liter. The reaction system was placed at 30 degrees Celsius and reacted for 60 minutes. After the reaction, add 5 microliters of ADP-Glo reagent to each well and continue the reaction at 30 degrees Celsius for 40 minutes. After the reaction, add 10 microliters of kinase detection reagent to each well. After 30 minutes at 30 degrees Celsius, use the PerkinElmer Envision multi-label analyzer to read Chemiluminescence, integration time 0.5 seconds.
p38α：p38α:
将待测化合物用排枪进行5倍稀释至第8个浓度，即从50微摩尔每升稀释至0.65纳摩尔每升，二甲亚砜终浓度为5％，设置双复孔实验。向微孔板中加入1微升抑制剂各浓度梯度，2微升p38α酶(4纳克)，2微升底物和ATP的混合物(150微摩尔每升ATP，0.2微克每微升p38肽)，此时化合物终浓度梯度为10微摩尔每升稀释至0.13纳摩尔每升。反应体系置于30摄氏度反应60分钟。反应结束后，每孔加入5微升ADP-Glo试剂，30摄氏度继续反应40分钟，结束反应后每孔加入10微升的激酶检测试剂，30摄氏度反应30分钟后采用PerkinElmer Envision多标记分析仪读数化学发光，积分时间0.5秒。The compound to be tested was diluted 5 times to the eighth concentration with an exhaust gun, that is, from 50 micromoles per liter to 0.65 nanomoles per liter, and the final concentration of dimethyl sulfoxide was 5%. A double-multiwell experiment was set up. Add 1 μl inhibitor concentration gradient to the microplate, 2 μl p38α enzyme (4 ng), 2 μl substrate and ATP mixture (150 μmol / L ATP, 0.2 μg / L p38 peptide) ), At this time the final concentration gradient of the compound is diluted from 10 micromoles per liter to 0.13 nanomoles per liter. The reaction system was placed at 30 degrees Celsius and reacted for 60 minutes. After the reaction, add 5 microliters of ADP-Glo reagent to each well and continue the reaction at 30 degrees Celsius for 40 minutes. After the reaction, add 10 microliters of kinase detection reagent to each well. After 30 minutes at 30 degrees Celsius, use the PerkinElmer Envision multi-label analyzer to read Chemiluminescence, integration time 0.5 seconds.
数据分析：data analysis:
原始数据换算成抑制率，IC 50的值即可通过四参数进行曲线拟合得出。表1提供了本发明实施例化合物对TGF-βR1、TGF-βR1(T204D)、LCK、p38α酶学抑制活性。 The original data is converted into the inhibition rate, and the IC 50 value can be obtained by curve fitting through four parameters. Table 1 provides enzymatic inhibitory activities of the compounds of the examples of the present invention on TGF-βR1, TGF-βR1 (T204D), LCK, and p38α.
实验结果：见表1：Experimental results: see table 1:
表1Table 1
Figure PCTCN2019114347-appb-000104
Figure PCTCN2019114347-appb-000104
--表示未测试-Means not tested
结论：本发明化合物对TGF-βR1、TGF-βR1(T204D)具有优异的体外抑制活性，同时对LCK、p38α的体外抑制活性弱，展现出了极佳的体外活性及选择性。Conclusion: The compound of the present invention has excellent in vitro inhibitory activity against TGF-βR1, TGF-βR1 (T204D), and weak in vitro inhibitory activity against LCK and p38α, exhibiting excellent in vitro activity and selectivity.
2.pSMAD响应单元抑制实验2. pSMAD response unit suppression experiment
实验原理：Experimental principle:
该实验将一个SMAD-响应元素控制的荧光素酶基因整合到HEK293细胞中。该细胞系通过人类TGFβ1的刺激响应和TGFβ/SMAD信号通路抑制剂的处理而得到验证。This experiment integrated a SMAD-responsive element controlled luciferase gene into HEK293 cells. This cell line was verified by the stimulation response of human TGFβ1 and the treatment of TGFβ / SMAD signaling pathway inhibitors.
实验操作：Experimental operation:
1.生长培养基中培养HEK293细胞。在96孔板中，每个孔均放置约35000个种子细胞和100微升无基因霉素的生长介质。并在37摄氏度孵化。1. Culture HEK293 cells in growth medium. In a 96-well plate, each well contains approximately 35,000 seed cells and 100 microliters of gene-free growth medium. And incubate at 37 degrees Celsius.
2.24小时后，向孔中分别加入三倍浓度梯度的待测物的90微升分析培养基溶液。在一个二氧化碳培养箱中于37摄氏度孵化4-5个小时。对照组只加入不含待测物的90微升分析培养基。2. After 24 hours, add 90 microliters of the analysis medium solution of the analyte with a three-fold concentration gradient to the wells. Incubate for 4-5 hours at 37 degrees Celsius in a carbon dioxide incubator. The control group only added 90 microliters of analysis medium without test substance.
3.每一个浓度点需包含三组实验。a.在培养基中加入人源TGFβ1的10微升分析培养基溶液(最后的TGFβ1浓度为20纳克/毫升)。b.在无刺激组加入10微升空白培养基。c.在无细胞组加入100微升空白培养基。3. Each concentration point should include three sets of experiments. a. Add 10 microliters of human-derived TGFβ1 analysis medium solution to the medium (the final TGFβ1 concentration is 20 ng / ml). b. Add 10 μl of blank culture medium to the unstimulated group. c. Add 100 μl of blank medium to the cell-free group.
4.在二氧化碳培养箱中于37摄氏度下孵化18小时。4. Incubate in a carbon dioxide incubator at 37 degrees Celsius for 18 hours.
5.使用荧光素酶分析系统进行测定：在室温下每个孔中加入100微升ONE-Step TM荧光素酶试剂，室温放置15到30分钟。使用光度计测量荧光。 5. Measurement using a luciferase analysis system: add 100 μl of ONE-Step ™ luciferase reagent to each well at room temperature and leave at room temperature for 15 to 30 minutes. Use a photometer to measure fluorescence.
6.数据分析：从所有孔的发光读数中减去平均背景发光(无细胞组孔)。6. Data analysis: Subtract the average background luminescence from all luminescence readings (without cell group wells).
实验结果：见表2：Experimental results: see table 2:
表2Table 2
实施例Examples pSmad抑制IC 50(微摩尔每升) pSmad inhibits IC 50 (micromoles per liter)
11 0.020.02
22 0.040.04
66 0.060.06
77 0.050.05
88 0.040.04
99 0.01270.0127
1212 0.010.01
结论：本发明化合物具有优异的pSmad抑制活性。证明本发明化合物能够起到抑制TGF-β/SMAD信号通路的作用。Conclusion: The compound of the present invention has excellent pSmad inhibitory activity. It is proved that the compound of the present invention can play a role in inhibiting the TGF-β / SMAD signaling pathway.
3.小鼠结肠癌CT-26细胞BALB/c小鼠皮下同种移植瘤模型的体内抗肿瘤药效研究3. In vivo antitumor drug efficacy of mouse colon cancer CT-26 cell BALB / c mouse subcutaneous allograft model
实验目的：Purpose:
本研究主要的目的是在CT26小鼠同种移植瘤模型上研究受测化合物的抗肿瘤药效。The main purpose of this study is to study the anti-tumor efficacy of the tested compounds on the CT26 mouse allograft model.
实验操作：Experimental operation:
细胞培养：小鼠结肠癌CT-26细胞(中国科学院典型培养物保藏委员会细胞库)体外单层培养，培养条件为RPMI-1640培养基(生产厂家：Gibco)中加10％胎牛血清，37摄氏度5％二氧化碳孵箱培养。一周两次用胰酶-EDTA(Ethylene DiamineTetraaceticAcid，乙二胺四乙酸)进行常规消化处理传代。当细胞饱和度为80％-90％，数量到达要求时，收取细胞，计数，接种。Cell culture: Mouse colon cancer CT-26 cells (Typical Culture Depository of the Chinese Academy of Sciences Cell Bank) in vitro monolayer culture, culture conditions are RPMI-1640 medium (manufacturer: Gibco) plus 10% fetal bovine serum, 37 Cultivation in 5% carbon dioxide incubator. Twice a week, try digestion with EDTA (Ethylene Diamine Tetraacetic Acid). When the cell saturation is 80% -90%, and the quantity reaches the requirement, collect the cells, count and inoculate.
动物：BALB/c小鼠，雌性，6～8周龄。Animals: BALB / c mice, female, 6-8 weeks old.
肿瘤接种：将0.1毫升含3×10 5个CT26细胞的DPBS细胞悬液皮下接种于每只小鼠的右侧腹股沟处，接种当天开始给药。 Tumor inoculation: 0.1 ml of a DPBS cell suspension containing 3 × 10 5 CT26 cells was subcutaneously inoculated into the right groin of each mouse, and the administration started on the day of inoculation.
实验指标：实验指标是考察肿瘤生长是否被抑制、延缓或治愈。每周两次用游标卡尺测量肿瘤直径。肿瘤体积的计算公式为：V＝0.5L×W  2，L和W分别表示肿瘤的长径和短径。 Experimental index: The experimental index is to investigate whether the tumor growth is inhibited, delayed or cured. The diameter of the tumor was measured with vernier calipers twice a week. The calculation formula of the tumor volume is: V = 0.5L × W 2 , L and W represent the long and short diameters of the tumor, respectively.
实验结果：化合物肿瘤抑制效果见表3。Experimental results: The compound's tumor suppressing effect is shown in Table 3.
表3 CT26同种异位移植实验结果Table 3 Experimental results of CT26 allotransplantation
Figure PCTCN2019114347-appb-000105
Figure PCTCN2019114347-appb-000105
实验结论：本发明化合物在小鼠结肠癌CT-26细胞BALB/c小鼠皮下同种移植瘤模型中具有明显的体内抗肿瘤药效。在相同剂量下(50毫克每公斤，每天两次)下表现出比化合物A明显更优的肿瘤抑制效果。Experimental conclusion: The compound of the invention has obvious antitumor effect in vivo in mouse colon cancer CT-26 cell BALB / c mouse subcutaneous allograft model. At the same dose (50 mg per kg, twice a day), it showed a significantly better tumor suppressive effect than compound A.

Claims (30)

  1. 式(I)所示化合物、其药学上可接受的盐或其异构体，The compound represented by formula (I), its pharmaceutically acceptable salt or its isomer,
    Figure PCTCN2019114347-appb-100001
    Figure PCTCN2019114347-appb-100001
    其中，环A为5-6元杂芳基、苯基、C 5-6环烷基或5-6元杂环烷基； Among them, ring A is 5-6 membered heteroaryl, phenyl, C 5-6 cycloalkyl or 5-6 membered heterocycloalkyl;
    R 1、R 2和R 3各自独立地为H、F、Cl、Br、I、-CN、-OH、C 1-6烷氧基、C 1-6烷基、C 2-4烯基、C 2-4炔基或C 3-6环烷基，其中所述C 1-6烷氧基、C 1-6烷基、C 2-4烯基、C 2-4炔基和C 3-6环烷基任选被1、2或3个独立选自F、Cl、Br、CN、-OH、-CH 3、-OCH 3和-NH 2的取代基所取代； R 1 , R 2 and R 3 are each independently H, F, Cl, Br, I, -CN, -OH, C 1-6 alkoxy, C 1-6 alkyl, C 2-4 alkenyl, C 2-4 alkynyl or C 3-6 cycloalkyl, wherein the C 1-6 alkoxy, C 1-6 alkyl, C 2-4 alkenyl, C 2-4 alkynyl and C 3- 6 cycloalkyl groups are optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, CN, -OH, -CH 3 , -OCH 3 and -NH 2 ;
    m为1或2；m is 1 or 2;
    R 4为5-6元杂芳基或苯基，其中所述5-6元杂芳基和苯基任选被1、2或3个R a所取代； R 4 is 5-6 membered heteroaryl or phenyl, wherein the 5-6 membered heteroaryl and phenyl are optionally substituted with 1, 2 or 3 R a ;
    T 1为-O-或-CH 2-； T 1 is -O- or -CH 2- ;
    T 2为N或C(R 5)； T 2 is N or C (R 5 );
    R 5为H、F、Cl、Br、-CN、-OH、-NH 2、-OCH 3或-CH 3； R 5 is H, F, Cl, Br, -CN, -OH, -NH 2 , -OCH 3 or -CH 3 ;
    各R a独立地为-CN、-C(＝O)NR bR c、C 1-6烷氧基或任选被1、2或3个独立选自F、Cl、Br、I、-OH、-OCH 3、-CN和NH 2的取代基所取代的C 1-6烷基； Each R a is independently -CN, -C (= O) NR b R c , C 1-6 alkoxy or optionally 1, 2, or 3 independently selected from F, Cl, Br, I, -OH , -OCH 3 , -CN and NH 2 are substituted by C 1-6 alkyl;
    或R 4和R 5与其连接的吡啶基和
    Figure PCTCN2019114347-appb-100002
    一起形成式(B)所示结构：     Or the pyridyl group to which R 4 and R 5 are linked
    Figure PCTCN2019114347-appb-100002
    Together form the structure shown in formula (B):
    Figure PCTCN2019114347-appb-100003
    Figure PCTCN2019114347-appb-100003
    L为单键、
    Figure PCTCN2019114347-appb-100004
    L is a single key,
    Figure PCTCN2019114347-appb-100004
    R 6独立地为H、-CN、-C(＝O)NR bR c、C 1-4烷基、C 3-6环烷基或5-6元杂芳基，其中所述C 1-4烷基、C 3-6环烷基和5-6元杂芳基任选被1、2或3个R d所取代； R 6 is independently H, -CN, -C (= O) NR b R c , C 1-4 alkyl, C 3-6 cycloalkyl, or 5-6 membered heteroaryl, wherein the C 1- 4 alkyl, C 3-6 cycloalkyl and 5-6 membered heteroaryl are optionally substituted with 1, 2 or 3 Rd ;
    各R d独立地为F、Cl、Br、I、-OH、-CN、-NH 2、-OCH 3、-OCH 2CH 3、-CH 3、-CF 3或-CH 2CH 3； Each R d is independently F, Cl, Br, I, -OH, -CN, -NH 2, -OCH 3, -OCH 2 CH 3, -CH 3, -CF 3 or -CH 2 CH 3;
    R b和R c各自独立地为H、-CH 3、-CH 2CH 3、-CH 2CH 2CH 3、-CH 2(CH 3) 2或环丙基； R b and R c are each independently H, -CH 3 , -CH 2 CH 3 , -CH 2 CH 2 CH 3 , -CH 2 (CH 3 ) 2 or cyclopropyl;
    所述5-6元杂环烷基和5-6元杂芳基分别包含1、2、3或4个独立选自N、-O-、-S-和-NH-的杂原子或杂原子团。The 5-6 membered heterocycloalkyl group and 5-6 membered heteroaryl group respectively contain 1, 2, 3 or 4 heteroatoms or heteroatom groups independently selected from N, -O-, -S- and -NH- .
  2. 根据权利要求1所述化合物、其药学上可接受的盐或其异构体，其具有式(I-A)所示结构：The compound according to claim 1, a pharmaceutically acceptable salt thereof, or an isomer thereof, which has a structure represented by formula (I-A):
    Figure PCTCN2019114347-appb-100005
    Figure PCTCN2019114347-appb-100005
    其中，环A、R 1、R 2、R 3、R 4、T 1和m如权利要求1所定义。 Among them, rings A, R 1 , R 2 , R 3 , R 4 , T 1 and m are as defined in claim 1.
  3. 根据权利要求1所述化合物、其药学上可接受的盐或其异构体，其具有式(I-B)所示结构：The compound according to claim 1, a pharmaceutically acceptable salt thereof, or an isomer thereof, which has a structure represented by formula (I-B):
    Figure PCTCN2019114347-appb-100006
    Figure PCTCN2019114347-appb-100006
    其中，环A、R 1、R 2、R 3、R 4、R 5、T 1和m如权利要求1所定义。 Wherein rings A, R 1 , R 2 , R 3 , R 4 , R 5 , T 1 and m are as defined in claim 1.
  4. 根据权利要求3所述化合物、其药学上可接受的盐或其异构体，其中所述R 4为5-6元杂芳基或苯基，其中所述5-6元杂芳基和苯基任选被1、2或3个R a所取代；R 5为H、F、Cl、Br、-CN、-OH、-OCH 3、-NH 2或-CH 3。 The compound according to claim 3, a pharmaceutically acceptable salt or isomer thereof, wherein R 4 is a 5-6 membered heteroaryl or phenyl group, wherein the 5-6 membered heteroaryl group and benzene group optionally substituted by one, two or three R a; R 5 is H, F, Cl, Br, -CN, -OH, -OCH 3, -NH 2 or -CH 3.
  5. 根据权利要求1所述化合物、其药学上可接受的盐或其异构体，其具有式(I-C)所示结构：The compound according to claim 1, a pharmaceutically acceptable salt thereof, or an isomer thereof, which has a structure represented by formula (I-C):
    Figure PCTCN2019114347-appb-100007
    Figure PCTCN2019114347-appb-100007
    其中，环A、R 1、R 2、R 3、T 1、m、R 6和L如权利要求1所定义。 Among them, rings A, R 1 , R 2 , R 3 , T 1 , m, R 6 and L are as defined in claim 1.
  6. 根据权利要求1所述化合物、其药学上可接受的盐或其异构体，其中所述各R a独立地为CN、-OCH 3、
    Figure PCTCN2019114347-appb-100008
    The compound according to claim 1, its pharmaceutically acceptable salt or isomer thereof, wherein each R a is independently CN, -OCH 3,
    Figure PCTCN2019114347-appb-100008
  7. 根据权利要求1～4或6任一项所述化合物、其药学上可接受的盐或其异构体，其中所述R 4为吡咯基、吡唑基、吡啶基、嘧啶基或苯基，其中所述吡咯基、吡唑基、吡啶基、嘧啶基和苯基任选被1、2或3个R a所取代。 The compound according to any one of claims 1 to 4 or 6, a pharmaceutically acceptable salt thereof, or an isomer thereof, wherein R 4 is pyrrolyl, pyrazolyl, pyridyl, pyrimidinyl, or phenyl, wherein said pyrrolyl, pyrazolyl, pyridyl, pyrimidinyl and phenyl is optionally substituted with 1, 2, or 3 R a.
  8. 根据权利要求7所述化合物、其药学上可接受的盐或其异构体，其中所述R 4为
    Figure PCTCN2019114347-appb-100009
    Figure PCTCN2019114347-appb-100010
    The compound according to claim 7, a pharmaceutically acceptable salt or isomer thereof, wherein R 4 is
    Figure PCTCN2019114347-appb-100009
    Figure PCTCN2019114347-appb-100010
  9. 根据权利要求8所述化合物、其药学上可接受的盐或其异构体，其中所述R 4为
    Figure PCTCN2019114347-appb-100011
    Figure PCTCN2019114347-appb-100012
    The compound according to claim 8, a pharmaceutically acceptable salt or isomer thereof, wherein R 4 is
    Figure PCTCN2019114347-appb-100011
    Figure PCTCN2019114347-appb-100012
  10. 根据权利要求8所述化合物、其药学上可接受的盐或其异构体，其具有式(I-A1)～(I-A4)所示结构：The compound according to claim 8, a pharmaceutically acceptable salt or isomer thereof, which has a structure represented by formulas (I-A1) to (I-A4):
    Figure PCTCN2019114347-appb-100013
    Figure PCTCN2019114347-appb-100013
    其中，环A、R 1、R 2、R 3、T 1和m如权利要求1所定义；R a如权利要求1或6所定义。 Wherein Ring A, R 1, R 2, R 3, T 1 and m are as defined as claimed in claim 1; 6 1 or R a as defined in the claims.
  11. 根据权利要求10所述化合物、其药学上可接受的盐或其异构体，其具有式(I-A5)～(I-A12)所示结构：The compound according to claim 10, a pharmaceutically acceptable salt thereof, or an isomer thereof, which has a structure represented by formulas (I-A5) to (I-A12):
    Figure PCTCN2019114347-appb-100014
    Figure PCTCN2019114347-appb-100014
    Figure PCTCN2019114347-appb-100015
    Figure PCTCN2019114347-appb-100015
    其中，环A、R 1、R 2、R 3和R a如权利要求10所定义。 Wherein Ring A, R 1, R 2, R 3 and R a are defined as claimed in claim 10.
  12. 根据权利要求8所述化合物、其药学上可接受的盐或其异构体，其具有式(I-B1)～(I-B4)所示结构：The compound according to claim 8, a pharmaceutically acceptable salt or isomer thereof, which has a structure represented by formula (I-B1) to (I-B4):
    Figure PCTCN2019114347-appb-100016
    Figure PCTCN2019114347-appb-100016
    其中，环A、R 1、R 2、R 3、R 5、T 1和m如权利要求1所定义；R a如权利要求1或6所定义。 Wherein Ring A, R 1, R 2, R 3, R 5, T 1 and m are as defined as claimed in claim 1; 6 1 or R a as defined in the claims.
  13. 根据权利要求12所述化合物、其药学上可接受的盐或其异构体，其具有式(I-B5)～(I-B12)所示结构：The compound according to claim 12, a pharmaceutically acceptable salt thereof, or an isomer thereof, which has a structure represented by formulas (I-B5) to (I-B12):
    Figure PCTCN2019114347-appb-100017
    Figure PCTCN2019114347-appb-100017
    其中，环A、R 1、R 2、R 3、R 5和R a如权利要求12所定义。 Among them, ring A, R 1 , R 2 , R 3 , R 5 and R a are as defined in claim 12.
  14. 根据权利要求5所述化合物、其药学上可接受的盐或其异构体，其具有式(I-C1)或(I-C2)所示结构：The compound according to claim 5, a pharmaceutically acceptable salt or isomer thereof, which has a structure represented by formula (I-C1) or (I-C2):
    Figure PCTCN2019114347-appb-100018
    Figure PCTCN2019114347-appb-100018
    其中，环A、R 1、R 2、R 3、R 6和L如权利要求5所定义。 Among them, rings A, R 1 , R 2 , R 3 , R 6 and L are as defined in claim 5.
  15. 根据权利要求1～5或10～14任一项所述化合物、其药学上可接受的盐或其异构体，其中所述R 1、R 2和R 3各自独立地为H、F、Cl、Br、I、-CN、-OH、-OCH 3、-OCH 2CH 3、-CH 3、-CH 2CH 3、乙烯基、乙炔基或环丙基，其中所述-OCH 3、-OCH 2CH 3、-CH 3、-CH 2CH 3、乙烯基、乙炔基和环丙基任选被1、2或3个独立 选自F、Cl、Br、CN、-OH、-CH 3、-OCH 3和-NH 2的取代基所取代。 The compound according to any one of claims 1 to 5 or 10 to 14, a pharmaceutically acceptable salt thereof, or an isomer thereof, wherein each of R 1 , R 2 and R 3 is independently H, F and Cl , Br, I, -CN, -OH, -OCH 3 , -OCH 2 CH 3 , -CH 3 , -CH 2 CH 3 , vinyl, ethynyl, or cyclopropyl, wherein -OCH 3 , -OCH 2 CH 3 , -CH 3 , -CH 2 CH 3 , vinyl, ethynyl and cyclopropyl are optionally selected from 1, 2, or 3 independently selected from F, Cl, Br, CN, -OH, -CH 3 , -OCH 3 and -NH 2 are substituted.
  16. 根据权利要求15所述化合物、其药学上可接受的盐或其异构体，其中所述R 1、R 2和R 3各自独立地为H、F、Cl、Br、-CN、-OH、-OCH 3、-CH 3、-CH 2CH 3或-CF 3。 The compound according to claim 15, a pharmaceutically acceptable salt thereof, or an isomer thereof, wherein each of R 1 , R 2 and R 3 is independently H, F, Cl, Br, -CN, -OH, -OCH 3 , -CH 3 , -CH 2 CH 3 or -CF 3 .
  17. 根据权利要求1～5或10～14任一项所述化合物、其药学上可接受的盐或其异构体，其中所述环A为噻吩基、吡咯基、吡啶基、嘧啶基、苯基或四氢-2H-吡喃基。The compound according to any one of claims 1 to 5 or 10 to 14, a pharmaceutically acceptable salt thereof, or an isomer thereof, wherein the ring A is thienyl, pyrrolyl, pyridyl, pyrimidinyl, or phenyl Or tetrahydro-2H-pyranyl.
  18. 根据权利要求1～5、10～14或16任一项所述化合物、其药学上可接受的盐或其异构体，其中所述
    Figure PCTCN2019114347-appb-100019
    为
    Figure PCTCN2019114347-appb-100020
    The compound according to any one of claims 1 to 5, 10 to 14 or 16, a pharmaceutically acceptable salt or isomer thereof, wherein the
    Figure PCTCN2019114347-appb-100019
    for
    Figure PCTCN2019114347-appb-100020
  19. 根据权利要求18所述化合物、其药学上可接受的盐或其异构体，其中所述
    Figure PCTCN2019114347-appb-100021
    为
    Figure PCTCN2019114347-appb-100022
    Figure PCTCN2019114347-appb-100023
    The compound according to claim 18, a pharmaceutically acceptable salt or isomer thereof, wherein the
    Figure PCTCN2019114347-appb-100021
    for
    Figure PCTCN2019114347-appb-100022
    Figure PCTCN2019114347-appb-100023
  20. 根据权利要求19所述化合物、其药学上可接受的盐或其异构体，其中所述
    Figure PCTCN2019114347-appb-100024
    为
    Figure PCTCN2019114347-appb-100025
    Figure PCTCN2019114347-appb-100026
    The compound according to claim 19, a pharmaceutically acceptable salt or isomer thereof, wherein the
    Figure PCTCN2019114347-appb-100024
    for
    Figure PCTCN2019114347-appb-100025
    Figure PCTCN2019114347-appb-100026
  21. 根据权利要求19所述化合物、其药学上可接受的盐或其异构体，其具有式(I-A13)～(I-A16)所示结构：The compound according to claim 19, a pharmaceutically acceptable salt thereof, or an isomer thereof, which has a structure represented by formulas (I-A13) to (I-A16):
    Figure PCTCN2019114347-appb-100027
    Figure PCTCN2019114347-appb-100027
    其中，R 1如权利要求1或16所定义；R a如权利要求1所定义。 Wherein 1 or 16 R 1 is as defined in claim 1; a R a as defined in the claims.
  22. 根据权利要求19所述化合物、其药学上可接受的盐或其异构体，其具有式(I-B13)～(I-B36)所示结构：The compound according to claim 19, a pharmaceutically acceptable salt thereof, or an isomer thereof, which has a structure represented by formulas (I-B13) to (I-B36):
    Figure PCTCN2019114347-appb-100028
    Figure PCTCN2019114347-appb-100028
    Figure PCTCN2019114347-appb-100029
    Figure PCTCN2019114347-appb-100029
    其中，R 1如权利要求1或16所定义；R a如权利要求1所定义。 Wherein 1 or 16 R 1 is as defined in claim 1; a R a as defined in the claims.
  23. 根据权利要求19所述化合物、其药学上可接受的盐或其异构体，其具有式(I-C3)～(I-C5)所示结构：The compound according to claim 19, a pharmaceutically acceptable salt thereof, or an isomer thereof, which has a structure represented by formulas (I-C3) to (I-C5):
    Figure PCTCN2019114347-appb-100030
    Figure PCTCN2019114347-appb-100030
    其中，R 1如权利要求1或16所定义；L和R 6如权利要求1所定义。 Wherein R 1 is as defined in claim 1 or 16; L and R 6 are as defined in claim 1.
  24. 根据权利要求23所述化合物、其药学上可接受的盐或其异构体，其具有式(I-C6)～(I-C9)所示结构：The compound according to claim 23, a pharmaceutically acceptable salt thereof, or an isomer thereof, which has a structure represented by formulas (I-C6) to (I-C9):
    Figure PCTCN2019114347-appb-100031
    Figure PCTCN2019114347-appb-100031
    其中，R 1和R 6如权利要求23所定义。 Here, R 1 and R 6 are as defined in claim 23.
  25. 根据权利要求1、5、14、23或24任一项所述化合物、其药学上可接受的盐或其异构体，其中所述R 6独立地为H、-CN、-C(＝O)NH 2、异丙基、环丙基、环己基、吡唑基或吡啶基，其中所述异丙基、环丙基、环己基、吡唑基和吡啶基任选被1、2或3个R d所取代。 The compound according to any one of claims 1, 5, 14, 23, or 24, a pharmaceutically acceptable salt thereof, or an isomer thereof, wherein R 6 is independently H, -CN, -C (= O ) NH 2 , isopropyl, cyclopropyl, cyclohexyl, pyrazolyl or pyridyl, wherein the isopropyl, cyclopropyl, cyclohexyl, pyrazolyl and pyridyl are optionally 1, 2 or 3 Replaced by Rd .
  26. 根据权利要求25所述化合物、其药学上可接受的盐或其异构体，其中所述R 6独立地为H、-CN、-C(＝O)NH 2、
    Figure PCTCN2019114347-appb-100032
    The compound according to claim 25, a pharmaceutically acceptable salt or isomer thereof, wherein R 6 is independently H, -CN, -C (= O) NH 2 ,
    Figure PCTCN2019114347-appb-100032
  27. 根据权利要求26所述化合物、其药学上可接受的盐或其异构体，其中所述R 6独立地为H、-CN、-C(＝O)NH 2、
    Figure PCTCN2019114347-appb-100033
    The compound according to claim 26, a pharmaceutically acceptable salt thereof, or an isomer thereof, wherein R 6 is independently H, -CN, -C (= O) NH 2 ,
    Figure PCTCN2019114347-appb-100033
  28. 下式化合物，其药学上可接受的盐或其异构体：The compound of the formula, its pharmaceutically acceptable salt or its isomer:
    Figure PCTCN2019114347-appb-100034
    Figure PCTCN2019114347-appb-100034
    Figure PCTCN2019114347-appb-100035
    Figure PCTCN2019114347-appb-100035
  29. 根据权利要求1～28任一项所述化合物、其药学上可接受的盐或其异构体在制备TGF-βR1抑制剂药物中的应用。Use of the compound according to any one of claims 1 to 28, its pharmaceutically acceptable salt, or its isomer in the preparation of TGF-βR1 inhibitor drugs.
  30. 根据权利要求1～28任一项所述化合物、其药学上可接受的盐或其异构体在制备实体瘤药物中的应用。Use of the compound according to any one of claims 1 to 28, its pharmaceutically acceptable salt, or its isomer in the preparation of a solid tumor drug.
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113698395A (en) * 2020-05-22 2021-11-26 成都倍特药业股份有限公司 Transforming growth factor receptor antagonist, preparation method and application thereof
CN115340497A (en) * 2022-08-29 2022-11-15 安徽医科大学 Diaryl pyrimidine amide compound or pharmaceutically acceptable salt thereof, pharmaceutical composition and application thereof

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1951939A (en) * 2001-05-24 2007-04-25 伊莱利利公司 Novel pyrrole derivatives as pharmaceutical agents
WO2014072517A1 (en) * 2012-11-12 2014-05-15 Institució Catalana De Recerca I Estudis Avançats Methods and kits for the prognosis of colorectal cancer
CN106795139A (en) * 2014-10-07 2017-05-31 伊莱利利公司 Aminopyridine base epoxide pyrazole compound
WO2017215506A1 (en) * 2016-06-13 2017-12-21 南京明德新药研发股份有限公司 Benzotriazole-derived α and β unsaturated amide compound used as tgf-βr1 inhibitor
WO2018171611A1 (en) * 2017-03-22 2018-09-27 江苏恒瑞医药股份有限公司 6-pyrazole-[1,2,4]triazolo[4,3-a]pyridine-3-amide derivative, preparation method therefor and use thereof in medicine

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1951939A (en) * 2001-05-24 2007-04-25 伊莱利利公司 Novel pyrrole derivatives as pharmaceutical agents
WO2014072517A1 (en) * 2012-11-12 2014-05-15 Institució Catalana De Recerca I Estudis Avançats Methods and kits for the prognosis of colorectal cancer
CN106795139A (en) * 2014-10-07 2017-05-31 伊莱利利公司 Aminopyridine base epoxide pyrazole compound
WO2017215506A1 (en) * 2016-06-13 2017-12-21 南京明德新药研发股份有限公司 Benzotriazole-derived α and β unsaturated amide compound used as tgf-βr1 inhibitor
WO2018171611A1 (en) * 2017-03-22 2018-09-27 江苏恒瑞医药股份有限公司 6-pyrazole-[1,2,4]triazolo[4,3-a]pyridine-3-amide derivative, preparation method therefor and use thereof in medicine

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113698395A (en) * 2020-05-22 2021-11-26 成都倍特药业股份有限公司 Transforming growth factor receptor antagonist, preparation method and application thereof
CN113698395B (en) * 2020-05-22 2023-12-08 赛诺哈勃药业（成都）有限公司 Transforming growth factor receptor antagonist, preparation method and application thereof
CN115340497A (en) * 2022-08-29 2022-11-15 安徽医科大学 Diaryl pyrimidine amide compound or pharmaceutically acceptable salt thereof, pharmaceutical composition and application thereof
CN115340497B (en) * 2022-08-29 2023-12-05 安徽医科大学 Diaryl pyrimidine amide compound or pharmaceutically acceptable salt thereof, pharmaceutical composition and application thereof

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