WO2020035065A1 - 作为ret抑制剂的吡唑衍生物 - Google Patents

作为ret抑制剂的吡唑衍生物 Download PDF

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Publication number
WO2020035065A1
WO2020035065A1 PCT/CN2019/101121 CN2019101121W WO2020035065A1 WO 2020035065 A1 WO2020035065 A1 WO 2020035065A1 CN 2019101121 W CN2019101121 W CN 2019101121W WO 2020035065 A1 WO2020035065 A1 WO 2020035065A1
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Prior art keywords
compound
added
pharmaceutically acceptable
isomer
acceptable salt
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PCT/CN2019/101121
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English (en)
French (fr)
Inventor
陈正霞
张杨
戴美碧
李婕
龚珍
黎健
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 CN201980053423.0A priority Critical patent/CN112566907A/zh
Publication of WO2020035065A1 publication Critical patent/WO2020035065A1/zh

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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
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
    • 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
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing three or more hetero rings

Definitions

  • the present invention relates to a class of pyrazole derivatives and its application in the preparation of a medicament for treating diseases related to RET kinase inhibitors. Specifically, it relates to a compound represented by formula (III) and a pharmaceutically acceptable salt thereof.
  • RET protein is a receptor tyrosine kinase RTK, and also a transmembrane glycoprotein, expressed by the proto-oncogene RET (REarranged during Transfection) on chromosome 10, during the development of the kidney and enteric nervous system in the embryonic stage It plays an important role, and in addition, homeostasis is also critical in a variety of tissues, such as neurons, neuroendocrine, hematopoietic tissue, and male germ cells. Unlike other RTKs, RET does not directly bind to ligand molecules: such as neuroetin (artemin), glial cell line-derived neurotrophic factor (GDNF), neuroturin, and persephin.
  • ligand molecules such as neuroetin (artemin), glial cell line-derived neurotrophic factor (GDNF), neuroturin, and persephin.
  • GFLs GNDF family
  • GFLs GNDF family receptor ⁇
  • GFR ⁇ GDNF family receptor ⁇
  • Recruitment of related linker proteins, activation of cascades of signaling such as cell proliferation, related signaling pathways include MAPK, PI3K, JAK-STAT, PKA, PKC, etc.
  • RET oncogenic activation There are two main mechanisms of RET oncogenic activation: one is the rearrangement of chromosomes to generate new fusion proteins, usually the fusion of the RET kinase domain and the protein containing the self-dimerization domain; the second is the direct or indirect RET mutation Activated RET's kinase activity. These changes in somatic or germ cell levels are involved in the pathogenesis of many cancers. 5% to 10% of patients with papillary thyroid cancer have RET chromosome rearrangement; while 60% of myeloid thyroid medullary cancers have RET point mutations; about 1-2% of all NSCLC patients have RET fusion protein, KIF5B-RET is the most common.
  • RET inhibitors have potential clinical value in tumor or intestinal disorders.
  • the present invention provides a compound represented by formula (III), an isomer thereof, or a pharmaceutically acceptable salt thereof,
  • a ring is selected from Said Optionally substituted with 1, 2 or 3 R g ;
  • R 1 is selected from H, F, Cl, Br, I, OH, NH 2 , CN, C 1-3 alkyl, C 1-3 alkoxy and C 3-6 cycloalkyl, said C 1-3 alkyl group, C 1-3 alkoxy and C 3-6 cycloalkyl optionally substituted with 1,2 or 3 substituents R a;
  • R 2 is selected from H, F, Cl, Br, I, OH, NH 2 , CN, C 1-3 alkyl and C 1-3 alkoxy, said C 1-3 alkyl and C 1-3 alkoxy
  • the oxy group is optionally substituted with 1, 2 or 3 R b ;
  • R 4 is selected from H, F, Cl, Br, I, OH, NH 2 , CN, C 1-5 alkyl, C 1-5 alkoxy, C 1-5 alkylamino, and 5-6 membered heteroaryl ring the C 1- 5 alkyl, C 1-5 alkoxy, C 1-5 alkylamino, and 5-6 membered heteroaromatic ring optionally substituted with 1, 2 or 3 R d;
  • R a and R b are independently selected from F, Cl, Br, I, OH, and NH 2 ;
  • R d is selected from H, F, Cl, Br, I, OH, NH 2 , CN, C 1-3 alkyl, C 1-3 alkoxy, and C 1-4 alkylamino;
  • T 1 , T 2 , T 3 and T 5 are independently selected from CH and N;
  • T 4 is selected from CR 5 and N;
  • L 2 is selected from NH and O;
  • L 3 is selected from -CH (R 6 )-, -CH (R 6 ) CH 2- , -CHCH 2 CH 2- , -CH (R 6 ) -O-, -CHCH 2 -O-, -CH 2- N (R 6 )-, -CH 2 CH 2 -N (R 6 )-;
  • R 5 is selected from H
  • R 6 is selected from H and CH 3 ;
  • R g is selected from H, F, Cl, Br, I, OH, NH 2 and CN;
  • the 5- to 6-membered heteroaryl group includes 1, 2, 3, or 4 heteroatoms or heteroatom groups independently selected from -NH-, -O-, -S-, and N, respectively.
  • the present invention also provides a compound represented by formula (II), an isomer thereof, or a pharmaceutically acceptable salt thereof,
  • R 1 is selected from H, F, Cl, Br, I, OH, NH 2 , CN, C 1-3 alkyl and C 1-3 alkoxy, said C 1-3 alkyl and C 1-3 alkoxy
  • the oxy group is optionally substituted with 1, 2 or 3 R a ;
  • R 2 is selected from H, F, Cl, Br, I, OH, NH 2 , CN, C 1-3 alkyl and C 1-3 alkoxy, said C 1-3 alkyl and C 1-3 alkoxy
  • the oxy group is optionally substituted with 1, 2 or 3 R b ;
  • R 3 is selected from H, F, Cl, Br, I, OH, NH 2 , CN, C 1-3 alkyl and C 1-3 alkoxy, said C 1-3 alkyl and C 1-3 alkoxy
  • the oxy group is optionally substituted by 1, 2 or 3 R c ;
  • the carbon atoms with "*" are chiral carbon atoms and exist in the form of (R) or (S) single enantiomer or are rich in one enantiomer;
  • R 4 is selected from H, F, Cl, Br, I, OH, NH 2 , CN, C 1-5 alkyl, C 1-5 alkoxy, C 1-5 alkylamino, and 5-6 membered heteroaryl ring the C 1- 5 alkyl, C 1-5 alkoxy, C 1-5 alkylamino, and 5-6 membered heteroaromatic ring optionally substituted with 1, 2 or 3 R d;
  • R a , R b and R c are each independently selected from F, Cl, Br, I, OH and NH 2 ;
  • R d is selected from H, F, Cl, Br, I, OH, NH 2 , CN, C 1-3 alkyl, C 1-3 alkoxy, and C 1-4 alkylamino;
  • T 1 , T 2 , T 3 , T 4 and T 5 are independently selected from CH and N;
  • a ring is selected from Said Optionally substituted with 1, 2 or 3 R g ;
  • L 1 and L 2 are independently selected from NH and O;
  • R g is selected from H, F, Cl, Br, I, OH, NH 2 and CN;
  • the 5- to 6-membered heteroaryl group includes 1, 2, 3, or 4 heteroatoms or heteroatom groups independently selected from -NH-, -O-, -S-, and N, respectively.
  • the present invention also provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,
  • R 1 is selected from H, F, Cl, Br, I, OH, NH 2 , CN, C 1-3 alkyl and C 1-3 alkoxy, said C 1-3 alkyl and C 1-3 alkoxy
  • the oxy group is optionally substituted with 1, 2 or 3 R a ;
  • R 2 is selected from H, F, Cl, Br, I, OH, NH 2 , CN, C 1-3 alkyl and C 1-3 alkoxy, said C 1-3 alkyl and C 1-3 alkoxy
  • the oxy group is optionally substituted with 1, 2 or 3 R b ;
  • R 3 is selected from H, F, Cl, Br, I, OH, NH 2 , CN, C 1-3 alkyl and C 1-3 alkoxy, said C 1-3 alkyl and C 1-3 alkoxy
  • the oxy group is optionally substituted by 1, 2 or 3 R c ;
  • the carbon atoms with "*" are chiral carbon atoms and exist in the form of (R) or (S) single enantiomer or are rich in one enantiomer;
  • R 4 is selected from H, F, Cl, Br, I, OH, NH 2 , CN, C 1-5 alkyl, C 1-5 alkoxy, C 1-5 alkylamino, and 5-6 membered heteroaryl ring the C 1- 5 alkyl, C 1-5 alkoxy, C 1-5 alkylamino, and 5-6 membered heteroaromatic ring optionally substituted with 1, 2 or 3 R d;
  • R a , R b and R c are each independently selected from F, Cl, Br, I, OH and NH 2 ;
  • R d is selected from H, F, Cl, Br, I, OH, NH 2 , CN, C 1-3 alkyl, C 1-3 alkoxy, and C 1-4 alkylamino;
  • T 1 , T 2 , T 3 , T 4 and T 5 are independently selected from CH and N;
  • a ring is selected from Said Optionally substituted with 1, 2 or 3 R g ;
  • R g is selected from H, F, Cl, Br, I, OH, NH 2 and CN;
  • the 5- to 6-membered heteroaryl group includes 1, 2, 3, or 4 heteroatoms or heteroatom groups independently selected from -NH-, -O-, -S-, and N, respectively.
  • R 1 is selected from the group consisting of H, F, Cl, Br, I, OH, NH 2 , CN, CH 3 , CH 2 CH 3 , Said CH 3 , CH 2 CH 3 , It is optionally substituted with 1, 2 or 3 R a .
  • R 1 is selected from the group consisting of H, F, Cl, Br, I, OH, NH 2 , CN, CH 3 , CH 2 CH 3 and Said CH 3 , CH 2 CH 3 and Optionally substituted with 1,2 or 3 substituents R a, the other variables are as defined in the present invention.
  • R 1 is selected from the group consisting of H, F, Cl, Br, I, OH, NH 2 , CN, CH 3 , CH 2 F, CHF 2 , CF 3 , CH 2 CH 3 and Other variables are as defined in the present invention.
  • R 1 is selected from CH 3 and Other variables are as defined in the present invention.
  • R 1 is selected from CH 3 and other variables are as defined in the invention.
  • the R 2 is selected from the group consisting of H, F, Cl, Br, I, OH, NH 2 , CN, CH 3 , CH 2 CH 3 and Said CH 3 , CH 2 CH 3 and It is optionally substituted by 1, 2 or 3 Rb , other variables are as defined in the present invention.
  • the R 2 is selected from CH 3 and other variables are as defined in the invention.
  • R 3 is selected from the group consisting of H, F, Cl, Br, I, OH, NH 2 , CN, CH 3 , CH 2 CH 3 and Said CH 3 , CH 2 CH 3 and It is optionally substituted by 1, 2 or 3 R c and other variables are as defined in the present invention.
  • the R 3 is selected from the group consisting of H, F, Cl, Br, I, OH, NH 2 , CN, CH 3 , CH 2 F, CHF 2 , CF 3 , CH 2 CH 3 and Other variables are as defined in the present invention.
  • R 3 is selected from CH 3 and other variables are as defined in the invention.
  • the R d is selected from H, F, Cl, Br, I, OH, NH 2, CN, CH 3 , and Other variables are as defined in the present invention.
  • the R 4 is selected from the group consisting of: H, F, Cl, Br, I, OH, NH 2 , CN, CH 3 , pyrazolyl, isoxazolyl, imidazolyl, triazolyl , Oxazolyl,
  • R 4 is selected from: Other variables are as defined in the present invention.
  • the structural unit From Other variables are as defined in the present invention.
  • the A ring is selected from Other variables are as defined in the present invention.
  • the A ring is selected from Other variables are as defined in the present invention.
  • the structural unit From Other variables are as defined in the present invention.
  • the structural unit From Other variables are as defined in the present invention.
  • the structural unit From Other variables are as defined in the present invention.
  • the compound, an isomer thereof, or a pharmaceutically acceptable salt thereof is selected from the group consisting of
  • R 1 , R 2 , R 3 , R 4 , L 2 , L 3 , T 1 , T 2 , T 3 , T 4 and T 5 are as defined in the present invention.
  • the present invention also has some solutions from any combination of the above variables.
  • the present invention provides a compound represented by the following formula, an isomer thereof, or a pharmaceutically acceptable salt thereof, which is selected from:
  • the compound, an isomer thereof, or a pharmaceutically acceptable salt thereof is selected from:
  • the present invention also provides a pharmaceutical composition containing a therapeutically effective amount of the above-mentioned compound, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient and a pharmaceutically acceptable carrier.
  • the present invention also provides the use of the aforementioned compound, an isomer thereof, or a pharmaceutically acceptable salt thereof in the preparation of an RET kinase inhibitor.
  • the invention also provides the application of the above composition in the preparation of a RET kinase inhibitor.
  • pharmaceutically acceptable refers to those compounds, materials, compositions, and / or dosage forms that are within the scope of sound 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 a sufficient amount of a base with a neutral form of such compounds 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 the neutral form of 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, Hydrogen sulfate, 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, Similar acids such as fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; also include salts of amino acids (such as arginine, etc.) , And salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain basic and acidic functional groups
  • the pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing an acid group or a base by a conventional chemical method. Generally, such salts are prepared by reacting these compounds in the form of a free acid or base with a stoichiometric appropriate base or acid in water or an organic solvent or a mixture of the two.
  • the compounds of the invention may exist in specific geometric or stereoisomeric forms.
  • This invention contemplates all such compounds, including cis and trans isomers, (-)-and (+)-enantiomers, (R)-and (S) -enantiomers, diastereomers Isomers, (D) -isomers, (L) -isomers, and racemic and other mixtures thereof, such as enantiomeric or diastereomeric enriched mixtures, all of which belong to the present invention Within the scope of the invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl. All these isomers and their mixtures are included in the scope of the present invention.
  • enantiomers or “optical isomers” refer to stereoisomers in mirror image relationship to each other.
  • cis-trans isomer or “geometric isomer” are caused by the inability of a double bond or a single bond of a ring-forming carbon atom to rotate freely.
  • diastereomer refers to a stereoisomer in which a molecule has two or more centers of chirality and is in a non-mirror relationship between molecules.
  • wedge solid line key And wedge dashed keys Represents the absolute configuration of a solid center, using straight solid line keys And straight dashed keys Represents the relative configuration of the solid center, with wavy lines Represents a wedge solid line key Or wedge-shaped dotted key Or with wavy lines Represents a straight solid line key And straight dashed keys
  • tautomer or “tautomeric form” means that at room temperature, the isomers of different functional groups are in dynamic equilibrium and can be quickly converted to each other. If tautomers are possible (eg in solution), the chemical equilibrium of the tautomers can be reached.
  • proton tautomers also known as prototropic tautomers
  • proton migration such as keto-enol isomerization and imine-ene Amine isomerization.
  • Valence tautomers include recombination of some bonding electrons for mutual conversion.
  • a specific example of the 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”, “enriched in isomers”, “enriched in one enantiomer” or “enantiomerically enriched” refer to one of the isomers or the The enantiomeric content is less than 100%, and the content of the isomer or enantiomer is 60% or more, or 70% or more, or 80% or more, or 90% or more, or 95% or more, or 96% or more, or 97% or more, or 98% or more, or 99% or more, or 99.5% or more, or 99.6% or more, or 99.7% or more, or 99.8% or more, or more 99.9%.
  • the terms “isomer excess” or “enantiomeric excess” refer to the difference between the two isomers or the relative percentages of the 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%, the isomer or enantiomeric excess (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, in which the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide pure The desired enantiomer.
  • a diastereomeric salt is formed with an appropriate optically active acid or base, and then by a conventional method known in the art Diastereomeric resolution is performed and the pure enantiomer is recovered.
  • Diastereomeric resolution is performed and the pure enantiomer is recovered.
  • the separation of enantiomers and diastereoisomers is usually accomplished by using chromatography that employs a chiral stationary phase and optionally is combined with chemical derivatization (such as the generation of amino groups from amines) Formate).
  • the compounds of the invention may contain atomic isotopes in unnatural proportions on one or more of the atoms constituting the compound.
  • compounds such as tritium ( 3 H), iodine-125 ( 125 I) or C-14 ( 14 C) can be labeled with radioisotopes.
  • deuterated drugs can be replaced by heavy hydrogen. The bond between deuterium and carbon is stronger than the bond between ordinary hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs have reduced side effects and increased drug stability. , Enhance efficacy, extend the biological half-life of drugs and other advantages. Transformations of all isotopic compositions of the compounds of the invention, whether radioactive or not, are included within the scope of the invention.
  • substituted refers to the replacement of any one or more hydrogen atoms on a specific atom with a substituent, and can include deuterium and hydrogen variants, as long as the valence of the specific atom is normal and the substituted compound is stable of.
  • O oxygen
  • Oxygen substitution does not occur on aromatic groups.
  • optionally substituted means that it may or may not be substituted, and unless otherwise specified, the kind and number of substituents may be arbitrary on the basis of chemically achievable.
  • any variable such as R
  • its definition in each case is independent.
  • the group may be optionally substituted with at most two R, and R in each case has independent options.
  • combinations of substituents and / or variants are only permitted if such combinations result in stable compounds.
  • linking group When the number of a linking group is 0, such as-(CRR) 0- , the linking group is a single bond.
  • a substituent When a substituent is vacant, it means that the substituent does not exist. For example, when X is vacant in AX, it means that the structure is actually A.
  • the listed linking group does not indicate its connection direction, its connection direction is arbitrary, for example, The intermediate linking group L is -MW-. At this time, -MW- can be connected to ring A and ring B in the same direction as the reading order from left to right. You can also connect ring A and ring B in the opposite direction from the reading order from left to right. Combinations of the linking groups, substituents, and / or variants thereof are permitted only if such combinations result in stable compounds.
  • C 1-5 alkyl is used to indicate a straight or branched chain saturated hydrocarbon group consisting of 1 to 5 carbon atoms.
  • the C 1-5 alkyl group includes C 1-4 , C 1-3 , C 1-2 , C 2-5 , C 2-4 and C 5 alkyl, etc .; it may be monovalent (such as methyl) , Divalent (such as methylene) or polyvalent (such as methine).
  • C 1-5 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) and the like.
  • C 1-3 alkyl is used to indicate a straight or branched chain saturated hydrocarbon group consisting of 1 to 3 carbon atoms.
  • the C 1-3 alkyl group includes C 1-2 and C 2-3 alkyl groups, and the like; it may be monovalent (such as methyl), divalent (such as methylene), or polyvalent (such as methine).
  • Example C 1- 3 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n- propyl and isopropyl) and the like.
  • C 2-8 alkenyl is used to indicate a straight or branched hydrocarbon group consisting of 2 to 8 carbon atoms containing at least one carbon-carbon double bond. It can be located anywhere on this group.
  • the C 2-8 alkenyl includes C 2-6 , C 2-4 , C 2-3 , C 4 , C 3, and C 2 alkenyl, and the like; it may be monovalent, divalent, or polyvalent.
  • Examples of C 2-8 alkenyl include, but are not limited to, vinyl, propenyl, butenyl, pentenyl, hexenyl, butadienyl, pentadienyl, hexadienyl, and the like.
  • C 1-5 alkoxy refers to those alkyl groups containing 1 to 5 carbon atoms that are connected to the rest of the molecule through one oxygen atom.
  • the C 1-5 alkoxy group includes C 1-5 , C 1-4 , C 1-3 , C 1-2 , C 2-5 , C 2-4 , C 5 , C 4 , C 3 and C 2 alkoxy and the like.
  • C 1- 5 alkoxy groups 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), and the like.
  • C 1-3 alkoxy refers to those alkyl groups containing 1 to 3 carbon atoms that are attached to the rest of the molecule through one oxygen atom.
  • the C 1-3 alkoxy group includes C 1-2 , C 2-3 , C 3 and C 2 alkoxy, and the like.
  • Examples of C 1-3 alkoxy include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.
  • C 1-5 alkylamino refers to those alkyl groups containing 1 to 5 carbon atoms that are attached to the rest of the molecule through an amino group.
  • the C 1-6 alkylamino group includes C 1-5 , C 1-4 , C 1-3 , C 1-2 , C 2-5 , C 2-4 , C 5 , C 4 , C 3 and C 2 Alkylamino, etc.
  • C 1-5 alkylamino examples include, but are not limited to, -NHCH 3 , -N (CH 3 ) 2 , -NHCH 2 CH 3 , -N (CH 3 ) CH 2 CH 3 , -N (CH 2 CH 3 ) ( CH 2 CH 3 ), -NHCH 2 CH 2 CH 3 , -NHCH 2 (CH 3 ) 2 , -NHCH 2 CH 2 CH 2 CH 3, and the like.
  • C 1-4 alkylamino refers to those alkyl groups containing 1 to 4 carbon atoms attached to the rest of the molecule through an amino group.
  • the C 1-4 alkylamino group includes C 1-3 , C 1-2 , C 2-4 , C 4 , C 3 and C 2 alkylamino, and the like.
  • C 1-4 alkylamino examples include, but are not limited to, -NHCH 3 , -N (CH 3 ) 2 , -NHCH 2 CH 3 , -N (CH 3 ) CH 2 CH 3 , -N (CH 2 CH 3 ) ( CH 2 CH 3 ), -NHCH 2 CH 2 CH 3 , -NHCH 2 (CH 3 ) 2 , -NHCH 2 CH 2 CH 2 CH 3, and the like.
  • C 3-6 cycloalkyl means a saturated cyclic hydrocarbon group consisting of 3 to 6 carbon atoms, which is a monocyclic and bicyclic system.
  • the C 3-6 cycloalkyl includes C 3-5 , C 4-5 and C 5-6 cycloalkyl and the like; it may be monovalent, divalent or polyvalent.
  • Examples of C 3-6 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.
  • the terms “5-6 membered heteroaryl ring” and “5-6 membered heteroaryl group” in the present invention are used interchangeably, and the term “5-6 membered heteroaryl group” means from 5 to 6 ring atoms A single-ring group consisting of a conjugated ⁇ -electron system.
  • One, two, three, or four ring atoms are heteroatoms independently selected from O, S, and N, and the rest are carbon atoms.
  • the nitrogen and sulfur heteroatoms can be optionally oxidized (ie NO and S (O) p , p is 1 or 2).
  • the 5- to 6-membered heteroaryl can be attached to the rest of the molecule through a heteroatom or a carbon atom.
  • the 5- to 6-membered heteroaryl includes 5- and 6-membered heteroaryl.
  • Examples of the 5- to 6-membered heteroaryl include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrrolyl 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-triazo
  • 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 include 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.
  • n yuan to n + m means that the number of atoms on the ring is n to n + m.
  • 3-12-membered rings include 3-, 4-, 5-, 6-, 7-, 8-, and 9-membered rings , 10-membered ring, 11-membered ring, and 12-membered ring, 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 Ring, 5-7 member ring, 6-7 member ring, 6-8 member ring, and 6-10 member ring, etc.
  • halogen refers to a fluorine, chlorine, bromine or iodine atom by itself or as part of another substituent.
  • haloalkyl is intended to include monohaloalkyl and polyhaloalkyl.
  • halo (C 1 -C 4 ) alkyl is intended to include, but not limited to, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like Wait.
  • examples of haloalkyl include, but are not limited to, trifluoromethyl, trichloromethyl, pentafluoroethyl, and pentachloroethyl.
  • 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-toluenesulfonic acid. Esters, etc .; acyloxy, such as acetoxy, trifluoroacetoxy and the like.
  • 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-butyldi
  • hydroxy-protecting group refers to a protecting group suitable for preventing a hydroxyl side reaction.
  • Representative hydroxy protecting groups include, but are not limited to: alkyl groups such as methyl, ethyl, and tert-butyl; acyl groups such as alkanoyl (such as acetyl); aryl methyl groups such as benzyl (Bn), p-formyl Oxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl groups such as trimethylsilyl (TMS) and tert-butyl Dimethylsilyl (TBS) and more.
  • alkyl groups such as methyl, ethyl, and tert-butyl
  • acyl groups such as alkanoyl (such as acetyl)
  • aryl methyl groups such as benzyl (Bn), p-formyl 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 combining them with other chemical synthesis methods, and those familiar to those skilled in the art. Equivalent alternatives, preferred embodiments include, but are not limited to, the embodiments of the present invention.
  • the compounds of the invention may have a variety of uses or indications, including but not limited to the specific uses or indications enumerated herein.
  • the solvent used in the present invention is commercially available.
  • the present invention uses the following abbreviations: aq stands for water; HATU stands for O- (7-azabenzotriazol-1-yl) -N, N, N ', N'-tetramethylurea hexafluorophosphate ; EDC stands for N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride; m-CPBA stands for 3-chloroperoxybenzoic acid; eq stands for equivalent, equivalent; M stands for mol / L; CDI stands for carbonyldiimidazole; DCM stands for dichloromethane; PE stands for petroleum ether; DIAD stands for diisopropyl azodicarboxylate; DMF stands for N, N-dimethylformamide; DMSO stands for dimethylene Sulfone; EtOAc for ethyl acetate; EtOH for ethanol; MeOH for m
  • the compound of the present invention has excellent inhibitory activity on RET and its mutation RET V804M, and will have excellent therapeutic effect in patients with RET abnormal tumors.
  • a catalytic amount of DMF (2.32 g, 31.74 mmol) was added to a mixed solution of 6-chloronicotinic acid (50 g, 317.35 mmol) containing sulfoxide (150 mL), and refluxed at 85 ° C for 3 hours under a nitrogen atmosphere. After spinning off the solution, it was dissolved in DCM (1200 mL), methoxy-N-methyl-amino hydrochloride (46.43 g, 476.03 mmol) was added, cooled to 0 ° C, and triethylamine (96.34 g, 952.05 mmol), and stirred at 25 ° C for 16 hours. The reaction solution was added with a saturated sodium bicarbonate solution, and the layers were separated to obtain an organic phase. The organic phase was dried over sodium sulfate and concentrated under reduced pressure to give 1a.
  • 2,4-dichloro-6-methyl-pyrimidine (10 g, 61.35 mmol), 5-methyl-3-amino-1H-pyrazole (6.55 g, 67.48 mmol), diisopropylethylamine (11.89 g, 92.02 mmol, 16.03 mL) was added to DMSO (30 mL), and the reaction was stirred at 60 ° C for 16 hours.
  • the reaction solution was cooled to 20-30 ° C, poured into 200 mL of ice water, stirred at 15-20 ° C for 1 hour, filtered, and the filter cake was dried under reduced pressure at 40-50 ° C to obtain 1f.
  • Triphosgene (23.95 mg, 80.70 ⁇ mol) was added to DCM (4 mL), and diisopropylethylamine (62.58 mg,
  • Example 1 The crude product was separated and purified by a preparative chromatography (HCl system) to obtain the hydrochloride of Example 1.
  • the hydrochloride of Example 1 was added to a sodium bicarbonate solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate. Concentrated under reduced pressure to obtain Example 1.
  • the crude product is sent to preparative chromatography (chromatographic column: Xtimate 150 ⁇ 25mm ⁇ 5 ⁇ m; mobile phase: [water (0.075% trifluoroacetic acid) -acetonitrile]; B%: 10% -30%, 7min), and then sent to SFC for resolution (Chromatography column: DAICEL CHIRALPAK AS-H (250mm ⁇ 30mm, 5 ⁇ m); mobile phase: [0.1% ammonia water EtOH]; B%: 45% -45%)
  • Example 2 was obtained (retention time: 4.957 min).
  • Example 4 The crude product was separated by a preparative chromatographic column (column: Xtimate 150 ⁇ 25 mm ⁇ 5 ⁇ m; mobile phase: [water (0.225% trifluoroacetic acid) -acetonitrile]; B%: 25% -45%, 7min).
  • the trifluoroacetate of Example 4 was obtained.
  • the trifluoroacetate of Example 4 was added to a sodium bicarbonate solution, extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain Example 4.
  • the crude product was separated using a preparative chromatographic column (chromatographic column: ⁇ timate, C18, 150 ⁇ 25 mm ⁇ 5 ⁇ m; mobile phase: [water (0.075% trifluoroacetic acid) -acetonitrile]; B%: 15% -35%, 8min) to obtain 6d.
  • chromatographic column ⁇ timate, C18, 150 ⁇ 25 mm ⁇ 5 ⁇ m; mobile phase: [water (0.075% trifluoroacetic acid) -acetonitrile]; B%: 15% -35%, 8min
  • the reaction solution was spin-dried to obtain a crude product, which was purified by a preparative column (column: Xtimate, C18 150 ⁇ 25mm ⁇ 5 ⁇ m; mobile phase: [water (0.075% trifluoroacetic acid) -acetonitrile]; B%: 32% -46%, 7min)
  • the product 10a is obtained.
  • Compound 10a was subjected to chiral resolution.
  • the SFC resolution conditions were a chromatographic column: YMC CHIRAL Amylose-C (250mm ⁇ 30mm, 10 ⁇ m; mobile phase: [0.1% ammonia water isopropanol]; B%: 50% -50%, min.
  • reaction solution 1 1e (23.90mg, 115.88 ⁇ mol) was added to DCM (5mL), triphosgene (19.10mg, 64.38 ⁇ mol) was added to the reaction solution, and DIEA (49.92mg, 386.26 ⁇ mol, 67.28 ⁇ L) was added dropwise to the reaction solution.
  • the reaction temperature was lowered to 0 ° C, and was designated as reaction liquid 2.
  • the reaction liquids 1 and 2 were stirred at 0 ° C for 10 minutes.
  • the reaction liquid 1 was slowly added to the reaction liquid 2 at 0 ° C, and the reaction was stirred at 0 ° C. 10min. 10 mL of water was added to the reaction solution, followed by extraction with DCM (5 mL ⁇ 2).
  • the organic phases were combined, the organic phases were dried over anhydrous sodium sulfate, and dried under reduced pressure at 40-45 ° C to obtain a crude product.
  • the crude product was passed through a thin layer to prepare a chromatography plate. Purification yielded Example 12.
  • Example 13 Water (2 mL) was added to the reaction solution to quench the reaction, and concentrated to obtain a crude product.
  • the crude product was purified by preparative chromatography (column: Boston Green ODS 150 ⁇ 30mm 5 ⁇ m; mobile phase: [water (0.075% trifluoroacetic acid) -acetonitrile]; B%: 29% -45%, 9min) to obtain the trifluoro of Example 13 Acetate, the trifluoroacetate of Example 13 was added to a sodium bicarbonate solution, extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, and concentrated to obtain Example 13.
  • Example 14 The reaction solution was directly spin-dried to obtain a crude product, and then separated by a preparative chromatography column (chromatographic column: Boston, Green, ODS, 150 ⁇ 30 mm, 5 ⁇ m; mobile phase: [water (0.075% trifluoroacetic acid) -acetonitrile]; B%: 23% -53%, 8min), and the trifluoroacetate of Example 14 was purified.
  • the trifluoroacetate of Example 14 was added to a sodium bicarbonate solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain Example 14.
  • 15a (6.5 g) was separated with SFC (chromatographic column: Phenomenex-Amylose-1 (250 mm ⁇ 30 mm, 5 ⁇ m); mobile phase: [0.1% ammonia water ethanol]; B%: 25% -25%, min). 15b1 (retention time: 1.882min) and 15b2 (retention time: 2.010min) were obtained.
  • Example 15 The reaction solution was directly sent to a preparative chromatographic column (chromatographic column: Welch Xtimate C18 150 * 25mm * 5um; mobile phase: [water (0.075% trifluoroacetic acid) -acetonitrile]; B%: 24% -44%, 12min) was purified and implemented Trifluoroacetate of Example 15.
  • the trifluoroacetate of Example 15 was added to a sodium bicarbonate solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain Example 15.
  • Example 16 Using 15b2 as a raw material, the trifluoroacetate of Example 16 was obtained in the same manner. The trifluoroacetate of Example 16 was added to a sodium bicarbonate solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain Example 16.
  • Example 17 Trifluoroacetate of Example 17.
  • the trifluoroacetate of Example 17 was added to a sodium bicarbonate solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain Example 17.
  • the compound 17c (1.00 g, 1.21 mmol, TFA) was dissolved in dichloromethane (1 mL), and N, N-diisopropylethylamine (470.04 mg, 3.64 mmol, 633.48 uL) was added.
  • Example 18 The crude product was separated by preparative HPLC (column: YMC-Triart Prep C18 150 * 40mm * 7um; : [Water (0.1% trifluoroacetic acid) -acetonitrile]; B%: 25% -45%, 10min) to obtain the trifluoroacetate salt of Example 18.
  • the trifluoroacetate of Example 18 was added to a sodium bicarbonate solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain Example 18.
  • the trifluoroacetate of Example 19 was added to a sodium bicarbonate solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain Example 19.
  • Example 20 1 mL of water was added to the reaction solution to quench the reaction, and then 5 mL of water and 5 mL of dichloromethane were added for extraction. The layers were separated, and the aqueous phase was extracted with 5 mL of dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, and evaporated to dryness. The crude product was purified by preparation (column: Boston, Green ODS 150 * 30, 5u; mobile phase: [water (0.075% trifluoroacetic acid) -acetonitrile]; B%: 33% -53%, 7min) to obtain three of Example 20. Fluoroacetate. The trifluoroacetate of Example 20 was added to a sodium bicarbonate solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain Example 20.
  • Example 20 The trifluoroacetate of Example 20 (60mg, 118.92umol) was sent to SFC for separation (column: DAICELCHIRALPAK AS-H (250mm * 30mm, 5um); mobile phase: [0.1% ammonia water ethanol]; B%: 50% -50%, min). Finally, Example 21 (retention time: 3.391min) and Example 22 (retention time: 4.092min)
  • Example 23 Send the crude product for preparation and separation (chromatographic column: Boston, Green, ODS, 150 * 30mm * 5um; mobile phase: [water (0.075% trifluoroacetic acid) -acetonitrile]; B%: 30% -60%, 9min) Fluoroacetate.
  • the trifluoroacetate of Example 23 was added to a sodium bicarbonate solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain Example 23.
  • Potassium permanganate (45.60 g, 288.54 mmol) was added to a mixed solution of compound 24a (7 g, 48.09 mmol) in water (60 mL) and pyridine (60 mL), and stirred at 100 ° C for 3 hours.
  • the reaction solution was directly filtered, washed with 100 mL of methanol, and the filtrate was concentrated under reduced pressure to obtain compound 24b.
  • SFC chromatographic column: DAICEL CHIRALPAK AD-H ( 250mm * 30mm, 5 ⁇ m); mobile phase: [0.1% ammonia water methanol]; B%: 35% -35%, min) separated to 24g1 (retention time: 4.177min) and 24g2 (retention time: 4.980min).
  • Example 24 Trifluoroacetate.
  • the trifluoroacetate of Example 24 was added to a sodium bicarbonate solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain Example 24.
  • Example 25 Using the compound 24g2, the trifluoroacetate of Example 25 was prepared in the same manner. The trifluoroacetate of Example 25 was added to a sodium bicarbonate solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain Example 25.
  • Example 28a (130mg, 257.15umol) was sent to SFC for separation.
  • the SFC conditions are (chromatographic column: DAICELCHIRALPAKAD (250mm * 30mm, 10um); mobile phase: [0.1% NH3H2O, IPA]; B%: 60% -60%, min).
  • Example 28 (retention time: 1.499min) and Example 29 (retention time: 2.252min) were obtained.
  • Z′-LYTE TM kinase test was used to test the enzyme activity, and the compound's IC 50 value was used as an index to evaluate the inhibitory effect of the compound on RET and RET (V804M) kinase.
  • the compounds used in the test of RET and RET (V804M) kinase were diluted three-fold to a concentration of 3 ⁇ M to 0.152 nM 10 concentrations; the content of DMSO in the detection reaction was 1%.
  • Basic reaction buffer 20 mM Hydroxyethylpiperazine Ethionate (pH 7.5) buffer, 10 mM MgCl 2 , 1 mM ethylene glycol bisaminoethyl ether tetraacetic acid (EGTA), 0.02% polyoxyethylene twelve Alkyl ether (Brij35), 0.02 mg / ml bovine serum albumin (), 0.1 mM Na 3 VO 4 , 2 mM dithiothreitol (DTT), 1% DMSO.
  • EGTA ethylene glycol bisaminoethyl ether tetraacetic acid
  • Brij35 polyoxyethylene twelve Alkyl ether
  • DTT dithiothreitol
  • test compound was dissolved in 100% DMSO system and diluted to 10 mM for use.
  • the solution was diluted using Integra Viaflo Assist.
  • General enzyme reaction process :
  • the concentration of RET enzyme is 3 ⁇ M, the concentration of peptide substrate CHKtide is 1000 ⁇ M, the concentration of ATP is 20 ⁇ M; the concentration of RET (V804M) enzyme is 80 ⁇ M, the concentration of substrate peptide is 1000 ⁇ M, and the concentration of ATP is 30 ⁇ M.
  • Reaction process Prepare the kinase / polypeptide solution according to the test conditions. Compound solutions of different concentrations were added, and incubated at room temperature for 20 minutes, and corresponding concentrations of 33P-ATP were added, and the reaction was incubated at room temperature for 120 minutes. Radioactivity was detected by the filter-binding method.
  • the reaction was stopped by adding 0.5% phosphoric acid to the kinase reaction solution, and the plate was read by the Envision instrument.
  • the data were converted into phosphorylation rate and inhibition rate, and the parameter IC 50 was obtained by parameter curve fitting (GraphPad Software).
  • the compound of the present application has excellent inhibitory activity on RET and its mutation RET V804M, and will have excellent therapeutic effect in patients with RET abnormal tumors.

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Abstract

一系列带有吡唑结构的化合物，及其在制备RET激酶抑制剂中的应用。所述化合物具体为式(III)所示的衍生化合物、其异构体或其药学上可接受的盐。

Description

作为RET抑制剂的吡唑衍生物
相关申请的引用
本申请主张如下优先权：
CN201810947337.1，申请日2018-08-17；
CN201811322012.0，申请日2018-11-07；
CN201811501055.5，申请日2018-12-07；
CN201910482535.X，申请日2019-06-04；
CN201910487945.3，申请日2019-06-05。
技术领域
本发明涉及一类吡唑衍生物及其在制备治疗与RET激酶抑制剂相关疾病的药物中的应用。具体涉及式(III)所示化合物及其药学上可接受的盐。
背景技术
RET蛋白是一个受体酪氨酸激酶RTK，同时也是一个跨膜的糖蛋白，由位于10号染色体上的原癌基因RET(REarranged during Transfection)表达，在胚胎阶段的肾脏和肠神经系统的发育中起着重要作用，另外在多种组织内稳态也很关键，如神经元、神经内分泌、造血组织和男性生殖细胞等。和其他的RTK不同，RET并不是直接结合到配体分子：如神经导向素(artemin)、胶质细胞源性神经营养因子(GDNF)、neurturin和persephin，这些都是属于GNDF家族配体(GFLs)。这些配体GFLs通常结合到GDNF家族受体α(GFRα)，形成的GFLs-GFRα复合物介导了RET蛋白的自二聚化，引起胞内结构域上酪氨酸的反式自磷酸化反应，招募相关接头蛋白，激活细胞增殖等信号传导的级联反应，相关的信号通路包括MAPK、PI3K、JAK-STAT、PKA、PKC等等。
RET的致癌激活机制主要有两个：一是染色体的重排产生了新的融合蛋白，通常是RET的激酶结构域和包含自二聚化结构域的蛋白融合；二就是RET突变直接或间接的激活了RET的激酶活性。这些体细胞或生殖细胞水平的改变涉及多种癌症的发病机制。5％-10％的乳头状甲状腺癌患者存在RET染色体重排；而在髓样性甲状腺髓样癌中发现有60％存在RET点突变；在所有NSCLC患者中，大概有1-2％的具有RET融合蛋白，其中KIF5B-RET最为多见。
总之，在多种肿瘤和胃肠道紊乱如过敏性肠道综合症中均发现异常的RET表达或激活。因此RET抑制剂在肿瘤或肠道紊乱疾病中具有潜在的临床价值。
发明内容
本发明提供式(III)所示化合物、其异构体或其药学上可接受的盐，
Figure PCTCN2019101121-appb-000001
其中，
A环选自
Figure PCTCN2019101121-appb-000002
所述
Figure PCTCN2019101121-appb-000003
任选被1、2或3个R g取代；
R 1选自H、F、Cl、Br、I、OH、NH 2、CN、C 1-3烷基、C 1-3烷氧基和C 3-6环烷基，所述C 1-3烷基、C 1-3烷氧基和C 3-6环烷基任选被1、2或3个R a取代；
R 2选自H、F、Cl、Br、I、OH、NH 2、CN、C 1-3烷基和C 1-3烷氧基，所述C 1-3烷基和C 1-3烷氧基任选被1、2或3个R b取代；
R 4选自H、F、Cl、Br、I、OH、NH 2、CN、C 1-5烷基、C 1-5烷氧基、C 1-5烷氨基和5-6元杂芳环，所述C 1- 5烷基、C 1-5烷氧基、C 1-5烷氨基和5-6元杂芳环任选被1、2或3个R d取代；
R a和R b分别独立地选自F、Cl、Br、I、OH和NH 2；
R d选自H、F、Cl、Br、I、OH、NH 2、CN、C 1-3烷基、C 1-3烷氧基和C 1-4烷氨基；
T 1、T 2、T 3和T 5分别独立地选自CH和N；
T 4选自CR 5和N；
L 2选自NH和O；
L 3选自-CH(R 6)-、-CH(R 6)CH 2-、-CHCH 2CH 2-、-CH(R 6)-O-、-CHCH 2-O-、-CH 2-N(R 6)-、-CH 2CH 2-N(R 6)-；
R 5选自H；
R 6选自H和CH 3；
或者R 5、R 6连接在一起使结构单元
Figure PCTCN2019101121-appb-000004
选自
Figure PCTCN2019101121-appb-000005
R g选自H、F、Cl、Br、I、OH、NH 2和CN；
所述5～6元杂芳基分别包含1、2、3或4个独立选自-NH-、-O-、-S-和N的杂原子或杂原子团。
本发明还提供了式(II)所示化合物、其异构体或其药学上可接受的盐，
Figure PCTCN2019101121-appb-000006
其中，
R 1选自H、F、Cl、Br、I、OH、NH 2、CN、C 1-3烷基和C 1-3烷氧基，所述C 1-3烷基和C 1-3烷氧基任选被1、2或3个R a取代；
R 2选自H、F、Cl、Br、I、OH、NH 2、CN、C 1-3烷基和C 1-3烷氧基，所述C 1-3烷基和C 1-3烷氧基任选被1、2或3个R b取代；
R 3选自H、F、Cl、Br、I、OH、NH 2、CN、C 1-3烷基和C 1-3烷氧基，所述C 1-3烷基和C 1-3烷氧基任选被1、2或3个R c取代；
带“*”碳原子为手性碳原子，以(R)或(S)单一对映体形式或富含一种对映体形式存在；
R 4选自H、F、Cl、Br、I、OH、NH 2、CN、C 1-5烷基、C 1-5烷氧基、C 1-5烷氨基和5-6元杂芳环，所述C 1- 5烷基、C 1-5烷氧基、C 1-5烷氨基和5-6元杂芳环任选被1、2或3个R d取代；
R a、R b和R c分别独立地选自F、Cl、Br、I、OH和NH 2；
R d选自H、F、Cl、Br、I、OH、NH 2、CN、C 1-3烷基、C 1-3烷氧基和C 1-4烷氨基；
T 1、T 2、T 3、T 4和T 5分别独立地选自CH和N；
A环选自
Figure PCTCN2019101121-appb-000007
所述
Figure PCTCN2019101121-appb-000008
任选被1、2或3个R g取代；
L 1和L 2分别独立地选自NH和O；
R g选自H、F、Cl、Br、I、OH、NH 2和CN；
所述5～6元杂芳基分别包含1、2、3或4个独立选自-NH-、-O-、-S-和N的杂原子或杂原子团。
本发明还提供了式(Ⅰ)所示化合物或其药学上可接受的盐，
Figure PCTCN2019101121-appb-000009
其中，
R 1选自H、F、Cl、Br、I、OH、NH 2、CN、C 1-3烷基和C 1-3烷氧基，所述C 1-3烷基和C 1-3烷氧基任选被1、2或3个R a取代；
R 2选自H、F、Cl、Br、I、OH、NH 2、CN、C 1-3烷基和C 1-3烷氧基，所述C 1-3烷基和C 1-3烷氧基任选被1、2或3个R b取代；
R 3选自H、F、Cl、Br、I、OH、NH 2、CN、C 1-3烷基和C 1-3烷氧基，所述C 1-3烷基和C 1-3烷氧基任选被1、2或3个R c取代；
带“*”碳原子为手性碳原子，以(R)或(S)单一对映体形式或富含一种对映体形式存在；
R 4选自H、F、Cl、Br、I、OH、NH 2、CN、C 1-5烷基、C 1-5烷氧基、C 1-5烷氨基和5-6元杂芳环，所述C 1- 5烷基、C 1-5烷氧基、C 1-5烷氨基和5-6元杂芳环任选被1、2或3个R d取代；
R a、R b和R c分别独立地选自F、Cl、Br、I、OH和NH 2；
R d选自H、F、Cl、Br、I、OH、NH 2、CN、C 1-3烷基、C 1-3烷氧基和C 1-4烷氨基；
T 1、T 2、T 3、T 4和T 5分别独立地选自CH和N；
A环选自
Figure PCTCN2019101121-appb-000010
所述
Figure PCTCN2019101121-appb-000011
任选被1、2或3个R g取代；
R g选自H、F、Cl、Br、I、OH、NH 2和CN；
所述5～6元杂芳基分别包含1、2、3或4个独立选自-NH-、-O-、-S-和N的杂原子或杂原子团。
在本发明的一些方案中，所述R 1选自H、F、Cl、Br、I、OH、NH 2、CN、CH 3、CH 2CH 3、
Figure PCTCN2019101121-appb-000012
所述CH 3、CH 2CH 3、
Figure PCTCN2019101121-appb-000013
任选被1、2或3个R a取代。
在本发明的一些方案中，所述R 1选自H、F、Cl、Br、I、OH、NH 2、CN、CH 3、CH 2CH 3和
Figure PCTCN2019101121-appb-000014
所述CH 3、CH 2CH 3和
Figure PCTCN2019101121-appb-000015
任选被1、2或3个R a取代，其他变量如本发明所定义。
在本发明的一些方案中，所述R 1选自H、F、Cl、Br、I、OH、NH 2、CN、CH 3、CH 2F、CHF 2、CF 3、CH 2CH 3和
Figure PCTCN2019101121-appb-000016
其他变量如本发明所定义。
在本发明的一些方案中，所述R 1选自CH 3和
Figure PCTCN2019101121-appb-000017
其他变量如本发明所定义。
在本发明的一些方案中，所述R 1选自CH 3，其他变量如本发明所定义。
在本发明的一些方案中，所述R 2选自H、F、Cl、Br、I、OH、NH 2、CN、CH 3、CH 2CH 3和
Figure PCTCN2019101121-appb-000018
所述CH 3、CH 2CH 3和
Figure PCTCN2019101121-appb-000019
任选被1、2或3个R b取代，其他变量如本发明所定义。
在本发明的一些方案中，所述R 2选自CH 3，其他变量如本发明所定义。
在本发明的一些方案中，所述R 3选自H、F、Cl、Br、I、OH、NH 2、CN、CH 3、CH 2CH 3和
Figure PCTCN2019101121-appb-000020
所述CH 3、CH 2CH 3和
Figure PCTCN2019101121-appb-000021
任选被1、2或3个R c取代，其他变量如本发明所定义。
在本发明的一些方案中，所述R 3选自H、F、Cl、Br、I、OH、NH 2、CN、CH 3、CH 2F、CHF 2、CF 3、CH 2CH 3和
Figure PCTCN2019101121-appb-000022
其他变量如本发明所定义。
在本发明的一些方案中，所述R 3选自CH 3，其他变量如本发明所定义。
在本发明的一些方案中，所述R d选自H、F、Cl、Br、I、OH、NH 2、CN、CH 3和
Figure PCTCN2019101121-appb-000023
其他变量如本发明所定义。
在本发明的一些方案中，所述R 4选自：H、F、Cl、Br、I、OH、NH 2、CN、CH 3、吡唑基、异噁唑基、咪唑基、三唑基、噁唑基、
Figure PCTCN2019101121-appb-000024
所述CH 3、吡唑基、异噁唑基、咪唑基、三唑基、噁唑基、
Figure PCTCN2019101121-appb-000025
任选被1、2或3个R d取代，其他变量如本发明所定义。
在本发明的一些方案中，所述R 4选自：
Figure PCTCN2019101121-appb-000026
Figure PCTCN2019101121-appb-000027
其他变量如本发明所定义。
在本发明的一些方案中，所述结构单元
Figure PCTCN2019101121-appb-000028
选自
Figure PCTCN2019101121-appb-000029
Figure PCTCN2019101121-appb-000030
其他变量如本发明所定义。
在本发明的一些方案中，所述A环选自
Figure PCTCN2019101121-appb-000031
其他变量如本发明所定义。
在本发明的一些方案中，所述A环选自
Figure PCTCN2019101121-appb-000032
其他变量如本发明所定义。
在本发明的一些方案中，所述结构单元
Figure PCTCN2019101121-appb-000033
选自
Figure PCTCN2019101121-appb-000034
Figure PCTCN2019101121-appb-000035
其他变量如本发明所定义。
在本发明的一些方案中，所述结构单元
Figure PCTCN2019101121-appb-000036
选自
Figure PCTCN2019101121-appb-000037
Figure PCTCN2019101121-appb-000038
其他变量如本发明所定义。
在本发明的一些方案中，所述结构单元
Figure PCTCN2019101121-appb-000039
选自
Figure PCTCN2019101121-appb-000040
其他变量如本发明所定义。
在本发明的一些方案中，所述化合物、其异构体或其药学上可接受的盐，其选自
Figure PCTCN2019101121-appb-000041
Figure PCTCN2019101121-appb-000042
其中，R 1、R 2、R 3、R 4、L 2、L 3、T 1、T 2、T 3、T 4和T 5如本发明所定义。
本发明还有一些方案由上述变量任意组合而来。
本发明提供下式所示化合物、其异构体或其药学上可接受的盐，其选自：
Figure PCTCN2019101121-appb-000043
Figure PCTCN2019101121-appb-000044
Figure PCTCN2019101121-appb-000045
在本发明的一些方案中，所述化合物、其异构体或其药学上可接受的盐，其选自：
Figure PCTCN2019101121-appb-000046
Figure PCTCN2019101121-appb-000047
Figure PCTCN2019101121-appb-000048
Figure PCTCN2019101121-appb-000049
本发明还提供一种药物组合物，其含有治疗有效量的上述的化合物、其异构体或其药学上可接受的盐作为活性成分以及药学上可接受的载体。
本发明还提供上述化合物、其异构体或其药学上可接受的盐在制备RET激酶抑制剂的应用。
本发明还提供上述组合物在制备RET激酶抑制剂的应用。
定义和说明
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。
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。
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的中性形式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的中性形式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。
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。
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。
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。
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。
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。
除非另有说明，“(D)”或者“(+)”表示右旋，“(L)”或者“(-)”表示左旋，“(DL)”或者“(±)”表示外消旋。
除非另有说明，用楔形实线键
Figure PCTCN2019101121-appb-000050
和楔形虚线键
Figure PCTCN2019101121-appb-000051
表示一个立体中心的绝对构型，用直形实线键
Figure PCTCN2019101121-appb-000052
和直形虚线键
Figure PCTCN2019101121-appb-000053
表示立体中心的相对构型，用波浪线
Figure PCTCN2019101121-appb-000054
表示楔形实线键
Figure PCTCN2019101121-appb-000055
或楔形虚线键
Figure PCTCN2019101121-appb-000056
或用波浪线
Figure PCTCN2019101121-appb-000057
表示直形实线键
Figure PCTCN2019101121-appb-000058
和直形虚线键
Figure PCTCN2019101121-appb-000059
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。
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％。
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％。
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)。
本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。
“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。
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。
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。
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。
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。
当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 PCTCN2019101121-appb-000060
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 PCTCN2019101121-appb-000061
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 PCTCN2019101121-appb-000062
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。
除非另有规定，术语“C 1-5烷基”用于表示直链或支链的由1至5个碳原子组成的饱和碳氢基团。所述C 1-5烷基包括C 1-4、C 1-3、C 1-2、C 2-5、C 2-4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 1-5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)等。
除非另有规定，术语“C 1-3烷基”用于表示直链或支链的由1至3个碳原子组成的饱和碳氢基团。所述C 1-3烷基包括C 1-2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- 3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)等。除非另有规定，“C 2-8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8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 2-8烯基包括C 2-6、C 2-4、C 2-3、C 4、C 3和C 2烯基等；其可以是一价、二价或者多价。C 2-8烯基的实例包括但不限于乙烯基、丙烯基、丁烯基、戊烯基、己烯基、丁间二烯基、戊间二烯基、己间二烯基等。
除非另有规定，术语“C 1-5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5个碳原子的烷基基团。所述C 1-5烷氧基包括C 1-5、C 1-4、C 1-3、C 1-2、C 2-5、C 2-4、C 5、C 4、C 3和C 2烷氧基等。C 1- 5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)等。
除非另有规定，术语“C 1-3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3个碳原子的烷基基团。所述C 1-3烷氧基包括C 1-2、C 2-3、C 3和C 2烷氧基等。C 1-3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)等。
除非另有规定，术语“C 1-5烷氨基”表示通过氨基连接到分子的其余部分的那些包含1至5个碳原子的烷基基团。所述C 1-6烷氨基包括C 1-5、C 1-4、C 1-3、C 1-2、C 2-5、C 2-4、C 5、C 4、C 3和C 2烷氨基等。C 1-5烷氨基的实例包括但不限于-NHCH 3、-N(CH 3) 2、-NHCH 2CH 3、-N(CH 3)CH 2CH 3、-N(CH 2CH 3)(CH 2CH 3)、-NHCH 2CH 2CH 3、-NHCH 2(CH 3) 2、-NHCH 2CH 2CH 2CH 3等。
除非另有规定，术语“C 1-4烷氨基”表示通过氨基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烷氨基包括C 1-3、C 1-2、C 2-4、C 4、C 3和C 2烷氨基等。C 1-4烷氨基的实例包括但不限于-NHCH 3、-N(CH 3) 2、-NHCH 2CH 3、-N(CH 3)CH 2CH 3、-N(CH 2CH 3)(CH 2CH 3)、-NHCH 2CH 2CH 3、-NHCH 2(CH 3) 2、-NHCH 2CH 2CH 2CH 3等。
除非另有规定，“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。
除非另有规定，本发明术语“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)。
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。
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。此外，术语“卤代烷基”意在包括单卤代烷基和多卤代烷基。例如，术语“卤代(C 1-C 4)烷基”意在包括但不仅限于三氟甲基、2，2，2-三氟乙基、4-氯丁基和3-溴丙基等等。除非另有规定，卤代烷基的实例包括但不仅限于：三氟甲基、三氯甲基、五氟乙基，和五氯乙基。
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。
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。
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。
本发明的化合物可以有多种用途或适应症，包括但不限于本申请所列举的具体用途或适应症。
本发明所使用的溶剂可经市售获得。本发明采用下述缩略词：aq代表水；HATU代表O-(7-氮杂苯并三唑-1-基)-N，N，N'，N'-四甲基脲六氟磷酸盐；EDC代表N-(3-二甲基氨基丙基)-N'-乙基碳二亚胺盐酸盐；m-CPBA代表3-氯过氧苯甲酸；eq代表当量、等量；M代表mol/L；CDI代表羰基二咪唑；DCM代表二氯甲烷；PE代表石油醚；DIAD代表偶氮二羧酸二异丙酯；DMF代表N，N-二甲基甲酰胺；DMSO代表二甲亚砜；EtOAc代表乙酸乙酯；EtOH代表乙醇；MeOH代表甲醇；CBz代表苄氧羰基，是一种胺保护基团；BOC代表叔丁氧羰基是一种胺保护基团；HOAc代表乙酸；N乙腈BH 3代表氰基硼氢化钠；r.t.代表室温；O/N代表过夜；THF代表四氢呋喃；Boc 2O代表二-叔丁基二碳酸酯；三氟乙酸代表三氟乙酸；DIPEA代表二异丙基乙基胺；SOCl 2代表氯化亚砜；CS 2代表二硫化碳；TsOH代表对甲苯磺酸；NFSI代表N-氟-N-(苯磺酰基)苯磺酰胺；NCS代表1-氯吡咯烷-2,5-二酮；n-Bu 4NF代表氟化四丁基铵；iPrOH代表2-丙醇；mp代表熔点；LDA代表二异丙基胺基锂。
化合物依据本领域常规命名原则或者使用
Figure PCTCN2019101121-appb-000063
软件命名，市售化合物采用供应商目录名称。
技术效果
本发明化合物对RET及其突变RET V804M具有优异的抑制剂活性，将会在RET异常肿瘤患者中具有优异的治疗效果。
具体实施方式
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。
实施例1
Figure PCTCN2019101121-appb-000064
步骤一
将催化量DMF(2.32g,31.74mmol)加入含氯化亚砜(150mL)的6-氯烟酸(50g,317.35mmol)混合液中，氮气保护下85℃下回流3小时。旋去溶液后再溶于DCM(1200mL)中，加入甲氧基-N-甲基-氨基盐酸盐(46.43g,476.03mmol)，冷却至0℃，再滴加三乙胺(96.34g,952.05mmol)，25℃搅拌16小时。反应液加入饱和碳酸氢钠溶液，分层得有机相。有机相经硫酸钠干燥，再减压浓缩得1a。
LCMS(ESI)m/z:200.9[M+1] +
步骤二
0℃下向1a(59.71g,297.60mmol)的THF(500mL)溶液滴加甲基溴化镁(3M,119.04mL)，25℃搅拌2小时。向反应液中加入饱和氯化铵(500mL)溶液淬灭，再加入EtOAc萃取(500mL×2)，合并有机相并用无水硫酸钠干燥，减压浓缩得粗品。粗品经快速硅胶柱分离(PE:EtOAC＝3:1)得1b。
步骤三
将4-氟-吡唑(4.98g,57.85mmol)，1b(9g,57.85mmol)的DMF(80mL)溶液中加入碳酸钾(19.99g,144.62mmol)，100℃搅拌16小时。向反应液中加入水(400mL)，再加入EtOAc(400mL×2)萃取，合并有机相。有机相依次用水(600mL)洗，饱和碳酸钠水溶液(600mL)。最后有机相用无水硫酸钠干燥，减压浓缩得粗品。粗品经快速硅胶柱分离(PE:EtOAC＝3:1)得1c。
LCMS(ESI)m/z:205.9[M+1] +
步骤四
向1c(1g,4.87mmol)的THF(10mL)溶液中加入(R)-叔丁基亚磺酰胺(590.68mg,4.87mmol)，再加入四乙基氧化钛(2.62g,9.75mmol,2.38mL,85％纯度)，75℃搅拌20小时，再冷却至-78℃，滴加三仲丁基硼氢化锂(1M,14.62mL)溶液，-78℃搅拌30分钟。-78℃下向反应液中加入MeOH 5mL淬灭反应，再加入水(20mL)，加入EtOAc(20mL×2)萃取。得有机相，用无水硫酸钠干燥，减压浓缩得粗品，粗品经快速硅胶柱分离(DCM:MeOH＝10:1)得1d。
LCMS(ESI)m/z:310.9[M+1] +,333.1[M+23] +
步骤五
向1d(8g,25.77mmol)中加入MeOH(67.30g,2.10mol,85mL)和氯化氢的二氧六环溶液(4M,85mL,13.19eq)，30℃搅拌2小时。反应液经减压浓缩旋干得粗品，再加水稀释，用饱和碳酸钠溶液调节pH到8。加入二氯甲烷萃取，得有机相。有机相经减压浓缩得粗品。粗品经经快速硅胶柱分离(DCM:MeOH＝10:1)得1e。经过高效液相色谱手性拆分(柱子:DAICEL CHIRALPAK AD-H(250mm×50mm,10μm)；流动相:[0.1％氨水EtOH]；B％:35％-35％,保留时间：5.386分钟，收集主峰2)，得1e(99.85％ee％)。
1H NMR(400MHz,DMSO-d 6)δ8.66(d,J＝4.02Hz,1H),8.42(d,J＝2.01Hz,1H),7.99(dd,J＝2.01,8.53Hz,1H),7.87-7.91(m,1H),7.84(d,J＝8.53Hz,1H),4.08(q,J＝6.53Hz,1H),1.72-2.34(m,2H),1.29(d,J＝6.53Hz,3H).
步骤六
2,4-二氯-6-甲基-嘧啶(10g,61.35mmol)，5-甲基-3-胺基-1H-吡唑(6.55g,67.48mmol)，二异丙基乙胺(11.89g,92.02mmol,16.03mL)加入DMSO(30mL)中，反应60℃搅拌16小时。反应液冷却至20～30℃，倒入200mL冰水中，15～20度搅拌1小时，过滤，滤饼40～50℃减压旋干得到1f。
LCMS(ESI)m/z:224.0[M+1] +，225.9[M+3] +
1H NMR(400MHz,DMSO-d 6)δppm 12.08(s,1H)10.14(s,1H)6.50-7.68(m,1H)5.55-6.45(m,1H)2.25(s,3H)2.20(s,3H).
步骤七
1f(30mg,134.13μmol)，叔丁氧羰基哌啶(44.83mg,240.69μmol)，二异丙基乙胺(69.34mg,536.52μmol,93.45μL)加入到N-甲基吡咯烷酮(2mL)中。反应90℃搅拌16小时，反应液冷却至室温，加入5mL水，然后用EtOAc(5mL×2)萃取，有机相用无水硫酸钠干燥，减压下浓缩得1g。
LCMS(ESI)m/z:374.7[M+1] +,318.2[M-55] +
步骤八
1g(200mg,535.54μmol)加入到EtOAc(5mL)中，将氯化氢的乙酸乙酯溶液(4M,267.77μL)滴加到反应液中，反应20～30℃搅拌16小时。反应液直接40～50℃减压旋干得1h。
LCMS(ESI)m/z:274.0[M+1] +
1H NMR(400MHz,DMSO-d6)δppm 11.18(m,1H)9.81(s,2H)6.18-6.43(m,1H)4.21-4.33(m,4H)3.25(m,4H)2.42(s,3H)2.28-2.31(m,1H)2.26(s,3H).
步骤九
三光气(23.95mg,80.70μmol)加入到DCM(4mL)中，搅拌下将二异丙基乙胺(62.58mg,
484.19μmol,84.33μL)加入到反应液中，反应温度降至0～5℃搅拌下将1e(26.63mg,129.12μmol)加入到反应液中，反应在0～5℃搅拌10分钟，然后将1h(50mg,161.40μmol)和二异丙基乙胺(62.58mg,484.19μmol,84.33μL)加入到反应液中，体系在该0～5℃下搅拌1小时。反应液直接减压浓缩，得粗品。粗品通过制备色谱分析仪分离纯化(HCl体系)，得到实施例1的盐酸盐，实施例1的盐酸盐加入碳酸氢钠溶液中，乙酸乙酯萃取，有机相用无水硫酸钠干燥，减压下浓缩得到实施例1。
LCMS(ESI)m/z:506.1[M+1] +
1H NMR(400MHz,MeOH-d 4)δppm 8.52(d,J＝4.27Hz,1H),8.44(s,1H),7.98-8.06(m,1H),7.89-7.97(m,1H),7.72(d,J＝4.02Hz,1H),6.19-6.65(m,2H),5.00-5.08(m,1H),3.91(m,4H),3.69(m,4H),2.46(s,3H),2.41(s,3H),1.57(d,J＝7.03Hz,3H).
实施例2、3
Figure PCTCN2019101121-appb-000065
步骤一
将化合物2a(500mg,3.21mmol)，二甲胺盐酸盐(1.81g,16.07mmol)加入到乙醇(5mL)中，在80℃条件下搅拌回流2小时。反应液冷却至25℃后旋干，旋干后加入叔丁基甲基醚溶解，用10％的柠檬酸洗涤，再用1mol/L的氢氧化钠将pH调到10～11，再用20mL叔丁基甲基醚萃取3次，收集有机相并用无水硫酸钠干燥，过滤旋干得2b。
LCMS(ESI)m/z:164.8[M+1] +
1H NMR(400MHz,CDCl 3)δppm 8.67-8.86(m,1H),7.90-8.11(m,1H),6.36-6.66(m,1H),3.18(s,6H),2.49(s,3H).
步骤二
将2b(220mg,1.34mmol)加入MeOH(10mL)中，然后加入乙酸铵(1.03g,13.40mmol)，在65℃条件下搅拌1小时，溶解完全后加入氰基硼氢化钠(252.59mg,4.02mmol)，在65℃条件下搅拌过夜。原料反应完全反应液直接旋干得到粗品。粗品经制备层析板(DCM/MeOH＝10/1,Rf＝0.2.3)纯化得到2c。
LCMS(ESI)m/z:165.8[M+1] +
1H NMR(400MHz,CDCl 3)δppm 8.11(d,J＝2.00Hz,1H),7.59(dd,J＝8.80,2.64Hz,1H),6.55(d,J＝9.20Hz,1H),4.27-4.29(m,1H),2.93-3.23(m,16H),1.57(d,J＝6.80Hz,3H).
步骤三
在0℃条件下将2c(57mg,344.96μmol)DIEA(133.75mg,1.03mmol,180.25μL,3eq),三光气(51.18mg,172.48μmol)加入DCM(2mL)中，搅拌10min，将0℃的化合物1h(113.15mg,413.95μmol)，DIEA(133.75mg,1.03mmol)，DCM(2mL)混合物加入上述反应体系，在0℃条件下反应20min。反应液直接旋干得粗品。粗品送制备色谱(色谱柱:Xtimate C18 150×25mm×5μm；流动相:[水(0.075％三氟乙酸)-乙腈]；B％:10％-30％,7min)分离纯化后送SFC拆分(色谱柱:DAICEL CHIRALPAK AS-H(250mm×30mm,5μm)；流动相:[0.1％氨水EtOH]；B％:45％-45％)
得到实施例2(保留时间：4.957min)。
LCMS(ESI)m/z:465.4[M+1] +
1H NMR(400MHz,MeOH-d 4)δppm 8.03(d,J＝2.80Hz,1H),7.55(dd,J＝8.80,2.51Hz,1H),6.65(d,J＝8.80Hz,1H),6.11-6.12(m,2H),3.74-3.78(m,4H),3.48-3.52(m,4H),3.01(s,6H),2.28(s,3H),2.21(s,3H).
得到实施例3(保留时间：5.737min)。
LCMS(ESI)m/z:465.4[M+1] +
1H NMR(400MHz,MeOH-d 4)δppm 8.03(d,J＝2.80Hz,1H),7.56(dd,J＝8.80,2.51Hz,1H),6.64(d,J＝8.80Hz,1H),6.11-6.12(m,2H),3.75-3.79(m,4H),3.48-3.51(m,4H),3.04(s,6H),2.28(s,3H),2.21(s,3H).
实施例4
Figure PCTCN2019101121-appb-000066
步骤一
将2,6-二氯,4-甲基吡啶(5g,30.86mmol)，N-Boc哌嗪(6.32g,33.95mmol)和碳酸铯(30.17g,92.58mmol)一起加到DMF(100mL)中，然后油浴加热到120℃下搅拌反应16小时。反应液直接过滤，收集滤液直接旋干得到粗品。粗品通过快速硅胶柱(PE/EtOAc＝5/1,Rf＝0.5)纯化得到4b。
1H NMR(400MHz,CDCl3)δ6.49(s,1H),6.30(s,1H),3.51(s,8H),2.24(s,3H),1.48(s,9H).
步骤二
将4b(373.76mg,3.85mmol)溶解到1,4-二氧六环(10mL)中，然后将Pd 2(dba) 3(146.84mg,160.36μmol)，5-甲基-3-胺基-1H-吡唑(1g,3.21mmol)，2-二环己基磷-2,4,6-三异丙基联苯(229.33mg,481.07μmol)和碳酸铯(3.13g,9.62mmol)一起加到反应液中，通过微波合成仪加热到140℃并且搅拌反应1小时。反应液直接过滤，收集滤液旋干得到粗品。粗品通过快速硅胶柱(PE/EtOAc＝1/1，Rf＝0.2)纯化得到产物4c。
LCMS(ESI)m/z:373.3[M+H] +
步骤三
将4c(100mg,268.48μmol)溶解到EtOAc(5mL)中，然后滴加HCl/EtOAc(4M,10mL)，最后反应液在20-30℃下搅拌反应2小时。随着反应的进行，反应液中的固体逐渐增多。反应液继续搅拌反应16小时。LCMS检测显示原料消失，主峰为产物峰。直接将反应液浓缩至干得到4d。
LCMS(ESI)m/z:273.2[M+1] +
步骤四
0℃下，将化合物4d(40mg,146.87μmol)和N,N-二异丙基乙胺(56.95mg,440.61μmol,76.75μL)加到二氯甲烷(5mL)中，随后加入化合物1e(36.35mg,176.25μmol)，N,N-二异丙基乙胺(56.95mg,440.61μmol)和三光气(21.79mg,73.44μmol)搅拌10分钟后的混合液，反应液在25-30℃下搅拌反应10分钟。将反应液直接20-30℃旋干得到粗品。粗品送制备色谱柱分离(column:Xtimate C18 150×25mm×5μm；流动相:[水(0.225％三氟乙酸)-乙腈]；B％:25％-45％,7min)。得到实施例4的三氟乙酸盐，实施例4的三氟乙酸盐加入碳酸氢钠溶液中，乙酸乙酯萃取，有机相用无水硫酸钠干燥，减压下浓缩得到实施例4。
LCMS(ESI)m/z:505.3[M+1] +
1H NMR(400MHz,MeOH-d 4)δ8.47(br s,2H),7.98～7.90(m,1H),7.84(s,1H),7.69(s,1H),6.34(s,1H),6.23(s,1H),5.83(s,1H),5.02～4.99(m,1H),3.66-3.74(m,8H),2.37(s,3H),2.33(s,3H),1.57(d,J＝6.8Hz,3H).
实施例6、7
Figure PCTCN2019101121-appb-000067
步骤一
用DMF(110mL)将4-吡唑-1-苯甲酸(7g,37.20mmol)，甲氧基甲基胺盐酸盐(5.44g,55.80mmol)溶解，加入HATU(14.14g,37.20mmol)、DIPEA(24.04g,185.99mmol,32.40mL)，30℃搅拌16小时。LCMS表明有产物生成。反应液加水，用乙酸乙酯(100ml×3)萃取，合并有机相用水洗(100ml×2)， 饱和氯化钠溶液(100ml×2)洗涤，无水硫酸钠干燥，减压浓缩得到6a。
1H NMR(400MHz,CDCl 3-d 6)δ7.95(d,J＝4Hz,1H),7.74-7.81(m,2H),7.58-7.73(m,3H),6.36-6.49(m,1H),3.51(s,3H),3.32(s,3H).
步骤二
0℃下向化合物6a(9.4g,40.65mmol)的THF(90mL)溶液中滴加甲基溴化镁(3M,27.10mL),慢慢升至30℃搅拌1小时。在反应液中加入饱和的氯化铵溶液进行淬灭，然后用乙酸乙酯(150ml×3)萃取，合并有机层，用饱和氯化钠溶液洗涤(200ml×3)，并用无水硫酸钠干燥，最后减压旋除溶剂得粗品。硅胶柱层析(PE:EA＝3:1,Rf＝0.57)，发现产物易析出，最后将过柱得到的交叉产物加入少量的乙酸乙酯溶解后加入大量的石油醚，有白色固体洗出，过滤，得产物6b。
步骤三
向6b(4.75g,25.51mmol)中加入MeOH(50mL)溶解，并加入乙酸铵(19.66g,255.09mmol)，60℃搅拌1h后加入氰基硼氢化钠(4.81g,76.53mmol)，保持60℃继续搅拌3h。TLC监测表明有产物生成。旋干反应液得粗品，经硅胶柱层析(DCM:MeOH＝10:1，Rf＝0.22)分离得产物6c。
1H NMR(400MHz,DMSO-d 6)δ8.50(d,J＝2.4Hz,1H),7.87(d,J＝8.8Hz,2H),7.75(d,J＝1.6Hz,1H),7.56(d,J＝8.8Hz,2H),6.49-6.62(m,1H),4.36(q,J＝6.8Hz,1H),1,45(d,J＝6.8Hz,3H).
步骤四
0℃，将化合物1h(72.99mg,267.04μmol)溶于DCM(1mL)中，并加入DIEA(103.54mg,801.11μmol)，向其加入化合物6c，DIEA(103.54mg,801.11μmol,139.54μL)，三光气(39.62mg,133.52μmol)搅拌10min的混合液。保持0℃搅拌10min。LCMS表明有产物生成。将反应液旋干得粗品。粗品使用制备色谱柱进行分离(色谱柱:×timate C18 150×25mm×5μm；流动相:[水(0.075％三氟乙酸)-乙腈]；B％:15％-35％,8min)得6d。
LCMS(ESI)m/z:487.2[M+1] +
步骤五
将化合物6d(28mg,57.55μmol)进行手性拆分，SFC拆分条件为色谱柱:DAICEL CHIRALPAK AD(250mm×30mm,10μm)；流动相:[0.1％氨水EtOH]；B％:55％-55％]
收集峰1(保留时间：2.111min)，得实施例6。
LCMS(ESI)m/z:487.2[M+1] +
1H NMR(400MHz,CDCl 3-d 6)δ7.88(d,J＝2.4Hz,1H),7.69(s,1H),7.63(d,J＝8.4Hz,2H),7.42(d,J＝8.8Hz,2H),6.43(d,J＝1.6Hz,1H),5.95-6.13(m.2H),5.01-5.10(m,1H),4.85(s,1H),3.74-3.8(m,4H),3.43-3.45(m,4H),2.27(s,3H),2.22(s,4H),1.51(d,J＝7.2Hz,3H).
收集峰2(保留时间：3.756min),得实施例7。
LCMS(ESI)m/z:487.2[M+1] +
1H NMR(400MHz,CDCl 3-d 6)δ7.89(d,J＝2.4Hz,1H),7.71(d,J＝1.2Hz,1H),7.64(d,J＝8.4Hz,2H),7.43(d,J＝8.4Hz,2H),6.45(t,J＝2Hz,1H),6.11(s,1H),6.01(s,1H),5.03-5.11(m,1H),4.76(d,J＝7.2Hz,1H),3.77-3.86(m,4H),3.43-3.48(m,4H),2.29(s,3H),2.23(s,3H),1.53(d,J＝7.2Hz,3H).
实施例8、9
Figure PCTCN2019101121-appb-000068
步骤一
将化合物8a(500mg,3.03mmol)和乙酸铵(2.33g,30.27mmol)加入到甲醇(7mL)中，将反应65℃搅拌1小时，然后将氰基硼氢化钠(570.64mg,9.08mmol)分批加入到反应液中，反应继续65℃搅拌12个小时。反应液直接40～50℃减压旋干，得粗品，通过薄层制备层析板(PE/EtOAc＝1/1)纯化得到8b。
1H NMR(400MHz,CDCl 3)δppm 8.19(d,J＝2.51Hz,1H),7.79(dd,J＝8.78,2.51Hz,1H),6.80(s,1H),4.25(q,J＝7.03Hz,2H),3.48(s,3H),1.37(t,J＝7.03Hz,3H).
步骤二
将化合物1h(100mg,365.85μmol)和DIEA(141.85mg,1.10mmol,191.17μL)加入到盛有DCM(2mL)反应液中，搅拌10min。0～5℃下加入NN-二异丙基乙胺(141.85mg,1.10mmol,191.17μL)，8b(60.81mg,365.85μmol)及三光气(54.28mg,182.92μmol)搅拌好的DCM(2mL)溶液，反应在0～5℃搅拌20min，反应液直接减压浓缩，得粗品，通过制备色谱柱(色谱柱:Xtimate C18 150×25mm×5μm；流动相:[水(0.075％三氟乙酸)-乙腈]；B％:17％-27％,9min)纯化得到8c。
LCMS(ESI)m/z 466.3[M+1] +
步骤三
化合物8c(28mg,60.14μmol)通过SFC拆分(色谱柱:DAICEL CHIRALPAK IC(250mm×30mm,10μm)；流动相:[0.1％氨水EtOH]；B％:50％)。
收集峰1(保留时间5.211min)，得实施例8。
LCMS(ESI)m/z:466.2[M+1] +
1H NMR(400MHz,MeOH-d 4)δppm 8.09(d,J＝2.51Hz,1H)7.70(dd,J＝8.78,2.51Hz,1H)6.76(d,J＝8.53Hz,1H)6.13(s,2H)4.07-4.18(m,1H)3.77-3.80(m,4H)3.49-3.53(m,4H)3.49-3.52(m,1H)2.29(s,3H)2.22(s,3H)1.50(d,J＝4.0Hz,3H)1.37-1.40(m,3H).
收集峰2(保留时间6.498min)，得实施例9。
LCMS(ESI)m/z:466.2[M+1] +
1H NMR(400MHz,MeOH-d 4)δppm 8.09(d,J＝2.51Hz,1H)7.70(dd,J＝8.78,2.51Hz,1H)6.76(d,J＝8.53Hz,1H)6.13(s,2H)3.78-3.80(m,4H)3.49-3.52(m,4H)2.29(s,3H)2.22(s,3H)1.50(d,J＝4.0Hz,2H)1.48-1.51(m,1H)1.37-1.40(m,3H).
实施例10、11
Figure PCTCN2019101121-appb-000069
Figure PCTCN2019101121-appb-000070
步骤一
化合物4d(40mg,146.87μmol)溶解在DCM(1mL)，加入DIEA(56.95mg,440.61μmol,76.75μL)，0℃搅拌10min。向上述反应液滴加化合物6c(27.50mg,146.87μmol)、DIEA(56.95mg,440.61μmol)、三光气(21.79mg,73.44μmol)0℃搅拌10分钟后得到的DCM(1mL)溶液。在0℃条件下搅拌10min。旋干反应液得粗品，经制备色谱柱纯化(column:Xtimate C18 150×25mm×5μm；流动相:[水(0.075％三氟乙酸)-乙腈]；B％:32％-46％,7min)得产物10a。
LCMS(ESI)m/z:486.4[M+1] +
步骤二
将化合物10a进行手性拆分。SFC拆分条件为色谱柱:YMC CHIRAL Amylose-C(250mm×30mm,10μm；流动相:[0.1％氨水异丙醇]；B％:50％-50％,min。
收集峰1(保留时间1.642min)，得实施例10。
LCMS(ESI)m/z:486.4[M+1] +
1H NMR(400MHz,CDCl 3)δ7.90(d,J＝2.4Hz,1H),7.71(d,J＝1.6Hz,1H),7.66(d,J＝8.8Hz,2H),7.43(d,J＝8.8Hz,2H),6.52(s,1H),6.45(t,J＝1.6Hz,1H),6.18(s,1H),5.95(s,1H),5.83(s,1H),5.02-5.16(m,1H),4.70(d,J＝6.8Hz,1H),3.47-3.54(m,8H),2.28(s,3H),2.21(s,3H),1.36-1.45(m,3H).
收集峰2(保留时间2.516min)，得实施例11。
LCMS(ESI)m/z:486.4[M+1] +
1H NMR(400MHz,CDCl 3)δ7.88(d,J＝2.8Hz,1H),7.70(d,J＝1.2Hz,1H),7.63(d,J＝8.4Hz,2H),7.42(d,J＝8.8Hz,2H),6.75(s,1H),6.43-6.46(m,1H),6.11(s,1H),5.94(s,1H),5.82(s,1H),5.02-5.14(m,1H),3.49-3.52(m,8H),2.26(s,4H),2.20(s,3H),1.36-1.38(m,3H).
实施例12
Figure PCTCN2019101121-appb-000071
步骤一
化合物12a(200mg,1.23mmol)和3-甲基-3-吡唑啉-5-酮(144.44mg,1.47mmol)加入到DMSO(2mL)中，然后将DIEA(237.86mg,1.84mmol,320.57μL)滴加到反应液中，反应在60℃搅拌16hr。反应液冷却至20～30℃，然后滴加到20mL冰水中，有淡黄色固体析出，减压抽滤得到化合物12b。
LCMS(ESI)m/z 224.6[M+1] +
步骤二
化合物12b(100mg,445.14μmol)和N-BOC哌嗪(165.82mg,890.29μmol)加入到NMP(2mL)中，然后将DIEA(230.13mg,1.78mmol)滴加到反应液中，反应在100℃搅拌1hr。反应液冷却至室温，然后加入10mL水，然后用乙酸乙酯(10mL×2)萃取，合并有机相，有机相用无水硫酸钠干燥，40～50℃减压旋干，得到粗品，粗品通过柱层析纯化(PE/EA＝3/1～1/1)纯化得到化合物12c。
LCMS(ESI)m/z 375.2[M+1] +
步骤三
化合物12c(45mg,120.18μmol)加入到DCM(4mL)中，然后将TFA(2.74g,24.04mmol,1.78mL)加入到反应液中，反应20～30℃反应20min。反应液直接40～50℃减压旋干，得到化合物12d的TFA盐。
LCMS(ESI)m/z 274.8[M+1]+
步骤四
化合物12d的TFA盐(50mg,128.75μmol)加入到DCM(5mL)中，然后将DIEA(49.92mg,386.26μmol,67.28μL)加入到反应液中，反应温度降至0℃，定为反应液1，1e(23.90mg,115.88μmol)加入到DCM(5mL)中，将三光气(19.10mg,64.38μmol)加入到反应液中，DIEA(49.92mg,386.26μmol,67.28μL)滴加到反应液中，反应温度降至0℃，定为反应液2，反应液1和2分别在0℃搅拌10min，在0℃条件下将反应液1慢慢滴加到反应液2中，反应在0℃搅拌10min。反应液中加入10mL水，然后用DCM(5mL×2)萃取，合并有机相，有机相用无水硫酸钠干燥，40～45℃减压旋干，得粗品，粗品通过薄层制备层析板纯化得到实施例12。
LCMS(ESI)m/z 507.3[M+1] +
1H NMR(400MHz,DMSO-d 6)δ12.19(s,1H)8.58-8.76(m,1H)8.41(d,J＝2.01Hz,1H)7.93-7.97(m,1H)7.91(d,J＝4.27Hz,1H)7.85-7.89(m,1H)6.99(d,J＝7.53Hz,1H)6.01(s,1H)5.83(s,1H)4.92(quin,J＝7.03Hz,1H)3.59-3.67(m,4H)3.37-3.42(m,4H)2.23(s,6H)1.43(d,J＝7.03Hz,3H).
实施例13
Figure PCTCN2019101121-appb-000072
步骤一
将化合物13a的TFA盐(0.06g,154.90μmol)溶到无水甲醇(1mL)中，0℃下加入硼氢化钠(121.25mg,3.21mmol)，25℃下搅拌2hr。将反应液加到水(10mL)中，乙酸乙酯萃取(10mL×3)，合并有机相，无水硫酸钠干燥，过滤减压浓缩得粗品。将粗品用板层析(石油醚：乙酸乙酯＝1:1)纯化得白色油状物实施例13b。
LCMS(ESI)m/z:190.1[M+1] +
步骤二
将化合物13b(0.06g,154.90μmol,TFA盐)和DIEA(60.06mg,464.70μmol,80.94μL)加到干燥得二氯甲烷(1mL)中，搅拌10mins，形成溶液1，然后在0℃下，将三光气(137.90mg,464.70μmol)加到化合物1h和DIEA(60.06mg,464.70μmol,80.94μL,3eq)的二氯甲烷(1mL)溶液中，0℃下反应10mins，然后将溶液1滴加到反应液中，0℃下反应20mins。将水(2mL)加到反应液中淬灭反应，浓缩得粗品。粗品通过制备色谱(column:Boston Green ODS 150×30mm 5μm；流动相:[水(0.075％三氟乙酸)-乙腈]；B％:29％-45％,9min)纯化得实施例13的三氟乙酸盐,实施例13的三氟乙酸盐加入碳酸氢钠溶液中，乙酸乙酯萃取，有机相用无水硫酸钠干燥，浓缩得到实施例13。
LCMS(ESI)m/z:475.2[M+1] +
1H NMR(400MHz,MeOH-d 4)δ＝8.61(d,J＝2.4Hz,1H),8.49(s,1H),8.07-7.99(m,1H),7.99-7.91(m,1H),7.78(s,1H),6.56(br s,1H),6.44-6.14(m,1H),5.93(q,J＝6.6Hz,1H),3.96-3.49(m,9H),2.40(s,3H),2.34(s,3H),1.67(d,J＝6.4Hz,3H).
实施例14
Figure PCTCN2019101121-appb-000073
步骤一
将2-溴-5-羟基吡啶(1g,5.75mmol)溶于丙酮(10mL)中，加入氯乙酸甲酯(748.46mg,6.90mmol,608.50uL)、碳酸钾(794.31mg,5.75mmol)，60℃搅拌16小时。过滤除去不溶固体，旋干滤液得粗品。粗品经快速硅胶柱纯化(PE:EA＝3:1)，得化合物14a(1.1g,4.47mmol)。
LCMS(ESI)m/z:245.4[M+1] +,247.4[M+3] +
步骤二
将化合物14a(50mg,203.20umol)溶于甲苯(1mL)中，加入4-氟-1氢-吡唑(14.57mg,169.34umol)、 碘化亚铜(3.23mg,16.93umol)、碳酸钾(46.81mg,338.67umol)、N1,N2-二甲基环己二胺(4.82mg,33.87umol)，110℃搅拌16小时。过滤除去不溶物质，旋干滤液得粗品。粗品经制备层析板纯(PE:EA＝1:1)，得化合物14b。
1H NMR(400MHz,CDCl 3)δppm8.31(d,J＝4.4Hz,1H),8.07(d,J＝2.8Hz,1H),7.88(d,J＝8.8Hz,1H),7.55(d,J＝4.4Hz,1H),7.37(dd,J＝8.8,2.8Hz,1H),4.71(s,2H),3.83(s,3H).
步骤三
将化合物14b(10mg,39.81umol)溶于四氢呋喃(0.3mL)/水(0.1mL)溶液中，加入氢氧化锂一水合物(5.01mg,119.42umol)，16℃搅拌5小时。加入4M盐酸调pH至2-3，乙酸乙酯(2×10mL)萃取，旋干溶剂得粗品化合物14c。
LCMS(ESI)m/z:237.8[M+1] +
步骤四
将化合物14c(10mg,42.16umol)溶于N,N二甲基甲酰胺(0.5mL)中，加入化合物1h(16.33mg,42.16umol,TFA)、HATU(24.05mg,63.24umol)、N,N-二异丙基乙胺(16.35mg,126.48umol,22.03uL)，16℃搅拌3小时。直接旋干反应液得粗品，再经制备色谱柱分离(色谱柱:Boston Green ODS 150×30mm5μm；流动相:[水(0.075％三氟乙酸)-乙腈]；B％:23％-53％,8min)，纯化得实施例14的三氟乙酸盐。实施例14的三氟乙酸盐加入碳酸氢钠溶液中，乙酸乙酯萃取，有机相用无水硫酸钠干燥，减压下浓缩得到实施例14。
LCMS(ESI)m/z:493.1[M+1] +
1H NMR(400MHz,METHANOL-d4)δppm 8.43(d,J＝4.02Hz,1H),8.19(d,J＝3.04Hz,2H),7.89(d,J＝9.20Hz,1H),7.66(d,J＝4.02Hz,1H),7.62(d,J＝3.04Hz,1H),7.60(d,J＝2.76Hz,1H),5.06(s,2H),3.99(s,2H),3.91(s,2H),3.83(s,4H),2.42(s,3H),34(s,3H).
实施例15、16
Figure PCTCN2019101121-appb-000074
Figure PCTCN2019101121-appb-000075
步骤一
将化合物1c(6.5g,31.68mmol)加到甲醇(50mL)中，0℃下加入硼氢化钠(3.60g,95.03mmol)，0℃下搅拌1hr。将反应液加水(100mL),乙酸乙酯萃取(100mL*3)，合并有机相，无水硫酸钠干燥，减压浓缩得化合物15a。
1H NMR(400MHz,DMSO-d6)δ8.68(d,J＝4.5Hz,1H),8.42(d,J＝2.3Hz,1H),7.98-7.95(m,1H),7.91(d,J＝4.5Hz,1H),7.89-7.84(m,1H),5.41(d,J＝4.5Hz,1H),4.90-4.79(m,1H),1.38(d,J＝6.5Hz,3H).
步骤二
将15a(6.5g)用SFC(色谱柱:Phenomenex-Amylose-1(250mm×30mm,5μm)；流动相:[0.1％氨水乙醇]；B％:25％-25％,min)分离。得15b1(保留时间：1.882min)和15b2(保留时间：2.010min)。
步骤三
将15b1(882.61mg,4.26mmol),N,N-二琥珀酰亚胺基碳酸酯(1.49g,5.81mmol)和4-N,N-二甲基氨基吡啶(236.54mg,1.94mmol)，三乙胺(391.85mg,3.87mmol,538.99uL)加到DMF(1mL)中，然后在60℃下搅拌0.5hr，中间态生成。25℃下加入三乙胺(391.85mg,3.87mmol,538.99uL)，4-N,N-二甲基氨基吡啶(236.54mg,1.94mmol)和化合物1h(1.5g,3.87mmol,三氟乙酸盐)的DMF(1mL)混合溶液，25℃下反应0.5hr。反应液直接送制备色谱柱(色谱柱:Welch Xtimate C18 150*25mm*5um；流动相:[水(0.075％三氟乙酸)-乙腈]；B％:24％-44％,12min)纯化得到实施例15的三氟乙酸盐。实施例15的三氟乙酸盐加入碳酸氢钠溶液中，乙酸乙酯萃取，有机相用无水硫酸钠干燥，减压下浓缩得到实施例15。
LCMS(ESI)m/z:507.0[M+1] +
1H NMR(400MHz,METHANOL-d4)δ＝8.52(d,J＝4.4Hz,1H),8.48(s,1H),8.07-7.93(m,2H),7.72(d,J＝4.2Hz,1H),6.23(br s,1H),5.92(q,J＝6.8Hz,1H),4.01-3.60(m,8H),2.40(s,3H),2.34(s,3H),1.66(d,J＝6.8Hz,3H).
以15b2为原料，同样方法得到实施例16的三氟乙酸盐。实施例16的三氟乙酸盐加入碳酸氢钠溶液中，乙酸乙酯萃取，有机相用无水硫酸钠干燥，减压下浓缩得到实施例16。
LCMS(ESI)m/z:507.2[M+1] +
1H NMR(400MHz,METHANOL-d4)δ＝8.53(d,J＝4.4Hz,1H),8.48(s,1H),8.07-7.95(m,2H),7.72(d,J＝4.2Hz,1H),6.23(br s,1H),5.91(q,J＝6.8Hz,1H),3.98-3.65(m,8H),2.40(s,3H),2.34(s,3H),1.66(d,J＝6.8Hz,3H).
实施例17、18
Figure PCTCN2019101121-appb-000076
步骤一
将2,4-二氯-6-甲基嘧啶(2g,12.27mmol)、3-环丙基-5-氨基-1H-吡唑(1.66g,13.50mmol溶于二甲亚砜(30mL)中，加入N,N-二异丙基乙胺(2.38g,18.40mmol,3.21mL)，50℃搅拌4小时。将反应液加入500mL水中，搅拌30分钟后有固体析出,过滤得到化合物17a。
LCMS(ESI)m/z:250.3[M+1] +
1H NMR(400MHz,DMSO-d6)δppm 12.15(s,1H),10.14(s,1H),7.55-6.85(m,1H),6.76(s,1H),6.22-5.49(m,1H),2.67(d,J＝1.76Hz,1H),2.26(s,3H),1.97-1.79(m,1H),1.00-0.83(m,2H),0.77-0.60(m,2H)
步骤二
将化合物17a(825mg,3.30mmol)、N-Boc哌嗪(923.05mg,4.96mmol)溶于二甲亚砜(15mL)中，加入N,N-二异丙基乙胺(1.28g,9.91mmol,1.73mL)，140℃搅拌5小时。将反应液倒入250ml水中，有固体析出，继续搅拌30分钟，过滤得到化合物17b。.
LCMS(ESI)m/z:400.2[M+1] +
1H NMR(400MHz,DMSO-d6)δppm 11.91(s,1H),9.26(s,1H),6.30-5.92(m,2H),3.72-3.62(m,4H),3.38(s,4H),2.12(s,3H),1.95-1.78(m,1H),1.46-1.42(m,9H),0.99-0.81(m,2H),0.76-0.59(m,2H).
步骤三
在化合物17b(1.3g,3.25mmol)中加入三氟乙酸(7.42g,65.08mmol,4.82mL)/二氯甲烷(5mL)混合溶液，16℃搅拌8小时。旋干溶剂得化合物17c(3g,粗品,三氟乙酸盐)。
步骤四
将15b1(50mg,241.31umol)溶于N,N二甲基甲酰胺(1mL)中，加入N,N-二琥珀酰亚胺基碳酸酯(84.29mg,329.06umol)，60℃搅拌30分钟，,在25℃条件下，加入化合物17c(181.37mg,219.37umol,三氟乙酸盐)、三乙胺(22.20mg,219.37umol,30.53uL)、4-N,N-二甲基氨基吡啶(13.40mg,109.69umol)的N,N-二甲基甲酰胺(1mL)溶液，保持25℃搅拌30分钟。LCMS显示有产物生成。将反应液直接进行制备色谱柱分离(column:Boston Green ODS 150*30mm*5um；流动相:[水(0.075％三氟乙酸)-乙腈]；B％:25％-55％,10min)得到实施例17的三氟乙酸盐。实施例17的三氟乙酸盐加入碳酸氢钠溶液中，乙酸乙酯萃取，有机相用无水硫酸钠干燥，减压下浓缩得到实施例17。
LCMS(ESI)m/z:533.1[M+1] +
1H NMR(400MHz,MeOH-d4)δppm 8.51(d,J＝4.60Hz,1H),8.46(d,J＝1.52Hz,1H),8.02-7.92(m,2H),7.70(d,J＝4.02Hz,1H),6.20(s,1H),5.90(q,J＝6.64Hz,1H),3.97-3.50(m,10H),2.38(s,3H),1.97-1.88(m,1H),1.64(d,J＝6.64Hz,3H),1.07-0.97(m,2H),0.81-0.70(m,2H).
步骤五
在0℃条件下，将化合物17c(1.00g,1.21mmol,TFA)溶于二氯甲烷(1mL)中，加入N,N-二异丙基乙 胺(470.04mg,3.64mmol,633.48uL)，搅拌10分钟制成溶液1，同时将化合物1e(200mg,969.84umol)溶于二氯甲烷(1mL)中，加入三光气(179.87mg,606.15umol)、N,N-二异丙基乙胺(470.04mg,3.64mmol,633.48uL)，搅拌10分钟，制成溶液2，然后在0℃条件下，将溶液2加入溶液1中，搅拌10分钟。反应液加水0.5ml进行淬灭，旋干反应液，加入甲醇溶解，过滤去除不溶物质，旋干滤液得粗品，粗品经制备HPLC分离(column:YMC-Triart Prep C18 150*40mm*7um；流动相:[水(0.1％三氟乙酸)-乙腈]；B％:25％-45％,10min),得实施例18的三氟乙酸盐。实施例18的三氟乙酸盐加入碳酸氢钠溶液中，乙酸乙酯萃取，有机相用无水硫酸钠干燥，减压下浓缩得到实施例18。
LCMS(ESI)m/z:532.3[M+1] +
1H NMR(400MHz,METHANOL-d4)δppm 8.49(d,J＝4.52Hz,1H),7.94-7.87(m,2H),8.41(s,1H),7.68(d,J＝4.28Hz,1H),6.26(s,1H),5.01(q,J＝7.02Hz,1H),3.91-3.81(m,4H),3.69-3.60(m,4H),2.38(s,3H),1.97-1.90(m,1H),1.54(d,J＝7.02Hz,3H),1.05-0.96(m,2H),0.77-0.71(m,2H).
实施例19
Figure PCTCN2019101121-appb-000077
步骤一
向化合物19a(1.9g,8.88mmol)，溴乙酸乙酯(14.08g,84.32mmol,9.33mL)的DMF(20mL)溶液中加入碳酸铯(7.23g,22.19mmol)，110℃搅拌20小时。LCMS表明原料已反应完全，有产物生成。反应液经硅藻土抽滤，得滤液，再经油泵减压浓缩得粗品。粗品经快速硅胶柱分离(PE:EA＝3:1,Rf＝0.61)分离得化合物19b。
LCMS(ESI)m/z:299.7[M+1] +,301.7[M+3] +
步骤二
向化合物19b(2g,6.66mmol)的THF(10mL)/MeOH(10mL)/H2O(10mL)溶液中加入一水合氢氧化锂(2.80g,66.63mmol)，20℃搅拌16小时。反应液浓缩除去四氢呋喃和甲醇，再加入水(20mL)，再用乙酸乙酯萃取(20mL*2)除去杂质，水相再用4N HCl调pH至3后，加入乙酸乙酯(20mL*2)萃取，用无水硫酸钠干燥后减压浓缩得化合物19c。
LCMS(ESI)m/z:271.8[M+1] +,273.8[M+3] +
1H NMR(400MHz,DMSO-d6)δ12.68(br s,1H),6.73-6.93(m,2H),6.50(d,J＝8.53Hz,1H),4.14-4.25(m,2H),4.06(s,2H),3.37-3.43(m,2H).
步骤三
向化合物19c(0.1g,367.52umol)，化合物1h(341.67mg,882.05umol,三氟乙酸盐)的DMF(3mL)溶液中加入HATU(209.61mg,551.28umol)，N,N-二异丙基乙胺(142.50mg,1.10mmol,192.05uL)。20℃搅拌16小时。向反应液中加入水(15mL)，再加入乙酸乙酯(10mL*2)萃取，合并有机相并用无水硫酸钠干燥，减压浓缩得粗品。粗品经薄层层析板(PE:EA＝0:1,Rf＝0.44)分离得化合物19d。
LCMS(ESI)m/z:527.0[M+1] +,529.0[M+1] +
步骤四
向化合物19d(100mg,189.60umol)，4-氟-1氢-吡唑(16.32mg,189.60umol)的DMF(4mL)溶液中加入碘化亚铜(3.61mg,18.96umol)，碳酸铯(123.55mg,379.21umol)，(1S,2S)-N1,N2-二甲基环己二胺(5.39mg,37.92umol)。氮气保护下110℃搅拌16小时。反应液经过滤收集滤液。滤液送制备色谱柱(色谱柱:Gemini 150*25mm 5μm；流动相:[水(0.075％三氟乙酸)-乙腈]；B％:17％-47％,11.5min)分离得白色固体产物实施例19的三氟乙酸盐。实施例19的三氟乙酸盐加入碳酸氢钠溶液中，乙酸乙酯萃取，有机相用无水硫酸钠干燥，减压下浓缩得到实施例19。
LCMS(ESI)m/z:533.1[M+1] +
1H NMR(400MHz,DMSO-d6)δ12.32(s,1H),11.01(s,1H),8.46(d,J＝4.52Hz,1H),7.68(d,J＝4.27Hz,1H),7.06-7.09(m,1H),7.10(s,1H),6.65(d,J＝8.78Hz,1H),4.36(s,2H),4.21-4.29(m,2H),3.89(br s,2H),3.79(br s,2H),3.70(br s,2H),3.62-3.66(m,2H),3.42(br d,J＝4.27Hz,2H),2.33(s,3H),2.25(s,3H).
实施例20、21、22
Figure PCTCN2019101121-appb-000078
Figure PCTCN2019101121-appb-000079
步骤一
将化合物20a(10g,50.19mmol)溶解到四氢呋喃(50mL)中，-78℃下将二(三甲基硅)氨基锂(8.40g,50.19mmol,1M)滴加到反应液中搅拌反应0.5小时后，将N-苯基双(三氟甲烷磺酰)亚胺(17.93g,50.19mmol)溶解到THF(50mL)中滴加到上述反应液中。将反应液升温到0℃并且搅拌反应3小时。向反应液中加入3mL饱和氯化铵水溶液淬灭反应，然后直接加入无水硫酸钠干燥，过滤，收集滤液旋蒸蒸干，得到粗品。粗品通过硅胶柱(PE/EA＝10/1,Rf＝0.8)纯化得到化合物20b1和20b2的混合物，摩尔比例约3:1。
1H NMR(400MHz,CHLOROFORM-d)δ7.69(s,1H),5.93(s,0.35H),4.05～4.03m,0.7H),3.57～3.43(m,2.7H),2.45～2.42(m,2H),1.96～1.89(m,2.7H),1.49(s,9H),1.47(s,3H).
步骤二
将化合物20b1(6g,18.11mmol)，化合物20b2(2g,6.04mmol)的混合物，双联频哪醇硼酸酯(7.36g,28.98mmol)，Pd(dppf)Cl 2(1.06g,1.45mmol)和醋酸钾(7.11g,72.44mmol)一起加到二氧六环(160mL)中，然后氮气保护下通过油浴加热到80℃并且搅拌反应16小时。将反应液过滤，收集滤液，旋干得到粗品。粗品通过快速硅胶柱(PE/EA＝10/1，Rf1＝0.76&Rf2＝0.72)纯化得到化合物20c1和20c2的混合物，摩尔比例接近3:1。
1H NMR(400MHz,CHLOROFORM-d)δ7.79(m,1H),6.62(s,1H),3.96(s,2H),3.51(s,3H),2.19～2.16(m,3H),1.80～1.72(m,3H),1.45(s,13H),1.32(s,5H),1.23(s,16H).
步骤三
将化合物20c1(1g,3.23mmol)，化合物20c2(300.00mg,970.22umol)的混合物，化合物1f(940.34mg,4.20mmol)，Pd(PPh 3) 4(373.72mg,323.41umol)和K 2CO 3(1.79g,12.94mmol)一起加到二氧六环(12mL)和水(4mL)中，然后氮气保护下将反应液通过微波合成仪加热到125℃并且搅拌反应0.5小时。向反应液中加入5mL水和5mL乙酸乙酯萃取，分液，水相再用5mL乙酸乙酯萃取，合并有机相，无水硫酸钠干燥，旋蒸蒸干得到粗品，通过快速硅胶柱(二氯甲烷/甲醇＝10/1，Rf＝0.43)纯化得到化合物20d1和20d2的混合物。
LCMS(ESI)m/z:371.3[M+1] +
步骤四
将化合物20d1(100mg,269.94umol)，化合物20d2(100.00mg,269.94umol)的混合物，甲酸铵(170.21mg,2.70mmol)和Pd/C(50mg,10％含量)一起加到乙醇(5mL)中，然后油浴条件下加热到70℃并且搅拌反应4小时。反应液直接过滤，收集滤液，减压旋蒸得到粗品，通过制备层析板(二氯甲烷/甲醇＝20/1，Rf＝0.4)纯化，得到化合物20e。
LCMS(ESI)m/z:373.3[M+1] +
步骤五
将化合物20e(60mg,161.09umol)和氯化氢/乙酸乙酯(4M,2mL)一次加到EtOAc(2mL)中，然后在20-30℃下搅拌反应16小时。将反应液直接通过旋蒸蒸干。得到化合物20f(盐酸盐)，直接用于下一步。
LCMS(ESI)m/z:273.2[M+1] +
步骤六
将化合物20f(25mg,80.96umol)和N,N-二异丙基乙胺(31.39mg,242.88umol)加到DCM(2mL)中，随后将化合物1e(16.70mg,80.96umol)，N,N-二异丙基乙胺(31.39mg,242.88umol,42.31uL)和三光气(12.01mg,40.48umol)加在一起搅拌10分钟后0℃下加到搅拌中的反应液中，反应液在25-30℃下搅拌反应10分钟。向反应液中加入1mL水淬灭反应，然后加入5mL水和5mL二氯甲烷萃取，分液，水相再用5mL二氯甲烷萃取，合并有机相，无水硫酸钠干燥，旋蒸蒸干得到粗品，通过制备分离(column:Boston Green ODS 150*30 5u；流动相:[水(0.075％三氟乙酸)-乙腈]；B％:33％-53％,7min)纯化得到实施例20的三氟乙酸盐。实施例20的三氟乙酸盐加入碳酸氢钠溶液中，乙酸乙酯萃取，有机相用无水硫酸钠干燥，减压下浓缩得到实施例20。
LCMS(ESI)m/z:505.3[M+1] +
1H NMR(400MHz,METHANOL-d4)δ8.50～8.49(m,1H),8.39(d,J＝8.4Hz,1H),7.90(d,J＝8.4Hz,2H),7.70(t,J＝3.6Hz,1H),7.56(s,1H),6.61(s,1H),5.02～4.97(m,1H),4.30～4.27(m,1H),4.05～3.97(m,1H),3.27～3.10(m,2H),2.96～2.87(m,1H),2.48(d,J＝6.8Hz,3H),2.33(d,J＝4Hz,3H),2.17(s,1H),2.04～1.91(m,2H),1.63(m,1H),1.54(d,J＝6.8Hz,3H).
步骤七
将实施例20(60mg,118.92umol)的三氟乙酸盐送SFC分离(column:DAICEL CHIRALPAK AS-H(250mm*30mm,5um)；流动相:[0.1％氨水乙醇]；B％:50％-50％,min)。最终得到实施例21(保留时间：3.391min)和实施例22保留时间：4.092min)
实施例21：
LCMS(ESI)m/z:505.2[M+1] +
1H NMR(400MHz,METHANOL-d4)δ8.48(d,J＝4.4Hz,1H),8.38(s,1H),7.87～7.85(m,2H),7.68(d,J＝4Hz,1H),6.74(s,1H),6.19(s,1H),5.03～4.98(m,1H),4.23(d,J＝13.2Hz,1H),4.11(d,J＝13.2Hz,1H),3.25～3.19(m,1H),2.88～2.81(m,2H),2.33(s,3H),2.29(s,3H),2.12(s,1H),1.89～1.81(m,2H),1.61～1.60(m,1H),1.52(d,J＝7.2Hz,3H)
实施例22：
LCMS(ESI)m/z:505.2[M+1] +
1H NMR(400MHz,METHANOL-d4)δ8.36(d,J＝4Hz,1H),8.28(d,J＝1.6Hz,1H),7.81～7.75(m,2H),7.84(s,1H),7.55(d,J＝4Hz,1H),6.62(s,1H),6.08(s,1H),4.90～4.85(m,1H),4.11(d,J＝13.2Hz,1H),4.00(d,J＝13.2Hz,1H),3.11～3.05(m,1H),2.74～2.70(m,2H),2.22(s,3H),2.17(s,3H),1.99(s,1H),1.77～1.66(m,2H),1.47(m,1H),1.39(d,J＝7.2Hz,3H)
实施例23
Figure PCTCN2019101121-appb-000080
Figure PCTCN2019101121-appb-000081
步骤一
将化合物1e(200mg,969.84umol)和Boc 2O(222.25mg,1.02mmol)一起加到THF(4mL)和H 2O(4mL)中，然后在10℃下搅拌反应16小时。将反应液直接旋蒸蒸干得到粗品化合物23a，直接用于下一步。
LCMS(ESI)m/z:307.3[M+1] +
步骤二
将化合物23a(250mg,816.10umol)溶解到THF(5mL)中，然后将钠氢(65.28mg,1.63mmol,60％)加到反应液中，搅拌十分钟后将碘甲烷(231.67mg,1.63mmol)加到搅拌中的反应液中，最后反应液在20℃下搅拌反应16小时。向反应液中加入5mL水淬灭反应，然后加20mL乙酸乙酯萃取，分液，水相再用20mL乙酸乙酯萃取，合并有机相，无水硫酸钠干燥，旋蒸蒸干得到化合物23b。
LCMS(ESI)m/z:320.9[M+1] +
步骤三
将化合物23b(200mg,624.30umol)溶解到乙酸乙酯(5mL)中，然后将氯化氢/乙酸乙酯(4M,4.00mL)加到反应液中，然后20℃搅拌反应16小时。将反应液直接过滤，滤饼即为化合物23c。
LCMS(ESI)m/z:220.9[M+1] +
步骤四
将化合物1h(140mg,453.35umol,HCl)和N,N-二异丙基乙胺(175.77mg,1.36mmol)加到二氯甲烷(5mL)中，随后将化合物23c(99.85mg,453.35umol)，N,N-二异丙基乙胺(175.77mg,1.36mmol,236.89uL)和三光气(67.27mg,226.68umol)加在一起搅拌10分钟后0℃下加到搅拌中的反应液中，反应液在15-20℃下搅拌反应10分钟。将反应液直接旋蒸蒸干得到粗品。将粗品送制备分离(色谱柱:Boston Green ODS 150*30mm*5um；流动相:[水(0.075％三氟乙酸)-乙腈]；B％:30％-60％,9min)得到实施23的三氟乙酸 盐。实施例23的三氟乙酸盐加入碳酸氢钠溶液中，乙酸乙酯萃取，有机相用无水硫酸钠干燥，减压下浓缩得到实施例23。
LCMS(ESI)m/z:519.1[M+1] +
1H NMR(400MHz,METHANOL-d 4)δ8.51(br d,J＝3.8Hz,1H),8.44(br s,1H),7.95(br s,2H),7.71(br s,1H),6.37(s,1H),6.24(s,1H),5.83(s,1H),5.33(br d,J＝6.8Hz,1H),3.70(br s,4H),3.55(br s,4H),2.79(s,3H),2.39(s,3H),2.34(s,3H),1.67(br d,J＝6.8Hz,3H)
实施例24,25
Figure PCTCN2019101121-appb-000082
步骤一
将高锰酸钾(45.60g,288.54mmol)加入到含化合物24a(7g,48.09mmol)的水(60mL)，吡啶(60mL)混合溶液中，100℃搅拌3小时。反应液直接过滤，用甲醇100mL洗涤，滤液经减压浓缩即得化合物24b。
1H NMR(400MHz,DMSO-d 6)δ8.60(s,1H),8.02(dd,J＝1.64,9.16Hz,1H).
步骤二
将化合物24b(4g,22.79mmol)，N,O-二甲基羟胺盐酸盐(1.39g,22.79mmol)的DMF(100mL)溶液中加入HATU(13.00g,34.18mmol)，N,N-二异丙基乙胺(5.89g,45.57mmol,7.94mL),30℃搅拌2小时。反应液加入水100mL，再加入乙酸乙酯(100mL*2)萃取，合并有机相并用无水硫酸钠干燥，减压浓缩得粗品。粗品经(PE:EA＝4:3，Rf＝0.42)分离得化合物24c。
LCMS(ESI)m/z:218.6[M+1] +
1H NMR(400MHz,CHLOROFORM-d)δ8.56(d,J＝1.76Hz,1H),7.83(dd,J＝1.88,8.42Hz,1H),3.52-3.57(m,3H),3.30-3.39(m,3H).
步骤三
0℃下向化合物24c(3g,13.72mmol)的THF(30mL)溶液中滴加甲基溴化镁(3M,5.49mL)，25℃搅拌2小时。向反应液中加入饱和氯化铵溶液(50mL)，再加入乙酸乙酯(50mL*2)萃取，合并有机相并用无水硫酸钠干燥，减压浓缩得粗品。粗品经快速硅胶柱分离(PE:EA＝3:1,Rf＝0.43)得到化合物24d。
LCMS(ESI)m/z:173.8[M+1] +
1H NMR(400MHz,CHLOROFORM-d)δ8.75(d,J＝1.52Hz,1H),7.99(dd,J＝1.64,8.16Hz,1H),2.65(s,3H).
步骤四
向化合物24d(1.3g,7.49mmol)，1氢-吡唑(509.88mg,7.49mmol)的DMF(20mL)溶液中加入K 2CO 3(3.11g,22.47mmol)，100℃搅拌16小时。反应液经硅藻土抽滤，滤液经油泵减压浓缩得粗品。粗品经制备层析板(PE:EA＝3:1,Rf＝0.38)分离得化合物24e。
LCMS(ESI)m/z:205.8[M+1] +
1H NMR(400MHz,CHLOROFORM-d)δ8.84(s,1H),8.46(d,J＝2.76Hz,1H),8.15(dd,J＝1.76,11.54Hz,1H),7.89(d,J＝1.00Hz,1H),6.52-6.61(m,1H),2.66(s,3H).
步骤五
向化合物24e(0.60g,2.92mmol)的甲醇(10mL)溶液中加入乙酸铵(2.25g,29.24mmol)，65℃搅拌1小时后，再加入NaBH 3CN(551.26mg,8.77mmol,，65℃搅拌3小时。反应液直接浓缩得粗品。粗品经制备层析板(DCM:MeOH＝10:1,Rf＝0.20)分离得产物24f，再进一步通过SFC分离(色谱柱:DAICEL CHIRALPAK AD-H(250mm*30mm,5μm)；流动相:[0.1％氨水甲醇]；B％:35％-35％,min)分离得24g1(保留时间：4.177min)和24g2(保留时间：4.980min)。
24g1:
LCMS(ESI)m/z:206.8[M+1] +
1H NMR(400MHz,CHLOROFORM-d)δ8.28(s,2H),7.83(d,J＝1.00Hz,1H),7.70(dd,J＝1.63,11.92Hz, 1H),6.46-6.55(m,1H),4.28(q,J＝6.70Hz,1H),1.43(d,J＝6.54Hz,3H).
24g2:
LCMS(ESI)m/z:206.8[M+1] +
1H NMR(400MHz,CHLOROFORM-d)δ8.28(s,2H),7.82(d,J＝1.26Hz,1H),7.69(dd,J＝1.64,11.92Hz,1H),6.47-6.51(m,1H),4.27(q,J＝6.62Hz,1H),1.42(d,J＝6.54Hz,3H).
步骤六
0℃化合物24g1(59.63mg,289.14umol)的DCM(2mL)溶液中加入N,N-二异基丙胺(140.13mg,1.08mmol,188.86uL)，三光气(53.63mg,180.71umol)，0℃下搅拌10分钟，再将反应液滴入到含化合物1h(140mg,361.42umol,三氟乙酸盐)，N,N-二异基丙胺(140.13mg,1.08mmol,188.86uL)，DCM(2mL)的单口瓶中，0℃下继续搅拌10分钟。向反应液中加入3mL水，再用二氯甲烷(3mL*3)萃取，合并有机相减压浓缩得粗品。粗品送制备色谱柱分离(色谱柱:Boston Green ODS 150*30mm 5μm；流动相:[水(0.075％三氟乙酸)-乙腈]；B％:21％-41％,7min)得实施例24的三氟乙酸盐。实施例24的三氟乙酸盐加入碳酸氢钠溶液中，乙酸乙酯萃取，有机相用无水硫酸钠干燥，减压下浓缩得到实施例24。
LCMS(ESI)m/z:506.2[M+1] +
1H NMR(400MHz,DMSO-d 6)δ12.39(br s,1H),11.72(br s,1H),11.05(br s,1H),8.31-8.43(m,1H),7.90-7.99(m,1H),7.80-7.87(m,1H),7.09(d,J＝7.54Hz,1H),6.58(t,J＝2.02Hz,1H),6.09-6.45(m,2H),4.98(q,J＝7.10Hz,1H),3.78(br s,4H),3.54(br s,4H),2.33(s,3H),2.25(s,3H),1.46(d,J＝7.03Hz,3H).用化合物24g2以同样的方法制备得到实施例25的三氟乙酸盐。实施例25的三氟乙酸盐加入碳酸氢钠溶液中，乙酸乙酯萃取，有机相用无水硫酸钠干燥，减压下浓缩得到实施例25。
LCMS(ESI)m/z:506.2[M+1] +
1H NMR(400MHz,DMSO-d 6)δ12.09-12.66(m,1H),11.45-12.01(m,1H),11.04(br s,1H),8.30-8.43(m,1H),7.95(dd,J＝1.52,11.80Hz,1H),7.83(d,J＝1.52Hz,1H),7.08(d,J＝7.54Hz,1H),6.58(t,J＝2.14Hz,1H),6.37(br s,1H),6.18(br s,1H),4.98(q,J＝7.04Hz,1H),3.78(br s,5H),3.53(br s,3H),2.32(s,3H),2.24(s,3H),1.46(d,J＝7.04Hz,3H).
实施例26、27
Figure PCTCN2019101121-appb-000083
Figure PCTCN2019101121-appb-000084
步骤一
将化合物1f(3.56g,15.90mmol)溶于1-甲基吡咯烷酮(15mL)中，加入3-甲酸乙酯哌啶(5g,31.80mmol)、N,N-二异丙基乙胺(6.17g,47.71mmol,8.31mL)，90℃搅拌16小时。反应液加水40mL，用乙酸乙酯(40mL*3)萃取，合并有机相，用水(100mL*5)洗涤，饱和氯化钠溶液(100mL*1)洗涤，有机相无水硫酸钠干燥，过滤，旋干溶液得粗品。粗品经快速硅胶柱纯化(PE:EA＝1:1)得化合物26a。
LCMS(ESI)m/z:345.2[M+1] +
1H NMR(400MHz,DMSO-d6)δppm 9.23(s,1H),11.85(s,1H),6.15(s,1H),4.67(d,J＝10.79Hz,1H),4.43(d,J＝12.80Hz,1H),4.13-4.08(m,2H),3.17(d,J＝5.26Hz,1H),3.06(t,J＝11.54Hz,1H),2.99-2.89(m,1H),2.47-2.41(m,1H),2.19(s,3H),2.11(s,3H),2.03–1.96(m,1H),1.74-1.57(m,2H),1.45-1.36(m,1H),1.19(t,J＝7.16Hz,4H).
步骤二
将化合物26a(100mg,290.35umol)溶于四氢呋喃(1mL)/水(0.3mL)溶液中，加入氢氧化锂一水合物(36.55mg,871.05umol)，16℃搅拌2小时。用4N盐酸调反应液pH至2-3，乙酸乙酯萃取(10mL*2)，旋干溶剂得化合物26b。.
LCMS(ESI)m/z:316.9[M+1] +
步骤三
在化合物26b(90mg,284.49umol)溶于N,N二甲基甲酰胺(1mL)中，加入化合物1e(58.67mg,284.49umol)、HATU(162.26mg,426.73umol)、N,N-二异丙基乙胺(110.30mg,853.46umol,148.66uL)，16℃搅拌3小时，旋干溶剂得粗品。粗品经制备色谱柱分离纯化(色谱柱:Xtimate C18 150*25mm*5um；流动相:[水(0.1％三氟乙酸)-乙腈]；B％:30％-40％,11min)得到的产物再经SFC拆分(column:DAICEL  CHIRALCEL OJ-H(250mm*30mm,5um)；mobile phase:[0.1％氨水乙醇]；B％:30％-30％,min)得到实施例26(保留时间：2.947min)，实施例27(保留时间：3.504min)。
实施例26：
LCMS(ESI)m/z:505.7[M+1] +
1H NMR(400MHz,METHANOL-d4)δppm 8.44(dd,J＝4.52,0.80Hz,1H),8.30(d,J＝2.00Hz,1H),7.84-7.79(m,1H),7.78-7.73(m,1H),7.66(d,J＝4.28Hz,1H),6.11(s,1H),6.02(s,1H),5.07(q,J＝6.80Hz,1H),4.29(dd,J＝13.60,3.20Hz,1H),4.21-4.11(m,1H),3.57-3.51(m,2H),3.40-3.33(m,1H),2.58-2.45(m,1H),2.17(d,J＝9.06Hz,6H),2.00–1.89(m,2H),1.71-1.63(m,1H),1.45(d,J＝7.04Hz,3H).
实施例27：
LCMS(ESI)m/z:505.6[M+1] +
1H NMR(400MHz,METHANOL-d4)δppm 8.46(d,J＝4.02Hz,1H),8.35(s,1H),7.94-781(m,2H),7.67(d,J＝4.28Hz,1H),5.05(q,J＝7.04Hz,1H),4.38(d,J＝12.00Hz,1H),4.21(d,J＝13.80Hz,1H)3.54-3.49(m,2H),3.35(s,1H),2.65-2.54(m,1H),2.35-2.27(m,6H),2.08-1.98(m,1H),1.87-1.78(m,2H),1.70-1.58(m,1H),1.50(d,J＝7.20Hz,3H).
实施例28、29
Figure PCTCN2019101121-appb-000085
步骤一
将化合物15b1(335.49mg,1.62mmol)，N,N-二琥珀酰亚胺基碳酸酯(622.15mg,2.43mmol)和4-N,N-二甲氨基吡啶(98.90mg,809.56umol)以及三乙胺(163.84mg,1.62mmol,225.36uL)一起加到DMF(5mL)中，然后在60℃下搅拌反应1小时，然后25℃将化合物20f(500mg,1.62mmol,HCl)和三乙胺(163.84mg,1.62mmol)的DMF(5mL)溶液加到反应液中，最后反应液在25℃下搅拌反应0.5小时。将反应液直接通过减压浓缩得粗品。送制备色谱柱分离(色谱柱:Phenomenex Gemini-NX 150*30mm*5um；mobile phase:[水(0.04％氨水+10mM碳酸氢铵)-乙腈]；B％:39％-69％,8min)纯化。再次送制备色谱柱分离(色谱 柱:Boston Green ODS 150*30mm*5um；mobile phase:[水(0.075％三氟乙酸)-乙腈]；B％:25％-55％,7min)，得到化合物28a。
LCMS(ESI)m/z:506.2[M+1] +
步骤二
将28a(130mg,257.15umol)送SFC分离。SFC条件为(色谱柱:DAICEL CHIRALPAK AD(250mm*30mm,10um)；流动相:[0.1％NH3H2O,IPA]；B％:60％-60％,min)。最终得到实施例28(保留时间：1.499min)和实施例29(保留时间：2.252min)。
实施例28
LCMS(ESI)m/z:506.1[M+1] +
1H NMR(400MHz,METHANOL-d4)δ8.46-8.21(m,2H),7.90-7.71(m,2H),7.57(br s,1H),6.73-5.99(m,1H),5.73(q,J＝6.5Hz,1H),5.19-5.00(m,1H),4.24-4.02(m,2H),2.90-2.55(m,2H),2.21(br s,3H),2.16(s,3H),1.99(br d,J＝11.8Hz,1H),1.75(br s,2H),1.55-1.37(m,5H).
实施例29
LCMS(ESI)m/z:506.6[M+1] +
1H NMR(400MHz,METHANOL-d4)δ＝8.49(d,J＝4.4Hz,1H),8.43(br s,1H),7.97-7.89(m,2H),7.68(d,J＝4.3Hz,1H),6.78-6.16(m,1H),5.85(br s,1H),5.32-5.19(m,1H),4.36-4.29(m,2H),3.04-2.72(m,2H),2.30-2.25(m,1H),2.28(s,2H),2.05-2.03(m,4H),1.84(br d,J＝11.0Hz,2H),1.61-1.53(m,5H).
生物测试数据：
实验例1：本发明化合物的体外酶活性测试
实验目的
通过Z′-LYTE TM激酶测试实验检测酶活性,以化合物的IC 50值为指标，来评价化合物对RET和RET(V804M)激酶的抑制作用。
实验方法
RET和RET(V804M)激酶的测试所用化合物进行3倍浓度稀释，浓度为3μM到0.152nM 10个浓度；DMSO在检测反应中的含量为1％。
试剂：
基本反应缓冲液，20mM羟乙基哌嗪乙硫磺酸(Hepes)(pH 7.5)缓冲液,10mM MgCl 2,1mM乙二醇双氨乙基醚四乙酸(EGTA),0.02％聚氧乙烯十二烷醚(Brij35),0.02mg/ml牛血清蛋白(),0.1mM Na 3VO 4,2mM二硫苏糖醇(DTT),1％DMSO。
化合物：
待测化合物溶解在100％的DMSO体系中稀释成10mM待用。溶液稀释采用Integra Viaflo Assist。通用酶的反应过程：
测试条件：RET酶的浓度3μM，肽底物CHKtide浓度为1000μM，ATP浓度为20μM；RET(V804M)酶的浓度为80μM，底物肽的浓度为1000μM,ATP浓度为30μM。
反应过程：按照测试条件，配制激酶/多肽溶液。加入不同浓度的化合物溶液，室温孵育20分钟，加入对应浓度的33P-ATP，室温孵育反应120分钟。通过filter-binding方法检测到放射活性。
反应检测：
激酶反应液中添加0.5％浓度的磷酸终止反应，Envision仪器读板。
数据分析
将数据转化为磷酸化率和抑制率，参数曲线拟合(GraphPad Software)得到化合物IC 50数据。
实验结果见表1：
表1 实施例1激酶活性IC 50测试结果
  RET酶IC 50(nM) RET V804M IC 50(nM)
实施例1的盐酸盐 1.1 7.86
实施例2 293 609
实施例3 62.1 204
实施例4的三氟乙酸盐 0.5 2.58
实施例6 60.7 69.6
实施例7 4.2 11.7
实施例8 50.4 134
实施例9 201 250
实施例10 23.9 31.7
实施例11 4.76 11.6
实施例12 68.2 83.3
实施例13的三氟乙酸盐 6.73 9.16
实施例14的三氟乙酸盐 4.57 8.24
实施例15的三氟乙酸盐 4.00 9.00
实施例16的三氟乙酸盐 7.3 11.0
实施例17的三氟乙酸盐 4.3 2.74
实施例18的三氟乙酸盐 1.06 2.36
实施例19的三氟乙酸盐 44.9 35.6
实施例20的三氟乙酸盐 4.31 4.19
实施例21 96.4 91.4
实施例22 0.9 1.62
实施例23的三氟乙酸盐 1.35 12.8
实施例24的三氟乙酸盐 22.6 24.7
实施例25的三氟乙酸盐 5.67 14.4
实施例26 N/A N/A
实施例27 N/A N/A
实施例28 231 131
实施例29 2.40 4.77
N/A代表未检测
实验结论：本申请化合物对RET及其突变RET V804M具有优异的抑制剂活性，将会在RET异常肿瘤患者中具有优异的治疗效果。

Claims (19)

  1. 式(III)所示化合物、其异构体或其药学上可接受的盐，
    Figure PCTCN2019101121-appb-100001
    其中，
    A环选自
    Figure PCTCN2019101121-appb-100002
    所述
    Figure PCTCN2019101121-appb-100003
    任选被1、2或3个R g取代；
    R 1选自H、F、Cl、Br、I、OH、NH 2、CN、C 1-3烷基、C 1-3烷氧基和C 3-6环烷基，所述C 1-3烷基、C 1-3烷氧基和C 3-6环烷基任选被1、2或3个R a取代；
    R 2选自H、F、Cl、Br、I、OH、NH 2、CN、C 1-3烷基和C 1-3烷氧基，所述C 1-3烷基和C 1-3烷氧基任选被1、2或3个R b取代；
    R 4选自H、F、Cl、Br、I、OH、NH 2、CN、C 1-5烷基、C 1-5烷氧基、C 1-5烷氨基和5-6元杂芳环，所述C 1- 5烷基、C 1-5烷氧基、C 1-5烷氨基和5-6元杂芳环任选被1、2或3个R d取代；
    R a和R b分别独立地选自F、Cl、Br、I、OH和NH 2；
    R d选自H、F、Cl、Br、I、OH、NH 2、CN、C 1-3烷基、C 1-3烷氧基和C 1-4烷氨基；
    T 1、T 2、T 3和T 5分别独立地选自CH和N；
    T 4选自CR 5和N；
    L 2选自NH和O；
    L 3选自-CH(R 6)-、-CH(R 6)CH 2-、-CHCH 2CH 2-、-CH(R 6)-O-、-CHCH 2-O-、-CH 2-N(R 6)-、-CH 2CH 2-N(R 6)-；
    R 5选自H；
    R 6选自H和CH 3；
    或者R 5、R 6连接在一起使结构单元
    Figure PCTCN2019101121-appb-100004
    选自
    Figure PCTCN2019101121-appb-100005
    R g选自H、F、Cl、Br、I、OH、NH 2和CN；
    所述5～6元杂芳基分别包含1、2、3或4个独立选自-NH-、-O-、-S-和N的杂原子或杂原子团。
  2. 根据权利要求1所述化合物、其异构体或其药学上可接受的盐，其中，R 1选自H、F、Cl、Br、I、OH、NH 2、CN、CH 3、CH 2CH 3、
    Figure PCTCN2019101121-appb-100006
    所述CH 3、CH 2CH 3、
    Figure PCTCN2019101121-appb-100007
    任选被1、2或3个R a取代。
  3. 根据权利要求2所述化合物、其异构体或其药学上可接受的盐，其中，R 1选自CH 3和
    Figure PCTCN2019101121-appb-100008
  4. 根据权利要求1～3任意一项所述化合物、其异构体或其药学上可接受的盐，其中，R 2选自H、F、Cl、Br、I、OH、NH 2、CN、CH 3、CH 2CH 3和
    Figure PCTCN2019101121-appb-100009
    所述CH 3、CH 2CH 3和
    Figure PCTCN2019101121-appb-100010
    任选被1、2或3个R b取代。
  5. 根据权利要求4所述化合物、其异构体或其药学上可接受的盐，其中，R 2选自CH 3。
  6. 根据权利要求1～3任意一项所述化合物、其异构体或其药学上可接受的盐，其中R d选自H、F、Cl、Br、I、OH、NH 2、CN、CH 3和
    Figure PCTCN2019101121-appb-100011
  7. 根据权利要求1～3任意一项所述化合物、其异构体或其药学上可接受的盐，其中，R 4选自：H、F、Cl、Br、I、OH、NH 2、CN、CH 3、吡唑基、异噁唑基、咪唑基、三唑基、噁唑基、
    Figure PCTCN2019101121-appb-100012
    所述CH 3、吡唑基、异噁唑基、咪唑基、三唑基、噁唑基、
    Figure PCTCN2019101121-appb-100013
    任选被1、2或3个R d取代。
  8. 根据权利要求7所述化合物、其异构体或其药学上可接受的盐，其中，R 4选自：
    Figure PCTCN2019101121-appb-100014
    Figure PCTCN2019101121-appb-100015
  9. 根据权利要求1～3任意一项所述化合物、其异构体或其药学上可接受的盐，其中，结构单元
    Figure PCTCN2019101121-appb-100016
    选自
    Figure PCTCN2019101121-appb-100017
  10. 根据权利要求1～3任意一项所述化合物、其异构体或其药学上可接受的盐，其中，A环选自
    Figure PCTCN2019101121-appb-100018
    Figure PCTCN2019101121-appb-100019
  11. 根据权利要求1～3任意一项所述化合物、其异构体或其药学上可接受的盐，其中，结构单元
    Figure PCTCN2019101121-appb-100020
    选自
    Figure PCTCN2019101121-appb-100021
  12. 根据权利要求11任意一项所述化合物、其异构体或其药学上可接受的盐，其中，结构单元
    Figure PCTCN2019101121-appb-100022
    选自
    Figure PCTCN2019101121-appb-100023
  13. 根据权利要求1～3任意一项所述化合物、其异构体或其药学上可接受的盐，其中，结构单元
    Figure PCTCN2019101121-appb-100024
    选自
    Figure PCTCN2019101121-appb-100025
  14. 根据权利要求1～9、10～13任意一项所述化合物、其异构体或其药学上可接受的盐，其选自
    Figure PCTCN2019101121-appb-100026
    其中，R 1、R 2、R 4、L 2、L 3、T 1、T 2、T 3、T 4和T 5如权利要求1～9、11～13任意一项所定义。
  15. 下式所示化合物、其异构体或其药学上可接受的盐，其选自：
    Figure PCTCN2019101121-appb-100027
    Figure PCTCN2019101121-appb-100028
    Figure PCTCN2019101121-appb-100029
  16. 根据权利要求15所述的下式所示化合物、其异构体或其药学上可接受的盐，其选自：
    Figure PCTCN2019101121-appb-100030
    Figure PCTCN2019101121-appb-100031
    Figure PCTCN2019101121-appb-100032
    Figure PCTCN2019101121-appb-100033
  17. 一种药物组合物，包括治疗有效量的根据权利要求1～16任意一项所述的化合物、其异构体或其药学上可接受的盐作为活性成分以及药学上可接受的载体。
  18. 根据权利要求1～16任意一项所述的化合物、其异构体或其药学上可接受的盐在制备制备RET激酶抑制剂的应用。
  19. 根据权利要求17所述的组合物在制备RET激酶抑制剂的应用。
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