WO2021197467A1 - Multi-target anti-tumor compound, preparation method therefor and use thereof - Google Patents
Multi-target anti-tumor compound, preparation method therefor and use thereof Download PDFInfo
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- WO2021197467A1 WO2021197467A1 PCT/CN2021/085239 CN2021085239W WO2021197467A1 WO 2021197467 A1 WO2021197467 A1 WO 2021197467A1 CN 2021085239 W CN2021085239 W CN 2021085239W WO 2021197467 A1 WO2021197467 A1 WO 2021197467A1
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- amino
- pyran
- tetrahydro
- benzamide
- indazol
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- 0 C*C(CC(C)(C(C*(C)[Cn])CC1C(C2)C2C*1)C(CC1)C*1C(*C)C(C(C)CC*)C(C)N)C(C)C Chemical compound C*C(CC(C)(C(C*(C)[Cn])CC1C(C2)C2C*1)C(CC1)C*1C(*C)C(C(C)CC*)C(C)N)C(C)C 0.000 description 15
- UAIVFDJJMVMUGY-UHFFFAOYSA-N CC1N(C)CCN(C)C1 Chemical compound CC1N(C)CCN(C)C1 UAIVFDJJMVMUGY-UHFFFAOYSA-N 0.000 description 2
- VVRPLPXESFSMPF-UHFFFAOYSA-N CC(c1cc(F)cc(F)c1)c(nc1)cc2c1[nH]nc2NC(c(ccc(N1CCN(C)CC1)c1)c1NC1CCOCC1)=O Chemical compound CC(c1cc(F)cc(F)c1)c(nc1)cc2c1[nH]nc2NC(c(ccc(N1CCN(C)CC1)c1)c1NC1CCOCC1)=O VVRPLPXESFSMPF-UHFFFAOYSA-N 0.000 description 1
- CLPZHEDSMNQBPP-UHFFFAOYSA-N CC1NC(C)CN(C)C1 Chemical compound CC1NC(C)CN(C)C1 CLPZHEDSMNQBPP-UHFFFAOYSA-N 0.000 description 1
- BAUOUXYQYYCEBR-UHFFFAOYSA-N CCN(C)C(C)=C Chemical compound CCN(C)C(C)=C BAUOUXYQYYCEBR-UHFFFAOYSA-N 0.000 description 1
- SZLAPTPUVHWDDJ-UHFFFAOYSA-N CCN(CCN1CCCC1)c(cc1)cc(NC2CCOCC2)c1C(Nc1n[nH]c2c1cc(Cc1cccc(F)c1)cc2)=O Chemical compound CCN(CCN1CCCC1)c(cc1)cc(NC2CCOCC2)c1C(Nc1n[nH]c2c1cc(Cc1cccc(F)c1)cc2)=O SZLAPTPUVHWDDJ-UHFFFAOYSA-N 0.000 description 1
- OXRJBMLZPOYATC-UHFFFAOYSA-N CCOC(c(c(Br)c1)ccc1I)=O Chemical compound CCOC(c(c(Br)c1)ccc1I)=O OXRJBMLZPOYATC-UHFFFAOYSA-N 0.000 description 1
- LSDHHVOALFWKDJ-UHFFFAOYSA-N CCOC(c1ccc(CC(C(C2)CC(C3)C4)(C3C24C(OCc2ccccc2)=O)O)cc1Br)=O Chemical compound CCOC(c1ccc(CC(C(C2)CC(C3)C4)(C3C24C(OCc2ccccc2)=O)O)cc1Br)=O LSDHHVOALFWKDJ-UHFFFAOYSA-N 0.000 description 1
- QCAOGKWLNFBEQF-UHFFFAOYSA-N CN(C)CCSc(cc1)cc(NC2CCOCC2)c1C(Nc1n[nH]c2ccc(Cc3cc(F)cc(F)c3)cc12)=O Chemical compound CN(C)CCSc(cc1)cc(NC2CCOCC2)c1C(Nc1n[nH]c2ccc(Cc3cc(F)cc(F)c3)cc12)=O QCAOGKWLNFBEQF-UHFFFAOYSA-N 0.000 description 1
- WWLPOKCLCOLCKR-UHFFFAOYSA-N CN(CC1)CCN1c(cc1)cc(NC2CCOCC2)c1C(Nc1n[nH]c2c1cc(C(CF)c1cc(F)ccc1)cc2)=O Chemical compound CN(CC1)CCN1c(cc1)cc(NC2CCOCC2)c1C(Nc1n[nH]c2c1cc(C(CF)c1cc(F)ccc1)cc2)=O WWLPOKCLCOLCKR-UHFFFAOYSA-N 0.000 description 1
- MQORKDBNAWGHIZ-UHFFFAOYSA-N CN(CC1)CCN1c1ccc(C(Nc2n[nH]c3cnc(Cc4cc(F)cc(F)c4)cc23)=O)c(NC2CCOCC2)c1 Chemical compound CN(CC1)CCN1c1ccc(C(Nc2n[nH]c3cnc(Cc4cc(F)cc(F)c4)cc23)=O)c(NC2CCOCC2)c1 MQORKDBNAWGHIZ-UHFFFAOYSA-N 0.000 description 1
- XNLRUDFBVYFTKV-UHFFFAOYSA-N CN1CC2(CC2)N(C)CC1 Chemical compound CN1CC2(CC2)N(C)CC1 XNLRUDFBVYFTKV-UHFFFAOYSA-N 0.000 description 1
- RXYPXQSKLGGKOL-UHFFFAOYSA-N CN1CCN(C)CC1 Chemical compound CN1CCN(C)CC1 RXYPXQSKLGGKOL-UHFFFAOYSA-N 0.000 description 1
- RONBDFSZIHJBMI-UHFFFAOYSA-N NC1(CC2CC(C3)C1)C3C2c1ccc(C(Nc2n[nH]c3c2cc(Cc2cc(F)cc(F)c2)cc3)=O)c(NC2CCNCC2)c1 Chemical compound NC1(CC2CC(C3)C1)C3C2c1ccc(C(Nc2n[nH]c3c2cc(Cc2cc(F)cc(F)c2)cc3)=O)c(NC2CCNCC2)c1 RONBDFSZIHJBMI-UHFFFAOYSA-N 0.000 description 1
- JMJRYTGVHCAYCT-UHFFFAOYSA-N O=C1CCOCC1 Chemical compound O=C1CCOCC1 JMJRYTGVHCAYCT-UHFFFAOYSA-N 0.000 description 1
- APGWJGNOWJHSLW-UHFFFAOYSA-N OC(c(c(Br)c1)ccc1I)=O Chemical compound OC(c(c(Br)c1)ccc1I)=O APGWJGNOWJHSLW-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- 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
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene
-
- 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
- C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
- C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
- C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
Definitions
- the invention belongs to the field of medicinal chemistry, and specifically relates to a new type of small molecule compound with multiple kinase inhibitory activity and a preparation method thereof, as well as the use of this type of compound in preventing and treating tumors.
- Gene fusion is a driving factor for many malignant tumors. Structurally, they fuse with upstream genes and maintain a complete tyrosine kinase domain, independent of ligand dimerization, leading to continuous activation of downstream signals to promote tumor cell growth and proliferation. With the continuous maturity and improvement of technologies such as second-generation sequencing and RT-PCR, many clinically relevant fusion genes have been detected. Including NTRK1, NTRK2, NTRK3 (encoding TRKA, TRKB and TRKC protein), ALK, ROS1, etc.
- Tropomyosin receptor kinase Tropomyosin receptor kinase
- TRK Tropomyosin receptor kinase
- NTRK1, NTRK2 and NTRK3 genes which are found in neuronal tissues It is widely expressed and participates in the maintenance, signal transduction and survival of neuronal cells.
- TRK is a transmembrane receptor composed of extracellular ligand binding domain, transmembrane and intracellular ATP binding domain.
- TRKA, TRKB and TRKC show a high degree of structural similarity, including two immunoglobulin-like motifs (Ig1 and Ig2), which are considered to contain ligand binding sites, and three rich The 24-residue motif containing leucine (LRR1-3 motif), two cysteine clusters (C1 and C2) flanking the LRR1-3 motif are specific to TRK protein, Not found in other subfamily of amino acid kinases.
- the TRKA is a high-affinity receptor for nerve growth factor (hereinafter referred to as NGF), and the TRKB is a high-affinity receptor for brain-derived neurotrophic factor (BDNF) and neurotrophic factor (hereinafter referred to as NT)-4/5. Affinity receptor, and the TRKC is a high affinity receptor for NT-3.
- NGF nerve growth factor
- BDNF brain-derived neurotrophic factor
- NT neurotrophic factor-4/5.
- Affinity receptor NT-4/5
- the TRKC is a high affinity receptor for NT-3. It is known that the activation of TRKA in peripheral nerves by NGF causes hyperalgesia, including nociceptive pain, neuropathic pain, and osteoarthritis, chronic low back pain, rheumatoid arthritis, fractures, interstitial cystitis, and chronic pancreatitis. In cancer pain accompanied by two of the above-mentioned pain types.
- the literature shows that the activation of the BDNF/TrkB pathway is also involved in various types of pain, including inflammatory pain, neuropathic pain and surgical pain, so TRKA kinase and other TRK kinase inhibitors can provide chronic pain states. Effective treatment.
- changes in the TRK signaling pathway including gene fusion, protein overexpression, or single nucleotide changes, have been found to be associated with many cancers.
- the cancer includes neuroblastoma, ovarian cancer, breast cancer, prostate cancer, pancreatic cancer, multiple myeloma, astrocytoma and medulloblastoma, glioma, melanoma, thyroid cancer, lung gland Cancer, large cell neuroendocrine tumors, and colorectal cancer.
- NTRK gene fusion is the most clear cause of cancer.
- TRK fusion protein can drive the spread and growth of TRK fusion tumors by activating PI3K, RAS/MAPK/ERK and PLC- ⁇ pathways and triggering a permanent signal cascade reaction independently of ligand dimerization and phosphorylation.
- the NTRK gene fusion can occur in any part of the body, so TRK fusion cancer may appear in a variety of adult and child solid tumors, including breast analog secreting carcinoma (MASC), colon cancer, lung cancer, pancreatic cancer, thyroid cancer, And various sarcomas.
- MSC breast analog secreting carcinoma
- NTRK gene fusions in lung cancer About 1 to 3% of NTRK gene fusions in lung cancer; 0.5% to 1% of NTRK gene fusions in colon cancer, breast cancer, thyroid cancer, and melanoma; salivary gland cancer, a juvenile breast-secreting cancer and infant fiber
- the incidence of NTRK gene fusion in sarcoma exceeds 90%. Therefore, inhibitors of the TRK kinase family have utility in the treatment of cancer.
- Anaplastic lymphoma kinase (hereinafter referred to as ALK) is a 200kd receptor tyrosine kinase encoded by the ALK gene on chromosome 2p23.
- ALK belongs to the insulin receptor superfamily. ALK plays an important role in fetal development, but it is not expressed in adult tissues except brain tissue. ALK is considered to be an oncogene. Studies have found that it is abnormally expressed in B-cell lymphoma, inflammatory myofibroblastoma, some non-small cell lung cancer, kidney cancer, colorectal cancer, and neuroblastoma.
- the ALK gene rearrangement allows it to get rid of the control of the silent promoter, accompanied by the transcription and translation activation protein of the fusion gene, and the ALK fusion protein is mainly expressed in the cytoplasm, which transduce mitosis, cell survival and anti-apoptotic signals to the nucleus through intracellular pathways. effect.
- chromosomal translocation causes NPM-ALK gene fusion.
- NPM small cell lung cancer
- ALK abnormal expression tumors respond well to targeted small molecule inhibitors, such as crizotinib (crizotinib) has been approved for clinical treatment of advanced and metastatic ALK-positive non-small cell lung cancer, and in inflammatory myofibroblastoma and The effect is good in anaplastic large cell lymphoma. Therefore, small molecule inhibitors of ALK are beneficial for therapeutic intervention in ALCL, NSCLC, neuroblastoma, and other cancers that rely on ALK for growth and survival.
- crizotinib crizotinib
- ROS1 The c-ROS proto-oncogene 1 (hereinafter referred to as ROS1) was first discovered to be the transformation sequence of the proto-oncogene and the UR2 avian sarcoma virus.
- the ROS1 gene is located on chromosome 6q22.1, with 46 exons and a total length of 127kb.
- the protein encoded by this gene is a type I integral membrane protein with tyrosine kinase activity, belonging to the receptor tyrosine kinase family (RTK). ) Member, the protein contains 2347 amino acids and functions as a growth or differentiation factor receptor.
- ROS1 protein is highest in the kidney, followed by the cerebellum, peripheral nerve tissue, stomach, small intestine and colon, and lower expression in other tissues.
- ROS1 is a new tumor driver mutation gene, and its fusion is designated as a new molecular subtype in non-small cell lung cancer.
- the ROS1 gene rearrangement was first identified in human glioma cell lines, and subsequent ROS1 gene rearrangements were also found in several other malignant tumors, such as cholangiocarcinoma, ovarian cancer, gastric cancer and non-small cells Lung cancer, where the mutation frequency in non-small cell lung cancer is 1%-2%.
- ROS1 fuses with other genes, the extracellular region is lost, the transmembrane region and the intracellular tyrosine kinase region are retained, and the rearrangement sites mainly occur in exons 32 to 36 of the ROS1 gene.
- the ROS1 gene rearrangement produces a fusion protein with a constitutively active kinase domain that activates downstream signaling pathways in cells that lead to oncogenic properties, including uncontrolled proliferation and prolonged tumor cell survival resistance Cell death. These pathways include the Ras-ERK pathway for cell proliferation, as well as the JAK-STAT pathway and PI3K/AKT pathway, which regulate cell survival (anti-apoptosis) and proliferation.
- the ROS1 fusion protein can also activate the mTOR pathway, which is essential for regulating protein translation. Cancers that have these pathways activated tend to be more aggressive, and invasion and metastasis lead to poorer survival rates for patients.
- the ROS1 gene can undergo fusion mutations with multiple genes, and the most important fusion partner is CD74. For example, in NSCLC, the ROS1 gene is mainly fused with SLC34A2 and CD74 and continuously activates the ROS1 tyrosine kinase domain and downstream signaling pathways, which in turn causes tumors.
- Inhibitors of RTK have the potential to cause the death of cancer cells that depend on dysregulation of RTK activity without damaging normal tissues.
- TRK family protein tyrosine kinase inhibitors can be used to treat pain, inflammation, cancer and certain infectious diseases. Therefore, small molecule inhibitors targeting TRK, ALK, and ROS1 have promising therapeutic prospects.
- the present invention relates to a class of small molecule inhibitors targeting TRK, ALK, and ROS1, and specifically relates to a class of compounds represented by formula II or their tautomers, mesosomes, racemates, and enantiomers Isomers, diastereomers, and mixtures thereof, and pharmaceutically acceptable salts, polymorphs, solvates, prodrugs, metabolites, isotope derivatives, and compounds containing said compounds
- the pharmaceutical composition can be used to prevent or treat various types of cancer,
- Y 1 is carbon
- Y 3 is nitrogen
- Y 2 is carbon
- Y 4 is carbon
- Y 5 is carbon
- Y 1 is nitrogen
- Y 2 is nitrogen
- Y 3 is carbon
- Y 4 is carbon
- Y 5 is carbon
- X is carbon or nitrogen
- R 1 is -CF 2 -R 6 ⁇
- R 2 is -NR a R b , -SO 2 -NR a R b , -S(O)-NR a R b , -S(O)-(CH 2 ) 2 -NR a R b , 4-10 yuan Heterocyclic group, -C 4-10 cycloalkyl group, wherein said 4-10 membered heterocyclic group or said -C 4-10 cycloalkyl group includes monocyclic, bicyclic, tricyclic, bridged ring or spiro ring Structure of the compound group;
- the 4-10 membered heterocyclic group or -C 4-10 cycloalkyl group may be mono- or multi-substituted by the following groups: -OH, -C 1-3 alkyl, -C 3-6 cycloalkyl, 4-6 membered heterocyclic group, 5-10 membered heteroaryl group, -OC 1-4 alkyl, -NR c R d , -NH 2 , halogen, -C 1-3 haloalkyl, phenyl or cyano;
- R 2 is preferably
- R a and R b are independently hydrogen, -C 1-3 alkyl, -C 3-6 cycloalkyl, -C 1-4 alkylene-OH, -C 1-3 alkylene-C 3 -9 heterocycloalkyl, -C 1-3 alkylene-C 5-6 oxoheterocycloalkyl , -C 1-3 alkylene-C 4-6 cycloalkyl, -C(O)- (CH 2 ) n -T, wherein the -C 3-9 heterocycloalkyl group includes a compound group with a monocyclic, bicyclic, tricyclic, bridged ring or spiro ring structure, wherein the -C 3-9
- the heterocycloalkyl group may be mono- or poly-substituted by the following groups: -C 1-3 alkyl, halogen, -C 1-3 haloalkyl, -OH, oxo or cyano;
- R a and R b or together with the nitrogen atom to which they are attached form a 4-9 membered heterocycloalkyl which is unsubstituted or substituted with a T group or -NR c R d , and T is -C 1-4 alkyl , -C 2-4 alkyl substituted by -C 1-3 alkyl, -C 3-6 cycloalkyl, 4-6 membered heterocyclic group, -C 1-3 haloalkyl, -(CH 2 ) n -NR c R d ;
- R c and R d are independently hydrogen, -C 1-3 alkyl, -C 1-4 alkylene-OH, -C 2-4 alkylene -OCH 3 , -(CH 2 ) 2 -OC 1-3 alkyl;
- R 3 is -NH-C 4-8 cycloalkyl, -NH-C 3-8 heterocycloalkyl, -NH-CH 2 -C 3-8 heterocycloalkyl or -OC 1-4 alkyl; R 3 is preferably -OC 1-4 alkyl, -NH-CH 2 -C 4-5 heterocycloalkyl; T is -C 1-4 alkyl; T 1 is -O, -S, -NH, -N-CH 3 ;
- R 4 is phenyl, heteroaryl, -NH-phenyl, wherein the phenyl group may be substituted by R 2 and/or R k , and the heteroaryl group may be substituted by R 2 ;
- R k is halogen or -OC 1-4 alkyl;
- R 4 is preferably X 1 is carbon or nitrogen;
- R 6 is a phenyl group or a 4-6 membered heterocycloalkyl group, wherein the phenyl group or a 4-6 membered heterocycloalkyl group is mono- or di-substituted by the following groups: halogen, -OH, -C 1-3 Alkyl, -C(CH 3 ) 2 -OH or cyano; R 6 is preferably
- R 7 is hydrogen, -C 1-3 alkyl, -(CH 2 ) n -OH, C 1-3 haloalkyl, -(CH 2 ) n -OC 1-3 alkyl, -(CH 2 ) n- OC(O)-C 1-3 alkyl; R 7 is preferably hydrogen, -(CH 2 ) n -OH, -(CH 2 ) n -F, -(CH 2 ) n -OC 1-3 alkyl;
- n 1 or 2.
- the present invention provides a method for preparing a compound represented by general formula II, which is prepared by the following scheme:
- the oxidizing agent is preferably Dess-Martin oxidizing agent, PCC, PDC and other oxidizing agents
- the fluorinating agent is preferably BAST, DAST and other fluorinating agents.
- the amide forming method is preferably carboxylic acid reacting with acid chloride to amine to make amide, or carboxylic acid After 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI)/1-hydroxybenzotriazole (HOBT), cyclohexylcarbodiimide (DCC), 2 -(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU) and other condensing agents are activated to form amides with amines;
- EDCI 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride
- HOBT cyclohexy
- the oxidizing agent is preferably Dess-Martin oxidant, PCC, PDC and other oxidants; the method of forming the three-membered ring is preferably diethyl zinc/diiodomethane, trimethyl sulfoxide iodide or trimethyl sulfoxide bromide Salt/sodium hydrogen; the boronating reagent is preferably borane, 9-BBN, etc.; the oxidizing reagent is preferably hydrogen peroxide; the fluorinating reagent is preferably fluorinating reagents such as BAST and DAST; To amide, or carboxylic acid through 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI)/1-hydroxybenzotriazole (HOBT), cyclohexyl carbodiimide Amine (DCC), 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea
- halogenation is actually preferably bromine, iodine, N-bromosuccinimide, N-iodosuccinimide, etc., and the coupling reaction is preferably Stille reaction;
- the nitrating reagent is preferably nitric acid, potassium nitrate, sodium nitrate, etc.
- the reducing reagent is preferably iron powder, zinc powder, stannous chloride, hydrogen/palladium on carbon, hydrogen/nickel, etc.
- the amide formation method is preferably carboxylic acid through acid chloride to amine Reaction to produce amide, or carboxylic acid through 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI)/1-hydroxybenzotriazole (HOBT), cyclohexyl carbon Diimine (DCC), 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU) and other condensing agents are activated to form amides with amines;
- EDCI 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydro
- the pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts thereof.
- Suitable acid addition salts are formed by acids that form non-toxic salts, examples include but are not limited to hydrochloride, sulfate/bisulfate, nitrate, phosphate/hydrogen phosphate/dihydrogen phosphate, hydrobromic acid Salt, hydroiodide, acetate, lactate, methanesulfonate, citrate, malate, maleate, fumarate, tartrate, salicylate, stearate , And its similar salt.
- Suitable base addition salts are formed from bases that form non-toxic salts, for example as alkali metal salts, alkaline earth metal salts, or as ammonium salts. Examples include, but are not limited to, sodium, potassium, calcium, magnesium, or with ammonia Or organic amines such as ethylamine, ethanolamine, triethanolamine or salts of amino acids.
- the present invention also includes internal salts or betaines (zwitterions) in addition to the salt forms mentioned.
- the compound of formula (II) may exist in crystalline or amorphous form.
- certain crystal forms of the compound of formula (II) may exist in polymorphic forms, which are included in the scope of the present invention.
- Many conventional analysis techniques can be used including but not limited to single crystal X-ray powder diffraction (XRPD) patterns, infrared (IR) spectroscopy, Raman spectroscopy, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA) and solids Nuclear magnetic resonance (ssNMR) characterization to distinguish the polymorphic forms of the compound.
- the compounds of the present invention also include tautomeric forms.
- the tautomeric form is produced by the exchange of a single bond with an adjacent double bond and the accompanying migration of protons.
- Tautomeric forms include proton transfer tautomers, which are isomeric protonated states with the same empirical formula and total charge. Examples of proton transfer tautomers include keto-enol pairings, amide-imine pairings, lactam-lactam pairings, enamine-imine pairings, and two in which protons can occupy the heterocyclic ring system.
- Tautomeric forms can be in equilibrium or sterically fixed to one form by appropriate substitution.
- the present invention further relates to the compound represented by the general formula (II) or its tautomer, meso, racemate, enantiomer, diastereomer, and mixtures thereof, and
- the pharmaceutically acceptable salt or the pharmaceutical composition containing the same is further used in combination with another one or more agents for regulating the immune system of mammals, anticancer agents or anti-inflammatory agents.
- the present invention further relates to the compound represented by the general formula (II) or its tautomer, meso, racemate, enantiomer, diastereomer, and mixtures thereof, and Its pharmaceutically acceptable salt or pharmaceutical composition containing it is used to treat or prevent the following cancers, including solid tumors (such as prostate cancer, kidney cancer, liver cancer, pancreatic cancer, stomach cancer, breast cancer, lung cancer, head and neck cancer, Thyroid cancer, glioblastoma, melanoma, etc.), blood cancer (such as lymphoma, leukemia, etc.), skin cancer (such as skin T-cell lymphoma, skin B-cell lymphoma), etc.
- solid tumors such as prostate cancer, kidney cancer, liver cancer, pancreatic cancer, stomach cancer, breast cancer, lung cancer, head and neck cancer, Thyroid cancer, glioblastoma, melanoma, etc.
- blood cancer such as lymphoma, leukemia, etc.
- skin cancer such as skin T-cell lymph
- the present invention also relates to a method for treating or preventing cancer, which comprises administering to a patient a therapeutically effective amount of a compound represented by general formula (II) or its tautomers, mesosomes, racemates, Enantiomers, diastereomers, and mixtures thereof, and pharmaceutically acceptable salts thereof, or pharmaceutical compositions containing them, wherein the cancer includes, for example, solid tumors (such as prostate cancer, renal cancer) , Liver cancer, pancreatic cancer, stomach cancer, breast cancer, lung cancer, head and neck cancer, thyroid cancer, glioblastoma, melanoma, etc.), blood cancer (such as lymphoma, leukemia, etc.), skin cancer (such as skin T- Cell lymphoma, skin B-cell lymphoma) and so on.
- solid tumors such as prostate cancer, renal cancer
- Liver cancer pancreatic cancer
- stomach cancer breast cancer
- lung cancer head and neck cancer
- thyroid cancer glioblastoma, melanoma,
- composition of the present invention can be formulated in a conventional manner using one or more pharmaceutically acceptable carriers. Therefore, the active compound of the present invention can be formulated into a dosage form for oral, buccal administration, intranasal, parenteral (for example, intravenous, intramuscular or subcutaneous) or rectal administration, or suitable for administration by inhalation or insufflation.
- the dosage form of the medicine can also be formulated into sustained release dosage forms.
- the pharmaceutical composition can be made into various types of administration unit dosage forms, such as tablets, pills, powders, liquids, suspensions, emulsions, granules, capsules, suppositories, and injections (solutions and suspensions). Or it is suitable for a dosage form for administration by inhalation or insufflation.
- any excipient known and widely used in the art can be used.
- carriers such as lactose, white sugar, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose and silicic acid, etc.
- binders such as water, ethanol, propanol, ordinary syrup, glucose solution, starch Solution, gelatin solution, carboxymethyl cellulose, shellac, methyl cellulose and potassium phosphate, polyvinylpyrrolidone, etc.
- disintegrating agent such as dry starch, sodium alginate, agar powder and kelp powder, sodium bicarbonate, carbonic acid Fatty acid esters of calcium, polyethylene sorbitan, sodium lauryl sulfate, monoglyceride stearate, starch and lactose, etc.
- disintegration inhibitors such as white sugar, glyceryl tristearate, coconut oil and hydrogenated Oil
- Adsorption promoters such as quaternary sulfate, sodium lauryl
- any known and widely used excipients in the art can be used, for example, carriers such as lactose, starch, coconut oil, hardened vegetable oil, kaolin and talc, etc.; binders , Such as gum arabic powder, gum tragacanth powder, gelatin and ethanol; disintegrants such as agar and kelp powder.
- carriers such as lactose, starch, coconut oil, hardened vegetable oil, kaolin and talc, etc.
- binders Such as gum arabic powder, gum tragacanth powder, gelatin and ethanol
- disintegrants such as agar and kelp powder.
- any excipient known and widely used in the art can be used, for example, polyethylene glycol, coconut oil, higher alcohols, esters of higher alcohols, gelatin and semi-synthetic glycerides Wait.
- the solution or suspension can be sterilized (preferably by adding an appropriate amount of sodium chloride, glucose or glycerol, etc.) to prepare an injection that is isotonic with blood.
- any carriers commonly used in the art can also be used.
- usual dissolving agents, buffering agents and analgesics can also be added.
- the active compound of the present invention is suitably released in the form of a solution or suspension from a pump spray container received or squeezed or sucked, or in the form of a spray. Release from a pressurized container or aerosol, and use a suitable propellant, such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gases.
- a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gases.
- the dosage unit can be determined by a valve that provides a metered release.
- the pressurized container or nebulizer may contain a solution or suspension of the active compound.
- Capsules or cartridges (for example made of gelatin) used in inhalers or insufflators can be formulated as a powder mixture containing the compound of the invention and a suitable powder
- stereoisomer means an isomer formed due to at least one asymmetric center. In compounds with one or more (for example, one, two, three or four) asymmetric centers, it can produce racemates, racemic mixtures, mesosomes, single enantiomers , Diastereomeric mixtures and individual diastereomers. Certain individual molecules can also exist as geometric isomers (cis/trans).
- solvate refers to a form of a compound that is generally physically combined with a solvent through a solvolysis reaction. This physical bond includes hydrogen bonding.
- solvents include water, ethanol, methanol, acetic acid and the like.
- the compound of formula (II) can be prepared in crystalline form and can be in the form of a solvate (e.g., a hydrated form).
- Suitable solvates include pharmaceutically acceptable solvates (such as hydrates), and further include stoichiometric solvates and non-stoichiometric solvates. In some cases, such as when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid, the solvate will be able to dissociate.
- “Solvate” encompasses both solution-phase and dissociable solvates. Representative solvates include hydrates, ethanolates, methanolates, and the like.
- prodrug refers to a derivative that is converted into a compound of the present invention by reaction with enzymes, gastric acid, etc. in the living body under physiological conditions, for example, through oxidation, reduction, hydrolysis, etc., respectively, catalyzed by enzymes.
- metabolite refers to all molecules derived from any compound of the present invention in a cell or organism, preferably a human.
- isotopic derivative refers to a compound that contains isotopes in unnatural proportions at one or more of the atoms constituting the compound.
- deuterium 2 H or D
- tritium 3 H or T
- carbon-13 13 C
- nitrogen-15 15 N
- oxygen-18 18 O
- pharmaceutical composition means a mixture containing one or more of the compounds of the present invention or their physiologically/pharmaceutically acceptable salts or prodrugs and other chemical components, such as physiologically/pharmaceutically acceptable Carriers and excipients.
- the purpose of the pharmaceutical composition is to promote the administration to the organism, which is beneficial to the absorption of the active ingredient and thus the biological activity.
- the carrier includes all pharmaceutical preparations in the pharmaceutical field that can be used for injection and non-injection administration routes, such as diluents, wetting agents, fillers, adhesives, slip agents, disintegrants, absorption enhancers, Surfactants, retarders, adsorbents, suspending agents, flocculants, deflocculants, emulsifiers, commonly used bases, solubilizers, co-solvents, latent solvents, preservatives, flavoring agents, coloring agents, antioxidants , Buffers, bacteriostatic agents, isotonic regulators, pH regulators, metal ion complexing agents, hardeners, thickeners, etc.
- diluents wetting agents, fillers, adhesives, slip agents, disintegrants, absorption enhancers, Surfactants, retarders, adsorbents, suspending agents, flocculants, deflocculants, emulsifiers, commonly used bases, solubilizers, co-solvents, latent
- the novel RTK inhibitor provided in the present application has the potential to cause the death of cancer cells that depend on the imbalance of RTK activity without damaging normal tissues at the same time.
- the TRK family protein tyrosine kinase inhibitors provided in this application can be used to treat pain, inflammation, cancer and certain infectious diseases.
- the inventors unexpectedly discovered that the RTK inhibitor provided in the present application can effectively inhibit the activities of tropomyosin receptor kinase A (TRKA), anaplastic lymphoma kinase (ALK), and proto-oncogene tyrosine kinase (ROS1).
- TRKA tropomyosin receptor kinase A
- ALK anaplastic lymphoma kinase
- ROS1 proto-oncogene tyrosine kinase
- Tumor cells, cancer cell proliferation and tumor growth have strong inhibitory effects, and the inhibitory activity is better than the prior art Entrectinib (Entrectinib).
- the first step is the preparation of 4-fluoro-2-nitro-benzoic acid tert-butyl ester I-1b
- the second step is the preparation of tert-butyl 4-(4-methylpiperazin-1-yl)-2-nitrobenzoate I-1c
- the third step is the preparation of tert-butyl 4-(4-methylpiperazin-1-yl)-2-aminobenzoate I-1d
- the fourth step is the preparation of tert-butyl 4-(4-methylpiperazin-1-yl)-2-(tetrahydropyran-4-yl)aminobenzoate I-1e
- the sixth step 4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydropyran-4-yl)acetamido)benzoic acid 1- 1 preparation
- the first step is the preparation of (3-cyano-4-fluorophenyl)-(3,5-difluorophenyl)methanol I-2c
- the second step is the preparation of (3-cyano-4-fluorophenyl)-(3,5-difluorophenyl)methane I-2d
- the first step is the preparation of 5-chloro-3-nitropyrazolo[1,5-a]pyrimidine 1b
- 5-Chloropyrazolo[1,5-a]pyrimidine 1a (2g, 13mmol) was dissolved in H 2 SO 4 (8mL), KNO 3 (2g, 19.6mmol) was added under ice bath, and the temperature was slowly heated to Room temperature. The mixture was slowly poured into ice water, a yellow solid precipitated out, filtered, the solid was collected and dried to obtain 5-chloro-3-nitropyrazolo[1,5-a]pyrimidine 1b (2g, 10.1mmol), the yield was 77 %.
- the second step is the preparation of 5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)-3-nitropyrazolo[1,5-a]pyrimidine 1d
- the third step is the preparation of 5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-amine 1e
- the first step Preparation of tert-butyl-3-[bis(tert-butoxycarbonyl)amino]-5-bromo-1H-indazole-1-carboxylate 3b
- the third step is the preparation of 5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)-1H-indazol-3-amine 3d
- the first step is the preparation of 5-bromo-1-triphenyl-1H-indazole 4b
- the second step is the preparation of (3,5-difluorophenyl)(1-triphenyl-1H-indazol-5-yl)methanol 4c
- reaction solution was quenched with saturated ammonium chloride solution and extracted with dichloromethane (100ml*2).
- the organic phases were combined, dried with anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography to obtain the title compound 4c (6.5 g, 12.9 mmol) with a yield of 43%.
- the third step is the preparation of (3,5-difluorophenyl)-(1H-indazol-5-yl)methanol 4d
- Step 9 (5-(3,5-Difluorophenyl)-3-(2'-(4-methylpiperazin-1-yl)-[2,4'-bipyridyl]-6-yl) Preparation of -1H-indazol-1-yl)methanol 4j
- the first step is the preparation of 1-(3-bromo-4-fluorophenyl)-1-(3,5-difluorophenyl)ethyl-1-ol 6c
- the second step is the preparation of 2-bromo-4-(1-(3,5-difluorophenyl)vinyl)-1-fluorobenzene 6d
- the third step is the preparation of 2-bromo-4-(1-(3,5-difluorophenyl)cyclopropyl)-1-fluorobenzene 6e
- the fourth step is the preparation of 5-(1-(3,5-difluorophenyl)cyclopropyl)-2-fluorobenzonitrile 6f
- the fifth step is the preparation of 5-(1-(3,5-difluorophenyl)cyclopropyl)-1H-indazole-3-amine 6g
- the first step is the preparation of 5-(3,5-difluorobenzoyl)-2-fluorobenzonitrile 7a
- the second step is the preparation of 5-((3,5-difluorophenyl)difluoromethyl)-2-fluorobenzonitrile 7b
- the third step is the preparation of 5-((3,5-difluorophenyl)difluoromethyl)-1H-indazol-3-amine 7c
- the first step is the preparation of ethyl 4-fluoro-2-nitrobenzoate 8b
- the second step is the preparation of ethyl 4-((3-((2-ethylhexyl)oxo)-3-carbonylpropyl)thio)-2-nitrobenzoate 8d
- the fourth step is the preparation of compound ethyl 4-(chlorosulfonyl)-2-nitrobenzoate 8f
- the seventh step is the preparation of ethyl 4-((4-methylpiperazin-1-yl)sulfonyl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzoate 8i
- the eighth step is the preparation of 4-((4-methylpiperazin-1-yl)sulfonyl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzoic acid 8j
- Step 9 4-((4-Methylpiperazin-1-yl)sulfonyl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetyl (Amino) benzoic acid 8k preparation
- reaction solution was allowed to warm up naturally, and LCMS showed that the main peak was the product point.
- the second step is the preparation of 5-(3,5-difluorobenzyl)pyrazolo[1,5-a]pyrimidine 9d
- the third step is the preparation of 5-(3,5-difluorobenzyl)-3-iodopyrazolo[1,5-a]pyrimidine 9e
- the first step is the preparation of (2-(3-bromo-4-fluorophenyl)-2-(3,5-difluorophenyl)ethyl)boronic acid 10a
- reaction solution was stirred at room temperature for 2 hours, and then the reaction solution Ice water (150mL) was added, the reaction solution was stirred at room temperature for 3 hours, the reaction solution was concentrated and then extracted with ethyl acetate (200mL), the organic phase was washed with saturated sodium bicarbonate solution and brine, dried and spin-dried to obtain After purification by column chromatography, the crude product of compound 10a (10g) MS m/z(ESI): 383[M+23] + was obtained .
- the second step is the preparation of (2-(3-bromo-4-fluorophenyl)-2-(3,5-difluorophenyl)ethyl-1-ol 10b
- the third step is the preparation of 4-(2-(benzyloxy)-1-(3,5-difluorophenyl)ethyl)-2-bromo-1-fluorobenzene 10c
- the fourth step is the preparation of 5-(2-(benzyloxy)-1-(3,5-difluorophenyl)ethyl)-2-fluorobenzonitrile 10d
- Step 6 N-(5-(2-(Benzyloxy)-1-(3,5-difluorophenyl)ethyl)-1H-indazol-3-yl)-4-(4- Preparation of methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 10f
- Step 8 N-(5-(1-(3,5-Difluorophenyl)-2-hydroxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazine-1 -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 10
- the first step is the preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-fluoro-2-nitrobenzamide 11b
- the first step is the synthesis of 5-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-c]pyridine 17b
- the third step is the synthesis of 5-(3,5-difluorobenzyl)-1H-pyrazolo[3,4-c]pyridine 17e
- Dissolve compound 17d (400mg, 1.1mmol, 1.0eq) in 1,4-dioxane (30ml), add 4N HCl (20ml, 20.0mmol, 19.0eq), react at 70°C for 5h, and evaporate to dryness under reduced pressure. 17e, unpurified, used directly in the next step.
- the first step is the preparation of 2-fluoro-5-hydroxybenzonitrile 19a
- the second step is the preparation of 2-fluoro-5-bromomethylbenzonitrile 19b
- Dissolve 19a (700mg, 4.6mmol) in dichloromethane (20mL), add phosphorus tribromide (800mg, 2.96mmol) under ice bath, stir for 2 hours at room temperature and then ice bath, add water (20mL), continue stirring for 30 After 5 minutes, let stand for layering, add anhydrous Na2SO4 to the organic phase, dry, filter, and concentrate to obtain the crude title compound 19b and use it directly in the next step.
- the third step is the preparation of 2-fluoro-5-((4-(2-hydroxypropane-2-yl)piperidin-1-yl)methyl)benzonitrile 19d
- the fourth step is the preparation of 2-(1-((3-amino-1-hydroindazol-5-yl)methyl)piperidin-4-yl)-2-propanol 19e
- the first step is the preparation of 5-(1-(3,5-difluorophenyl)-2-hydroxyethyl)-2-fluorobenzeneacetonitrile 21a
- the second step is the preparation of 5-(1-(3,5-difluorophenyl)-2-fluoroethyl)-2-fluorobenzeneacetonitrile 21b
- the third step is the preparation of 5-(1-(3,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-amine 21c
- the first step is the preparation of 1-(3-methoxy-4-nitrophenyl)-4-methylpiperazine 22b
- the fourth step is the preparation of tert-butyl 5-bromo-1H-indazole-3-carboxylate 22f
- the seventh step is the preparation of tert-butyl 5-(3,5-difluorophenyl)-1-trityl-1H-indazole-3-carboxylate 22i
- Step 8 Preparation of tert-butyl 5-(3,5-difluorophenyl)-1-trityl-1H-indazole-3-carboxylate 22j
- Trifluoroacetic acid (2mL) was added to the dichloromethane (8mL) solution of compound 22i (200mg, 0.34mmol). The reaction solution was stirred at room temperature for 2 hours. The reaction solution was concentrated and spin-dried to obtain compound 22j (60mg, 0.20mmol). The rate is 61%.
- the first step is the preparation of 2-bromo-4-(1-(3,5-difluorophenyl)-2-methoxyethyl)-1-fluorobenzene 23a
- the second step is the preparation of 5-(1-(3,5-difluorophenyl)-2-methoxyethyl)-2-fluorobenzonitrile 23b
- the third step is the preparation of 5-(1-(3,5-difluorophenyl)-2-methoxyethyl)-1H-indazol-3-amine 23c
- the first step 1- (6-bromo-1,2,3,4-tetrahydroisoquinolin-2-yl)-2,2,2-trifluoroethane-1-one 24b
- the third step is the preparation of 7-nitro-2-(2,2,2-trifluoroacetyl)-1,2,3,4-tetrahydroisoquinoline-6-carbonitrile 24d
- the fifth step is the preparation of 2-methyl-7-nitro-1,2,3,4-tetrahydroisoquinoline-6-carbonitrile 24f
- Step 8 7-Amino-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-2-methyl-1,2,3,4-tetrahydroiso Preparation of quinoline-6-carboxamide 24i
- the second step is the preparation of benzyl (1s, 3R, 5S, 7s)-4-(3-bromo-4-(ethoxycarbonyl)phenyl)-4-hydroxyadamantane-1-carboxylate 25d
- the third step is the preparation of benzyl (1s, 3R, 5S, 7s)-4-(3-bromo-4-(ethoxycarbonyl)phenyl)adamantane-1-carboxylate 25e
- Step 6 Ethyl 4-((1R,3S,5s,7s)-5-(((benzyloxy)carbonyl)amino)adamantan-2-yl)-2-((tert-butoxycarbonyl) Preparation of amino) benzoate 25h
- the seventh step is the preparation of ethyl 2-amino-4-((1R,3S,5s,7s)-5-(((benzyloxy)carbonyl)amino)adamantan-2-yl)benzoate 25i
- Step 8 Ethyl 4-((1R,3S,5s,7s)-5-(((benzyloxy)carbonyl)amino)adamantan-2-yl)-2-((tetrahydro-2H-pyran) Preparation of -4-yl)amino)benzoate 25j
- Step 9 4-((1R,3S,5s,7s)-5-(((benzyloxy)carbonyl)amino)adamantan-2-yl)-2-((tetrahydro-2H-pyran-4 -(Yl)amino)benzoic acid 25k preparation
- Step 10 4-((1R,3S,5s,7s)-5-(((benzyloxy)carbonyl)amino)adamantan-2-yl)-2-(2,2,2-trifluoro-N -(Tetrahydro-2H-pyran-4-yl)acetamido)benzoic acid 25l
- Step 12 Benzyl ((1s,3R,5S,7s)-4-(4-((5-(3,5-difluorobenzyl)-1H-indazol-3-yl)carboxamide) Preparation of -3-((tetrahydro-2H-pyran-4-yl)amine)phenyl)adamantan-1-yl)carbamate 25n
- the first step is the preparation of 4-((5-benzyloxycarbonylamino)adamantane)-2-yl)-2-((N-benzyloxycarbonylpiperidin-4-yl)amino)-benzoic acid ethyl ester 26a
- 26a (900 mg, 1.35 mmol) was dissolved in 15 mL of ethanol, and 8 mL of 4M sodium hydroxide aqueous solution was added. The system was refluxed for 16 hours. LCMS monitors the reaction. After the reaction was completed, the solvent was rotated to remove most of the ethanol, and after adding 10 mL of water, the pH was adjusted to 4, and a solid was precipitated. The solid was filtered out and washed with water to obtain target compound 26b (780 mg) as a grayish solid with a yield of 91%. It was used directly in the next step without further purification.
- Step 4 4-((5-Benzyloxycarbonylamino)adamantane)-2-yl)-2-(N-(1-benzyloxycarbonylpiperidin-4-yl)-2,2,2-trifluoro Preparation of acetamido)-benzoic acid 26c
- 26b (780mg, 1.22mmol) was dissolved in 20mL of dichloromethane, and trifluoroacetic anhydride (1.2eq) and triethylamine (1.5eq) were added at 0°C. After the addition, stir at room temperature for three hours. Rotate the solvent to dryness, and the crude product obtained is 26c. It was not purified and used directly in the next step.
- the first step 5-(2-(benzyloxy)-1-(3,5-difluorophenyl)ethyl)-3-(4-(4-methylpiperazin-1-yl)-2- ((Tetrahydro-2H-pyran-4)tert-butyl)amino)benzamido)-1H-indazole-1-carboxylic acid tert-butyl ester 29a
- the third step 5-(2-acetoxy-1-(3,5-difluorophenyl)ethyl)-3-(4-(4-methylpiperazin-1-yl)-2-(( Tetrahydro-2H-pyran-4-yl)tert-butyl)amino)benzamido)-1H-indazole-1-carboxylate 29c
- reaction solution was added with water and extracted three times with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography to obtain the title compound 35c (3.4 g, 14.847 mmol).
- the fifth step 5-(1-(2,5-difluorophenyl)vinyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c] Pyridine 35f
- reaction solution was extracted with water and ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography to obtain the title compound 35f (1 g, 2.932 mmol).
- the first step is the synthesis of tert-butyl 4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-nitrobenzoate 43a
- the second step is the synthesis of tert-butyl 2-amino-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)benzoate 43b
- Tetramethyltriacetoxyammonium borohydride (1130mg, 4.32mmol) was slowly added to a solution of compound 43b (360mg, 1.08mmol) in DCM (10mL) and trifluoroacetic acid (1mL), and the mixture was stirred at room temperature for 2 hours .
- the reaction was quenched by adding saturated aqueous NaHCO 3 solution.
- the fifth step 4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4- (4) Acetylamino) benzoic acid 43e synthesis
- Step 8 N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino )-2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide 43
- compound 50 can be obtained.
- the first step Dimethyl 2-((tetrahydro-2H-pyran-4-yl)amino)terephthalate 60b
- the second step Dimethyl 2-bromo-5-((tetrahydro-2H-pyran-4-yl)amino)terephthalate 60c
- the third step Dimethyl 2-cyano-5-((tetrahydro-2H-pyran-4-yl)amino)terephthalate 60d
- Step 7 2-(2-(Dimethylamino)ethyl)-1-carbonyl-6-((tetrahydro-2H-pyran-4-yl)amino)isoindoline-5-carboxylic acid 60h
- Step 8 2-(2-(Dimethylamino)ethyl)-1-carbonyl-6-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido )Isoindole-5-carboxylic acid 60i
- Step 9 N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-2-(2-(dimethylamino)ethyl)-1-carbonyl-6- (2,2,2-Trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)isoindole-5-carboxamide 60j
- Step 10 N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-2-(2-(dimethylamino)ethyl)-1-carbonyl-6- ((Tetrahydro-2H-pyran-4-yl)amino)isoindole-5-carboxamide 60
Abstract
Disclosed is a new class of small molecule compounds having multiple kinase inhibitory activities, which compounds have the structure as shown in general formula II; and a compound as shown in general formula II has good efficacy and safety in the prevention or treatment of multiple cancer indications such lung cancer and lymphoma.
Description
本申请要求于2020年04月02日提交中国国家知识产权局、申请号为202010252669.5发明名称为“多靶点的的抗肿瘤化合物及其制备方法和应用”的中国专利申请的优先权，其全部内容通过引用结合在本申请中。This application claims the priority of the Chinese patent application filed with the State Intellectual Property Office of China with the application number 202010252669.5 and the invention titled "Multi-targeted antitumor compound and its preparation method and application" on April 02, 2020, all of which The content is incorporated in this application by reference.
本发明属于医药化学领域，具体涉及一类新的具有多重激酶抑制活性的小分子化合物及其制备方法，以及这类化合物在防治肿瘤方面的用途。The invention belongs to the field of medicinal chemistry, and specifically relates to a new type of small molecule compound with multiple kinase inhibitory activity and a preparation method thereof, as well as the use of this type of compound in preventing and treating tumors.
基因融合是很多恶性肿瘤的驱动因素。结构上，他们与上游基因融合并保持完整的酪氨酸激酶结构域，不依赖配体二聚化，导致下游信号持续活化促进肿瘤细胞生长及增殖。随着二代测序、RT-PCR等技术的不断成熟完善，很多与临床相关的融合基因被检测出来。包括NTRK1、NTRK2、NTRK3(编码TRKA、TRKB及TRKC蛋白)、ALK、ROS1等。Gene fusion is a driving factor for many malignant tumors. Structurally, they fuse with upstream genes and maintain a complete tyrosine kinase domain, independent of ligand dimerization, leading to continuous activation of downstream signals to promote tumor cell growth and proliferation. With the continuous maturity and improvement of technologies such as second-generation sequencing and RT-PCR, many clinically relevant fusion genes have been detected. Including NTRK1, NTRK2, NTRK3 (encoding TRKA, TRKB and TRKC protein), ALK, ROS1, etc.
原肌球蛋白受体激酶(Tropomyosin receptor kinase，以下简称为TRK)是一类神经生长因子受体，包含NTRK1、NTRK2和NTRK3基因编码的TRKA、TRKB和TRKC三种受体，在神经元组织中广泛表达并且参与神经元细胞的维持、信号传导和存活。TRK是由细胞外配体结合域，跨膜和细胞内ATP结合域组成的跨膜受体。TRKA，TRKB和TRKC的细胞外结构域表现出高度的结构相似性，包括两个免疫球蛋白样基序(Ig1和Ig2)，它被认为是包含配体结合位点的基序，三个富含亮氨酸的24-残基基序(LRR1-3基序)，LRR1-3基序两侧有两个半胱氨酸簇(C1和C2)是TRK蛋白特异性的，在受体酪氨酸激酶的其他亚家族中没有发现。Tropomyosin receptor kinase (Tropomyosin receptor kinase, hereinafter referred to as TRK) is a type of nerve growth factor receptor, including three receptors of TRKA, TRKB and TRKC encoded by NTRK1, NTRK2 and NTRK3 genes, which are found in neuronal tissues It is widely expressed and participates in the maintenance, signal transduction and survival of neuronal cells. TRK is a transmembrane receptor composed of extracellular ligand binding domain, transmembrane and intracellular ATP binding domain. The extracellular domains of TRKA, TRKB and TRKC show a high degree of structural similarity, including two immunoglobulin-like motifs (Ig1 and Ig2), which are considered to contain ligand binding sites, and three rich The 24-residue motif containing leucine (LRR1-3 motif), two cysteine clusters (C1 and C2) flanking the LRR1-3 motif are specific to TRK protein, Not found in other subfamily of amino acid kinases.
所述TRKA为神经生长因子(以下简称为NGF)的高亲和性受体，所述TRKB 为脑源性神经营养因子(BDNF)和神经营养因子(以下简称为NT)-4/5的高亲和性受体，且所述TRKC为NT-3的高亲和性受体。已知NGF对周围神经中的TRKA的活化引发痛觉过敏，包括骨关节炎、慢性腰背痛、风湿性关节炎、骨折、间质性膀胱炎和慢性胰腺炎的伤害性疼痛、神经性疼痛以及伴有两种上述疼痛类型的癌症疼痛中。同时，文献表明BDNF/TrkB途径的激活也牵涉于各种类型的疼痛，所述疼痛包括炎症性疼痛、神经性疼痛和手术疼痛，所以TRKA激酶和其它TRK激酶的抑制剂可以为慢性疼痛状态提供有效的治疗。The TRKA is a high-affinity receptor for nerve growth factor (hereinafter referred to as NGF), and the TRKB is a high-affinity receptor for brain-derived neurotrophic factor (BDNF) and neurotrophic factor (hereinafter referred to as NT)-4/5. Affinity receptor, and the TRKC is a high affinity receptor for NT-3. It is known that the activation of TRKA in peripheral nerves by NGF causes hyperalgesia, including nociceptive pain, neuropathic pain, and osteoarthritis, chronic low back pain, rheumatoid arthritis, fractures, interstitial cystitis, and chronic pancreatitis. In cancer pain accompanied by two of the above-mentioned pain types. At the same time, the literature shows that the activation of the BDNF/TrkB pathway is also involved in various types of pain, including inflammatory pain, neuropathic pain and surgical pain, so TRKA kinase and other TRK kinase inhibitors can provide chronic pain states. Effective treatment.
此外，TRK信号通路的改变，包括基因融合、蛋白过度表达或单核苷酸改变，已经被发现与许多癌症相关。所述癌症包括神经母细胞瘤、卵巢癌、乳腺癌、***癌、胰腺癌、多发性骨髓瘤、星形细胞瘤和髓母细胞瘤、神经胶质瘤、黑素瘤、甲状腺癌、肺腺癌、大细胞神经内分泌肿瘤以及结肠直肠癌。其中NTRK基因的融合是目前最明确的致癌原因。TRK融合蛋白可不依赖配体二聚化、磷酸化，通过激活PI3K、RAS/MAPK/ERK和PLC-γ通路并引发永久性的信号级联反应，驱动TRK融合肿瘤的扩散和生长。所述NTRK基因融合可以发生在身体的任何部位，因此TRK融合癌症可能出现在多种成人和儿童实体瘤中，包括乳腺类似物分泌癌(MASC)、结肠癌、肺癌，胰腺癌、甲状腺癌、以及各种肉瘤。肺癌约有1～3％的NTRK基因融合；结肠癌，乳腺癌和甲状腺癌以及黑色素瘤中约有0.5％至1％的NTRK基因融合；唾液腺癌，一种青少年乳腺分泌性癌和婴幼儿纤维肉瘤中的NTRK基因融合发生率超过90％。因此，TRK激酶家族的抑制剂在治疗癌症中具有效用。In addition, changes in the TRK signaling pathway, including gene fusion, protein overexpression, or single nucleotide changes, have been found to be associated with many cancers. The cancer includes neuroblastoma, ovarian cancer, breast cancer, prostate cancer, pancreatic cancer, multiple myeloma, astrocytoma and medulloblastoma, glioma, melanoma, thyroid cancer, lung gland Cancer, large cell neuroendocrine tumors, and colorectal cancer. Among them, NTRK gene fusion is the most clear cause of cancer. TRK fusion protein can drive the spread and growth of TRK fusion tumors by activating PI3K, RAS/MAPK/ERK and PLC-γ pathways and triggering a permanent signal cascade reaction independently of ligand dimerization and phosphorylation. The NTRK gene fusion can occur in any part of the body, so TRK fusion cancer may appear in a variety of adult and child solid tumors, including breast analog secreting carcinoma (MASC), colon cancer, lung cancer, pancreatic cancer, thyroid cancer, And various sarcomas. About 1 to 3% of NTRK gene fusions in lung cancer; 0.5% to 1% of NTRK gene fusions in colon cancer, breast cancer, thyroid cancer, and melanoma; salivary gland cancer, a juvenile breast-secreting cancer and infant fiber The incidence of NTRK gene fusion in sarcoma exceeds 90%. Therefore, inhibitors of the TRK kinase family have utility in the treatment of cancer.
间变性淋巴瘤激酶(Anaplasticlymphomakinase，以下简称为ALK)是由染色体2p23上的ALK基因编码的200kd的受体酪氨酸激酶。ALK属于胰岛素受体超家族。ALK在胎儿发育中起到重要作用，但在成人除脑组织外其他组织中不表达。ALK被认为是一种致癌基因，研究发现在B细胞淋巴瘤、炎性肌纤维母细胞瘤、部分非小细胞肺癌、肾癌、结直肠癌、神经母细胞癌中异常表达。ALK基因重排可让其摆脱沉默启动子的控制，伴随融合基因转录翻译活化蛋白，并且ALK融合蛋白主要表达于细胞质，通过细胞内途径转导有丝***、细胞存活和抗凋亡信号到核内发挥作用。例如，在间变性大细胞淋巴瘤(ALCL)中，染色体易位引起NPM-ALK基因融合，当与ALK融合时，NPM的二聚化结构域引起ALK的 组成性二聚和激活，同时ALK募集若干接头蛋白，并刺激已知介导肿瘤细胞生长和存活的多个信号通路，包括STAT3、PLC-γ、RAS-ERK1,2和PI3K-AKT，发挥其致癌活性；同样在约5％的非小细胞肺癌(NSCLC)中检测到ALK易位，与ALK在ALCL中的易位相似，NSCLC中的融合蛋白显示出组成性ALK活性并推动肿瘤生长和存活。除了染色体重排之外，在偶发性和家族性神经母细胞瘤的亚群中已报道了激活点突变和扩增，这进一步扩大了依赖于ALK活性的肿瘤谱。ALK异常表达肿瘤对于靶向小分子抑制剂反应良好，如克唑替尼(crizotinib)已被批准用于临床治疗晚期和转移性ALK阳性非小细胞型肺癌，并且在炎性肌纤维母细胞瘤和间变性大细胞淋巴瘤中效果良好。因此，ALK的小分子抑制剂有益于ALCL、NSCLC、神经母细胞瘤和依赖于ALK进行生长和存活的其它癌症中的治疗干预。Anaplastic lymphoma kinase (hereinafter referred to as ALK) is a 200kd receptor tyrosine kinase encoded by the ALK gene on chromosome 2p23. ALK belongs to the insulin receptor superfamily. ALK plays an important role in fetal development, but it is not expressed in adult tissues except brain tissue. ALK is considered to be an oncogene. Studies have found that it is abnormally expressed in B-cell lymphoma, inflammatory myofibroblastoma, some non-small cell lung cancer, kidney cancer, colorectal cancer, and neuroblastoma. The ALK gene rearrangement allows it to get rid of the control of the silent promoter, accompanied by the transcription and translation activation protein of the fusion gene, and the ALK fusion protein is mainly expressed in the cytoplasm, which transduce mitosis, cell survival and anti-apoptotic signals to the nucleus through intracellular pathways. effect. For example, in anaplastic large cell lymphoma (ALCL), chromosomal translocation causes NPM-ALK gene fusion. When fused with ALK, the dimerization domain of NPM causes the constitutive dimerization and activation of ALK, and ALK recruits Several adaptor proteins, and stimulate multiple signaling pathways known to mediate tumor cell growth and survival, including STAT3, PLC-γ, RAS-ERK1,2 and PI3K-AKT, exert their carcinogenic activity; similarly in about 5% of non- ALK translocation is detected in small cell lung cancer (NSCLC). Similar to the translocation of ALK in ALCL, the fusion protein in NSCLC shows constitutive ALK activity and promotes tumor growth and survival. In addition to chromosomal rearrangements, activating point mutations and amplifications have been reported in subpopulations of incidental and familial neuroblastoma, which further expands the spectrum of tumors that depend on ALK activity. ALK abnormal expression tumors respond well to targeted small molecule inhibitors, such as crizotinib (crizotinib) has been approved for clinical treatment of advanced and metastatic ALK-positive non-small cell lung cancer, and in inflammatory myofibroblastoma and The effect is good in anaplastic large cell lymphoma. Therefore, small molecule inhibitors of ALK are beneficial for therapeutic intervention in ALCL, NSCLC, neuroblastoma, and other cancers that rely on ALK for growth and survival.
c-ROS原癌基因1(以下简称为ROS1)最初被发现是原癌基因和UR2禽类肉瘤病毒的转化序列。ROS1基因位于染色体6q22.1，共46个外显子，全长127kb，该基因编码的蛋白是一种具有酪氨酸激酶活性的I型整合膜蛋白，属于受体酪氨酸激酶家族(RTK)成员之一，该蛋白包含2347个氨基酸，作为生长或分化因子受体发挥作用，由含N-糖基化位点的胞外结构域、外显子34编码的跨膜结构域和外显子36-41编码的酪氨酸激酶结构域三个结构域组成。ROS1蛋白在肾脏中表达最高，其次是小脑、周围神经组织、胃、小肠和结肠，在其他组织中表达较低。The c-ROS proto-oncogene 1 (hereinafter referred to as ROS1) was first discovered to be the transformation sequence of the proto-oncogene and the UR2 avian sarcoma virus. The ROS1 gene is located on chromosome 6q22.1, with 46 exons and a total length of 127kb. The protein encoded by this gene is a type I integral membrane protein with tyrosine kinase activity, belonging to the receptor tyrosine kinase family (RTK). ) Member, the protein contains 2347 amino acids and functions as a growth or differentiation factor receptor. It is composed of an extracellular domain containing an N-glycosylation site, a transmembrane domain encoded by exon 34, and an exon The tyrosine kinase domain encoded by sub36-41 consists of three domains. The expression of ROS1 protein is highest in the kidney, followed by the cerebellum, peripheral nerve tissue, stomach, small intestine and colon, and lower expression in other tissues.
最新的研究表明ROS1是一种新的肿瘤驱动突变基因，其融合在非小细胞肺癌中被定为一种新的分子亚型。ROS1基因的重排最开始是在人脑胶质瘤细胞系里被鉴定出来，后续在其他几个恶性肿瘤里也发现了ROS1基因的重排，如胆管癌、卵巢癌、胃癌和非小细胞肺癌，其中在非小细胞肺癌里的突变频率为1％-2％。ROS1与其他基因发生融合时，丢失细胞外区域，保留跨膜区和胞内酪氨酸激酶区域，重排位点主要发生在ROS1基因的32～36外显子。ROS1基因重排产生具有组成型活性激酶结构域的融合蛋白，所述激酶结构域激活细胞中导致致癌特性的下游信号传导途径，所述致癌特性包括不受控制的增殖以及用延长肿瘤细胞存活抵抗细胞死亡。这些途径包括针对细胞增殖的Ras-ERK途径以及JAK-STAT途径和PI3K/AKT途径，其调节细胞存活(抗细胞凋亡)和增殖。ROS1融合蛋白还可以激活对调节蛋白翻译至关重要的mTOR途径。具有这些途径被激活的癌 症倾向于更具侵袭性，侵袭和转移导致患者具有较差的存活率。ROS1基因可与多个基因发生融合突变，其中最主要的融合伴侣是CD74。例如，在NSCLC中ROS1基因主要与SLC34A2、CD74发生融合并持续激活ROS1酪氨酸激酶区及下游信号通路，进而引起肿瘤的发生。The latest research shows that ROS1 is a new tumor driver mutation gene, and its fusion is designated as a new molecular subtype in non-small cell lung cancer. The ROS1 gene rearrangement was first identified in human glioma cell lines, and subsequent ROS1 gene rearrangements were also found in several other malignant tumors, such as cholangiocarcinoma, ovarian cancer, gastric cancer and non-small cells Lung cancer, where the mutation frequency in non-small cell lung cancer is 1%-2%. When ROS1 fuses with other genes, the extracellular region is lost, the transmembrane region and the intracellular tyrosine kinase region are retained, and the rearrangement sites mainly occur in exons 32 to 36 of the ROS1 gene. The ROS1 gene rearrangement produces a fusion protein with a constitutively active kinase domain that activates downstream signaling pathways in cells that lead to oncogenic properties, including uncontrolled proliferation and prolonged tumor cell survival resistance Cell death. These pathways include the Ras-ERK pathway for cell proliferation, as well as the JAK-STAT pathway and PI3K/AKT pathway, which regulate cell survival (anti-apoptosis) and proliferation. The ROS1 fusion protein can also activate the mTOR pathway, which is essential for regulating protein translation. Cancers that have these pathways activated tend to be more aggressive, and invasion and metastasis lead to poorer survival rates for patients. The ROS1 gene can undergo fusion mutations with multiple genes, and the most important fusion partner is CD74. For example, in NSCLC, the ROS1 gene is mainly fused with SLC34A2 and CD74 and continuously activates the ROS1 tyrosine kinase domain and downstream signaling pathways, which in turn causes tumors.
RTK的抑制剂具有使依赖于RTK活性失调的癌细胞死亡而同时不损伤正常组织的潜力。其中，TRK家族蛋白酪胺酸激酶抑制剂可用于治疗疼痛、炎症、癌症及某些传染性疾病。因此，以TRK、ALK、ROS1为靶点的小分子抑制剂具有令人期待的治疗前景。Inhibitors of RTK have the potential to cause the death of cancer cells that depend on dysregulation of RTK activity without damaging normal tissues. Among them, TRK family protein tyrosine kinase inhibitors can be used to treat pain, inflammation, cancer and certain infectious diseases. Therefore, small molecule inhibitors targeting TRK, ALK, and ROS1 have promising therapeutic prospects.
发明内容Summary of the invention
本发明涉及一类以TRK、ALK、ROS1为靶点的小分子抑制剂，具体涉及式II所示的一类化合物或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、及其混合物形式、及其药学上可接受的盐，多晶型物，溶剂合物，前药，代谢物，同位素衍生物，及包含所述化合物的药物组合物，可用于预防或治疗各类癌症，The present invention relates to a class of small molecule inhibitors targeting TRK, ALK, and ROS1, and specifically relates to a class of compounds represented by formula II or their tautomers, mesosomes, racemates, and enantiomers Isomers, diastereomers, and mixtures thereof, and pharmaceutically acceptable salts, polymorphs, solvates, prodrugs, metabolites, isotope derivatives, and compounds containing said compounds The pharmaceutical composition can be used to prevent or treat various types of cancer,
其中，Y
1为碳时，Y
3为氮、Y
2为碳、Y
4为碳、Y
5为碳；
Wherein, when Y 1 is carbon, Y 3 is nitrogen, Y 2 is carbon, Y 4 is carbon, and Y 5 is carbon;
Y
1为碳时，Y
2为碳、Y
3为氮、Y
4为碳、Y
5为氮；
When Y 1 is carbon, Y 2 is carbon, Y 3 is nitrogen, Y 4 is carbon, and Y 5 is nitrogen;
Y
1为氮时，Y
4为氮、Y
2为碳、Y
3为碳、Y
5为碳；
When Y 1 is nitrogen, Y 4 is nitrogen, Y 2 is carbon, Y 3 is carbon, and Y 5 is carbon;
Y
1为氮时，Y
2为氮、Y
3为碳、Y
4为碳、Y
5为碳；
When Y 1 is nitrogen, Y 2 is nitrogen, Y 3 is carbon, Y 4 is carbon, and Y 5 is carbon;
R
0为
R 0 is
X为碳或氮；X is carbon or nitrogen;
R
1为
-CF
2-R
6、
R 1 is -CF 2 -R 6 、
R
2为-NR
aR
b、-SO
2-NR
aR
b、-S(O)-NR
aR
b、-S(O)-(CH
2)
2-NR
aR
b、4-10元杂环基、-C
4-10环烷基，其中所述的4-10元杂环基或所述的-C
4-10环烷基包括单环、双环、三环、桥环或螺环结构的化合物基团；
R 2 is -NR a R b , -SO 2 -NR a R b , -S(O)-NR a R b , -S(O)-(CH 2 ) 2 -NR a R b , 4-10 yuan Heterocyclic group, -C 4-10 cycloalkyl group, wherein said 4-10 membered heterocyclic group or said -C 4-10 cycloalkyl group includes monocyclic, bicyclic, tricyclic, bridged ring or spiro ring Structure of the compound group;
所述的4-10元杂环基或-C
4-10环烷基可被以下基团单取代或多取代：-OH、-C
1-3烷基、-C
3-6环烷基、4-6元杂环基、5-10元杂芳基、-O-C
1-4烷基、-NR
cR
d、-NH
2、卤素、-C
1-3卤代烷基、苯基或氰基；
The 4-10 membered heterocyclic group or -C 4-10 cycloalkyl group may be mono- or multi-substituted by the following groups: -OH, -C 1-3 alkyl, -C 3-6 cycloalkyl, 4-6 membered heterocyclic group, 5-10 membered heteroaryl group, -OC 1-4 alkyl, -NR c R d , -NH 2 , halogen, -C 1-3 haloalkyl, phenyl or cyano;
R
2优选为
R 2 is preferably
R
a和R
b分别独立的为氢、-C
1-3烷基、-C
3-6环烷基、-C
1-4亚烷基-OH、-C
1-3亚烷基-C
3-9杂环烷基、-C
1-3亚烷基-C
5-6氧代杂环烷基、-C
1-3亚烷基-C
4-6环烷基、-C(O)-(CH
2)
n-T，其中所述的-C
3-9杂环烷基包括单环、双环、三环、桥环或螺环结构的化合物基团，其中所述的-C
3-9杂环烷基可被以下基团单取代或多取代：-C
1-3烷基、卤素、-C
1-3卤代烷基、-OH、氧代基或氰基；
R a and R b are independently hydrogen, -C 1-3 alkyl, -C 3-6 cycloalkyl, -C 1-4 alkylene-OH, -C 1-3 alkylene-C 3 -9 heterocycloalkyl, -C 1-3 alkylene-C 5-6 oxoheterocycloalkyl , -C 1-3 alkylene-C 4-6 cycloalkyl, -C(O)- (CH 2 ) n -T, wherein the -C 3-9 heterocycloalkyl group includes a compound group with a monocyclic, bicyclic, tricyclic, bridged ring or spiro ring structure, wherein the -C 3-9 The heterocycloalkyl group may be mono- or poly-substituted by the following groups: -C 1-3 alkyl, halogen, -C 1-3 haloalkyl, -OH, oxo or cyano;
R
a和R
b或是连同它们所连接的氮原子一起形成未被取代或被T基团或-NR
cR
d取代的4-9元杂环烷基，T为-C
1-4烷基、被-C
1-3烷基取代的-C
2-4烷基、-C
3-6环烷基、4-6元杂环基、-C
1-3卤代烷基、-(CH
2)
n-NR
cR
d；
R a and R b or together with the nitrogen atom to which they are attached form a 4-9 membered heterocycloalkyl which is unsubstituted or substituted with a T group or -NR c R d , and T is -C 1-4 alkyl , -C 2-4 alkyl substituted by -C 1-3 alkyl, -C 3-6 cycloalkyl, 4-6 membered heterocyclic group, -C 1-3 haloalkyl, -(CH 2 ) n -NR c R d ;
R
c和R
d分别独立的为氢、-C
1-3烷基、-C
1-4亚烷基-OH、-C
2-4亚烷基-OCH
3、-(CH
2)
2-OC
1-3烷基；
R c and R d are independently hydrogen, -C 1-3 alkyl, -C 1-4 alkylene-OH, -C 2-4 alkylene -OCH 3 , -(CH 2 ) 2 -OC 1-3 alkyl;
R
3为-NH-C
4-8环烷基、-NH-C
3-8杂环烷基、-NH-CH
2-C
3-8杂环烷基或-O-C
1-4烷基；R
3优选为
-O-C
1-4烷基、-NH-CH
2-C
4-5杂环烷基；T为-C
1-4烷基；T
1为-O、-S、-NH、-N-CH
3；
R 3 is -NH-C 4-8 cycloalkyl, -NH-C 3-8 heterocycloalkyl, -NH-CH 2 -C 3-8 heterocycloalkyl or -OC 1-4 alkyl; R 3 is preferably -OC 1-4 alkyl, -NH-CH 2 -C 4-5 heterocycloalkyl; T is -C 1-4 alkyl; T 1 is -O, -S, -NH, -N-CH 3 ；
R
4为苯基、杂芳基、-NH-苯基，其中所述的苯基可被R
2和/或R
k取代，所述的杂芳基可被R
2取代；R
k为卤素或-O-C
1-4烷基；R
4优选为
X
1为碳或氮；
R 4 is phenyl, heteroaryl, -NH-phenyl, wherein the phenyl group may be substituted by R 2 and/or R k , and the heteroaryl group may be substituted by R 2 ; R k is halogen or -OC 1-4 alkyl; R 4 is preferably X 1 is carbon or nitrogen;
R
6为苯基或4-6元杂环烷基，其中所述的苯基或4-6元杂环烷基被以下基团 单取代或双取代：卤素、-OH、-C
1-3烷基、-C(CH
3)
2-OH或氰基；R
6优选为
R 6 is a phenyl group or a 4-6 membered heterocycloalkyl group, wherein the phenyl group or a 4-6 membered heterocycloalkyl group is mono- or di-substituted by the following groups: halogen, -OH, -C 1-3 Alkyl, -C(CH 3 ) 2 -OH or cyano; R 6 is preferably
R
7为氢、-C
1-3烷基、-(CH
2)
n-OH、C
1-3卤代烷基、-(CH
2)
n-OC
1-3烷基、-(CH
2)
n-OC(O)-C
1-3烷基；R
7优选为氢、-(CH
2)
n-OH、-(CH
2)
n-F、-(CH
2)
n-OC
1-3烷基；
R 7 is hydrogen, -C 1-3 alkyl, -(CH 2 ) n -OH, C 1-3 haloalkyl, -(CH 2 ) n -OC 1-3 alkyl, -(CH 2 ) n- OC(O)-C 1-3 alkyl; R 7 is preferably hydrogen, -(CH 2 ) n -OH, -(CH 2 ) n -F, -(CH 2 ) n -OC 1-3 alkyl;
n为1或2。n is 1 or 2.
本发明所提供的化合物，具体结构如下所述：The specific structure of the compound provided by the present invention is as follows:
N-(5-(2-(2,5-二氟苯基)吡咯烷-1-基)吡唑并[1,5-a]嘧啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(2-(2,5-Difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiper (Azin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(6-(2-(2,5-二氟苯基)吡咯烷-1-基)咪唑并[1,2-b]哒嗪-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(6-(2-(2,5-Difluorophenyl)pyrrolidin-1-yl)imidazo[1,2-b]pyridazin-3-yl)-4-(4-methylpiper (Azin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(2-(2,5-二氟苯基)吡咯烷-1-基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(2-(2,5-Difluorophenyl)pyrrolidin-1-yl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
5-(3,5-二氟苯基)-3-(2’-(4-甲基哌嗪-1-基)-[2,4’-联吡啶]-6-基)-1H-吲唑；5-(3,5-Difluorophenyl)-3-(2'-(4-methylpiperazin-1-yl)-[2,4'-bipyridine]-6-yl)-1H-indyl Azole;
6-(5-(3,5-二氟苯基)-1H-吲唑-3-基)-N-(2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)吡啶-2胺；6-(5-(3,5-Difluorophenyl)-1H-indazol-3-yl)-N-(2-methoxy-4-(4-methylpiperazin-1-yl)benzene Base) pyridine-2 amine;
N-(5-(1-(3,5-二氟苯基)环丙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯并酰胺；N-(5-(1-(3,5-Difluorophenyl)cyclopropyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2- ((Tetrahydro-2H-pyran-4-yl)amino)benzoamide;
N-(5-((3,5-二氟苯基)二氟甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-((3,5-Difluorophenyl)difluoromethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-( (Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((4-甲基哌嗪-1-基)磺酰)-2-((四氢 -2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((4-methylpiperazin-1-yl)sulfonyl)-2-(( Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苯甲基)吡唑并[1,5-a]嘧啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiperazin-1-yl)-2- ((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(3,5-二氟苯基)-2-羟基乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(3,5-Difluorophenyl)-2-hydroxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-(噁丁环-3-基氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-(oxbutane-3 -Base amino) benzamide;
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(5-甲基六氢吡咯并[3,4-c]吡咯-2(1H)-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(5-methylhexahydropyrrolo[3,4-c]pyrrole-2(1H )-Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(甲基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidin-1-yl)ethyl)amino)-2 -((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
(R)-N-(5-(2-(2,5-二氟苯基)吡咯烷-1-基)吡唑并[1,5-a]嘧啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；(R)-N-(5-(2-(2,5-Difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4 -Methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(7-甲基-2,7-二氮杂螺[3.5]壬烷-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(7-methyl-2,7-diazaspiro[3.5]nonane-2 -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(6-甲基-2,6-二氮杂螺[3.3]庚烷-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(6-methyl-2,6-diazaspiro[3.3]heptane-2 -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苯甲基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(6-(3,5-二氟苄基)咪唑并[1,2-b]哒嗪-3-基)-4-(4-甲基哌嗪-1基)-2-((四氢吡喃-4-基)胺基)-苯甲酰胺；N-(6-(3,5-Difluorobenzyl)imidazo[1,2-b]pyridazin-3-yl)-4-(4-methylpiperazin-1 yl)-2-(( Tetrahydropyran-4-yl)amino)-benzamide;
N-(5-((4-(2-羟基丙烷-2-基)哌啶-1-基)甲基)-1H-吲唑-3-基)-4-(4-甲基哌啶-1-基)-2-((四氢吡喃-4-基)胺基)苯甲酰胺；N-(5-((4-(2-hydroxypropan-2-yl)piperidin-1-yl)methyl)-1H-indazol-3-yl)-4-(4-methylpiperidine- 1-yl)-2-((tetrahydropyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苯基)-1H-吲唑-3-基)-4-(5-(二甲基胺基)六氢环戊烷[c]并吡咯-2(1H)-基)-2((四氢-2H-吡喃-4-基)胺基)苯甲酰胺；N-(5-(3,5-Difluorophenyl)-1H-indazol-3-yl)-4-(5-(dimethylamino)hexahydrocyclopentane[c]pyrrole-2 (1H)-yl)-2((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(3,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(3,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
5-(3,5-二氟苯基)-N-(2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)-1H-吲唑-3-甲酰胺；5-(3,5-Difluorophenyl)-N-(2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)-1H-indazole-3-carboxamide;
N-(5-(1-(3,5-二氟苯基)-2-甲氧基乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(3,5-Difluorophenyl)-2-methoxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazine-1- Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-2-甲基-7-((四氢-2H-吡喃-4-基)氨基)-1,2,3,4-四氢异喹啉-6-甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-2-methyl-7-((tetrahydro-2H-pyran-4-yl)amino) -1,2,3,4-Tetrahydroisoquinoline-6-carboxamide;
4-((1R,3S,5s,7s)-5-氨基金刚烷-2-基)-N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺盐酸盐；4-((1R,3S,5s,7s)-5-aminoadamantan-2-yl)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide hydrochloride;
4-(5-氨基-金刚烷-2-基)-N-(5-(3,5-二氟苄基)-1H-吲哚-3-基)-2-(哌啶-4-基氨基)苯甲酰胺；4-(5-Amino-adamantan-2-yl)-N-(5-(3,5-difluorobenzyl)-1H-indol-3-yl)-2-(piperidin-4-yl) Amino) benzamide;
(R)-N-(5-(2-(2,5-二氟苯基)吡咯烷-1-基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；(R)-N-(5-(2-(2,5-Difluorophenyl)pyrrolidin-1-yl)-1H-indazol-3-yl)-4-(4-methylpiperazine- 1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(2,5-二氟苯基)-2-羟基乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-hydroxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
2-(3,5-二氟苯基)-2-(3-(4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰氨基)-1H-吲唑-5-基)乙酸乙酯；2-(3,5-Difluorophenyl)-2-(3-(4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl) Amino)benzoylamino)-1H-indazol-5-yl)ethyl acetate;
N-(5-(1-(3,5-二氟苯基)乙基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(3,5-Difluorophenyl)ethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazine -1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3-氟苄基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3-Fluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-( (Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(3-氟苯基)-2-羟基乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(3-Fluorophenyl)-2-hydroxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2- ((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(3-氟苯基)乙基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(3-Fluorophenyl)ethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazine-1- Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(2,5-二氟苯基)乙基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)ethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazine -1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(2,5-二氟苯基)-2-甲氧基乙基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-methoxyethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-( 4-Methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
2-(3-氟苯基)-2-(3-(4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)-1氢-吲唑-5-基)乙基异丁酯；2-(3-Fluorophenyl)-2-(3-(4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)- 1 Hydro-indazol-5-yl) ethyl isobutyl ester;
N-(5-(3-氟苄基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(5-甲基-2,5-二氮杂双环[2.2.1]庚-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3-Fluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(5-methyl-2,5-diazabicyclo[2.2 .1]hept-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(3-氟苯基)-2-甲氧基乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(3-Fluorophenyl)-2-methoxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)- 2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苄基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(5-甲基-2,5-二氮杂双环[2.2.1]庚-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(5-methyl-2,5-diazepine Bicyclo[2.2.1]hept-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(3-(二甲基氨基)氮杂环丁烷-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(3-(dimethylamino)azetidin-1-yl)-2- ((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(3-((2-甲氧基乙基)(甲基)氨基)氮杂环丁烷-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(3-((2-methoxyethyl)(methyl)amino)azacyclo Butane-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((2-(pyrrolidin-1-yl)ethyl)amino)-2-( (Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(乙基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2 -((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((2-(二甲氨基)乙基)硫代)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((2-(dimethylamino)ethyl)thio)-2-((tetra Hydrogen-2H-pyran-4-yl)amino)benzamide;
N-(5-(3-氟苯甲基)-1H-吲唑-3-基)-4-(乙基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3-fluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(( Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3-氟苯甲基)-1H-吲唑-3-基)-4-(甲基(2-(吡咯烷-1-基)乙基)氨基)-2-((四 氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3-Fluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(( Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((氧杂环丁烷-3-基甲基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((oxetane -3-ylmethyl)amino)benzamide;
N-(5-(3-氟苄基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((氧杂环丁烷-3-基甲基)氨基)苯甲酰胺；N-(5-(3-Fluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((oxetane-3- (Methyl)amino)benzamide;
N-(5-(2,5-二氟苄基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((氧杂环丁烷-3-基甲基)氨基)苯甲酰胺；N-(5-(2,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((oxetane -3-ylmethyl)amino)benzamide;
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(3-氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(3-fluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2- ((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-乙基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-ethylpiperazin-1-yl) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-***啉哌啶-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-morpholinepiperidin-1-yl) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-(四氢-2H-吡喃-4-基)哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-(tetrahydro-2H-pyran- 4-yl)piperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-环丙基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-cyclopropylpiperazin-1-yl )-2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-三氟甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-trifluoromethylpiperazine-1- Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(甲基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidine-1- (Yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苄基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-((1R，4R)-5-甲基-2,5-二氮杂双环[2.2.10.1]庚烷-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-((1R,4R)-5-methyl-2 ,5-Diazabicyclo[2.2.10.1]heptan-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苄基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-((1s，4s)-5-甲基-2,5-二氮杂双环[2.2.10.1]庚烷-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-((1s,4s)-5-methyl-2 ,5-Diazabicyclo[2.2.10.1]heptan-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-2-(2-(二甲氨基)乙基)-1-羰基-6-((四氢-2H-吡喃-4-基)氨基)异二氢吲哚-5-甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-2-(2-(dimethylamino)ethyl)-1-carbonyl-6-((tetra Hydrogen-2H-pyran-4-yl)amino)isoindole-5-carboxamide;
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(N-甲基-2-吗啉代乙酰氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(N-methyl-2-morpholinoacetamido)-2-((tetrahydro -2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(3,5-二氟苯基)-2,2,2-三氟-乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(3,5-Difluorophenyl)-2,2,2-trifluoro-ethyl)-1H-indazol-3-yl)-4-(4-methylpiper (Azin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(乙基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2 -((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(乙基(2-(3-氟吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(3-fluoropyrrolidin-1-yl)ethyl)amino) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((2-(吡咯烷-1-基)乙基)硫代)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((2-(pyrrolidin-1-yl)ethyl)thio)-2- ((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)--4-((2-(3,3-二氟吡咯烷-1-基)-乙基)-(乙基)-氨基)--2-((四氢-2H-吡喃-4-基)氨基)-苯甲酰胺；N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)--4-((2-(3,3-difluoropyrrolidin-1-yl)-ethyl )-(Ethyl)-amino)-2-((tetrahydro-2H-pyran-4-yl)amino)-benzamide;
N-(5-(3,5-二氟苄基)--1H-吲唑-3-基)--4-((2-(3,4-二氟吡咯烷-1-基)-乙基)(乙基)-氨基)--2-((四氢-2H-吡喃-4-基)-氨基)-苯甲酰胺；N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)--4-((2-(3,4-difluoropyrrolidin-1-yl)-ethyl Yl)(ethyl)-amino)-2-((tetrahydro-2H-pyran-4-yl)-amino)-benzamide;
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(3,4-二甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(3,4-dimethylpiperazine-1 -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(3-甲基-4-乙基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(3-methyl-4-ethylpiperazine -1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(2,4-二甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(2,4-dimethylpiperazine-1 -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苄基(-1H-吲唑-3-基)-4-((2-(3,3,4,4-四氟吡咯烷-1-基)-乙基)- (乙基)-氨基)--2-((四氢-2H-吡喃-4-基)-氨基)-苯甲酰胺；N-(5-(3,5-Difluorobenzyl(-1H-indazol-3-yl)-4-((2-(3,3,4,4-tetrafluoropyrrolidin-1-yl) -Ethyl)-(ethyl)-amino)-2-((tetrahydro-2H-pyran-4-yl)-amino)-benzamide;
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-2-((四氢-2H-吡喃-4-基)氨基)-4-(4-(2,2,2-三氟乙基)哌嗪-1-基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-2-((tetrahydro-2H-pyran-4- (Yl)amino)-4-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)benzamide;
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(2-甲基-4-乙基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(2-methyl-4-ethylpiperazine -1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
4-(环丙基(2-(吡咯烷-1-基)-乙基)-氨基)-N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)--2-((四氢-2H-吡喃-4-基)-氨基)-苯甲酰胺；4-(Cyclopropyl(2-(pyrrolidin-1-yl)-ethyl)-amino)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl) --2-((Tetrahydro-2H-pyran-4-yl)-amino)-benzamide;
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)--4-((2-(1,4-二氧-7-氮杂螺[4.4]壬烷-1-基)乙基)-(乙基)氨基)-2-((四氢-2H-吡喃-4-基)-氨基)-苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)--4-((2-(1,4-diox-7-azaspiro[4.4]non Alk-1-yl)ethyl)-(ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)-amino)-benzamide;
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-((2-(3-氧代吡咯烷-1-基)乙基)(乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((2-(3-oxopyrrolidin-1-yl)ethyl)(ethyl )Amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(2,5-二氟苯甲基)-1H-吲唑-3-基)-4-(甲基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(2,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidin-1-yl)ethyl)amino)-2 -((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(乙基(2-(氮杂环丁烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(azetidin-1-yl)ethyl)amino) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((2-(吡咯烷-1-基)乙基)亚硫酰基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((2-(pyrrolidin-1-yl)ethyl)sulfinyl)-2 -((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-甲基-4,7-二氮杂螺[2.5]辛烷-7-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methyl-4,7-diazepine Heterosspiro[2.5]octane-7-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(1-(2,5-二氟苯基(-2-氟乙基)-1H-吲唑-3-基)-4-(3,5-二甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl(-2-fluoroethyl)-1H-indazol-3-yl)-4-(3,5-dimethylpiperazine-1 -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(甲基(2-(3-氰基吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(3-cyanopyrrolidin-1-yl)ethyl)amino )-2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;
(R)-N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(甲基(2-(3-羟基吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；(R)-N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(3-hydroxypyrrolidin-1-yl)ethyl (Yl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;
(S)-N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(甲基(2-(3-羟基吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺。(S)-N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(3-hydroxypyrrolidin-1-yl)ethyl Yl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide.
本发明提供了制备通式II所示的化合物的方法，所述通式II所示的化合物通过以下方案制备：The present invention provides a method for preparing a compound represented by general formula II, which is prepared by the following scheme:
方案1，当其中，Y
1为碳时，Y
3为氮、Y
2为碳、Y
4为碳、Y
5为碳；R
1为-CF
2-R
6时，合成路线归纳如下，
Scheme 1, when Y 1 is carbon, Y 3 is nitrogen, Y 2 is carbon, Y 4 is carbon, and Y 5 is carbon; when R 1 is -CF 2 -R 6 , the synthetic route is summarized as follows:
其中所述氧化试剂优选为Dess-Martin氧化剂、PCC、PDC等氧化剂，氟化试剂优选为BAST、DAST等氟化试剂，成酰胺方法优选为羧酸经酰氯于胺反应制成酰胺，或羧酸经1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐(EDCI)/1-羟基苯并***(HOBT)、环己基碳二亚胺(DCC)、2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(HATU)等缩合剂活化与胺成酰胺；The oxidizing agent is preferably Dess-Martin oxidizing agent, PCC, PDC and other oxidizing agents, and the fluorinating agent is preferably BAST, DAST and other fluorinating agents. The amide forming method is preferably carboxylic acid reacting with acid chloride to amine to make amide, or carboxylic acid After 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI)/1-hydroxybenzotriazole (HOBT), cyclohexylcarbodiimide (DCC), 2 -(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU) and other condensing agents are activated to form amides with amines;
方案2，当其中，Y
1为碳时，Y
3为氮、Y
2为碳、Y
4为碳、Y
5为碳；R
1为
Scheme 2, when Y 1 is carbon, Y 3 is nitrogen, Y 2 is carbon, Y 4 is carbon, and Y 5 is carbon; R 1 is
合成路线归纳如下：The synthetic route is summarized as follows:
其中所述氧化试剂优选为Dess-Martin氧化剂、PCC、PDC等氧化剂；成三元环方法优选为二乙基锌/二碘甲烷、三甲基碘化亚砜盐或三甲基溴化亚砜盐/钠氢；硼化试剂优选为硼烷、9-BBN等；氧化试剂优选为双氧水；氟化试剂优选为BAST、DAST等氟化试剂，成酰胺方法优选为羧酸经酰氯于胺反应制成酰胺，或羧酸经1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐(EDCI)/1-羟基苯并***(HOBT)、环己基碳二亚胺(DCC)、2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(HATU)等缩合剂活化与胺成酰胺；The oxidizing agent is preferably Dess-Martin oxidant, PCC, PDC and other oxidants; the method of forming the three-membered ring is preferably diethyl zinc/diiodomethane, trimethyl sulfoxide iodide or trimethyl sulfoxide bromide Salt/sodium hydrogen; the boronating reagent is preferably borane, 9-BBN, etc.; the oxidizing reagent is preferably hydrogen peroxide; the fluorinating reagent is preferably fluorinating reagents such as BAST and DAST; To amide, or carboxylic acid through 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI)/1-hydroxybenzotriazole (HOBT), cyclohexyl carbodiimide Amine (DCC), 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU) and other condensing agents are activated to form amides with amines;
方案3，当其中，Y
1为碳时，Y
2为碳、Y
3为氮、Y
4为碳、Y
5为氮；或Y
1为氮时，Y
4为氮、Y
2为碳、Y
3为碳、Y
5为碳；或Y
1为氮时，Y
2为氮、Y
3为碳、Y
4为碳、Y
5为碳；R
1为-CH
2-R
6时；合成路线归纳如下：
Scheme 3, when Y 1 is carbon, Y 2 is carbon, Y 3 is nitrogen, Y 4 is carbon, and Y 5 is nitrogen; or when Y 1 is nitrogen, Y 4 is nitrogen, Y 2 is carbon, and Y 3 is carbon and Y 5 is carbon; or when Y 1 is nitrogen, Y 2 is nitrogen, Y 3 is carbon, Y 4 is carbon, and Y 5 is carbon; when R 1 is -CH 2 -R 6 ; synthetic route is summarized as follows:
其中卤代实际优选为溴、碘、N-溴代丁二酰亚胺、N-碘代丁二酰亚胺等，偶联反应优选为Stille反应；Wherein halogenation is actually preferably bromine, iodine, N-bromosuccinimide, N-iodosuccinimide, etc., and the coupling reaction is preferably Stille reaction;
方案4，当其中，Y
1为碳时，Y
2为碳、Y
3为氮、Y
4为碳、Y
5为碳；或Y
1
Scheme 4, when Y 1 is carbon, Y 2 is carbon, Y 3 is nitrogen, Y 4 is carbon, and Y 5 is carbon; or Y 1
其中硝化试剂优选为硝酸、硝酸钾、硝酸钠等；还原试剂优选为铁粉、锌粉、氯化亚锡、氢气/钯碳、氢气/镍等；成酰胺方法优选为羧酸经酰氯于胺反应制成酰胺，或羧酸经1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐(EDCI)/1-羟基苯并***(HOBT)、环己基碳二亚胺(DCC)、2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(HATU)等缩合剂活化与胺成酰胺；Among them, the nitrating reagent is preferably nitric acid, potassium nitrate, sodium nitrate, etc.; the reducing reagent is preferably iron powder, zinc powder, stannous chloride, hydrogen/palladium on carbon, hydrogen/nickel, etc.; the amide formation method is preferably carboxylic acid through acid chloride to amine Reaction to produce amide, or carboxylic acid through 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI)/1-hydroxybenzotriazole (HOBT), cyclohexyl carbon Diimine (DCC), 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU) and other condensing agents are activated to form amides with amines;
方案5，当其中，R
0为
R
1为-CH
2-R
6；合成路线归纳如下：
Scheme 5, when R 0 is R 1 is -CH 2 -R 6 ; the synthetic route is summarized as follows:
本发明的化合物的药学上可接受的盐包括其酸加成盐及碱加成盐。The pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts thereof.
适合的酸加成盐由形成无毒盐的酸来形成，实例包括但不限于盐酸盐、硫酸盐/硫酸氢盐、硝酸盐、磷酸盐/磷酸氢盐/磷酸二氢盐、氢溴酸盐、氢碘酸盐、醋酸盐、乳酸盐、甲磺酸盐、柠檬酸盐、苹果酸盐、马来酸盐、富马酸盐、酒石酸盐、水杨酸盐、硬脂酸盐，及其类似盐。Suitable acid addition salts are formed by acids that form non-toxic salts, examples include but are not limited to hydrochloride, sulfate/bisulfate, nitrate, phosphate/hydrogen phosphate/dihydrogen phosphate, hydrobromic acid Salt, hydroiodide, acetate, lactate, methanesulfonate, citrate, malate, maleate, fumarate, tartrate, salicylate, stearate , And its similar salt.
适合的碱加成盐由形成无毒盐的碱来形成，例如作为碱金属盐、碱土金属盐或作为铵盐，实例包括但不限于钠盐、钾盐、钙盐、镁盐，或与氨或有机胺如乙 胺、乙醇胺、三乙醇胺或氨基酸的盐。Suitable base addition salts are formed from bases that form non-toxic salts, for example as alkali metal salts, alkaline earth metal salts, or as ammonium salts. Examples include, but are not limited to, sodium, potassium, calcium, magnesium, or with ammonia Or organic amines such as ethylamine, ethanolamine, triethanolamine or salts of amino acids.
适合的盐的综述参见Stahl,P.H.及Wermuth,C.G.的“Handbook of Pharmaceutical Salts:Properties,Selection,and Use,2
nd Revised Edition”(Wiley-VCH,2011)。
For a review of suitable salts, see "Handbook of Pharmaceutical Salts: Properties, Selection, and Use, 2 nd Revised Edition" by Stahl, PH and Wermuth, CG (Wiley-VCH, 2011).
如果式(II)化合物在分子内同时包含酸性和碱性基团，本发明还包括除了提及的盐形式之外的内盐或内铵盐(两性离子)。If the compound of formula (II) contains both acidic and basic groups in the molecule, the present invention also includes internal salts or betaines (zwitterions) in addition to the salt forms mentioned.
用于制备本发明的化合物的药学上可接受的盐的方法为本领域技术人员已知的，例如通过使这些化合物与有机或无机酸或碱在溶剂或分散剂中接触获得，或通过与其他盐进行阴离子交换或阳离子交换获得。Methods for preparing pharmaceutically acceptable salts of the compounds of the present invention are known to those skilled in the art, for example, by contacting these compounds with organic or inorganic acids or bases in solvents or dispersants, or by contacting other The salt is obtained by anion exchange or cation exchange.
式(II)化合物可以以晶体或无定形形式存在。此外，式(II)化合物的某些晶体形式可以多晶型形式存在，其包括在本发明范围内。可以使用许多常规分析技术包括但不限于单晶X-射线粉末衍射(XRPD)图、红外(IR)光谱、拉曼光谱、差示扫描量热法(DSC)、热重分析(TGA)和固体核磁共振(ssNMR)表征来区分化合物的多晶型。The compound of formula (II) may exist in crystalline or amorphous form. In addition, certain crystal forms of the compound of formula (II) may exist in polymorphic forms, which are included in the scope of the present invention. Many conventional analysis techniques can be used including but not limited to single crystal X-ray powder diffraction (XRPD) patterns, infrared (IR) spectroscopy, Raman spectroscopy, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA) and solids Nuclear magnetic resonance (ssNMR) characterization to distinguish the polymorphic forms of the compound.
本发明的化合物也包括互变异构形式。互变异构形式由单键与相邻双键的对换以及伴随的质子迁移而产生。互变异构形式包括质子转移互变异构体，它们是具有相同经验式和总电荷的异构质子化状态。质子转移互变异构体的实例包括酮-烯醇配对、酰胺-亚胺酸配对、内酰胺-内酰亚胺配对、烯胺-亚胺配对，以及其中质子能够占据杂环体系的两个或多个位置的环状形式，例如，1H-和3H-咪唑，1H-、2H-和4H-1,2,4***，1H-和2H-异吲哚，以及1H-和2H-吡唑。互变异构形式可以通过适当的取代而处于平衡或空间上固定于一种形式。The compounds of the present invention also include tautomeric forms. The tautomeric form is produced by the exchange of a single bond with an adjacent double bond and the accompanying migration of protons. Tautomeric forms include proton transfer tautomers, which are isomeric protonated states with the same empirical formula and total charge. Examples of proton transfer tautomers include keto-enol pairings, amide-imine pairings, lactam-lactam pairings, enamine-imine pairings, and two in which protons can occupy the heterocyclic ring system. Or multiple positions of the cyclic form, for example, 1H- and 3H-imidazole, 1H-, 2H- and 4H-1,2,4 triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyridine Azole. Tautomeric forms can be in equilibrium or sterically fixed to one form by appropriate substitution.
本发明进一步涉及通式(II)所示的化合物或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、及其混合物形式、及其可药用盐或包含其的药物组合物，其进一步与另外一种或多种调节哺乳动物免疫***的试剂、抗癌剂或抗炎剂联合应用。The present invention further relates to the compound represented by the general formula (II) or its tautomer, meso, racemate, enantiomer, diastereomer, and mixtures thereof, and The pharmaceutically acceptable salt or the pharmaceutical composition containing the same is further used in combination with another one or more agents for regulating the immune system of mammals, anticancer agents or anti-inflammatory agents.
本发明进一步涉及通式(II)所示的化合物或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、及其混合物形式、及其可药用盐或包含其的 药物组合物，其用于治疗或预防下列癌症，包括实体瘤(如***癌、肾癌、肝癌、胰腺癌、胃癌、乳腺癌、肺癌、头颈部癌、甲状腺癌、胶质母细胞瘤、黑色素瘤等)、血液癌(如淋巴瘤、白血病等)、皮肤癌(如皮肤T-细胞淋巴瘤、皮肤B-细胞淋巴瘤)等。The present invention further relates to the compound represented by the general formula (II) or its tautomer, meso, racemate, enantiomer, diastereomer, and mixtures thereof, and Its pharmaceutically acceptable salt or pharmaceutical composition containing it is used to treat or prevent the following cancers, including solid tumors (such as prostate cancer, kidney cancer, liver cancer, pancreatic cancer, stomach cancer, breast cancer, lung cancer, head and neck cancer, Thyroid cancer, glioblastoma, melanoma, etc.), blood cancer (such as lymphoma, leukemia, etc.), skin cancer (such as skin T-cell lymphoma, skin B-cell lymphoma), etc.
本发明还涉及一种治疗或预防癌症的方法，其包括给予所需患者治疗有效量的通式(II)所示的化合物或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、及其混合物形式、及其可药用盐，或包含其的药物组合物，其中所述的癌症，包括例如实体瘤(如***癌、肾癌、肝癌、胰腺癌、胃癌、乳腺癌、肺癌、头颈部癌、甲状腺癌、胶质母细胞瘤、黑色素瘤等)、血液癌(如淋巴瘤、白血病等)、皮肤癌(如皮肤T-细胞淋巴瘤、皮肤B-细胞淋巴瘤)等。The present invention also relates to a method for treating or preventing cancer, which comprises administering to a patient a therapeutically effective amount of a compound represented by general formula (II) or its tautomers, mesosomes, racemates, Enantiomers, diastereomers, and mixtures thereof, and pharmaceutically acceptable salts thereof, or pharmaceutical compositions containing them, wherein the cancer includes, for example, solid tumors (such as prostate cancer, renal cancer) , Liver cancer, pancreatic cancer, stomach cancer, breast cancer, lung cancer, head and neck cancer, thyroid cancer, glioblastoma, melanoma, etc.), blood cancer (such as lymphoma, leukemia, etc.), skin cancer (such as skin T- Cell lymphoma, skin B-cell lymphoma) and so on.
本发明的组合物可以利用一种或多种可药用的载体按照常规的方式加以配制。因此，本发明的活性化合物可以被配制成口服、口腔含化给药、鼻内、肠胃外(例如静脉内、肌内或皮下)或直肠给药的剂型，或者适用于通过吸入或吹入给药的剂型。本发明的化合物也可以被配制成持续释放的剂型。The composition of the present invention can be formulated in a conventional manner using one or more pharmaceutically acceptable carriers. Therefore, the active compound of the present invention can be formulated into a dosage form for oral, buccal administration, intranasal, parenteral (for example, intravenous, intramuscular or subcutaneous) or rectal administration, or suitable for administration by inhalation or insufflation. The dosage form of the medicine. The compounds of the present invention can also be formulated into sustained release dosage forms.
根据治疗目的，可将药物组合物制成各种类型的给药单位剂型，如片剂、丸剂、粉剂、液体、悬浮液、乳液、颗粒剂、胶囊、栓剂和针剂(溶液及悬浮液)，或者适用于通过吸入或吹入给药的剂型等。According to the purpose of treatment, the pharmaceutical composition can be made into various types of administration unit dosage forms, such as tablets, pills, powders, liquids, suspensions, emulsions, granules, capsules, suppositories, and injections (solutions and suspensions). Or it is suitable for a dosage form for administration by inhalation or insufflation.
为了使片剂形式的药物组合物成形，可使用本领域任何己知并广泛使用的赋形剂。例如，载体，如乳糖、白糖、氯化钠、葡萄糖、尿素、淀粉、碳酸钙、高岭土、结晶纤维素和硅酸等；粘合剂，如水、乙醇、丙醇、普通糖浆、葡萄糖溶液、淀粉溶液、明胶溶液，羧甲基纤维素、紫胶、甲基纤维素和磷酸钾、聚乙烯吡咯烷酮等；崩解剂，如干淀粉、藻酸纳、琼脂粉和海带粉，碳酸氢纳、碳酸钙、聚乙烯脱水山梨醇的脂肪酸酯、十二烷基硫酸纳、硬脂酸单甘酯、淀粉和乳糖等；崩解抑制剂，如白糖、甘油三硬脂酸酯、椰子油和氢化油；吸附促进剂，如季胺碱和十二烷基硫酸纳等；润湿剂，如甘油、淀粉等；吸附剂，如淀粉、乳糖、高岭土、膨润土和胶体硅酸等；以及润滑剂，如纯净的滑石，硬脂酸盐、硼酸粉和聚乙二醇等。还可以根据需要选用通常的涂渍材料制成糖衣片剂、涂明胶膜片剂、肠衣片剂、涂膜片剂、双层膜片剂及多层片剂。In order to shape the pharmaceutical composition in the form of a tablet, any excipient known and widely used in the art can be used. For example, carriers such as lactose, white sugar, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose and silicic acid, etc.; binders such as water, ethanol, propanol, ordinary syrup, glucose solution, starch Solution, gelatin solution, carboxymethyl cellulose, shellac, methyl cellulose and potassium phosphate, polyvinylpyrrolidone, etc.; disintegrating agent, such as dry starch, sodium alginate, agar powder and kelp powder, sodium bicarbonate, carbonic acid Fatty acid esters of calcium, polyethylene sorbitan, sodium lauryl sulfate, monoglyceride stearate, starch and lactose, etc.; disintegration inhibitors such as white sugar, glyceryl tristearate, coconut oil and hydrogenated Oil; Adsorption promoters, such as quaternary amine bases and sodium lauryl sulfate, etc.; Wetting agents, such as glycerin, starch, etc.; Adsorbents, such as starch, lactose, kaolin, bentonite and colloidal silicic acid, etc.; and lubricants, Such as pure talc, stearate, boric acid powder and polyethylene glycol. Ordinary coating materials can be selected to make sugar-coated tablets, gelatin film-coated tablets, enteric-coated tablets, film-coated tablets, double-layer film tablets, and multi-layer tablets according to needs.
为了使丸剂形式的药物组合物成形，可使用本领域任何己知的并广泛使用的赋形剂，例如，载体，如乳糖，淀粉，椰子油，硬化植物油，高岭土和滑石粉等；粘合剂，如***树胶粉，黄蓍胶粉，明胶和乙醇等；崩解剂，如琼脂和海带粉等。In order to shape the pharmaceutical composition in the form of a pill, any known and widely used excipients in the art can be used, for example, carriers such as lactose, starch, coconut oil, hardened vegetable oil, kaolin and talc, etc.; binders , Such as gum arabic powder, gum tragacanth powder, gelatin and ethanol; disintegrants such as agar and kelp powder.
为了使栓剂形式的药物组合物成形，可使用本领域任何己知并广泛使用的赋形剂，例如，聚乙二醇，椰子油，高级醇，高级醇的酯，明胶和半合成的甘油酯等。In order to shape the pharmaceutical composition in the form of suppositories, any excipient known and widely used in the art can be used, for example, polyethylene glycol, coconut oil, higher alcohols, esters of higher alcohols, gelatin and semi-synthetic glycerides Wait.
为了制备针剂形式的药物组合物，可将溶液或悬浮液消毒后(最好加入适量的氯化钠，葡萄糖或甘油等)，制成与血液等渗压的针剂。在制备针剂时，也可使用本领域内任何常用的载体。例如，水，乙醇，丙二醇，乙氧基化的异硬脂醇，聚氧基化的异硬脂醇和聚乙烯脱水山梨醇的脂肪酸酯等。此外，还可加入通常的溶解剂、缓冲剂和止痛剂等。In order to prepare a pharmaceutical composition in the form of an injection, the solution or suspension can be sterilized (preferably by adding an appropriate amount of sodium chloride, glucose or glycerol, etc.) to prepare an injection that is isotonic with blood. When preparing injections, any carriers commonly used in the art can also be used. For example, water, ethanol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol and fatty acid esters of polyethylene sorbitan and the like. In addition, usual dissolving agents, buffering agents and analgesics can also be added.
为了鼻内给药或通过吸入给药，本发明的活性化合物适宜以溶液或悬液的形式而从收到或者挤压或抽吸的泵式喷雾容器内释放出来，或者以喷雾剂的形式从加压容器或喷雾器内释放出来，释放利用一种适合的推进剂，例如二氯二氟甲烷、三氯氟甲烷、二氯四氟乙烷、二氧化碳或其他适合的气体。在加压气雾剂的情况下，剂量单位可以通过提供计量释放的阀门加以确定。加压容器或喷雾器可以含有活性化合物的溶液或悬液。用在吸入器或吹入器内的胶囊或药筒(例如由明胶制成)可以被配制成含有本发明化合物与适合的粉末基质，例如乳糖或淀粉的粉末混合物。For intranasal administration or administration by inhalation, the active compound of the present invention is suitably released in the form of a solution or suspension from a pump spray container received or squeezed or sucked, or in the form of a spray. Release from a pressurized container or aerosol, and use a suitable propellant, such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gases. In the case of pressurized aerosols, the dosage unit can be determined by a valve that provides a metered release. The pressurized container or nebulizer may contain a solution or suspension of the active compound. Capsules or cartridges (for example made of gelatin) used in inhalers or insufflators can be formulated as a powder mixture containing the compound of the invention and a suitable powder base, such as lactose or starch.
除非有相反陈述，在说明书和权利要求书中使用的术语具有下述含义。Unless stated to the contrary, the terms used in the specification and claims have the following meanings.
术语“立体异构体”表示由于至少一个不对称中心形成的异构体。在具有一个或多个(例如一个、两个、三个或四个)不对称中心的化合物中，其可产生外消旋体、外消旋混合物、内消旋体、单一对映异构体、非对映异构体混合物和单独的非对映异构体。特定个别分子也可以几何异构体(顺式/反式)存在。The term "stereoisomer" means an isomer formed due to at least one asymmetric center. In compounds with one or more (for example, one, two, three or four) asymmetric centers, it can produce racemates, racemic mixtures, mesosomes, single enantiomers , Diastereomeric mixtures and individual diastereomers. Certain individual molecules can also exist as geometric isomers (cis/trans).
术语“溶剂合物”是指通常通过溶剂分解反应与溶剂物理结合的化合物形式。此物理结合包含氢键结合。常规溶剂包含水、乙醇、甲醇、乙酸等。式(II)化合 物可以结晶形式制备且可呈溶剂合物形式(例如水合形式)。适宜溶剂合物包含药学可接受的溶剂合物(例如水合物)，且进一步包含化学计量溶剂合物及非化学计量溶剂合物。在某些情形下，例如当一个或多个溶剂分子纳入结晶固体的晶格中时，溶剂合物将能够解离。“溶剂合物”涵盖溶液相及可解离溶剂合物。代表性溶剂合物包含水合物、乙醇合物及甲醇合物等。The term "solvate" refers to a form of a compound that is generally physically combined with a solvent through a solvolysis reaction. This physical bond includes hydrogen bonding. Conventional solvents include water, ethanol, methanol, acetic acid and the like. The compound of formula (II) can be prepared in crystalline form and can be in the form of a solvate (e.g., a hydrated form). Suitable solvates include pharmaceutically acceptable solvates (such as hydrates), and further include stoichiometric solvates and non-stoichiometric solvates. In some cases, such as when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid, the solvate will be able to dissociate. "Solvate" encompasses both solution-phase and dissociable solvates. Representative solvates include hydrates, ethanolates, methanolates, and the like.
术语“前药”是指通过与酶、胃酸等在生理条件下在活体内例如通过各自在酶催化下进行的氧化、还原、水解等反应转化为本发明化合物的衍生物。The term "prodrug" refers to a derivative that is converted into a compound of the present invention by reaction with enzymes, gastric acid, etc. in the living body under physiological conditions, for example, through oxidation, reduction, hydrolysis, etc., respectively, catalyzed by enzymes.
术语“代谢物”是指在细胞或有机体优选人中源自本发明任意化合物的所有分子。The term "metabolite" refers to all molecules derived from any compound of the present invention in a cell or organism, preferably a human.
术语“同位素衍生物”是指构成化合物的一或多个原子处，以非天然比例含有同位素的所述的化合物。例如氘(
2H或D)、氚(
3H或T)、碳－13(
13C)、氮－15(
15N)、氧－18(
18O)等。
The term "isotopic derivative" refers to a compound that contains isotopes in unnatural proportions at one or more of the atoms constituting the compound. For example, deuterium ( 2 H or D), tritium ( 3 H or T), carbon-13 ( 13 C), nitrogen-15 ( 15 N), oxygen-18 ( 18 O), etc.
术语“药物组合物”表示含有一种或多种本发明所述化合物或其生理学/药学上可接受的盐或前体药物与其他化学组分的混合物，其他组分例如生理学/药学上可接受的载体和赋形剂。药物组合物的目的是促进对生物体的给药，利于活性成分的吸收进而发挥生物活性。The term "pharmaceutical composition" means a mixture containing one or more of the compounds of the present invention or their physiologically/pharmaceutically acceptable salts or prodrugs and other chemical components, such as physiologically/pharmaceutically acceptable Carriers and excipients. The purpose of the pharmaceutical composition is to promote the administration to the organism, which is beneficial to the absorption of the active ingredient and thus the biological activity.
所述载体包括药学领域所有的可用于制成注射和非注射给药途径的药物制剂，例如稀释剂、润湿剂、填充剂、粘合剂、湿滑剂、崩解剂、吸收促进剂、表面活性剂、阻滞剂、吸附剂、助悬剂、絮凝剂、反絮凝剂、乳化剂、常用基质、增溶剂、助溶剂、潜溶剂、防腐剂、矫味剂、着色剂、抗氧剂、缓冲剂、抑菌剂、等渗调节剂、pH调节剂、金属离子络合剂、硬化剂、增稠剂等。The carrier includes all pharmaceutical preparations in the pharmaceutical field that can be used for injection and non-injection administration routes, such as diluents, wetting agents, fillers, adhesives, slip agents, disintegrants, absorption enhancers, Surfactants, retarders, adsorbents, suspending agents, flocculants, deflocculants, emulsifiers, commonly used bases, solubilizers, co-solvents, latent solvents, preservatives, flavoring agents, coloring agents, antioxidants , Buffers, bacteriostatic agents, isotonic regulators, pH regulators, metal ion complexing agents, hardeners, thickeners, etc.
本申请提供的新型RTK的抑制剂具有使依赖于RTK活性失调的癌细胞死亡而同时不损伤正常组织的潜力。并且，本申请提供的TRK家族蛋白酪胺酸激酶抑制剂可用于治疗疼痛、炎症、癌症及某些传染性疾病。发明人出人意料地发现，本申请提供的RTK抑制剂可以有效抑制原肌球蛋白受体激酶A(TRKA)、间变性淋巴瘤激酶(ALK)、原癌基因酪氨酸激酶(ROS1)活性，对瘤细胞、癌细胞的增殖以及肿瘤生长均有较强的抑制作用，并且抑制活性优于现有技术的恩曲替尼 (Entrectinib)。另外，本发明的化合物还具有较好的代谢特征及较好的生物利用度。The novel RTK inhibitor provided in the present application has the potential to cause the death of cancer cells that depend on the imbalance of RTK activity without damaging normal tissues at the same time. In addition, the TRK family protein tyrosine kinase inhibitors provided in this application can be used to treat pain, inflammation, cancer and certain infectious diseases. The inventors unexpectedly discovered that the RTK inhibitor provided in the present application can effectively inhibit the activities of tropomyosin receptor kinase A (TRKA), anaplastic lymphoma kinase (ALK), and proto-oncogene tyrosine kinase (ROS1). Tumor cells, cancer cell proliferation and tumor growth have strong inhibitory effects, and the inhibitory activity is better than the prior art Entrectinib (Entrectinib). In addition, the compounds of the present invention also have better metabolic characteristics and better bioavailability.
为使本发明的目的、技术方案、及优点更加清楚明白，以下结合实施例用于进一步描述本发明，但这些实施例并非限制着本发明的范围。显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。本领域普通技术人员基于本发明中的实施例所获得的所有其他实施例，都属于本发明保护的范围。本发明实施例中未注明具体条件的实验方法，通常按照常规条件，或按照原料或商品制造厂商所建议的条件。未注明具体来源的试剂，为市场购买的常规试剂。In order to make the objectives, technical solutions, and advantages of the present invention clearer, the following embodiments are used to further describe the present invention, but these embodiments do not limit the scope of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by a person of ordinary skill in the art based on the embodiments of the present invention fall within the protection scope of the present invention. The experimental methods that do not specify specific conditions in the embodiments of the present invention usually follow conventional conditions or the conditions recommended by raw material or commodity manufacturers. The reagents without specific sources are the conventional reagents purchased on the market.
中间体制备方法实施例：Examples of intermediate preparation methods:
一、4-(4-甲基哌嗪-1-基)-2-(2,2,2-三氟-N-(四氢吡喃-4-基)乙酰胺基)苯甲酸I-1的制备1. 4-(4-Methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydropyran-4-yl)acetamido)benzoic acid I-1 Preparation
第一步 4-氟-2-硝基-苯甲酸叔丁酯I-1b的制备The first step is the preparation of 4-fluoro-2-nitro-benzoic acid tert-butyl ester I-1b
化合物4-氟-2-硝基-苯甲酸I-1a(10g，54mmol)、Boc酸酐(23.6g，108mmol)和DMAP(2g，16.2mmol)溶于DCM(100mL)和叔丁醇(100mL)的混合溶液，室温下搅拌过夜，浓缩，加入500mL乙酸乙酯稀释，1N HCl(500mL*1)洗涤，水(500mL*1)洗涤，饱和食盐水(500mL*1)洗涤，无水Na
2SO
4干燥，过滤，浓缩等到标题化合物I-1b(13g，54mmol)，收率100％。
Compound 4-fluoro-2-nitro-benzoic acid I-1a (10g, 54mmol), Boc anhydride (23.6g, 108mmol) and DMAP (2g, 16.2mmol) were dissolved in DCM (100mL) and tert-butanol (100mL) The mixed solution was stirred overnight at room temperature, concentrated, diluted with 500mL ethyl acetate, washed with 1N HCl (500mL*1), washed with water (500mL*1), washed with saturated brine (500mL*1), anhydrous Na 2 SO 4 Dry, filter, concentrate and wait until the title compound I-1b (13 g, 54 mmol), the yield is 100%.
第二步 4-(4-甲基哌嗪-1-基)-2-硝基苯甲酸叔丁酯I-1c的制备The second step is the preparation of tert-butyl 4-(4-methylpiperazin-1-yl)-2-nitrobenzoate I-1c
将化合物2(13g，54mmol)加入到1-甲基哌嗪(17mL)中，室温反应过夜。加入400mL水继续搅拌1小时，过滤，加水洗涤，干燥，得到标题化合物I-1c(16g，49.8mmol)，收率92.4％。Compound 2 (13 g, 54 mmol) was added to 1-methylpiperazine (17 mL) and reacted at room temperature overnight. 400 mL of water was added to continue stirring for 1 hour, filtered, washed with water, and dried to obtain the title compound I-1c (16 g, 49.8 mmol) with a yield of 92.4%.
MS m/z(ESI):322[M+H]
+。
MS m/z (ESI): 322 [M+H] + .
第三步 4-(4-甲基哌嗪-1-基)-2-胺基苯甲酸叔丁酯I-1d的制备The third step is the preparation of tert-butyl 4-(4-methylpiperazin-1-yl)-2-aminobenzoate I-1d
化合物I-1c(16g，49.8mmol)溶于甲醇(200mL)中，依次加入水(200mL)，氯化铵(10.7g，200mmol)和锌粉(35g，538mmol)，室温反应过夜。加入800mL水稀释，加入乙酸乙酯(300mL╳2)萃取，和并有机相，无水硫酸钠干燥，过滤，浓缩，Flash柱层析纯化(PE/EA，0～60％)，得标题化合物I-1d(5.9g，20.3mmol)，收率40.7％。Compound I-1c (16g, 49.8mmol) was dissolved in methanol (200mL), water (200mL), ammonium chloride (10.7g, 200mmol) and zinc powder (35g, 538mmol) were added sequentially, and reacted overnight at room temperature. Dilute with 800mL water, add ethyl acetate (300mL╳2) for extraction, combine the organic phase, dry with anhydrous sodium sulfate, filter, concentrate, and purify by Flash column chromatography (PE/EA, 0～60%) to obtain the title compound I-1d (5.9g, 20.3mmol), the yield is 40.7%.
MS m/z(ESI):292[M+H]
+。
MS m/z (ESI): 292 [M+H] + .
第四步 4-(4-甲基哌嗪-1-基)-2-(四氢吡喃-4-基)胺基苯甲酸叔丁酯I-1e的制备The fourth step is the preparation of tert-butyl 4-(4-methylpiperazin-1-yl)-2-(tetrahydropyran-4-yl)aminobenzoate I-1e
化合物I-1d(5g，17.2mmol)溶于DCM(100mL)中，加入四氢吡喃-4-酮(5g，50mmol)和乙酸(5mL)，室温搅拌30分钟后，加入三乙酰氧基硼氢化钠(15g，71.1mmol)，室温搅拌过夜。浓缩，Flash柱层析纯化(DCM/MeOH，0～10％)，得标题化合物I-1e(6g，16mmol)，收率93.1％。Compound I-1d (5g, 17.2mmol) was dissolved in DCM (100mL), tetrahydropyran-4-one (5g, 50mmol) and acetic acid (5mL) were added, and after stirring for 30 minutes at room temperature, triacetoxyboron was added Sodium hydride (15 g, 71.1 mmol) was stirred at room temperature overnight. Concentrated and purified by Flash column chromatography (DCM/MeOH, 0-10%) to obtain the title compound I-1e (6 g, 16 mmol) with a yield of 93.1%.
MS m/z(ESI):376[M+H]
+。
MS m/z (ESI): 376 [M+H] + .
第五步 4-(4-甲基哌嗪-1-基)-2-(2，2，2-三氟-N-(四氢吡喃-4-基)乙酰胺基)苯甲酸叔丁酯I-1f的制备The fifth step 4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydropyran-4-yl)acetamido) tert-butyl benzoate Preparation of ester I-1f
化合物I-1e(5g，13.3mmol)溶于DCM(50mL)中，加入三乙胺(2.3g，23mmol)和三氟乙酸酐(4.2g，20mmol)，冰浴搅拌20分钟后，滴加30mL水，分液，有机相用饱和食盐水(30mL)洗涤，无水Na
2SO
4干燥，过滤，浓缩，Flash柱层析纯化(DCM/MeOH，0～5％)，得标题化合物I-1f(5.3g，11.2mmol)，收率84.4％。
Compound I-1e (5g, 13.3mmol) was dissolved in DCM (50mL), triethylamine (2.3g, 23mmol) and trifluoroacetic anhydride (4.2g, 20mmol) were added, and after stirring for 20 minutes in an ice bath, 30mL was added dropwise Separate the layers with water, wash the organic phase with saturated brine (30 mL), dry with anhydrous Na 2 SO 4 , filter, concentrate, and purify by Flash column chromatography (DCM/MeOH, 0～5%) to obtain the title compound I-1f (5.3g, 11.2mmol), the yield is 84.4%.
MS m/z(ESI):472[M+H]
+。
MS m/z (ESI): 472 [M+H] + .
第六步 4-(4-甲基哌嗪-1-基)-2-(2,2,2-三氟-N-(四氢吡喃-4-基)乙酰胺基)苯甲酸I-1的制备The sixth step 4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydropyran-4-yl)acetamido)benzoic acid 1- 1 preparation
化合物I-1f(5g，10.6mmol)溶于DCM(100mL)中，加入三氟乙酸(25mL)，室温反应2小时后，浓缩，加入甲基叔丁基醚(200mL)继续搅拌15分钟，过滤，甲基叔丁基醚洗涤，干燥，得标题化合物I-1(4.6g，11.1mmol)，收率97.3％.Compound I-1f (5g, 10.6mmol) was dissolved in DCM (100mL), trifluoroacetic acid (25mL) was added, and after reacting at room temperature for 2 hours, it was concentrated, and methyl tert-butyl ether (200mL) was added and stirred for 15 minutes, filtered , Washed with methyl tert-butyl ether and dried to obtain the title compound I-1 (4.6g, 11.1mmol), with a yield of 97.3%.
1H NMR(400MHz,DMSO-d
6)δ12.77(s,1H),10.03(s,1H),7.87(d,J＝8.9Hz,1H),7.08(dd,J
1＝8.9,J
2＝2.6Hz,1H),6.94(d,J＝2.6Hz,1H),4.44(m,1H),4.06(s,2H),3.79(m,2H),3.59–3.04(m,8H),2.83(s,3H),1.92(m,1H),1.56(m,1H),1.47(m,1H),1.01(m,1H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.77 (s, 1H), 10.03 (s, 1H), 7.87 (d, J = 8.9 Hz, 1H), 7.08 (dd, J 1 ＝8.9, J 2 =2.6Hz, 1H), 6.94 (d, J = 2.6Hz, 1H), 4.44 (m, 1H), 4.06 (s, 2H), 3.79 (m, 2H), 3.59–3.04 (m, 8H), 2.83 (s, 3H), 1.92 (m, 1H), 1.56 (m, 1H), 1.47 (m, 1H), 1.01 (m, 1H).
MS m/z(ESI):416[M+H]+。MS m/z(ESI):416[M+H]+.
二、3-氨基-5-((3,5-二氟苯基)甲基)-1H-吲唑I-2的制备2. Preparation of 3-amino-5-((3,5-difluorophenyl)methyl)-1H-indazole I-2
第一步 (3-氰基-4-氟苯基)-(3,5-二氟苯基)甲醇I-2c的制备The first step is the preparation of (3-cyano-4-fluorophenyl)-(3,5-difluorophenyl)methanol I-2c
3,5-二氟溴苯I-2b(3.8g,20mmol)分批滴加到回流的含有镁屑(0.48g,20mmol)的干燥四氢呋喃(30mL)悬浮液中，回流3小时后冷至室温，滴加I-2a(3g,20mmol)干燥四氢呋喃(30mL)溶液中，温度不超过-15℃，加完后搅拌30分钟，升至0℃，加入饱和氯化铵溶液猝灭反应，减压蒸去部分四氢呋喃，乙酸乙酯萃取，干燥、浓缩、柱层析得产物I-2c(3.6g,13.7mmol),收率64％。3,5-Difluorobromobenzene I-2b (3.8g, 20mmol) was added dropwise to the refluxing dry tetrahydrofuran (30mL) suspension containing magnesium chips (0.48g, 20mmol), refluxed for 3 hours and then cooled to room temperature , Add dropwise I-2a (3g, 20mmol) to dry tetrahydrofuran (30mL) solution, the temperature does not exceed -15℃, stir for 30 minutes after the addition, raise to 0℃, add saturated ammonium chloride solution to quench the reaction, reduce pressure Part of the tetrahydrofuran was evaporated, extracted with ethyl acetate, dried, concentrated, and column chromatography was used to obtain the product I-2c (3.6 g, 13.7 mmol) with a yield of 64%.
MS m/z(ESI):264[M+H]
+。
MS m/z(ESI): 264[M+H] + .
第二步 (3-氰基-4-氟苯基)-(3,5-二氟苯基)甲烷I-2d的制备The second step is the preparation of (3-cyano-4-fluorophenyl)-(3,5-difluorophenyl)methane I-2d
化合物I-2c(5.26g,20mmol)溶于乙腈(100mL)中，加入碘化钠(30g,200mmol)后加热至65℃，6小时内分批加入三甲基氯硅烷(21.7g,200mmol)，冷至室温，加入硫代硫酸钠溶液搅拌至无色，乙酸乙酯萃取，干燥浓缩的白色固体产物I-2d(3.2g,13mmol),收率65％。Compound I-2c (5.26g, 20mmol) was dissolved in acetonitrile (100mL), added sodium iodide (30g, 200mmol) and heated to 65°C, added trimethylchlorosilane (21.7g, 200mmol) in batches within 6 hours , Cooled to room temperature, added sodium thiosulfate solution and stirred until colorless, extracted with ethyl acetate, dried and concentrated the white solid product I-2d (3.2g, 13mmol), the yield was 65%.
MS m/z(ESI):248[M+H]
+。
MS m/z (ESI): 248 [M+H] + .
第三步 3-氨基-5-((3,5-二氟苯基)甲基)-1H-吲唑I-2的制备The third step Preparation of 3-amino-5-((3,5-difluorophenyl)methyl)-1H-indazole I-2
化合物I-2d(3g,12mmol)、80％的水合肼(3.8g,60mmol)加到N-甲基吡咯烷酮(15mL)中，150℃加热4小时，冷至室温，加入25mL水，搅拌至固体析出，过滤的白色固体产物I-2(2.2g,8.5mmol)，收率71％。Compound I-2d (3g, 12mmol) and 80% hydrazine hydrate (3.8g, 60mmol) were added to N-methylpyrrolidone (15mL), heated at 150°C for 4 hours, cooled to room temperature, added 25mL of water, and stirred until solid Precipitation, the filtered white solid product I-2 (2.2 g, 8.5 mmol), the yield was 71%.
MS m/z(ESI):260[M+H]
+。
MS m/z (ESI): 260 [M+H] + .
1H NMR(400MHz,DMSO-d
6)δ11.28(s,1H),7.49(s,1H),7.17–7.05(m,2H),6.99(tt,J＝9.4,2.4Hz,1H),6.94–6.85(m,2H),5.21(s,2H),3.96(s,2H).
1 H NMR(400MHz,DMSO-d 6 )δ11.28(s,1H),7.49(s,1H),7.17-7.05(m,2H),6.99(tt,J=9.4,2.4Hz,1H), 6.94--6.85(m,2H), 5.21(s,2H), 3.96(s,2H).
产物制备方法实施例：Examples of product preparation methods:
实施例1Example 1
N-(5-(2-(2,5-二氟苯基)吡咯烷-1-基)吡唑并[1,5-a]嘧啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺1的制备N-(5-(2-(2,5-Difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiper Preparation of azin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 1
第一步 5-氯-3-硝基吡唑并[1,5-a]嘧啶1b的制备The first step is the preparation of 5-chloro-3-nitropyrazolo[1,5-a]pyrimidine 1b
5-氯吡唑并[1,5-a]嘧啶1a(2g，13mmol)溶于H
2SO
4(8mL)，冰浴下加入KNO
3(2g，19.6mmol)，在搅拌下慢慢升温至室温。混合物慢慢倒入冰水中，有黄色固体析出，过滤，收集固体并干燥得5-氯-3-硝基吡唑并[1,5-a]嘧啶1b(2g，10.1mmol)，收率77％。
5-Chloropyrazolo[1,5-a]pyrimidine 1a (2g, 13mmol) was dissolved in H 2 SO 4 (8mL), KNO 3 (2g, 19.6mmol) was added under ice bath, and the temperature was slowly heated to Room temperature. The mixture was slowly poured into ice water, a yellow solid precipitated out, filtered, the solid was collected and dried to obtain 5-chloro-3-nitropyrazolo[1,5-a]pyrimidine 1b (2g, 10.1mmol), the yield was 77 %.
1H NMR(400MHz,DMSO-d
6)δ9.43(d,J＝7.2Hz,1H),9.07(s,1H),7.61(d,J＝7.2Hz,1H).
1 H NMR (400MHz, DMSO-d 6 ) δ 9.43 (d, J = 7.2 Hz, 1H), 9.07 (s, 1H), 7.61 (d, J = 7.2 Hz, 1H).
第二步 5-(2-(2,5-二氟苯基)吡咯烷-1-基)-3-硝基吡唑并[1,5-a]嘧啶1d的制备The second step is the preparation of 5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)-3-nitropyrazolo[1,5-a]pyrimidine 1d
将2-(2,5-二氟苯基)吡咯烷1c(600mg,3.27mmol)和三乙胺(990mg,9.81mmol)溶于THF中，在室温下滴加5-氯-3-硝基吡唑并[1,5-a]嘧啶1b(650mg,3.27mmol)的THF溶液。滴加完毕，在室温下继续搅拌2小时。加入淬灭，用乙酸乙酯萃取3次，合并有机相，有机相用饱和食盐水洗，用无水硫酸钠干燥，过滤，浓缩，粗品用柱层析纯化(PE：EA＝1:1)得到5-(2-(2,5-二氟苯基)吡咯烷-1-基)-3- 硝基吡唑并[1,5-a]嘧啶1d(0.9g,2.6mmol)呈黄色固体，收率79％。Dissolve 2-(2,5-difluorophenyl)pyrrolidine 1c (600mg, 3.27mmol) and triethylamine (990mg, 9.81mmol) in THF, add 5-chloro-3-nitro group dropwise at room temperature Pyrazolo[1,5-a]pyrimidine 1b (650 mg, 3.27 mmol) in THF. After the addition is complete, continue stirring at room temperature for 2 hours. It was quenched by addition, extracted 3 times with ethyl acetate, combined the organic phases, washed the organic phases with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography (PE:EA=1:1) to obtain 5-(2-(2,5-Difluorophenyl)pyrrolidin-1-yl)-3-nitropyrazolo[1,5-a]pyrimidine 1d (0.9g, 2.6mmol) is a yellow solid, The yield was 79%.
MS m/z(ESI):346[M+H]
+。
MS m/z (ESI): 346 [M+H] + .
第三步 5-(2-(2,5-二氟苯基)吡咯烷-1-基)吡唑并[1,5-a]嘧啶-3-胺1e的制备The third step is the preparation of 5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-amine 1e
将5-(2-(2,5-二氟苯基)吡咯烷-1-基)-3-硝基吡唑并[1,5-a]嘧啶1d(400mg,1.15mmol)和锌粉(376mg,5.79mmol)分散在THF(5mL)中，加入饱和氯化铵水溶液3mL。该混合物在室温下搅拌1小时，过滤，滤液用乙酸乙酯萃取2次，有机相用饱和食盐水洗，用无水硫酸钠干燥，过滤，浓缩，粗品用柱层析纯化(DCM:MeOH＝10:1)得到5-(2-(2,5-二氟苯基)吡咯烷-1-基)吡唑并[1,5-a]嘧啶-3-胺1e(200mg,0.63mmol)呈淡黄色固体，收率55％。Combine 5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)-3-nitropyrazolo[1,5-a]pyrimidine 1d (400mg, 1.15mmol) and zinc powder ( 376 mg, 5.79 mmol) was dispersed in THF (5 mL), and 3 mL of saturated aqueous ammonium chloride was added. The mixture was stirred at room temperature for 1 hour, filtered, the filtrate was extracted twice with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography (DCM:MeOH=10 : 1) Obtain 5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-amine 1e (200mg, 0.63mmol) in light Yellow solid, the yield is 55%.
MS m/z(ESI):316[M+H]
+。
MS m/z (ESI): 316 [M+H] + .
第四步 4-(4-甲基哌嗪-1-基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯甲酰氯1f的制备The fourth step 4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzyl Preparation of acid chloride 1f
将4-(4-甲基哌嗪-1-基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯甲酸I-1(150mg,0.36mmol)溶于DCM中，加入草酰氯(229mg,1.8mmol)和一滴DMF,该混合物在回流状态下搅拌1小时，反应液浓缩，粗品1f直接用于下一步反应。The 4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoic acid 1- 1 (150 mg, 0.36 mmol) was dissolved in DCM, oxalyl chloride (229 mg, 1.8 mmol) and a drop of DMF were added, the mixture was stirred under reflux for 1 hour, the reaction solution was concentrated, and the crude 1f was directly used in the next reaction.
第五步 N-(5-(2-(2,5-二氟苯基)吡咯烷-1-基)吡唑并[1,5-a]嘧啶-3-基)-4-(4-甲基哌嗪-1-基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯酰胺1g的制备Step 5 N-(5-(2-(2,5-Difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4- Preparation of methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 1g
将5-(2-(2,5-二氟苯基)吡咯烷-1-基)吡唑并[1,5-a]嘧啶-3-胺1e(100mg,0.31mmol)和三乙胺(93mg,0.92mmol)溶于DCM中，室温下滴加4-(4-甲基哌嗪-1-基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯甲酰氯1f(165mg,0.38mmol)的DCM溶液，反应混合物在室温下搅拌1小时，反应液浓缩，粗品1g直接用于下一步反应。Combine 5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-amine 1e (100mg, 0.31mmol) and triethylamine ( 93mg, 0.92mmol) was dissolved in DCM, 4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyridine) was added dropwise at room temperature A DCM solution of pyran-4-yl)acetamido)benzoyl chloride 1f (165mg, 0.38mmol), the reaction mixture was stirred at room temperature for 1 hour, the reaction solution was concentrated, and the crude product 1g was directly used in the next reaction.
第六步 N-(5-(2-(2,5-二氟苯基)吡咯烷-1-基)吡唑并[1,5-a]嘧啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺1的制备Step 6 N-(5-(2-(2,5-Difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4- Preparation of methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 1
将N-(5-(2-(2,5-二氟苯基)吡咯烷-1-基)吡唑并[1,5-a]嘧啶-3-基)-4-(4-甲基哌 嗪-1-基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯酰胺1g(150mg,0.21mmol)溶于甲醇中，加入5N K
2CO
3(4mL),该反应混合物在室温下搅拌2小时，反应液用乙酸乙酯萃取，有机相合并，用饱和食盐水洗，用无水硫酸钠干燥，过滤，浓缩，粗品用柱层析纯化(DCM：MeOH＝10:1)得到N-(5-(2-(2,5-二氟苯基)吡咯烷-1-基)吡唑并[1,5-a]嘧啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺1(60mg,0.097mmol)呈杏色固体，收率46％。
N-(5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methyl Piperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 1g (150mg, 0.21mmol) dissolved in methanol 5N K 2 CO 3 (4 mL) was added, the reaction mixture was stirred at room temperature for 2 hours, the reaction solution was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried with anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography (DCM:MeOH=10:1) to obtain N-(5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a ]Pyrimidin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 1 (60mg, 0.097mmol) It is an apricot solid with a yield of 46%.
MS m/z(ESI):617[M+H]
+
MS m/z(ESI):617[M+H] +
1H NMR(400MHz,DMSO-d6)δ9.03(br,1H),8.49(br,1H),8.15(br,1H),7.92(s,1H),7.59(br,1H),7.31–7.02(m,3H),6.83(br,1H),6.20(dd,J＝8.9,2.2Hz,1H),6.10(d,J＝2.3Hz,1H),5.34(br,1H),3.94–3.85(m,1H),3.78(dt,J＝11.6,3.9Hz,2H),3.68-3.53(m,2H),3.49–3.40(m,2H),3.21(t,J＝5.1Hz,4H),2.40(t,J＝5.0Hz,4H),2.19(s,3H),2.02-1.81(m,6H),1.37–1.25(m,2H).
1 H NMR(400MHz,DMSO-d6)δ9.03(br,1H), 8.49(br,1H), 8.15(br,1H), 7.92(s,1H), 7.59(br,1H), 7.31-7.02 (m, 3H), 6.83 (br, 1H), 6.20 (dd, J = 8.9, 2.2 Hz, 1H), 6.10 (d, J = 2.3 Hz, 1H), 5.34 (br, 1H), 3.94–3.85 ( m, 1H), 3.78 (dt, J = 11.6, 3.9 Hz, 2H), 3.68-3.53 (m, 2H), 3.49-3.40 (m, 2H), 3.21 (t, J = 5.1 Hz, 4H), 2.40 (t,J=5.0Hz,4H), 2.19(s,3H), 2.02-1.81(m,6H), 1.37-1.25(m,2H).
实施例2Example 2
N-(6-(2-(2,5-二氟苯基)吡咯烷-1-基)咪唑并[1,2-b]哒嗪-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺2的制备N-(6-(2-(2,5-Difluorophenyl)pyrrolidin-1-yl)imidazo[1,2-b]pyridazin-3-yl)-4-(4-methylpiper Preparation of azin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 2
采用与实施例1相同的方法制备，以6-氯-3-硝基咪唑并[1,2-b]哒嗪为原料，代替1b。It was prepared by the same method as in Example 1, using 6-chloro-3-nitroimidazo[1,2-b]pyridazine as a raw material instead of 1b.
MS m/z(ESI):617[M+H]+MS m/z(ESI):617[M+H]+
1H NMR(400MHz,DMSO-d6)δ9.23(s,1H),7.59(br,4H),7.21-7.05(m,3H),6.91–6.86(m,1H),6.16(br,1H),6.07(br,1H),5.26(d,J＝8.1Hz,1H),3.96– 3.85(s,1H),3.81(d,J＝11.4Hz,2H),3.68–3.58(m,2H),3.44(t,J＝10.7Hz,2H),3.20(t,J＝5.1Hz,4H),2.41(t,J＝5.1Hz,4H),2.19(s,3H),2.01–1.79(m,6H),1.52-1.30(m,2H).
1 H NMR(400MHz,DMSO-d6)δ9.23(s,1H),7.59(br,4H),7.21-7.05(m,3H),6.91-6.86(m,1H),6.16(br,1H) ,6.07(br,1H), 5.26(d,J=8.1Hz,1H), 3.96– 3.85(s,1H), 3.81(d,J=11.4Hz,2H), 3.68–3.58(m,2H), 3.44 (t, J = 10.7 Hz, 2H), 3.20 (t, J = 5.1 Hz, 4H), 2.41 (t, J = 5.1 Hz, 4H), 2.19 (s, 3H), 2.01–1.79 (m, 6H ), 1.52-1.30 (m, 2H).
实施例3Example 3
N-(5-(2-(2,5-二氟苯基)吡咯烷-1-基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺3的制备N-(5-(2-(2,5-Difluorophenyl)pyrrolidin-1-yl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl) Preparation of -2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 3
第一步 叔丁基-3-[二(叔丁氧基羰基)氨基]-5-溴-1H-吲唑-1-羧酸酯3b的制备The first step: Preparation of tert-butyl-3-[bis(tert-butoxycarbonyl)amino]-5-bromo-1H-indazole-1-carboxylate 3b
将化合物3a(1.0g，4.7mmol)，(Boc)
2O(3.1g，14.1mmol)和DMAP(0.57g，4.7mmol)投于DCM(20ml)中，室温反应过夜。减压蒸除溶剂，加入EA(50ml)，依次用satNH4Cl(15ml×2)和brine(20ml)洗涤，经无水硫酸钠干燥，浓缩，flash柱纯化(石油醚～石油醚/乙酸乙酯＝20:1)得白色泡沫状固体化合物3b(1.5g)，收率为63％。
Compound 3a (1.0 g, 4.7 mmol), (Boc) 2 O (3.1 g, 14.1 mmol) and DMAP (0.57 g, 4.7 mmol) were added to DCM (20 ml) and reacted at room temperature overnight. The solvent was evaporated under reduced pressure, EA (50ml) was added, washed with satNH4Cl (15ml×2) and brine (20ml) successively, dried over anhydrous sodium sulfate, concentrated, and purified by flash column (petroleum ether ~ petroleum ether/ethyl acetate = 20:1) A white foamy solid compound 3b (1.5g) was obtained with a yield of 63%.
MS m/z(ESI):534[M+Na]
+
MS m/z(ESI):534[M+Na] +
第二步 5-(2-(2,5-二氟苯基)吡咯烷-1-基)-1-(N-叔丁氧羰基)-吲唑-3-(二(叔丁 氧基羰基))胺3c的制备The second step 5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)-1-(N-tert-butoxycarbonyl)-indazole-3-(bis(tert-butoxycarbonyl) )) Preparation of amine 3c
将化合物3b(1.3g，2.5mmol)，化合物1c(0.38g，2.1mmol)和Cs
2CO
3(1.4g，4.2mmol)投于二氧六环(30ml)中，依次加入Pd
2(dba)
3(0.19g，0.21mmol)和Ruphos(0.20g，0.42mmol)，92℃反应4h。冷至室温，加入水(20ml)，用乙酸乙酯(50ml×2)萃取，合并有机层，用brine(20ml)洗涤，经无水硫酸钠干燥，浓缩，flash柱纯化(石油醚～石油醚/乙酸乙酯＝5:1)得白色泡沫状固体，化合物3c(0.8g)，收率为62％。
Put compound 3b (1.3g, 2.5mmol), compound 1c (0.38g, 2.1mmol) and Cs 2 CO 3 (1.4g, 4.2mmol) into dioxane (30ml), add Pd 2 (dba) in sequence 3 (0.19g, 0.21mmol) and Ruphos (0.20g, 0.42mmol), react at 92°C for 4h. Cool to room temperature, add water (20ml), extract with ethyl acetate (50ml×2), combine the organic layers, wash with brine (20ml), dry over anhydrous sodium sulfate, concentrate, and purify by flash column (petroleum ether ~ petroleum ether) /Ethyl acetate=5:1) to obtain a white foamy solid, compound 3c (0.8g), with a yield of 62%.
MS m/z(ESI):615[M+1]
+
MS m/z(ESI):615[M+1] +
第三步 5-(2-(2,5-二氟苯基)吡咯烷-1-基)-1H-吲唑-3-胺3d的制备The third step is the preparation of 5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)-1H-indazol-3-amine 3d
将化合物3c(0.8g，1.3mmol)投于4M HCl/二氧六环(20ml)中，室温反应过夜。减压蒸干得黄色固体，化合物3d(0.41g)，收率为100％。Compound 3c (0.8g, 1.3mmol) was added to 4M HCl/dioxane (20ml) and reacted at room temperature overnight. Evaporate to dryness under reduced pressure to obtain a yellow solid, compound 3d (0.41 g), with a yield of 100%.
MS m/z(ESI):315[M+1]
+
MS m/z(ESI):315[M+1] +
第四步 N-(5-(2-(2,5-二氟苯基)吡咯烷-1-基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯酰胺3e的制备The fourth step N-(5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)-1H-indazol-3-yl)-4-(4-methylpiperazine-1 -Yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 3e
将化合物3d(50mg，0.16mmol)和DIEA(83mg，0.64mmol)投于无水THF(5ml)中，冷制-20℃，将1f(100mg)溶于无水THF(1ml)中，用注射器取0.7ml滴加至上述溶液中，继续反应2h。加入水(10ml)，用乙酸乙酯(15ml×2)萃取，合并有机层，用brine(15ml)洗涤，经无水硫酸钠干燥，浓缩，flash柱纯化(DCM～DCM/MeOH＝6：1)得黄色固体，化合物3e(30mg)，收率为26％。Put compound 3d (50mg, 0.16mmol) and DIEA (83mg, 0.64mmol) in anhydrous THF (5ml), cool to -20℃, dissolve 1f (100mg) in anhydrous THF (1ml), use a syringe Take 0.7ml dropwise to the above solution, continue to react for 2h. Water (10ml) was added, extracted with ethyl acetate (15ml×2), the organic layers were combined, washed with brine (15ml), dried over anhydrous sodium sulfate, concentrated, and purified by flash column (DCM～DCM/MeOH=6:1 ) To obtain a yellow solid, compound 3e (30 mg), with a yield of 26%.
第五步 N-(5-(2-(2,5-二氟苯基)吡咯烷-1-基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺3的制备Step 5 N-(5-(2-(2,5-Difluorophenyl)pyrrolidin-1-yl)-1H-indazol-3-yl)-4-(4-methylpiperazine-1 -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 3
将化合物3e(30mg，0.042mmol)投于MeOH(6ml)和TEA(ml)中，回流反应2h，减压蒸干溶剂，加入乙酸乙酯(20ml)，依次用水(8ml)和brine(10ml)洗涤你，经无水硫酸钠干燥，浓缩，flash柱纯化(DCM～DCM/MeOH＝6：1)得近白色固体，终产物3(15mg)。Put compound 3e (30mg, 0.042mmol) in MeOH (6ml) and TEA (ml), reflux for 2h, evaporate the solvent under reduced pressure, add ethyl acetate (20ml), water (8ml) and then brine (10ml) Wash you, dry over anhydrous sodium sulfate, concentrate, and purify by flash column (DCM～DCM/MeOH=6:1) to obtain a nearly white solid, the final product 3 (15mg).
MS m/z(ESI):616[M+1]
+
MS m/z(ESI):616[M+1] +
1H NMR(400MHz,DMSO-d
6)δ12.32(s,1H),9.94(s,1H),8.28(d,J＝7.7Hz,1H),7.76(d,J＝9.0Hz,1H),7.31–7.19(m,2H),7.14–7.04(m,1H),6.87(m,1H),6.66(dd,J＝9.1,2.3Hz,1H),6.49(d,J＝2.2Hz,1H),6.22(dd,J＝9.0,2.3Hz,1H),6.14(d,J＝2.3Hz,1H),4.91(d,J＝8.3Hz,1H),3.83(m,2H),3.71(m,2H),3.56–3.45(m,2H),3.27(m,5H),2.48–2.36(m,4H),2.24(s,3H),2.05–1.80(m,5H),1.42–1.25(m,3H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.32 (s, 1H), 9.94 (s, 1H), 8.28 (d, J = 7.7 Hz, 1H), 7.76 (d, J = 9.0 Hz, 1H) ,7.31–7.19(m,2H),7.14–7.04(m,1H),6.87(m,1H),6.66(dd,J=9.1,2.3Hz,1H),6.49(d,J=2.2Hz,1H ), 6.22 (dd, J = 9.0, 2.3 Hz, 1H), 6.14 (d, J = 2.3 Hz, 1H), 4.91 (d, J = 8.3 Hz, 1H), 3.83 (m, 2H), 3.71 (m , 2H), 3.56–3.45(m, 2H), 3.27(m, 5H), 2.48–2.36(m, 4H), 2.24(s, 3H), 2.05–1.80(m, 5H), 1.42–1.25(m ,3H).
实施例4Example 4
5-(3,5-二氟苯基)-3-(2’-(4-甲基哌嗪-1-基)-[2,4’-联吡啶]-6-基)-1H-吲唑4的制备5-(3,5-Difluorophenyl)-3-(2'-(4-methylpiperazin-1-yl)-[2,4'-bipyridine]-6-yl)-1H-indyl Preparation of azole 4
第一步 5-溴-1-三苯基-1H-吲唑4b的制备The first step is the preparation of 5-bromo-1-triphenyl-1H-indazole 4b
化合物4a(23g，117mmol)和三乙胺(28g，280mmol)溶于二氯甲烷(200ml) 和THF(40mL)，向溶液中加入三苯基氯甲烷(39g，140mmol)。反应液在室温下搅拌过夜。反应液分别用水和饱和食盐水洗涤，有机相用无水硫酸钠干燥，过滤浓缩得到化合物4b(51g,117mmol)，收率100％。Compound 4a (23g, 117mmol) and triethylamine (28g, 280mmol) were dissolved in dichloromethane (200ml) and THF (40mL), and triphenylchloromethane (39g, 140mmol) was added to the solution. The reaction solution was stirred overnight at room temperature. The reaction solution was washed with water and saturated brine, and the organic phase was dried with anhydrous sodium sulfate, filtered and concentrated to obtain compound 4b (51 g, 117 mmol) with a yield of 100%.
MS m/z(ESI):243[M-195]
+。
MS m/z (ESI): 243[M-195] + .
第二步 (3,5-二氟苯基)(1-三苯基-1H-吲唑-5-基)甲醇4c的制备The second step is the preparation of (3,5-difluorophenyl)(1-triphenyl-1H-indazol-5-yl)methanol 4c
化合物4b(13.2g，30mmol)溶于四氢呋喃(100mL)，将溶液冷却至-78℃,向溶液中滴加正丁基锂(14.4ml，36mmol，2.5M in hexane)。反应液在-78℃下搅拌1h。然后向反应液中滴加3,5-二氟苯甲醛(4.7g,33mmol)的四氢呋喃(50ml)溶液。然后反应液在-78℃继续搅拌1h。反应液用饱合氯化铵溶液淬灭，用二氯甲烷(100ml*2)萃取。合并有机相，用无水硫酸钠干燥，过滤浓缩得到粗产品，经过柱层析纯化后得到标题化合物4c(6.5g,12.9mmol)，收率43％。Compound 4b (13.2g, 30mmol) was dissolved in tetrahydrofuran (100mL), the solution was cooled to -78°C, and n-butyllithium (14.4ml, 36mmol, 2.5M in hexane) was added dropwise to the solution. The reaction solution was stirred at -78°C for 1 h. Then, a solution of 3,5-difluorobenzaldehyde (4.7 g, 33 mmol) in tetrahydrofuran (50 ml) was added dropwise to the reaction solution. Then the reaction solution was stirred at -78°C for 1 h. The reaction solution was quenched with saturated ammonium chloride solution and extracted with dichloromethane (100ml*2). The organic phases were combined, dried with anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography to obtain the title compound 4c (6.5 g, 12.9 mmol) with a yield of 43%.
MS m/z(ESI):282[M-243+Na]
+。
MS m/z (ESI): 282 [M-243+Na] + .
第三步(3,5-二氟苯基)-(1H-吲唑-5-基)甲醇4d的制备The third step is the preparation of (3,5-difluorophenyl)-(1H-indazol-5-yl)methanol 4d
化合物4c(6g,12mmol)和碘化钠(9g,60mmol)加入到乙腈(150ml)中，溶液冷却至0℃，向溶液中分批加入三甲基氯硅烷(6.5g,60mmol)，反应液在常温下搅拌2小时，然后在0℃用亚硫酸氢钠淬灭，旋干后加入水(50ml)，用乙酸乙酯(100ml*2)萃取，合并有机相，用无水硫酸钠干燥，过滤浓缩得到粗产品，经过柱层析纯化后得到标题化合物4d(1.7g,33mmol)，收率55％。Compound 4c (6g, 12mmol) and sodium iodide (9g, 60mmol) were added to acetonitrile (150ml), the solution was cooled to 0°C, trimethylchlorosilane (6.5g, 60mmol) was added to the solution in batches, the reaction solution Stir at room temperature for 2 hours, then quench with sodium bisulfite at 0°C, spin dry, add water (50ml), extract with ethyl acetate (100ml*2), combine the organic phases, and dry with anhydrous sodium sulfate. The crude product was filtered and concentrated to obtain the title compound 4d (1.7 g, 33 mmol) after purification by column chromatography with a yield of 55%.
MS m/z(ESI):261[M+H]
+。
MS m/z (ESI): 261 [M+H] + .
第四步 5-(3,5-二氟苯基)-1H-吲唑4e的制备Step 4 Preparation of 5-(3,5-difluorophenyl)-1H-indazole 4e
化合物4d(1.7g，6.54mmol)溶于三乙基硅烷(5ml)和三氟乙酸(20ml)，反应液在60℃搅拌3小时，反应液旋干后溶于乙酸乙酯(80ml)，用碳酸氢钠溶液洗涤。有机相用无水硫酸钠干燥，过滤旋干得到化合物4e(915mg，3.75mmol)，收率57％。Compound 4d (1.7g, 6.54mmol) was dissolved in triethylsilane (5ml) and trifluoroacetic acid (20ml). The reaction solution was stirred at 60°C for 3 hours. The reaction solution was spin-dried and dissolved in ethyl acetate (80ml). Wash with sodium bicarbonate solution. The organic phase was dried with anhydrous sodium sulfate, filtered and spin-dried to obtain compound 4e (915 mg, 3.75 mmol) with a yield of 57%.
MS m/z(ESI):245[M+H]
+。
MS m/z (ESI): 245 [M+H] + .
第五步 3-溴-5-(3,5-二氟苯基)-1H-吲唑4f的制备Step 5 Preparation of 3-bromo-5-(3,5-difluorophenyl)-1H-indazole 4f
化合物1e(2.0g，8.2mmol)溶于乙腈(30ml)，向溶液中加入N-溴代丁二酰亚胺(2.9g,16.4mmol)，反应液在82℃搅拌2小时，冷却后反应液旋干后得到粗品，经过柱层析纯化后得到标题化合物4f(1.8g,5.57mmol)，收率68％。Compound 1e (2.0g, 8.2mmol) was dissolved in acetonitrile (30ml), and N-bromosuccinimide (2.9g, 16.4mmol) was added to the solution. The reaction solution was stirred at 82°C for 2 hours. After cooling, the reaction solution After rotary drying, the crude product was obtained. After purification by column chromatography, the title compound 4f (1.8 g, 5.57 mmol) was obtained with a yield of 68%.
MS m/z(ESI):325[M+H]
+。
MS m/z (ESI): 325 [M+H] + .
第六步 3-溴-5-(3,5-二氟苯基)-1-((2-(三甲基硅基)乙氧基)甲基)-1H-吲唑4g的制备Step 6 Preparation of 3-bromo-5-(3,5-difluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazole 4g
化合物4f(1.4g,4.33mmol)溶于DMF(10ml)中，溶液冷却至0℃，向溶液中加入钠氢(208mg,5.2mmol)，反应液在0℃搅拌15分钟，加入氯甲基三硅基乙基醚(866mg,5.2mmol)，然后在常温下继续搅拌2小时，向反应液中加入饱和氯化铵溶液，用乙酸乙酯(50ml)萃取两遍。有机相用无水硫酸钠干燥，过滤浓缩得到粗品，经过柱层析纯化后得到化合物4g(1.7g,3.75mmol)，收率87％。Compound 4f (1.4g, 4.33mmol) was dissolved in DMF (10ml), the solution was cooled to 0°C, sodium hydrogen (208mg, 5.2mmol) was added to the solution, the reaction solution was stirred at 0°C for 15 minutes, and chloromethyl trifluoroacetate was added. Silyl ethyl ether (866mg, 5.2mmol), then continue to stir at room temperature for 2 hours, add saturated ammonium chloride solution to the reaction solution, and extract twice with ethyl acetate (50ml). The organic phase was dried with anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography to obtain 4 g (1.7 g, 3.75 mmol) of compound with a yield of 87%.
MS m/z(ESI):453[M+H]
+。
MS m/z (ESI): 453 [M+H] + .
第七步 5-(3,5-二氟苯基)-1-((2-(三甲基硅基)乙氧基)甲基)-3-(三甲基锡基)-1H-吲唑4h的制备The seventh step 5-(3,5-difluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-3-(trimethylstannyl)-1H-indyl Preparation of azole 4h
化合物4g(500mg,1.1mmol),六甲基二锡(1.1g,3.3mmol)和四三苯基磷钯(127mg,0.11mmol)悬混于1,4-二氧六环(20ml)中，反应液在110℃搅拌18h，反应液冷却至室温，加入KF溶液(3.3ml,1M),用乙酸乙酯(50ml)萃取。有机相用无水硫酸钠干燥，过滤浓缩得到粗品，经过柱层析纯化后得到化合物4h(480mg,0.89mmol)，收率81％。Compound 4g (500mg, 1.1mmol), hexamethyl ditin (1.1g, 3.3mmol) and palladium tetrakistriphenylphosphorus (127mg, 0.11mmol) were suspended in 1,4-dioxane (20ml), The reaction solution was stirred at 110°C for 18h, the reaction solution was cooled to room temperature, KF solution (3.3ml, 1M) was added, and it was extracted with ethyl acetate (50ml). The organic phase was dried with anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography to obtain compound 4h (480 mg, 0.89 mmol) with a yield of 81%.
MS m/z(ESI):539[M+H]
+。
MS m/z (ESI): 539 [M+H] + .
第八步 5-(3,5-二氟苯基)-3-(2’-(4-甲基哌嗪-1-基)-[2,4’-联吡啶]-6-基)-1-((2-(三甲基硅基)乙氧基)甲基)-1H-吲唑4i的制备Step 8 5-(3,5-Difluorophenyl)-3-(2'-(4-methylpiperazin-1-yl)-[2,4'-bipyridine]-6-yl)- Preparation of 1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazole 4i
化合物4h(200mg,0.37mmol),6-溴-2’-(4-甲基哌嗪-1-基)-2,4’-二吡啶4k(124mg,0.37mmol)，四三苯基磷钯(46mg,0.04mmol)，碘化亚铜(15mg,0.08mmol)和三乙胺(75mg，0.74mmol)悬混于DMF(6ml)中，反应液在氮气保护下 在100℃搅拌2h，反应液冷却至室温，加入水(50ml),用二氯甲烷(30ml)萃取两遍。有机相用无水硫酸钠干燥，过滤浓缩得到粗品，经过柱层析纯化后得到化合物4i(140mg,0.22mmol)，收率60％。Compound 4h (200mg, 0.37mmol), 6-bromo-2'-(4-methylpiperazin-1-yl)-2,4'-dipyridine 4k (124mg, 0.37mmol), palladium tetrakistriphenylphosphorus (46mg, 0.04mmol), cuprous iodide (15mg, 0.08mmol) and triethylamine (75mg, 0.74mmol) were suspended in DMF (6ml), the reaction solution was stirred at 100℃ for 2h under the protection of nitrogen, the reaction solution Cool to room temperature, add water (50ml), and extract twice with dichloromethane (30ml). The organic phase was dried with anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography to obtain compound 4i (140 mg, 0.22 mmol) with a yield of 60%.
MS m/z(ESI):627[M+H]
+。
MS m/z (ESI): 627 [M+H] + .
第九步 (5-(3,5-二氟苯基)-3-(2’-(4-甲基哌嗪-1-基)-[2,4’-联吡啶]-6-基)-1H-吲唑-1-基)甲醇4j的制备Step 9 (5-(3,5-Difluorophenyl)-3-(2'-(4-methylpiperazin-1-yl)-[2,4'-bipyridyl]-6-yl) Preparation of -1H-indazol-1-yl)methanol 4j
化合物4i(120mg,0.19mmol)溶于二氯甲烷(8ml),向溶液中加入三氟乙酸(4ml)，反应液在常温下搅拌2小时，将反应液旋干，得到的产品4j直接用于下一步反应。Compound 4i (120mg, 0.19mmol) was dissolved in dichloromethane (8ml), trifluoroacetic acid (4ml) was added to the solution, the reaction solution was stirred at room temperature for 2 hours, the reaction solution was spin-dried, and the obtained product 4j was used directly Next reaction.
MS m/z(ESI):527[M+H]
+。
MS m/z (ESI): 527 [M+H] + .
第十步 5-(3,5-二氟苯基)-3-(2’-(4-甲基哌嗪-1-基)-[2,4’-联吡啶]-6-基)-1氢-吲唑4的制备The tenth step 5-(3,5-difluorophenyl)-3-(2'-(4-methylpiperazin-1-yl)-[2,4'-bipyridyl]-6-yl)- 1 Preparation of Hydro-indazole 4
化合物4j(100mg,0.19mmol)溶于四氢呋喃(8ml)，向溶液中加入2N LiOH(4ml)，反应液在常温下搅拌过夜。加入水(20ml),用二氯甲烷(50ml)萃取两遍。有机相用无水硫酸钠干燥，过滤浓缩得到粗品，经过柱层析纯化后得到化合物4(20mg,0.04mmol)，收率21％。Compound 4j (100mg, 0.19mmol) was dissolved in tetrahydrofuran (8ml), 2N LiOH (4ml) was added to the solution, and the reaction solution was stirred overnight at room temperature. Add water (20ml) and extract twice with dichloromethane (50ml). The organic phase was dried with anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography to obtain compound 4 (20 mg, 0.04 mmol) with a yield of 21%.
MS m/z(ESI):497[M+H]
+。
MS m/z (ESI): 497 [M+H] + .
1H NMR(400MHz,Chloroform-d)δ8.69–8.65(m,1H),8.33(dd,J＝5.3,0.8Hz,1H),8.20(dd,J＝7.9,1.0Hz,1H),7.86(t,J＝7.8Hz,1H),7.71(dd,J＝7.8,0.9Hz,1H),7.57–7.51(m,1H),7.47(dd,J＝8.6,0.8Hz,1H),7.26(dd,J＝5.3,1.3Hz,1H),7.20(dd,J＝8.6,1.6Hz,1H),6.74–6.66(m,2H),6.62(tt,J＝9.0,2.4Hz,1H),4.10(s,2H),3.73(t,J＝5.0Hz,4H),2.56(t,J＝5.0Hz,4H),2.36(s,3H)1H NMR(400MHz,Chloroform-d)δ8.69–8.65(m,1H), 8.33(dd,J=5.3,0.8Hz,1H), 8.20(dd,J=7.9,1.0Hz,1H),7.86( t,J=7.8Hz,1H),7.71(dd,J=7.8,0.9Hz,1H),7.57–7.51(m,1H),7.47(dd,J=8.6,0.8Hz,1H),7.26(dd ,J=5.3,1.3Hz,1H), 7.20(dd,J=8.6,1.6Hz,1H), 6.74–6.66(m,2H), 6.62(tt,J=9.0,2.4Hz,1H), 4.10( s, 2H), 3.73 (t, J = 5.0 Hz, 4H), 2.56 (t, J = 5.0 Hz, 4H), 2.36 (s, 3H)
实施例5Example 5
6-(5-(3,5-二氟苯基)-1H-吲唑-3-基)-N-(2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)吡啶-2胺5的制备6-(5-(3,5-Difluorophenyl)-1H-indazol-3-yl)-N-(2-methoxy-4-(4-methylpiperazin-1-yl)benzene Base) pyridine-2 amine 5 preparation
第一步 6-(5-(3,5-二氟苯基)-1-((2-(三甲基硅基)乙氧基)甲基)-1H-吲唑-3-基)-N-(2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)吡啶-2胺5b的制备The first step 6-(5-(3,5-difluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-3-yl)- Preparation of N-(2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)pyridin-2amine 5b
化合物4h(200mg,0.37mmol),6-溴-氮-(2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)吡啶-2-胺5a(140mg,0.37mmol)，四三苯基磷钯(46mg,0.04mmol)，碘化亚铜(15mg,0.08mmol)和三乙胺(75mg，0.74mmol)悬混于DMF(6ml)中，反应液在氮气保护下在100℃搅拌2h，反应液冷却至室温，加入水(50ml),用二氯甲烷(30ml)萃取两遍。有机相用无水硫酸钠干燥，过滤浓缩得到粗品，经过柱层析纯化后得到化合物5b(146mg,0.22mmol)，收率59％。Compound 4h (200mg, 0.37mmol), 6-bromo-nitrogen-(2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)pyridin-2-amine 5a (140mg, 0.37mmol) ), palladium tetrakistriphenylphosphorus (46mg, 0.04mmol), cuprous iodide (15mg, 0.08mmol) and triethylamine (75mg, 0.74mmol) were suspended in DMF (6ml), and the reaction solution was protected by nitrogen After stirring for 2 hours at 100°C, the reaction solution was cooled to room temperature, water (50ml) was added, and the mixture was extracted twice with dichloromethane (30ml). The organic phase was dried with anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography to obtain compound 5b (146 mg, 0.22 mmol) with a yield of 59%.
MS m/z(ESI):671[M+H]
+。
MS m/z (ESI): 671 [M+H] + .
第二步 (5-(3,5-二氟苯基)-3-(6-((2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)氨基)吡啶-2-基)-1H-吲唑-1-基)甲醇5c的制备The second step (5-(3,5-difluorophenyl)-3-(6-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyridine -2-yl)-1H-indazol-1-yl)methanol 5c
化合物5b(146mg,0.22mmol)溶于二氯甲烷(8ml),向溶液中加入三氟乙酸(2ml)，反应液在常温下搅拌2小时，将反应液旋干，得到的产品5c直接用于 下一步反应。Compound 5b (146mg, 0.22mmol) was dissolved in dichloromethane (8ml), trifluoroacetic acid (2ml) was added to the solution, the reaction solution was stirred at room temperature for 2 hours, the reaction solution was spin-dried, and the resulting product 5c was used directly Next reaction.
MS m/z(ESI):571[M+H]
+。
MS m/z (ESI): 571 [M+H] + .
第十步 6-(5-(3,5-二氟苯基)-1H-吲唑-3-基)-N-(2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)吡啶-2胺5的制备The tenth step 6-(5-(3,5-difluorophenyl)-1H-indazol-3-yl)-N-(2-methoxy-4-(4-methylpiperazine-1- (Phenyl) phenyl) pyridine-2 amine 5 preparation
化合物5c(100mg,0.18mmol)溶于四氢呋喃(10ml)，向溶液中加入2N LiOH(10ml)，反应液在常温下搅拌4小时。加入水(20ml),用二氯甲烷(50ml)萃取两遍。有机相用无水硫酸钠干燥，过滤浓缩得到粗品，经过柱层析纯化后得到化合物5(40mg,0.074mmol)，收率42％。Compound 5c (100mg, 0.18mmol) was dissolved in tetrahydrofuran (10ml), 2N LiOH (10ml) was added to the solution, and the reaction solution was stirred at room temperature for 4 hours. Add water (20ml) and extract twice with dichloromethane (50ml). The organic phase was dried with anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography to obtain compound 5 (40 mg, 0.074 mmol) with a yield of 42%.
MS m/z(ESI):541[M+H]
+。
MS m/z (ESI): 541 [M+H] + .
1H NMR(400MHz,DMSO-d6)δ13.18(s,1H),8.26(s,1H),7.67–7.56(m,1H),7.49(d,J＝8.5Hz,1H),7.42(d,J＝7.4Hz,1H),7.37–7.29(m,1H),7.01(d,J＝8.3Hz,3H),6.82(d,J＝8.5Hz,1H),6.65–6.55(m,1H),6.49–6.31(m,2H),4.53(s,1H),4.11(s,2H),3.77(s,3H),3.65(br,4H),3.07(br,4H),2.64(s,3H).1H NMR(400MHz,DMSO-d6)δ13.18(s,1H), 8.26(s,1H), 7.67–7.56(m,1H), 7.49(d,J=8.5Hz,1H), 7.42(d, J=7.4Hz,1H), 7.37–7.29(m,1H), 7.01(d,J=8.3Hz,3H), 6.82(d,J=8.5Hz,1H), 6.65–6.55(m,1H), 6.49--6.31(m,2H), 4.53(s,1H), 4.11(s,2H), 3.77(s,3H), 3.65(br,4H), 3.07(br,4H), 2.64(s,3H) .
实施例6Example 6
N-(5-(1-(3,5-二氟苯基)环丙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯并酰胺6的制备N-(5-(1-(3,5-Difluorophenyl)cyclopropyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2- Preparation of ((Tetrahydro-2H-pyran-4-yl)amino)benzoamide 6
第一步 1-(3-溴-4-氟苯基)-1-(3,5-二氟苯基)乙基-1-醇6c的制备The first step is the preparation of 1-(3-bromo-4-fluorophenyl)-1-(3,5-difluorophenyl)ethyl-1-ol 6c
化合物6a(10g，51.8mmol)溶解在四氢呋喃(150mL)中，将溶液冷却至-78℃,向溶液中滴加正丁基锂溶液(22.8mL，57mmol)，滴加完毕后在-78℃搅拌1小时，然后加入化合物6b(11.2g,51.8mmol)的四氢呋喃(50mL)，反应液在常温搅拌过夜，反应液用饱和氯化铵溶液淬灭，用乙酸乙酯(200mL)萃取，有机相干燥后旋干得到化合物6c的粗品(19g).Compound 6a (10g, 51.8mmol) was dissolved in tetrahydrofuran (150mL), the solution was cooled to -78°C, n-butyllithium solution (22.8mL, 57mmol) was added dropwise to the solution, and after the addition, stirred at -78°C After 1 hour, compound 6b (11.2g, 51.8mmol) in tetrahydrofuran (50mL) was added, the reaction solution was stirred at room temperature overnight, the reaction solution was quenched with saturated ammonium chloride solution, extracted with ethyl acetate (200mL), and the organic phase was dried Rotate to dryness to obtain the crude product (19g) of compound 6c.
MS m/z(ESI):313[M-17]
+。
MS m/z(ESI): 313[M-17] + .
第二步 2-溴-4-(1-(3,5-二氟苯基)乙烯基)-1-氟苯6d的制备The second step is the preparation of 2-bromo-4-(1-(3,5-difluorophenyl)vinyl)-1-fluorobenzene 6d
化合物6c(19g，57.5mmol)溶于乙酸(30mL)，冰浴下滴加浓硫酸(3mL)。反应液在常温下搅拌30分钟。将反应液加入冰水(200mL)中，用乙酸乙酯(200mL)萃取，有机相用饱和碳酸氢钠溶液和食盐水洗涤，干燥后用无水硫酸钠干燥，过滤旋干得到初产品，经过硅胶柱层析纯化后的到化合物6d(11.7g,37.4mmol)，收率65％。Compound 6c (19 g, 57.5 mmol) was dissolved in acetic acid (30 mL), and concentrated sulfuric acid (3 mL) was added dropwise under ice bath. The reaction solution was stirred at room temperature for 30 minutes. The reaction solution was added to ice water (200mL), extracted with ethyl acetate (200mL), the organic phase was washed with saturated sodium bicarbonate solution and brine, dried and dried over anhydrous sodium sulfate, filtered and spin-dried to obtain the initial product, which was passed through silica gel After purification by column chromatography, compound 6d (11.7 g, 37.4 mmol) was obtained with a yield of 65%.
第三步 2-溴-4-(1-(3,5-二氟苯基)环丙基)-1-氟苯6e的制备The third step is the preparation of 2-bromo-4-(1-(3,5-difluorophenyl)cyclopropyl)-1-fluorobenzene 6e
将NaH(392mg,9.8mmol)溶解在DMSO(10mL)中，冷却至0℃，加入(CH
3)
3SOI(5.87g,26.7mmol),反应液在常温下搅拌2小时，再向反应液中加入6d(2.8g,8.9mmol)的二甲亚砜(10mL)溶液，反应液在常温下搅拌1小时，用饱 和氯化铵溶液淬灭反应液，用乙酸乙酯(100mL)萃取，有机相用饱和食盐水洗涤，干燥旋干得到粗品，经过柱层析纯化后得到化合物6e(1.8g,5.5mmol),收率62％。
Dissolve NaH (392mg, 9.8mmol) in DMSO (10mL), cool to 0°C, add (CH 3 ) 3 SOI (5.87g, 26.7mmol), stir the reaction solution at room temperature for 2 hours, and then add it to the reaction solution Add 6d (2.8g, 8.9mmol) dimethyl sulfoxide (10mL) solution, the reaction solution was stirred at room temperature for 1 hour, the reaction solution was quenched with saturated ammonium chloride solution, extracted with ethyl acetate (100mL), the organic phase It was washed with saturated brine, dried and spin-dried to obtain a crude product. After purification by column chromatography, compound 6e (1.8 g, 5.5 mmol) was obtained with a yield of 62%.
第四步 5-(1-(3,5-二氟苯基)环丙基)-2-氟苯腈6f的制备The fourth step is the preparation of 5-(1-(3,5-difluorophenyl)cyclopropyl)-2-fluorobenzonitrile 6f
化合物6e(0.8g，2.45mmol),氰化锌(286mg,2.45mmol)和四三苯基磷钯(288mg,0.25mmol)混合在DMF(6mL)中,用氮气置换两次，反应液在微波上110℃反应1小时，冷却后将反应液倒入水(50mL)中，用乙酸乙酯(50mL)萃取，有机相干燥后旋干得到粗品，经过柱层析(PE/EA＝30：1)纯化后得到化合物6f(760mg)的粗品。Compound 6e (0.8g, 2.45mmol), zinc cyanide (286mg, 2.45mmol) and palladium tetrakistriphenylphosphorus (288mg, 0.25mmol) were mixed in DMF (6mL), replaced with nitrogen twice, the reaction solution was microwaved Reacted at 110°C for 1 hour. After cooling, the reaction solution was poured into water (50mL), extracted with ethyl acetate (50mL), the organic phase was dried and spin-dried to obtain the crude product, which was subjected to column chromatography (PE/EA=30:1) ) After purification, a crude product of compound 6f (760mg) was obtained.
MS m/z(ESI):274[M+H]
+。
MS m/z (ESI): 274 [M+H] + .
第五步 5-(1-(3,5-二氟苯基)环丙基)-1H-吲唑-3-胺6g的制备The fifth step is the preparation of 5-(1-(3,5-difluorophenyl)cyclopropyl)-1H-indazole-3-amine 6g
化合物6f(680mg,2.49mmol)和水合肼(1.25g,24.9mmol)混合在N-甲基吡咯烷酮(6mL)中，反应液在微波反应器上120℃反应1小时，冷却后加入到(30mL)中,用乙酸乙酯(50mL)萃取，有机相干燥后旋干得到粗品，经过柱层析(PE/EA＝30：1)纯化后得到化合物6g(500mg,1.75mmol),收率71％。Compound 6f (680mg, 2.49mmol) and hydrazine hydrate (1.25g, 24.9mmol) were mixed in N-methylpyrrolidone (6mL), the reaction solution was reacted on a microwave reactor at 120°C for 1 hour, and then added to (30mL) after cooling After extraction with ethyl acetate (50 mL), the organic phase was dried and then spin-dried to obtain a crude product. After purification by column chromatography (PE/EA=30:1), compound 6g (500 mg, 1.75 mmol) was obtained, with a yield of 71%.
MS m/z(ESI):286[M+H]
+。
MS m/z (ESI): 286 [M+H] + .
第六步 N-(5-(1-(3,5-二氟苯基)环丙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰)苯甲酰胺6h的制备Step 6 N-(5-(1-(3,5-Difluorophenyl)cyclopropyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl) Preparation of -2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetyl)benzamide 6h
将化合物6g(285mg,1.0mmol)和DIPEA(387mg,3.0mmol)的四氢呋喃(15mL)溶液滴加到上述1f的四氢呋喃(5mL)冷却液中，反应在-20℃搅拌2小时，然后在常温下继续搅拌2小时，反应液用二氯甲烷(50mL)稀释，用拌合食盐水洗涤，干燥旋干得到粗品，经过柱层析(DCM/MeOH＝10:1)纯化后得到化合物6h(300mg,0.44mmol)，收率44％。A solution of compound 6g (285mg, 1.0mmol) and DIPEA (387mg, 3.0mmol) in tetrahydrofuran (15mL) was added dropwise to the above 1f tetrahydrofuran (5mL) cooling liquid, and the reaction was stirred at -20°C for 2 hours, and then at room temperature Stirring was continued for 2 hours. The reaction solution was diluted with dichloromethane (50mL), washed with mixed brine, dried and spin-dried to obtain a crude product, which was purified by column chromatography (DCM/MeOH=10:1) to obtain compound 6h (300mg, 0.44mmol), the yield is 44%.
MS m/z(ESI):683[M+H]
+。
MS m/z (ESI): 683 [M+H] + .
第七步 N-(5-(1-(3,5-二氟苯基)环丙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺6The seventh step N-(5-(1-(3,5-difluorophenyl)cyclopropyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide 6
化合物6h(300mg,0.44mmol)溶于甲醇(10mL)和水(5mL)的混合溶剂中，加入碳酸钾(182mg,1.32mmol)，反应液在常温下搅拌3小时，加入水(20ml)，用二氯甲烷(50ml)萃取。有机相用无水硫酸钠干燥，过滤浓缩得到粗品，经过柱层析纯化后得到化合物6(40mg,0.075mmol)，收率16％。Compound 6h (300mg, 0.44mmol) was dissolved in a mixed solvent of methanol (10mL) and water (5mL), potassium carbonate (182mg, 1.32mmol) was added, the reaction solution was stirred at room temperature for 3 hours, and water (20ml) was added. Extract with dichloromethane (50ml). The organic phase was dried with anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography to obtain compound 6 (40 mg, 0.075 mmol) with a yield of 16%.
MS m/z(ESI):587[M+H]
+。
MS m/z (ESI): 587 [M+H] + .
1H NMR(400MHz,DMSO-d6)δ12.58(s,1H),10.05(s,1H),8.34(d,J＝7.7Hz,1H),7.76(d,J＝8.9Hz,1H),7.49(s,1H),7.40–7.21(m,3H),7.01(t,J＝8.0Hz,2H),6.21(dd,J＝9.2,2.2Hz,1H),6.11(d,J＝2.4Hz,1H),3.87–3.74(m,2H),3.75–3.65(m,1H),3.50(t,J＝10.6Hz,2H),3.23(d,J＝5.1Hz,4H),2.41(t,J＝4.8Hz,4H),2.20(s,3H),1.97–1.86(m,2H),1.36-1.14(m,6H).
1 H NMR(400MHz,DMSO-d6)δ12.58(s,1H),10.05(s,1H), 8.34(d,J=7.7Hz,1H), 7.76(d,J=8.9Hz,1H), 7.49(s,1H),7.40–7.21(m,3H),7.01(t,J=8.0Hz,2H), 6.21(dd,J=9.2,2.2Hz,1H),6.11(d,J=2.4Hz ,1H),3.87–3.74(m,2H),3.75–3.65(m,1H), 3.50(t,J＝10.6Hz,2H), 3.23(d,J＝5.1Hz,4H),2.41(t, J = 4.8Hz, 4H), 2.20 (s, 3H), 1.97-1.86 (m, 2H), 1.36-1.14 (m, 6H).
实施例7Example 7
N-(5-((3,5-二氟苯基)二氟甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺7的制备N-(5-((3,5-Difluorophenyl)difluoromethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-( Preparation of (Tetrahydro-2H-pyran-4-yl)amino)benzamide 7
第一步 5-(3,5-二氟苯甲酰)-2-氟苯甲腈7a的制备The first step is the preparation of 5-(3,5-difluorobenzoyl)-2-fluorobenzonitrile 7a
化合物5-((3,5-二氟苯基)(羟基)甲基)-2-氟苯甲腈I-2c(2g，7.6mmol)溶于DCM(20mL)，冰浴下加入Dess Martin(6.4g，15.2mmol)，在搅拌下慢慢升温 至室温。混合物在室温继续搅拌1小时。反应液过滤，滤液用饱和NaHCO
3水溶液中和，该混合物用EA萃取，有机相用饱和氯化钠溶液洗，无水硫酸钠干燥，过滤，浓缩，粗品用柱层析纯化(PE:EA＝4:1)得到5-(3,5-二氟苯甲酰)-2-氟苯甲腈7a(1.1g，4.2mmol)，收率55％。
Compound 5-((3,5-difluorophenyl)(hydroxy)methyl)-2-fluorobenzonitrile I-2c (2g, 7.6mmol) was dissolved in DCM (20mL), and Dess Martin( 6.4g, 15.2mmol), slowly warmed to room temperature under stirring. The mixture was stirred at room temperature for 1 hour. The reaction solution was filtered, the filtrate was neutralized with saturated NaHCO 3 aqueous solution, the mixture was extracted with EA, the organic phase was washed with saturated sodium chloride solution, dried with anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography (PE:EA= 4:1) 5-(3,5-difluorobenzoyl)-2-fluorobenzonitrile 7a (1.1 g, 4.2 mmol) was obtained with a yield of 55%.
MS m/z(ESI):262[M+H]
+。
MS m/z (ESI): 262 [M+H] + .
第二步 5-((3,5-二氟苯基)二氟甲基)-2-氟苯甲腈7b的制备The second step is the preparation of 5-((3,5-difluorophenyl)difluoromethyl)-2-fluorobenzonitrile 7b
将化合物5-(3,5-二氟苯甲酰)-2-氟苯甲腈7a(130mg,0.5mmol)和BAST(1mL)混合在一起，该混合物在110℃搅拌3小时，反应液冷却，加入水淬灭，用DCM萃取3次，合并有机相，有机相用饱和食盐水洗，用无水硫酸钠干燥，过滤，浓缩，粗品用柱层析纯化(PE：EA＝4:1)得到5-((3,5-二氟苯基)二氟甲基)-2-氟苯甲腈7b(100mg,0.35mmol)呈无色油状物，收率70％。The compound 5-(3,5-difluorobenzoyl)-2-fluorobenzonitrile 7a (130mg, 0.5mmol) and BAST (1mL) were mixed together, the mixture was stirred at 110°C for 3 hours, and the reaction solution was cooled , Quenched with water, extracted 3 times with DCM, combined the organic phases, washed the organic phases with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography (PE:EA=4:1) to obtain 5-((3,5-Difluorophenyl)difluoromethyl)-2-fluorobenzonitrile 7b (100mg, 0.35mmol) was a colorless oily substance with a yield of 70%.
MS m/z(ESI):284[M+H]
+。
MS m/z (ESI): 284 [M+H] + .
第三步 5-((3,5-二氟苯基)二氟甲基)-1H-吲唑-3-胺7c的制备The third step is the preparation of 5-((3,5-difluorophenyl)difluoromethyl)-1H-indazol-3-amine 7c
将5-((3,5-二氟苯基)二氟甲基)-2-氟苯甲腈7b(100mg,0.35mmol)和水合肼(100mg,3.5mmol)分散在THF(5mL)中，该混合物在45℃下搅拌2小时，反应液冷却后，用浓盐酸淬灭，蒸掉THF,加入水稀释后用氨水中和至中性，有黄色固体析出，过滤干燥得到黄色固体5-((3,5-二氟苯基)二氟甲基)-1H-吲唑-3-胺7c(100mg,0.33mmol)呈淡黄色固体，收率96％。Disperse 5-((3,5-difluorophenyl)difluoromethyl)-2-fluorobenzonitrile 7b (100mg, 0.35mmol) and hydrazine hydrate (100mg, 3.5mmol) in THF (5mL), The mixture was stirred at 45°C for 2 hours. After the reaction solution was cooled, it was quenched with concentrated hydrochloric acid. The THF was evaporated, diluted with water and neutralized with ammonia water. A yellow solid precipitated out, filtered and dried to obtain a yellow solid 5-( (3,5-Difluorophenyl)difluoromethyl)-1H-indazol-3-amine 7c (100mg, 0.33mmol) was a pale yellow solid with a yield of 96%.
MS m/z(ESI):296[M+H]
+。
MS m/z (ESI): 296 [M+H] + .
第四步 N-(5-((3,5-二氟苯基)二氟甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯酰胺7d的制备The fourth step N-(5-((3,5-difluorophenyl)difluoromethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)- Preparation of 2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 7d
将4-(4-甲基哌嗪-1-基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯甲酰氯1f(104mg,0.24mmol)的THF溶液在-20℃下滴加到5-((3,5-二氟苯基)二氟甲基)-1H-吲唑-3-胺7c(80mg,0.24mmol)和三乙胺(73mg,0.72mmol)的THF溶液中，滴加完毕反应混合物在-20℃下搅拌1小时，反应液升至室温，加入水淬灭，用乙酸乙酯萃取，有机相用饱和氯化钠洗，用无水硫酸钠干燥，过滤，浓 缩，粗品柱层析纯化得到N-(5-((3,5-二氟苯基)二氟甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯酰胺7d(60mg,0.08mmol)呈黄色固体,收率36％。Add 4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoyl chloride 1f (104mg, 0.24mmol) in THF was added dropwise to 5-((3,5-difluorophenyl)difluoromethyl)-1H-indazole-3-amine 7c (80mg, 0.24mmol) at -20°C ) And triethylamine (73mg, 0.72mmol) in THF solution, after the addition, the reaction mixture was stirred at -20°C for 1 hour, the reaction solution was warmed to room temperature, quenched by adding water, extracted with ethyl acetate, and the organic phase was used Wash with saturated sodium chloride, dry with anhydrous sodium sulfate, filter, concentrate, and purify the crude product by column chromatography to obtain N-(5-((3,5-difluorophenyl)difluoromethyl)-1H-indazole- 3-yl)-4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido) Benzamide 7d (60mg, 0.08mmol) was a yellow solid with a yield of 36%.
MS m/z(ESI):693[M+H]
+。
MS m/z (ESI): 693 [M+H] + .
第六步 N-(5-((3,5-二氟苯基)二氟甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺7的制备The sixth step N-(5-((3,5-difluorophenyl)difluoromethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)- Preparation of 2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 7
将N-(5-((3,5-二氟苯基)二氟甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯酰胺7d(60mg,0.08mmol)溶于甲醇(2mL)中，加入三乙胺(2mL,该反应混合物在室温下搅拌6小时，浓缩，粗品用柱层析纯化(DCM：MeOH＝10:1)得到N-(5-((3,5-二氟苯基)二氟甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺7(5mg,0.008mmol)呈浅黄色固体，收率10％。The N-(5-((3,5-difluorophenyl)difluoromethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2- (2,2,2-Trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 7d (60mg, 0.08mmol) was dissolved in methanol (2mL), and triethylamine ( 2mL, the reaction mixture was stirred at room temperature for 6 hours, concentrated, and the crude product was purified by column chromatography (DCM:MeOH=10:1) to obtain N-(5-((3,5-difluorophenyl)difluoromethyl )-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 7 (5mg , 0.008mmol) was a pale yellow solid with a yield of 10%.
MS m/z(ESI):597[M+H]
+
MS m/z(ESI):597[M+H] +
1H NMR(400MHz,DMSO-d6)δ10.05(s,1H),8.67(s,1H),8.09–8.01(m,1H),7.71(s,1H),7.64–7.52(m,2H),7.45–7.37(m,1H),7.35–7.29(m,2H),6.88(d,J＝9.0Hz,1H),6.21–6.13(m,1H),6.06(d,J＝2.2Hz,1H),3.80–3.72(m,2H),3.65-3.57(m,1H),3.43(t,J＝10.5Hz,2H),3.21(t,J＝5.1Hz,4H),2.38(t,J＝5.1Hz,4H),2.18(s,3H),1.86(d,J＝12.9Hz,2H),1.32–1.23(m,2H).
1 H NMR(400MHz,DMSO-d6)δ10.05(s,1H),8.67(s,1H),8.09-8.01(m,1H),7.71(s,1H),7.64-7.52(m,2H) ,7.45–7.37(m,1H),7.35–7.29(m,2H), 6.88(d,J=9.0Hz,1H), 6.21–6.13(m,1H), 6.06(d,J=2.2Hz,1H ), 3.80–3.72 (m, 2H), 3.65-3.57 (m, 1H), 3.43 (t, J = 10.5 Hz, 2H), 3.21 (t, J = 5.1 Hz, 4H), 2.38 (t, J = 5.1Hz, 4H), 2.18 (s, 3H), 1.86 (d, J = 12.9 Hz, 2H), 1.32-1.23 (m, 2H).
实施例8Example 8
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((4-甲基哌嗪-1-基)磺酰)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺8的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((4-methylpiperazin-1-yl)sulfonyl)-2-(( Preparation of tetrahydro-2H-pyran-4-yl)amino)benzamide 8
第一步 4-氟-2-硝基苯甲酸乙酯8b的制备The first step is the preparation of ethyl 4-fluoro-2-nitrobenzoate 8b
向干燥的反应瓶中加入8a(9.2g,50mmol)，碳酸钾(1.38g,100mmol)和DMF(50mL)，然后滴入碘乙烷(9.4g,60mmol)。加完后室温搅拌过夜。TLC(石油醚：乙酸乙酯＝2:1)显示反应已经完全。加入水和乙酸乙酯分液，乙酸乙酯层再用水洗多次，无水硫酸钠干燥，减压蒸干得化合物8b(9.6g，45mmol,90％)。Add 8a (9.2g, 50mmol), potassium carbonate (1.38g, 100mmol) and DMF (50mL) to the dry reaction flask, and then dropwise add iodoethane (9.4g, 60mmol). After the addition, stir at room temperature overnight. TLC (petroleum ether: ethyl acetate = 2:1) showed that the reaction was complete. Water and ethyl acetate were added for separation, the ethyl acetate layer was washed with water several times, dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure to obtain compound 8b (9.6 g, 45 mmol, 90%).
MS m/z(ESI):214[M+H]
+
MS m/z(ESI):214[M+H] +
第二步 乙基4-((3-((2-乙基己基)氧代)-3-羰基丙基)硫代)-2-硝基苯酸酯8d的制备The second step is the preparation of ethyl 4-((3-((2-ethylhexyl)oxo)-3-carbonylpropyl)thio)-2-nitrobenzoate 8d
向反应瓶中加入化合物8b(2.14g,10mmol)，化合物8c(2.6g,12mmol)，DMF(10mL)和三乙胺(2.0g，20mmol)。混合物在90～100℃下反应4小时，TLC(石油醚：乙酸乙酯＝2:1)显示原料反应完。倒入水中，加乙酸乙酯萃取，水洗多次，无水硫酸钠干燥后用flash纯化(石油醚：乙酸乙酯＝5：1)，得化合物8d(3.0g,7.5mmol，75％)。Compound 8b (2.14 g, 10 mmol), compound 8c (2.6 g, 12 mmol), DMF (10 mL) and triethylamine (2.0 g, 20 mmol) were added to the reaction flask. The mixture was reacted at 90-100°C for 4 hours, and TLC (petroleum ether: ethyl acetate = 2:1) indicated that the reaction of the raw materials was complete. Pour into water, add ethyl acetate for extraction, wash with water several times, dry with anhydrous sodium sulfate and purify with flash (petroleum ether: ethyl acetate = 5:1) to obtain compound 8d (3.0 g, 7.5 mmol, 75%).
MS m/z(ESI):434[M+Na]
+
MS m/z(ESI):434[M+Na] +
第三步 乙基4-巯基-2-硝基苯酸酯8e的制备The third step Preparation of ethyl 4-mercapto-2-nitrobenzoate 8e
向反应瓶中加入化合物8d(0.41g，1.0mmol)，乙醇(10ml)，溶液澄清后加入乙醇钠(0.200g,3mmol)。室温下搅拌4小时，TLC(石油醚：乙酸乙酯＝2:1)显示原料反应完,加入柠檬酸中和，乙酸乙酯萃取3次，饱和食盐水洗涤有机相，无 水硫酸钠干燥，减压旋干后得到化合物8e粗品0.4g，直接用于下一步。Compound 8d (0.41g, 1.0mmol) and ethanol (10ml) were added to the reaction flask, and sodium ethoxide (0.200g, 3mmol) was added after the solution was clear. Stir at room temperature for 4 hours, TLC (petroleum ether: ethyl acetate = 2:1) shows that the raw materials have reacted, add citric acid to neutralize, extract with ethyl acetate 3 times, wash the organic phase with saturated brine, and dry with anhydrous sodium sulfate. After rotary drying under reduced pressure, 0.4 g of crude compound 8e was obtained, which was directly used in the next step.
第四步 化合物乙基4-(氯磺酰)-2-硝基苯酸酯8f的制备The fourth step is the preparation of compound ethyl 4-(chlorosulfonyl)-2-nitrobenzoate 8f
将2M HCl和5倍体积的乙腈溶液混合，加入NCS(0.79g，6mmol)，并冰浴至10℃左右，溶液澄清后加入化合物8e(0.4g粗品)的乙腈溶液，10℃左右搅拌2小时，TLC(石油醚：乙酸乙酯＝2:1)显示原料反应完。加入***稀释反应液，加入水分液。有机相用少量稀碳酸氢钠水溶液洗涤，无水硫酸钠干燥，低温减压蒸干后得化合物8f(0.4g)的粗品。Mix 2M HCl and 5 times the volume of acetonitrile solution, add NCS (0.79g, 6mmol), and ice-bath to about 10℃, after the solution is clear, add the acetonitrile solution of compound 8e (0.4g crude product), stir at about 10℃ for 2 hours , TLC (petroleum ether: ethyl acetate=2:1) showed that the reaction of the raw materials was complete. Add ether to dilute the reaction solution and add water solution. The organic phase was washed with a small amount of dilute sodium bicarbonate aqueous solution, dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure at low temperature to obtain a crude product of compound 8f (0.4g).
第五步 乙基4-((4-甲基哌嗪-1-基)磺酰)-2-硝基苯酸酯8g的制备Step 5 Preparation of 8g ethyl 4-((4-methylpiperazin-1-yl)sulfonyl)-2-nitrobenzoate
化合物8f(0.4g)溶于二氯甲烷中，加入1.0Ml甲基哌嗪，室温搅拌1小时，TLC(二氯甲烷：甲醇＝10:1)显示反应完成，加入硅胶拌样，flash纯化(二氯甲烷：甲醇＝20:1)得化合物8g(0.22g，0.8mmol,61％)。Compound 8f (0.4g) was dissolved in dichloromethane, 1.0Ml methylpiperazine was added, and the mixture was stirred at room temperature for 1 hour. TLC (dichloromethane:methanol=10:1) showed that the reaction was complete. Add silica gel to mix the sample, and flash purification ( Dichloromethane: methanol = 20:1) to obtain compound 8g (0.22g, 0.8mmol, 61%).
MS m/z(ESI):358[M+H]
+
MS m/z(ESI):358[M+H] +
第六步 乙基2-氨基-4-((4-甲基哌嗪-1-基)磺酰)苯酸酯8h的制备Step 6 Preparation of ethyl 2-amino-4-((4-methylpiperazin-1-yl)sulfonyl) benzoate 8h
向反应瓶中加入化合物8g(0.36g,1.0mmol)，还原性铁粉(0.28g，5mmol)，THF(10ml)，AcOH(0.5mL)，水(0.5ml)。反应混合物在60℃下回流2小时，TLC(二氯甲烷：甲醇＝10:1)显示反应完全，加入大量乙酸乙酯稀释，过滤。滤液浓缩后加入乙酸乙酯，用碳酸氢钠水溶液洗涤，分液后用无水硫酸钠干燥有机相。减压浓缩后得化合物8h(0.26g，0.8mmol，80％)。Compound 8g (0.36g, 1.0mmol), reducing iron powder (0.28g, 5mmol), THF (10ml), AcOH (0.5mL), water (0.5ml) were added to the reaction flask. The reaction mixture was refluxed at 60°C for 2 hours. TLC (dichloromethane:methanol=10:1) showed that the reaction was complete. A large amount of ethyl acetate was added for dilution and filtered. After the filtrate was concentrated, ethyl acetate was added, washed with sodium bicarbonate aqueous solution, and the organic phase was dried with anhydrous sodium sulfate after liquid separation. After concentration under reduced pressure, compound 8h (0.26 g, 0.8 mmol, 80%) was obtained.
MS m/z(ESI):328[M+H]
+
MS m/z(ESI): 328[M+H] +
第七步 乙基4-((4-甲基哌嗪-1-基)磺酰)-2-((四氢-2H-吡喃-4-基)氨基)苯酸酯8i的制备The seventh step is the preparation of ethyl 4-((4-methylpiperazin-1-yl)sulfonyl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzoate 8i
向干燥的反应瓶中加入化合物8h(0.33g，1.0mmol)，TFA(1.0ml)和NaBH(OAc)
3(0.88g，4.0mmol)。室温搅拌10min后，缓慢加入化合物4-二氢吡喃酮(0.5g,5mmol)的二氯甲烷溶液，10min后TLC(二氯甲烷：甲醇＝10:1)显示已经反应完全。反应液倒入碳酸氢钠水溶液中，搅拌半个小时后分液,有机相分出后用食盐水洗涤，无水硫酸钠干燥，减压浓缩后得粗品用flash纯化(二氯甲烷： 甲醇＝10:1)得化合物8i(0.28g，0.7mmol,70％)。
Compound 8h (0.33 g, 1.0 mmol), TFA (1.0 ml) and NaBH(OAc) 3 (0.88 g, 4.0 mmol) were added to the dry reaction flask. After stirring at room temperature for 10 min, a dichloromethane solution of compound 4-dihydropyrone (0.5 g, 5 mmol) was slowly added. After 10 min, TLC (dichloromethane:methanol=10:1) showed that the reaction was complete. The reaction solution was poured into an aqueous sodium bicarbonate solution, stirred for half an hour and then separated. The organic phase was separated and washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product purified by flash (dichloromethane: methanol = 10:1) Compound 8i (0.28g, 0.7mmol, 70%) was obtained.
MS m/z(ESI):412[M+H]
+
MS m/z(ESI):412[M+H] +
第八步 4-((4-甲基哌嗪-1-基)磺酰)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酸8j的制备The eighth step is the preparation of 4-((4-methylpiperazin-1-yl)sulfonyl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzoic acid 8j
向反应瓶中加入化合物8i(4.11g,10.0mmol)，THF(50ml)，溶液澄清后加入LiOH(4M,10ml)和少量水。搅拌过夜后TLC(二氯甲烷：甲醇＝20:1)显示反应完成。加入柠檬酸中和，乙酸乙酯萃取2次，有机相合并后用食盐水洗，无水硫酸钠干燥，减压浓缩后得化合物8j(3.07g，8.0mmol,80％)。Compound 8i (4.11g, 10.0mmol), THF (50ml) were added to the reaction flask, LiOH (4M, 10ml) and a small amount of water were added after the solution was clarified. After stirring overnight, TLC (dichloromethane: methanol = 20:1) showed that the reaction was complete. It was neutralized by adding citric acid, extracted twice with ethyl acetate, the organic phases were combined and washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 8j (3.07 g, 8.0 mmol, 80%).
MS m/z(ESI):384[M+H]
+
MS m/z(ESI):384[M+H] +
第九步 4-((4-甲基哌嗪-1-基)磺酰)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯甲酸8k的制备Step 9 4-((4-Methylpiperazin-1-yl)sulfonyl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetyl (Amino) benzoic acid 8k preparation
将化合物8j(0.38g，1.0mmol)悬浮于二氯甲烷中，加入TFAA直至溶液澄清，然后减压浓缩反应液，所得固体用石油醚带干，过滤所得固体加水搅拌48小时，过滤，固体晾干后高温减压除水得化合物8k(0.33g，0.7mmol，70％)。Suspend compound 8j (0.38g, 1.0mmol) in dichloromethane, add TFAA until the solution is clear, and then concentrate the reaction solution under reduced pressure. The obtained solid is dried with petroleum ether. The obtained solid is filtered and stirred with water for 48 hours, filtered, and the solid is dried. After drying, water was removed under high temperature and reduced pressure to obtain compound 8k (0.33 g, 0.7 mmol, 70%).
第十步 N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((4-甲基哌嗪-1-基)磺酰)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯酰胺8l的制备Step 10 N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((4-methylpiperazin-1-yl)sulfonyl)-2 -(2,2,2-Trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido) benzamide 8l
氮气保护下，向干燥反应的瓶中加入化合物8k(48mg,0.1mmol)和二氯甲烷和一滴DMF，然后加入草酰氯(0.3ml)。室温搅拌4小时后LCMS显示原料完全转化，减压浓缩反应液，并用二氯甲烷带干，所得固体用THF溶解，氮气保护下冷却至-40℃(外温)。然后缓慢加入化合物I-2(26mg,0.1mmol)和DIPEA(0.1ml)的THF溶液，并保持外温处于-40～-30℃。加完后让反应液自然升温，LCMS显示主峰是产物点。反应液浓缩至较小体积，用PLC纯化(二氯甲烷：甲醇＝10:1)得化合物8l(30mg,0.04mmol)。Under the protection of nitrogen, compound 8k (48 mg, 0.1 mmol), dichloromethane and a drop of DMF were added to the dry reaction flask, and then oxalyl chloride (0.3 ml) was added. After stirring at room temperature for 4 hours, LCMS showed complete conversion of the raw materials. The reaction solution was concentrated under reduced pressure and dried with dichloromethane. The obtained solid was dissolved in THF and cooled to -40°C (external temperature) under nitrogen protection. Then slowly add compound 1-2 (26mg, 0.1mmol) and DIPEA (0.1ml) in THF solution, and keep the external temperature at -40～-30°C. After the addition, the reaction solution was allowed to warm up naturally, and LCMS showed that the main peak was the product point. The reaction solution was concentrated to a small volume and purified by PLC (dichloromethane: methanol = 10:1) to obtain compound 8l (30 mg, 0.04 mmol).
MS m/z(ESI):721[M+H]
+
MS m/z(ESI):721[M+H] +
第十一步 N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((4-甲基哌嗪-1-基)磺酰)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺8的制备The eleventh step N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((4-methylpiperazin-1-yl)sulfonyl)- Preparation of 2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 8
向反应瓶中加入化合物8l(30mg,0.04mmol)和甲醇，溶液澄清后加入少量碳酸钾水溶液，TLC(二氯甲烷：甲醇＝10:1)显示反应完成。加入乙酸乙酯稀释，少量食盐水洗涤有机相，无水硫酸钠干燥，减压浓缩后PLC纯化(二氯甲烷：甲醇＝10:1)，得目标产物8(20mg,0.03mmol，76％)。Compound 8l (30 mg, 0.04 mmol) and methanol were added to the reaction flask. After the solution was clarified, a small amount of potassium carbonate aqueous solution was added. TLC (dichloromethane: methanol = 10:1) showed that the reaction was complete. Dilute with ethyl acetate, wash the organic phase with a small amount of brine, dry with anhydrous sodium sulfate, concentrate under reduced pressure and purify by PLC (dichloromethane: methanol = 10:1) to obtain the target product 8 (20mg, 0.03mmol, 76%) .
MS m/z(ESI):625[M+H]
+
MS m/z(ESI): 625[M+H] +
1H NMR(400MHz,DMSO-d6)δ12.76(s,1H),10.75(s,1H),8.06(d,J＝8.2Hz,1H),7.94(d,J＝7.9Hz,1H),7.53(s,1H),7.41(d,J＝8.6Hz,1H),7.25(dd,J＝8.7,1.6Hz,1H),7.02–6.93(m,4H),6.89(dd,J＝8.2,1.7Hz,1H),4.02(s,2H),3.80(dt,J＝11.8,3.9Hz,2H),3.75-3.65(m,1H),3.55-3.42(m,2H),2.95(br,4H),2.35(br,4H),2.14(s,3H),1.94-1.85(m,2H),1.45-1.34(m,2H).
1 H NMR (400MHz, DMSO-d6) δ 12.76 (s, 1H), 10.75 (s, 1H), 8.06 (d, J = 8.2 Hz, 1H), 7.94 (d, J = 7.9 Hz, 1H), 7.53(s,1H),7.41(d,J=8.6Hz,1H),7.25(dd,J=8.7,1.6Hz,1H),7.02-6.93(m,4H),6.89(dd,J=8.2, 1.7Hz,1H),4.02(s,2H),3.80(dt,J=11.8,3.9Hz,2H),3.75-3.65(m,1H),3.55-3.42(m,2H),2.95(br,4H) ), 2.35 (br, 4H), 2.14 (s, 3H), 1.94-1.85 (m, 2H), 1.45-1.34 (m, 2H).
实施例9Example 9
N-(5-(3,5-二氟苯甲基)吡唑并[1,5-a]嘧啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺9的制备N-(5-(3,5-Difluorobenzyl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiperazin-1-yl)-2- ((Tetrahydro-2H-pyran-4-yl)amino)benzamide 9
第一步 4-(4-甲基哌嗪-1-基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基) 苯甲酰胺9a的制备The first step 4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido) benzyl Preparation of amide 9a
4-(4-甲基哌嗪-1-基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯甲酰氯1f(0.86g,2mmol)加到氨水(5mL)、二氯甲烷的混合溶剂中，搅拌过夜，分出二氯甲烷层，浓缩，柱层析(DCM:MeOH＝15:1)纯化的产物9a(0.25g，0.8mmol),收率39％。4-(4-Methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoyl chloride 1f( 0.86g, 2mmol) was added to a mixed solvent of ammonia (5mL) and dichloromethane, stirred overnight, the dichloromethane layer was separated, concentrated, and the purified product 9a(0.25) by column chromatography (DCM:MeOH=15:1) g, 0.8mmol), the yield is 39%.
MS m/z(ESI):319[M+H]
+
MS m/z(ESI): 319[M+H] +
第二步 5-(3,5-二氟苯甲基)吡唑并[1,5-a]嘧啶9d的制备The second step is the preparation of 5-(3,5-difluorobenzyl)pyrazolo[1,5-a]pyrimidine 9d
将锌粉(3g,46mmol)分散于THF(100mL)中，加入0.2mL的1,2-二氯乙烷，该混合物在氮气保护下在50m℃搅拌1小时，冷却后在0℃下加入1-(溴甲基)-3,5-二氟苯(7.2g,35mmol)，而后再加入0.2mL的TMSCl,该反应混合物继续在室温搅拌6小时得9c。将该反应液室温下慢慢滴入到5-氯吡唑并[1,5-a]嘧啶9b(1.5g,9.8mmol)的THF溶液中，加入Pd(pph
3)
4(1.1g,0.98mmol),该反应混合物在氮气保护下在60℃搅拌1小时，冷却后加入饱和氯化铵水溶液淬灭，用乙酸乙酯萃取3次，有机相用饱和氯化钠洗，用无水硫酸钠干燥，过滤，浓缩，粗品用柱层析纯化(PE:EA＝5:1)得到5-(3,5-二氟苯甲基)吡唑并[1,5-a]嘧啶9d(2.2g,8.9mmol)呈黄色固体，收率90％。
Disperse zinc powder (3g, 46mmol) in THF (100mL), add 0.2mL of 1,2-dichloroethane, and stir the mixture at 50m℃ for 1 hour under the protection of nitrogen. After cooling, add 1 at 0℃. -(Bromomethyl)-3,5-difluorobenzene (7.2g, 35mmol), then 0.2mL of TMSC1 was added, and the reaction mixture was stirred at room temperature for 6 hours to obtain 9c. The reaction solution was slowly dropped into the THF solution of 5-chloropyrazolo[1,5-a]pyrimidine 9b (1.5g, 9.8mmol) at room temperature, and Pd(pph 3 ) 4 (1.1g, 0.98) was added. mmol), the reaction mixture was stirred at 60°C for 1 hour under the protection of nitrogen, quenched by adding saturated aqueous ammonium chloride solution after cooling, extracted with ethyl acetate 3 times, the organic phase was washed with saturated sodium chloride, and anhydrous sodium sulfate Dry, filter, concentrate, and purify the crude product by column chromatography (PE:EA=5:1) to obtain 5-(3,5-difluorobenzyl)pyrazolo[1,5-a]pyrimidine 9d (2.2g) , 8.9mmol) was a yellow solid with a yield of 90%.
MS m/z(ESI):246[M+H]
+
MS m/z(ESI):246[M+H] +
第三步 5-(3,5-二氟苯甲基)-3-碘吡唑并[1,5-a]嘧啶9e的制备The third step is the preparation of 5-(3,5-difluorobenzyl)-3-iodopyrazolo[1,5-a]pyrimidine 9e
将NIS(918mg,4mmol)加入到5-(3,5-二氟苯甲基)-3-碘吡唑并[1,5-a]嘧啶9d(500mg,2mmol)的THF中，该反应混合物在RT反应2小时。用硫代硫酸钠水溶液淬灭，混合物用乙酸乙酯萃取，合并有机相，有机相用饱和食盐水洗，用无水硫酸钠干燥，过滤，浓缩，粗品用柱层析纯化得到5-(3,5-二氟苯甲基)-3-碘吡唑并[1,5-a]嘧啶9e(600mg,1.61mmol)呈黄色固体，收率80％。NIS (918mg, 4mmol) was added to 5-(3,5-difluorobenzyl)-3-iodopyrazolo[1,5-a]pyrimidine 9d (500mg, 2mmol) in THF, the reaction mixture React for 2 hours at RT. It was quenched with aqueous sodium thiosulfate solution, the mixture was extracted with ethyl acetate, the organic phases were combined, the organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography to obtain 5-(3, 5-Difluorobenzyl)-3-iodopyrazolo[1,5-a]pyrimidine 9e (600mg, 1.61mmol) was a yellow solid with a yield of 80%.
MS m/z(ESI):372[M+H]
+
MS m/z(ESI):372[M+H] +
第四步 N-(5-(3,5-二氟苯甲基)吡唑并[1,5-a]嘧啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺9的制备The fourth step N-(5-(3,5-difluorobenzyl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiperazin-1-yl) Preparation of -2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 9
将混合物5-(3,5-二氟苯甲基)-3-碘吡唑并[1,5-a]嘧啶9e(100mg,0.27mmol)，4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺9a(85mg,0.27mmol),CuI(25mg,0.13mmol),铜粉(8mg,0.13mmol),K
3PO
4(114mg,0.54mmol)和N1,N2-二甲基乙烷-1,2-二胺(11mg,0.13mmol)分散于4mL甲苯中，该混合物在110℃搅拌56小时。反应液冷却，过滤，滤饼用二氯甲烷洗3次，滤液合并后浓缩，粗品用柱层析纯化(DCM:MeOH＝10:1)得到N-(5-(3,5-二氟苯甲基)吡唑并[1,5-a]嘧啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺9(9mg,0.016mmol)呈浅黄色固体，收率6％。
The mixture 5-(3,5-difluorobenzyl)-3-iodopyrazolo[1,5-a]pyrimidine 9e (100mg, 0.27mmol), 4-(4-methylpiperazine-1- Base)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 9a (85mg, 0.27mmol), CuI (25mg, 0.13mmol), copper powder (8mg, 0.13mmol), K 3 PO 4 (114 mg, 0.54 mmol) and N1,N2-dimethylethane-1,2-diamine (11 mg, 0.13 mmol) were dispersed in 4 mL of toluene, and the mixture was stirred at 110°C for 56 hours. The reaction solution was cooled, filtered, the filter cake was washed 3 times with dichloromethane, the filtrates were combined and concentrated, and the crude product was purified by column chromatography (DCM:MeOH=10:1) to obtain N-(5-(3,5-difluorobenzene) Methyl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl )Amino)benzamide 9 (9mg, 0.016mmol) was a pale yellow solid with a yield of 6%.
MS m/z(ESI):562[M+H]
+
MS m/z(ESI):562[M+H] +
1H NMR(400MHz,DMSO-d6)δ9.56(s,1H),8.91(d,J＝7.2Hz,1H),8.27(s,1H),8.22(d,J＝7.7Hz,1H),7.74(d,J＝9.0Hz,1H),7.12–7.04(m,3H),6.90(d,J＝7.2Hz,1H),6.23(dd,J＝8.9,2.2Hz,1H),6.10(d,J＝2.3Hz,1H),4.14(s,2H),3.86–3.71(m,2H),3.68–3.57(m,1H),3.49–3.40(m,2H),3.22(t,J＝5.0Hz,4H),2.40(t,J＝5.1Hz,4H),2.19(s,3H),1.90(d,J＝12.7Hz,2H),1.38–1.28(m,2H).
1 H NMR(400MHz,DMSO-d6)δ9.56(s,1H), 8.91(d,J=7.2Hz,1H), 8.27(s,1H), 8.22(d,J=7.7Hz,1H), 7.74(d,J=9.0Hz,1H), 7.12–7.04(m,3H), 6.90(d,J=7.2Hz,1H), 6.23(dd,J=8.9,2.2Hz,1H), 6.10(d ,J=2.3Hz,1H), 4.14(s,2H), 3.86–3.71(m,2H), 3.68–3.57(m,1H), 3.49–3.40(m,2H), 3.22(t,J=5.0 Hz, 4H), 2.40 (t, J = 5.1 Hz, 4H), 2.19 (s, 3H), 1.90 (d, J = 12.7 Hz, 2H), 1.38-1.28 (m, 2H).
实施例10Example 10
N-(5-(1-(3,5-二氟苯基)-2-羟基乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺10的制备N-(5-(1-(3,5-Difluorophenyl)-2-hydroxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl) Preparation of -2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 10
第一步 (2-(3-溴-4-氟苯基)-2-(3,5-二氟苯基)乙基)硼酸10a的制备The first step is the preparation of (2-(3-bromo-4-fluorophenyl)-2-(3,5-difluorophenyl)ethyl)boronic acid 10a
化合物6d(8.9g，28.4mmol)的四氢呋喃(150mL)溶液在0℃滴加到硼烷四氢呋喃络合物(56.8mL,56.8mmol)中，反应液在常温下搅拌2小时，然后小心向反应液中加入冰水(150mL),反应液在常温下继续搅拌3小时，将反应液浓缩后用乙酸乙酯(200mL)萃取，有机相分别用饱和碳酸氢钠溶液和食盐水洗涤，干燥后旋干得到的粗品经过柱层析纯化后得到化合物10a的粗品(10g)MS m/z(ESI):383[M+23]
+。
A solution of compound 6d (8.9g, 28.4mmol) in tetrahydrofuran (150mL) was added dropwise to the borane tetrahydrofuran complex (56.8mL, 56.8mmol) at 0°C. The reaction solution was stirred at room temperature for 2 hours, and then the reaction solution Ice water (150mL) was added, the reaction solution was stirred at room temperature for 3 hours, the reaction solution was concentrated and then extracted with ethyl acetate (200mL), the organic phase was washed with saturated sodium bicarbonate solution and brine, dried and spin-dried to obtain After purification by column chromatography, the crude product of compound 10a (10g) MS m/z(ESI): 383[M+23] + was obtained .
第二步 (2-(3-溴-4-氟苯基)-2-(3,5-二氟苯基)乙基-1-醇10b的制备The second step is the preparation of (2-(3-bromo-4-fluorophenyl)-2-(3,5-difluorophenyl)ethyl-1-ol 10b
化合物10a(10g，27.8mmol)溶于四氢呋喃(60mL)中，加入3M氢氧化钠水溶液(60mL),反应液冷却到0℃，缓慢滴加30％的双氧水(30mL)。反应液在常 温下搅拌3小时，然后用饱和硫代硫酸钠溶液淬灭。用乙酸乙酯(200mL)萃取，有机相用饱和碳酸氢钠溶液和食盐水洗涤，干燥后用无水硫酸钠干燥，过滤旋干得到初产品，经过硅胶柱层析纯化后的到化合物10b(4g,12mmol)，收率43％。Compound 10a (10g, 27.8mmol) was dissolved in tetrahydrofuran (60mL), 3M aqueous sodium hydroxide solution (60mL) was added, the reaction solution was cooled to 0°C, and 30% hydrogen peroxide (30mL) was slowly added dropwise. The reaction solution was stirred at room temperature for 3 hours, and then quenched with saturated sodium thiosulfate solution. It was extracted with ethyl acetate (200 mL), the organic phase was washed with saturated sodium bicarbonate solution and brine, dried and dried over anhydrous sodium sulfate, filtered and spin-dried to obtain the initial product, which was purified by silica gel column chromatography to obtain compound 10b (4g , 12mmol), the yield is 43%.
MS m/z(ESI):315[M-17]
+。
MS m/z (ESI): 315[M-17] + .
第三步 4-(2-(苯甲基氧基)-1-(3,5-二氟苯基)乙基)-2-溴-1-氟苯10c的制备The third step is the preparation of 4-(2-(benzyloxy)-1-(3,5-difluorophenyl)ethyl)-2-bromo-1-fluorobenzene 10c
将化合物10b(1g,3mmol)溶解在四氢呋喃(15mL)中，冷却至0℃，加入钠氢(144mg,3.6mmol),反应液在0℃搅拌1小时，再向反应液中加入溴苄(615.6mg,3.6mmol)，反应液在70℃下搅拌5小时，冷却后反应液倒入水(50mL)中，用乙酸乙酯(50mL)萃取，有机相用饱和食盐水洗涤，干燥旋干得到粗品，经过柱层析纯化后得到化合物10c(320mg,0.76mmol)，收率25％。Compound 10b (1g, 3mmol) was dissolved in tetrahydrofuran (15mL), cooled to 0°C, sodium hydrogen (144mg, 3.6mmol) was added, the reaction solution was stirred at 0°C for 1 hour, and then benzyl bromide (615.6 mg, 3.6mmol). The reaction solution was stirred at 70°C for 5 hours. After cooling, the reaction solution was poured into water (50mL) and extracted with ethyl acetate (50mL). The organic phase was washed with saturated brine, dried and spin-dried to obtain a crude product. After purification by column chromatography, compound 10c (320 mg, 0.76 mmol) was obtained with a yield of 25%.
第四步 5-(2-(苯甲基氧基)-1-(3,5-二氟苯基)乙基)-2-氟苯腈10d的制备The fourth step is the preparation of 5-(2-(benzyloxy)-1-(3,5-difluorophenyl)ethyl)-2-fluorobenzonitrile 10d
化合物10c(160mg，0.38mmol)，氰化锌(45mg,0.38mmol)和四三苯基磷钯(46mg,0.04mmol)混合在DMF(3mL)中,用氮气置换两次，反应液在微波上110℃反应1小时，冷却后将反应液倒入水(30mL)中，用乙酸乙酯(30mL)萃取，有机相干燥后旋干得到粗品，经过柱层析(PE/EA＝30：1)纯化后得到化合物10d(113mg，0.31mmol)，收率81％。Compound 10c (160mg, 0.38mmol), zinc cyanide (45mg, 0.38mmol) and palladium tetrakistriphenylphosphorus (46mg, 0.04mmol) were mixed in DMF (3mL), replaced with nitrogen twice, and the reaction solution was microwaved Reacted at 110°C for 1 hour. After cooling, the reaction solution was poured into water (30mL), extracted with ethyl acetate (30mL), the organic phase was dried and spin-dried to obtain the crude product, which was subjected to column chromatography (PE/EA=30:1) After purification, compound 10d (113 mg, 0.31 mmol) was obtained with a yield of 81%.
MS m/z(ESI):368[M+H]
+。
MS m/z (ESI): 368 [M+H] + .
第五步 5-(2-(苯甲基氧基)-1-(3,5-二氟苯基)乙基)-1H-吲唑-3-胺10e的制备Step 5 Preparation of 5-(2-(benzyloxy)-1-(3,5-difluorophenyl)ethyl)-1H-indazole-3-amine 10e
化合物10d(230mg,0.63mmol)和水合肼(313mg,6.3mmol)混合在N-甲基吡咯烷酮(6mL)中，反应液在微波反应器上120℃反应1小时，冷却后加入到(30mL)中，用乙酸乙酯(50mL)萃取，有机相干燥后旋干得到粗品，经过硅胶柱层析纯化后得到化合物10e(400mg)的粗品。Compound 10d (230mg, 0.63mmol) and hydrazine hydrate (313mg, 6.3mmol) were mixed in N-methylpyrrolidone (6mL), the reaction solution was reacted on a microwave reactor at 120°C for 1 hour, and then added to (30mL) after cooling After extraction with ethyl acetate (50 mL), the organic phase was dried and then spin-dried to obtain a crude product, which was purified by silica gel column chromatography to obtain a crude product of compound 10e (400 mg).
MS m/z(ESI):380[M+H]
+。
MS m/z (ESI): 380 [M+H] + .
第六步 N-(5-(2-(苯甲基氧基)-1-(3,5-二氟苯基)乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯甲酰胺10f的制备Step 6 N-(5-(2-(Benzyloxy)-1-(3,5-difluorophenyl)ethyl)-1H-indazol-3-yl)-4-(4- Preparation of methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 10f
将化合物10e(400mg,1.05mmol)和DIPEA(387mg,3.0mmol)的四氢呋喃(15mL)溶液滴加到1f的四氢呋喃(5mL)冷却液中，反应在-20℃搅拌2小时，然后在常温下继续搅拌2小时，反应液用二氯甲烷(50mL)稀释，用拌合食盐水洗涤，干燥旋干得到粗品，经过柱层析(DCM/MeOH＝10:1)纯化后得到化合物10f(370mg,0.48mmol)，收率45％。The tetrahydrofuran (15mL) solution of compound 10e (400mg, 1.05mmol) and DIPEA (387mg, 3.0mmol) was added dropwise to 1f tetrahydrofuran (5mL) cooling liquid, and the reaction was stirred at -20°C for 2 hours, and then continued at room temperature After stirring for 2 hours, the reaction solution was diluted with dichloromethane (50mL), washed with mixed brine, dried and spin-dried to obtain a crude product, which was purified by column chromatography (DCM/MeOH=10:1) to obtain compound 10f (370mg, 0.48) mmol), the yield is 45%.
MS m/z(ESI):777[M+H]
+。
MS m/z (ESI): 777 [M+H] + .
第七步 N-(5-(2-(苯甲基氧基)-1-(3,5-二氟苯基)乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺10g的制备The seventh step N-(5-(2-(benzyloxy)-1-(3,5-difluorophenyl)ethyl)-1H-indazol-3-yl)-4-(4- Preparation of 10 g of methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide
化合物10f(170mg,0.22mmol)溶于甲醇(10mL)和水(5mL)的混合溶剂中，加入碳酸钾(90.7mg,0.66mmol)，反应液在常温下搅拌3小时，加入水(20ml)，用二氯甲烷(50ml)萃取。有机相用无水硫酸钠干燥，过滤浓缩得到粗品，经过硅胶柱层析纯化后得到化合物10g(82mg,0.12mmol)，收率55％。Compound 10f (170mg, 0.22mmol) was dissolved in a mixed solvent of methanol (10mL) and water (5mL), potassium carbonate (90.7mg, 0.66mmol) was added, the reaction solution was stirred at room temperature for 3 hours, and water (20ml) was added. Extract with dichloromethane (50ml). The organic phase was dried with anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by silica gel column chromatography to obtain 10 g (82 mg, 0.12 mmol) of compound, with a yield of 55%.
MS m/z(ESI):681[M+H]+。MS m/z(ESI): 681[M+H]+.
第八步 N-(5-(1-(3,5-二氟苯基)-2-羟基乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺10的制备Step 8 N-(5-(1-(3,5-Difluorophenyl)-2-hydroxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazine-1 -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 10
化合物10g(102mg,0.15mmol)溶于乙醇(10mL)中，加入10％钯炭(20mg)和甲酸铵(190mg，3mmol)，反应液在90℃搅拌24小时，冷却后过滤。滤液浓缩得到粗品，经过硅胶柱层析纯化后得到化合物10(12mg,0.02mmol)，收率16％。Compound 10g (102mg, 0.15mmol) was dissolved in ethanol (10mL), 10% palladium on carbon (20mg) and ammonium formate (190mg, 3mmol) were added, the reaction solution was stirred at 90°C for 24 hours, cooled and filtered. The filtrate was concentrated to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 10 (12 mg, 0.02 mmol) with a yield of 16%.
MS m/z(ESI):591[M+H]
+。
MS m/z (ESI): 591 [M+H] + .
1H NMR(400MHz,DMSO-d6)δ12.59(s,1H),10.06(s,1H),8.30(d,J＝7.7Hz,1H),7.75(d,J＝9.1Hz,1H),7.47(s,1H),7.39–7.34(m,1H),7.31–7.20(m,2H),7.00(t,J＝8.8Hz,2H),6.20(dd,J＝9.0,2.3Hz,1H),6.10(d,J＝2.3Hz,1H),4.92(t,J＝5.4Hz,1H),4.51(t,J＝7.8Hz,1H),4.06(t,J＝6.6Hz,2H),3.85–3.74(m,2H),3.72–3.64(m,1H),3.48(td,J＝11.7,10.9,2.5Hz,2H),3.23(t,J＝5.1Hz,4H),2.40(t,J＝5.0Hz,4H),2.19(s,3H),1.91(d,J＝13.2Hz,2H),1.41-1.25(m,2H).
1 H NMR(400MHz,DMSO-d6)δ12.59(s,1H), 10.06(s,1H), 8.30(d,J=7.7Hz,1H), 7.75(d,J=9.1Hz,1H), 7.47(s,1H),7.39–7.34(m,1H),7.31–7.20(m,2H), 7.00(t,J=8.8Hz,2H), 6.20(dd,J=9.0,2.3Hz,1H) ,6.10(d,J=2.3Hz,1H),4.92(t,J=5.4Hz,1H),4.51(t,J=7.8Hz,1H),4.06(t,J=6.6Hz,2H),3.85 –3.74(m,2H),3.72–3.64(m,1H), 3.48(td,J=11.7,10.9,2.5Hz,2H), 3.23(t,J=5.1Hz,4H), 2.40(t,J =5.0Hz,4H),2.19(s,3H),1.91(d,J=13.2Hz,2H),1.41-1.25(m,2H).
实施例11Example 11
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-(噁丁环-3-基氨基)苯甲酰胺11的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-(oxbutane-3 -Amino) benzamide 11
第一步 N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-氟-2-硝基苯甲酰胺11b的制备The first step is the preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-fluoro-2-nitrobenzamide 11b
化合物4-氟-2-硝基苯甲酸I-1a(3.7g，20mmol)悬浮于DCM(60mL)，冰水浴冷却下加入草酰氯(6.4g,50mmol)，搅拌3小时，减压浓缩的11a溶于四氢呋喃(40mL)中，再加入DIEA(5.16g,40mmol),冰水浴冷却下加入化合物I-2(3.9g,15mmol),搅拌过夜，加入乙酸乙酯、水萃取，乙酸乙酯层柱层析，得产物11b(1.2g，2.8mmol),收率14％。Compound 4-fluoro-2-nitrobenzoic acid I-1a (3.7g, 20mmol) was suspended in DCM (60mL), oxalyl chloride (6.4g, 50mmol) was added under ice-water bath cooling, stirred for 3 hours, 11a concentrated under reduced pressure Dissolve in tetrahydrofuran (40mL), then add DIEA (5.16g, 40mmol), add compound I-2 (3.9g, 15mmol) under ice-water bath cooling, stir overnight, add ethyl acetate, water for extraction, ethyl acetate layer column By chromatography, the product 11b (1.2 g, 2.8 mmol) was obtained with a yield of 14%.
MS m/z(ESI):427[M+H]
+。
MS m/z (ESI): 427 [M+H] + .
第二步 N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-硝基苯甲酰胺11c的制备The second step N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-nitrobenzene Preparation of formamide 11c
化合物N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-氟-2-硝基苯酰胺11b(200mg，0.47mmol),1-甲基哌嗪(93mg,0.93mmol)和DIPEA(181mg,1.41mmol)溶于 DMSO(4mL)。该混合物在100℃下搅拌3小时，冷却后加入水淬灭，用乙酸乙酯萃取，有机相合并，用饱和氯化钠水溶液洗2次，用无水硫酸钠干燥，过滤，浓缩，得到的粗品用柱层析纯化(DCM:MeOH＝10:1)得到200mg N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-硝基苯酰胺11c呈黄色固体,收率84％Compound N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-fluoro-2-nitrobenzamide 11b (200mg, 0.47mmol), 1-methyl Piperazine (93mg, 0.93mmol) and DIPEA (181mg, 1.41mmol) were dissolved in DMSO (4mL). The mixture was stirred at 100°C for 3 hours. After cooling, it was quenched by adding water, extracted with ethyl acetate, the organic phases were combined, washed twice with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain The crude product was purified by column chromatography (DCM:MeOH=10:1) to obtain 200mg N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methyl Piperazine-1-yl)-2-nitrobenzamide 11c is a yellow solid with a yield of 84%
MS m/z(ESI):507[M+H]
+
MS m/z(ESI): 507[M+H] +
第二步 2-氨基-N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)苯甲酰胺11d的制备The second step 2-amino-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)benzyl Preparation of amide 11d
将化合物N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-硝基苯酰胺11c(200mg,0.39mmol)溶于甲醇中，加入Pd/C(10％)，在氢气条件下室温搅拌过夜。反应液过滤，滤液浓缩，150mg粗品2-氨基-N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)苯酰胺11d直接用于下一步反应。收率79％。The compound N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-nitrobenzamide 11c (200 mg, 0.39 mmol) was dissolved in methanol, Pd/C (10%) was added, and the mixture was stirred overnight at room temperature under hydrogen conditions. The reaction solution was filtered, the filtrate was concentrated, and 150 mg of crude 2-amino-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazine- The 1-yl)benzamide 11d was directly used in the next reaction. The yield was 79%.
MS m/z(ESI):477[M+H]
+。
MS m/z (ESI): 477 [M+H] + .
第三步 N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-(噁丁环-3-基氨基)苯甲酰胺11的制备The third step N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-(oxbutan Preparation of Cyclo-3-ylamino)benzamide 11
将2-氨基-N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)苯酰胺11d(150mg,0.31mmol)和氧杂环丁酮(67mg,0.93mmol)分散在DCM(5mL)中，加入三氟乙酸(0.3mL),然后加入三乙酰氧基硼氢化四甲基铵(414mg,1.57mmol)。该混合物在室温下搅拌12小时，用饱和NaHCO
3水溶液淬灭，用DCM萃取，有机相用饱和氯化钠洗，用无水硫酸钠干燥，过滤，浓缩，粗品柱层析纯化得到2.5mg N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-(噁丁环-3-基氨基)苯酰胺11,收率1.5％。
The 2-amino-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)benzamide 11d( 150mg, 0.31mmol) and oxetanone (67mg, 0.93mmol) were dispersed in DCM (5mL), trifluoroacetic acid (0.3mL) was added, and then triacetoxytetramethylammonium borohydride (414mg, 1.57 mmol). The mixture was stirred at room temperature for 12 hours, quenched with saturated aqueous NaHCO 3 solution, extracted with DCM, the organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography to obtain 2.5 mg of N -(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-(oxbutane-3- (Amino) benzamide 11, the yield is 1.5%.
MS m/z(ESI):533[M+H]
+。
MS m/z (ESI): 533 [M+H] + .
1H NMR(400MHz,Methanol-d4)δ7.78(d,J＝8.9Hz,1H),7.60(s,1H),7.42(d,J＝8.6Hz,1H),7.23(dd,J＝8.6,1.6Hz,1H),6.84–6.78(m,2H),6.74–6.67(m,1H),6.39(dd,J＝8.9,2.4Hz,1H),5.83(d,J＝2.4Hz,1H),5.01(t,J＝6.6Hz,2H),4.74(p,J＝6.3Hz,1H),4.53(t,J＝6.1Hz,2H),4.06(s,2H),3.46(s,4H),3.00(s, 4H),2.65(s,3H).
1 H NMR(400MHz,Methanol-d4)δ7.78(d,J=8.9Hz,1H), 7.60(s,1H), 7.42(d,J=8.6Hz,1H), 7.23(dd,J=8.6 ,1.6Hz,1H), 6.84–6.78(m,2H), 6.74–6.67(m,1H), 6.39(dd,J=8.9,2.4Hz,1H), 5.83(d,J=2.4Hz,1H) ,5.01(t,J=6.6Hz,2H),4.74(p,J=6.3Hz,1H),4.53(t,J=6.1Hz,2H),4.06(s,2H),3.46(s,4H) , 3.00(s, 4H), 2.65(s, 3H).
实施例12Example 12
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(5-甲基六氢吡咯并[3,4-c]吡咯-2(1H)-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺12的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(5-methylhexahydropyrrolo[3,4-c]pyrrole-2(1H )-Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 12
第一步 N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(5-甲基六氢吡咯并[3,4-c]吡咯-2(1H)-基)-2-硝基苯甲酰胺12b的制备The first step N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(5-methylhexahydropyrrolo[3,4-c]pyrrole- Preparation of 2(1H)-yl)-2-nitrobenzamide 12b
化合物N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-氟-2-硝基苯酰胺11b(200mg，0.47mmol),2-甲基八氢吡咯并[3,4-c]吡咯12a(117mg,0.93mmol)和DIPEA(181mg,1.41mmol)溶于DMSO(4mL)。该混合物在150℃下搅拌3小时，冷却后加入水淬灭，用乙酸乙酯萃取，有机相合并，用饱和氯化钠水溶液洗2次，用无水硫酸钠干燥，过滤，浓缩，得到的粗品用柱层析纯化(DCM:MeOH＝10:1)得到200mg N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(5-甲基六氢吡咯并[3,4-c]吡咯-2(1H)-基)-2-硝基苯酰胺12b胺呈黄色固体。MS m/z(ESI):533[M+H]
+。收率84％
Compound N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-fluoro-2-nitrobenzamide 11b (200mg, 0.47mmol), 2-methyl Octahydropyrrolo[3,4-c]pyrrole 12a (117 mg, 0.93 mmol) and DIPEA (181 mg, 1.41 mmol) were dissolved in DMSO (4 mL). The mixture was stirred at 150°C for 3 hours. After cooling, it was quenched by adding water, extracted with ethyl acetate, the organic phases were combined, washed twice with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain The crude product was purified by column chromatography (DCM:MeOH=10:1) to obtain 200mg of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(5-methyl Hexahydropyrrolo[3,4-c]pyrrole-2(1H)-yl)-2-nitrobenzamide 12b amine is a yellow solid. MS m/z (ESI): 533 [M+H] + . Yield 84%
第二步 2-氨基-N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(5-甲基六氢吡咯并 [3,4-c]吡咯-2(1H)-基)苯甲酰胺12c的制备The second step 2-amino-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(5-methylhexahydropyrrolo[3,4- c) Preparation of pyrrole-2(1H)-yl)benzamide 12c
将化合物N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-硝基苯酰胺12b(200mg,0.37mmol)溶于甲醇中，加入Pd/C(10％)，在氢气条件下室温搅拌过夜。反应液过滤，滤液浓缩，150mg粗品2-氨基-N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(5-甲基六氢吡咯并[3,4-c]吡咯-2(1H)-基)苯酰胺12c直接用于下一步反应。收率80％。The compound N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-nitrobenzamide 12b (200 mg, 0.37 mmol) was dissolved in methanol, Pd/C (10%) was added, and the mixture was stirred overnight at room temperature under hydrogen conditions. The reaction solution was filtered and the filtrate was concentrated. 150mg of crude 2-amino-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(5-methylhexahydropyrrole) And [3,4-c]pyrrole-2(1H)-yl)benzamide 12c was directly used in the next reaction. The yield is 80%.
MS m/z(ESI):503[M+H]
+。
MS m/z (ESI): 503 [M+H] + .
第三步 N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(5-甲基六氢吡咯并[3,4-c]吡咯-2(1H)-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺12的制备The third step N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(5-methylhexahydropyrrolo[3,4-c]pyrrole- Preparation of 2(1H)-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 12
将2-氨基-N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(5-甲基六氢吡咯并[3,4-c]吡咯-2(1H)-基)苯酰胺12c(150mg,0.29mmol)和四氢-4H-吡喃-4-酮(87mg,0.87mmol)分散在DCM(5mL)中，加入三氟乙酸(0.3mL),然后加入四乙酰氧基硼氢化四甲基铵(414mg,1.57mmol)。该混合物在室温下搅拌12小时，用饱和NaHCO
3水溶液淬灭，用DCM萃取，有机相用饱和氯化钠洗，用无水硫酸钠干燥，过滤，浓缩，粗品柱层析纯化得到1.5mg N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(5-甲基六氢吡咯并[3,4-c]吡咯-2(1H)-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺12,收率1.5％。
The 2-amino-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(5-methylhexahydropyrrolo[3,4-c] Pyrrole-2(1H)-yl)benzamide 12c (150mg, 0.29mmol) and tetrahydro-4H-pyran-4-one (87mg, 0.87mmol) were dispersed in DCM (5mL), and trifluoroacetic acid (0.3 mL), and then add tetramethylammonium tetraacetoxyborohydride (414mg, 1.57mmol). The mixture was stirred at room temperature for 12 hours, quenched with saturated aqueous NaHCO 3 solution, extracted with DCM, the organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography to obtain 1.5 mg of N -(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(5-methylhexahydropyrrolo[3,4-c]pyrrole-2(1H) -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 12, the yield is 1.5%.
MS m/z(ESI):533[M+H]+。MS m/z(ESI): 533[M+H]+.
1H NMR(400MHz,Methanol-d4)δ7.70(d,J＝8.9Hz,1H),7.57(s,1H),7.40(d,J＝8.7Hz,1H),7.22(dd,J＝8.7,1.5Hz,1H),6.81(d,J＝7.0Hz,2H),6.75–6.66(m,1H),6.08(dd,J＝8.8,2.2Hz,1H),5.92(d,J＝2.1Hz,1H),4.05(s,2H),3.91(dt,J＝11.6,4.0Hz,2H),3.65–3.61(m,1H),3.59–3.52(m,2H),3.48–3.34(m,4H),3.10–3.06(m,2H),2.99–2.95(m,2H),2.62(d,J＝10.3Hz,2H),2.43(s,3H),2.03(d,J＝13.1Hz,2H),1.56–1.48(m,2H).
1 H NMR(400MHz,Methanol-d4)δ7.70(d,J=8.9Hz,1H), 7.57(s,1H), 7.40(d,J=8.7Hz,1H), 7.22(dd,J=8.7 ,1.5Hz,1H),6.81(d,J=7.0Hz,2H),6.75-6.66(m,1H),6.08(dd,J=8.8,2.2Hz,1H),5.92(d,J=2.1Hz ,1H),4.05(s,2H),3.91(dt,J=11.6,4.0Hz,2H), 3.65–3.61(m,1H),3.59–3.52(m,2H),3.48–3.34(m,4H) ), 3.10–3.06(m,2H), 2.99–2.95(m,2H), 2.62(d,J=10.3Hz,2H),2.43(s,3H),2.03(d,J=13.1Hz,2H) ,1.56–1.48(m,2H).
实施例13Example 13
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(甲基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺13的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidin-1-yl)ethyl)amino)-2 -((Tetrahydro-2H-pyran-4-yl)amino)benzamide 13
采用与实施例12相同的制备方法，以2-(吡咯烷-1-基)乙基氨为原料，可得到化合物13。Using the same preparation method as in Example 12, using 2-(pyrrolidin-1-yl)ethyl ammonia as a raw material, compound 13 can be obtained.
MS m/z(ESI):589[M+H]
+。
MS m/z (ESI): 589 [M+H] + .
1H NMR(400MHz,Methanol-d4)δ7.75(d,J＝8.9Hz,1H),7.55(s,1H),7.41(d,J＝8.6Hz,1H),7.27–7.22(m,1H),6.81(d,J＝6.9Hz,2H),6.74–6.67(m,1H),6.20(dd,J＝9.1,2.4Hz,1H),6.03(d,J＝2.3Hz,1H),4.06(s,2H),3.91(dt,J＝12.1,4.3Hz,2H),3.79–3.64(m,3H),3.62–3.53(m,2H),3.37–3.27(m,6H),3.05(s,3H),2.09–1.95(m,6H),1.58–1.47(m,2H).
1 H NMR(400MHz,Methanol-d4)δ7.75(d,J=8.9Hz,1H),7.55(s,1H),7.41(d,J=8.6Hz,1H),7.27–7.22(m,1H ), 6.81 (d, J = 6.9 Hz, 2H), 6.74–6.67 (m, 1H), 6.20 (dd, J = 9.1, 2.4 Hz, 1H), 6.03 (d, J = 2.3 Hz, 1H), 4.06 (s,2H),3.91(dt,J=12.1,4.3Hz,2H),3.79–3.64(m,3H),3.62–3.53(m,2H),3.37–3.27(m,6H),3.05(s ,3H),2.09-1.95(m,6H),1.58-1.47(m,2H).
实施例14Example 14
(R)-N-(5-(2-(2,5-二氟苯基)吡咯烷-1-基)吡唑并[1,5-a]嘧啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺14的制备(R)-N-(5-(2-(2,5-Difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4 -Methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 14
采用与实施例1相同的制备方法，以(R)-2-(2,5-二氟苯基)吡咯烷为原料，可得到化合物13。Using the same preparation method as in Example 1, using (R)-2-(2,5-difluorophenyl)pyrrolidine as a raw material, compound 13 can be obtained.
MS m/z(ESI):617[M+H]
+。
MS m/z (ESI): 617 [M+H] + .
1H NMR(400MHz,DMSO-d6)δ9.03(br,1H),8.49(br,1H),8.15(br,1H),7.92(s,1H),7.59(br,1H),7.31–7.02(m,3H),6.83(br,1H),6.20(dd,J＝8.9,2.2Hz, 1H),6.10(d,J＝2.3Hz,1H),5.34(br,1H),3.94–3.85(m,1H),3.78(dt,J＝11.6,3.9Hz,2H),3.68-3.53(m,2H),3.49–3.40(m,2H),3.21(t,J＝5.1Hz,4H),2.40(t,J＝5.0Hz,4H),2.19(s,3H),2.02-1.81(m,6H),1.37–1.25(m,2H).
1 H NMR(400MHz,DMSO-d6)δ9.03(br,1H), 8.49(br,1H), 8.15(br,1H), 7.92(s,1H), 7.59(br,1H), 7.31-7.02 (m, 3H), 6.83 (br, 1H), 6.20 (dd, J = 8.9, 2.2 Hz, 1H), 6.10 (d, J = 2.3 Hz, 1H), 5.34 (br, 1H), 3.94–3.85 ( m, 1H), 3.78 (dt, J = 11.6, 3.9 Hz, 2H), 3.68-3.53 (m, 2H), 3.49-3.40 (m, 2H), 3.21 (t, J = 5.1 Hz, 4H), 2.40 (t,J=5.0Hz,4H), 2.19(s,3H), 2.02-1.81(m,6H), 1.37-1.25(m,2H).
实施例15Example 15
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(7-甲基-2,7-二氮杂螺[3.5]壬烷-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺15的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(7-methyl-2,7-diazaspiro[3.5]nonane-2 -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 15
采用与实施例12相同的制备方法，以7-甲基-2,7-二氮杂螺[3.5]壬烷为原料，可得到化合物15。Using the same preparation method as in Example 12, using 7-methyl-2,7-diazaspiro[3.5]nonane as a raw material, compound 15 can be obtained.
MS m/z(ESI):601[M+1]+MS m/z(ESI):601[M+1]+
1H NMR(400MHz,DMSO-d
6)δ12.64(s,1H),10.03(s,1H),8.39(d,J＝7.5Hz,1H),7.78(d,J＝8.8Hz,1H),7.47(s,1H),7.40(d,J＝8.6Hz,1H),7.25(dd,J＝8.6,1.6Hz,1H),7.05–6.93(m,3H),5.71(dd,J＝8.7,2.1Hz,1H),5.61(d,J＝2.1Hz,1H),4.04(s,2H),3.82(dt,J＝11.6,4.0Hz,2H),3.66(s,4H),3.61–3.41(m,3H),2.78(br,4H),2.56(s,3H),1.97-1.85(m,6H),1.43–1.33(m,2H).
1 H NMR(400MHz,DMSO-d 6 )δ12.64(s,1H), 10.03(s,1H), 8.39(d,J=7.5Hz,1H), 7.78(d,J=8.8Hz,1H) ,7.47(s,1H),7.40(d,J=8.6Hz,1H),7.25(dd,J=8.6,1.6Hz,1H),7.05-6.93(m,3H),5.71(dd,J=8.7 ,2.1Hz,1H),5.61(d,J=2.1Hz,1H),4.04(s,2H),3.82(dt,J=11.6,4.0Hz,2H),3.66(s,4H),3.61-3.41 (m,3H), 2.78(br,4H), 2.56(s,3H), 1.97-1.85(m,6H), 1.43-1.33(m,2H).
实施例16Example 16
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(6-甲基-2,6-二氮杂螺[3.3]庚烷-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺16的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(6-methyl-2,6-diazaspiro[3.3]heptane-2 -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 16
采用与实施例12相同的制备方法，以6-甲基-2,6-二氮杂螺[3.5]庚烷为原料，可得到化合物16。Using the same preparation method as in Example 12, using 6-methyl-2,6-diazaspiro[3.5]heptane as a raw material, compound 16 can be obtained.
MS m/z(ESI):573[M+1]
+
MS m/z(ESI):573[M+1] +
1H NMR(400MHz,Chloroform-d)δ8.44(s,1H),8.19(d,J＝7.4Hz,1H),7.74(s,1H),7.50(d,J＝8.7Hz,1H),7.33(d,J＝8.6Hz,1H),7.16(d,J＝8.6Hz,1H),6.75–6.67(m,2H),6.61(tt,J＝9.0,2.3Hz,1H),5.72(d,J＝8.5Hz,1H),5.57(d,J＝2.1Hz,1H),4.09–3.90(m,8H),3.58-3.45(m,7H),2.42(s,3H),2.00(d,J＝13.0Hz,2H),1.66-1.53(m,2H).
1 H NMR(400MHz,Chloroform-d)δ8.44(s,1H), 8.19(d,J=7.4Hz,1H), 7.74(s,1H), 7.50(d,J=8.7Hz,1H), 7.33(d,J=8.6Hz,1H),7.16(d,J=8.6Hz,1H),6.75-6.67(m,2H),6.61(tt,J=9.0,2.3Hz,1H),5.72(d ,J=8.5Hz,1H),5.57(d,J=2.1Hz,1H),4.09–3.90(m,8H),3.58-3.45(m,7H),2.42(s,3H),2.00(d, J = 13.0Hz, 2H), 1.66-1.53 (m, 2H).
实施例17Example 17
N-(5-(3,5-二氟苯甲基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺17的制备N-(5-(3,5-Difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl) Preparation of -2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 17
第一步 5-溴-1-((2-(三甲基甲硅烷基)乙氧基)甲基)-1H-吡唑并[3,4-c]吡啶17b的合成The first step is the synthesis of 5-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-c]pyridine 17b
将5-溴-1H-吡唑并[3,4-c]吡啶17a(1.0g，5.0mmol，1.0eq)投于无水THF(20ml)中，冰水浴冷却下，分批加入NaH(0.4g，10.0mmol，2.0eq)，加毕，回流反应1h，冷至室温，加入SEMCl(1.3ml，7.5mmol，1.5eq)，继续反应2h。加入少量MeOH，淬灭，蒸除溶剂，加入EA(50ml)，依次用H2O(15ml)和brine(15ml)洗涤，经无水硫酸钠干燥，浓缩，flash柱纯化(PE～PE/EA＝1：1)得黄色产物17b(560mg)。Put 5-bromo-1H-pyrazolo[3,4-c]pyridine 17a (1.0g, 5.0mmol, 1.0eq) in anhydrous THF (20ml), cool in an ice-water bath, add NaH(0.4 g, 10.0mmol, 2.0eq). After the addition, the reaction was refluxed for 1h, cooled to room temperature, SEMCl (1.3ml, 7.5mmol, 1.5eq) was added, and the reaction was continued for 2h. Add a small amount of MeOH, quench, evaporate the solvent, add EA (50ml), wash with H2O (15ml) and brine (15ml) successively, dry over anhydrous sodium sulfate, concentrate, and purify by flash column (PE～PE/EA=1 :1) The yellow product 17b (560mg) was obtained.
MS m/z(ESI):330[M+1]
+
MS m/z(ESI):330[M+1] +
第二步 5-(3,5-二氟苯甲基)-1-((2-(三甲基甲硅烷基)乙氧基)甲基)-1H-吡唑并[3,4-c]吡啶17d的合成The second step 5-(3,5-difluorobenzyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-c ] Synthesis of pyridine 17d
将Zn粉(5.0g，76.5mmol，12.0eq)投于无水THF(50ml)中，加入1,2-二溴乙 烷(5滴)，N2换气5次，回流反应30min，冷至室温，加入TMSCl(4滴)，继续反应30min。冰水浴冷却下，滴加3,5-二氟溴苄(5.0ml，38.6mmol，1.0eq)，自然升至室温，反应过夜，得17c。Put Zn powder (5.0g, 76.5mmol, 12.0eq) into anhydrous THF (50ml), add 1,2-dibromoethane (5 drops), ventilate 5 times with N2, reflux for 30min, cool to room temperature , TMSCl (4 drops) was added, and the reaction was continued for 30 minutes. Under cooling in an ice-water bath, 3,5-difluorobenzyl bromide (5.0ml, 38.6mmol, 1.0eq) was added dropwise, and the mixture was naturally warmed to room temperature and reacted overnight to obtain 17c.
将化合物17b(200mg，0.61mmol，1.0eq)溶于无水THF(15ml)中，加入Pd(PPh3)4(76mg，0.061mmol，0.1eq)，N2气换气5次，滴加化合物17c的反应液(约10ml)，加毕，回流反应1h。减压蒸除溶剂，加入EA(30ml)和H
2O(15ml)，过滤不容物，分出EA层，用brine洗涤，无水硫酸钠干燥，浓缩，flash柱纯化(PE～PE/EA＝1：1)得黄色油状物17d(210mg)。
Compound 17b (200mg, 0.61mmol, 1.0eq) was dissolved in anhydrous THF (15ml), Pd(PPh3)4 (76mg, 0.061mmol, 0.1eq) was added, N2 gas was ventilated 5 times, and compound 17c was added dropwise Reaction solution (about 10ml), after addition, reflux for 1h. The solvent was evaporated under reduced pressure, EA (30ml) and H 2 O (15ml) were added, the insoluble matter was filtered, the EA layer was separated, washed with brine, dried with anhydrous sodium sulfate, concentrated, and purified by flash column (PE～PE/EA= 1:1) 17d (210mg) was obtained as a yellow oil.
MS m/z(ESI):376[M+1]
+
MS m/z(ESI):376[M+1] +
第三步 5-(3,5-二氟苯甲基)-1H-吡唑并[3,4-c]吡啶17e的合成The third step is the synthesis of 5-(3,5-difluorobenzyl)-1H-pyrazolo[3,4-c]pyridine 17e
将化合物17d(400mg，1.1mmol，1.0eq)溶于1,4-二氧六环(30ml)中，加入4N HCl(20ml，20.0mmol，19.0eq)，70℃反应5h，减压蒸干得17e，未纯化，直接用于下步反应。Dissolve compound 17d (400mg, 1.1mmol, 1.0eq) in 1,4-dioxane (30ml), add 4N HCl (20ml, 20.0mmol, 19.0eq), react at 70°C for 5h, and evaporate to dryness under reduced pressure. 17e, unpurified, used directly in the next step.
MS m/z(ESI):246[M+1]
+
MS m/z(ESI):246[M+1] +
第四步 5-(3,5-二氟苯甲基)-3-碘-1H-吡唑并[3,4-c]吡啶17f的合成Step 4 Synthesis of 5-(3,5-difluorobenzyl)-3-iodo-1H-pyrazolo[3,4-c]pyridine 17f
将17e溶于DMF(8ml)中，依次加入无水碳酸钾(0.54g，3.9mmol，3.0eq)和碘(0.69g，2.7mmol，2.0eq)，室温反应2h。加入EA(50ml)稀释，依次用H2O(15ml)和brine(15ml×2)洗涤，经无水硫酸钠干燥，浓缩，flash柱纯化(PE～PE/EA＝2：1)得黄色固体17f(220mg)。Dissolve 17e in DMF (8ml), add anhydrous potassium carbonate (0.54g, 3.9mmol, 3.0eq) and iodine (0.69g, 2.7mmol, 2.0eq) in sequence, and react at room temperature for 2h. Add EA (50ml) to dilute, wash sequentially with H2O (15ml) and brine (15ml×2), dry over anhydrous sodium sulfate, concentrate, flash column purification (PE～PE/EA=2:1) to obtain yellow solid 17f( 220mg).
MS m/z(ESI):372[M+1]
+
MS m/z(ESI):372[M+1] +
第五步 5-(3,5-二氟苯甲基)-3-碘-1-((2-(三甲基甲硅烷基)乙氧基)甲基)-1H-吡唑并[3,4-c]吡啶17g的合成Step 5 5-(3,5-Difluorobenzyl)-3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3 Synthesis of ,4-c]pyridine 17g
将化合物17f(220mg，0.59mmol，1.0eq)溶于THF(10ml)中，加入NaH(48mg，1.2mmol，2.0eq)，回流反应1h，冷至室温。冰水浴冷却下，加入SEMCl(0.12g，0.71mmol，1.2eq)，继续反应2h。flash柱纯化(PE～PE/EA＝1：1)得黄色油状物17g，放置后固化(150mg)。Compound 17f (220 mg, 0.59 mmol, 1.0 eq) was dissolved in THF (10 ml), NaH (48 mg, 1.2 mmol, 2.0 eq) was added, and the reaction was refluxed for 1 h, and then cooled to room temperature. Under ice-water bath cooling, SEMCl (0.12g, 0.71mmol, 1.2eq) was added, and the reaction was continued for 2h. Flash column purification (PE～PE/EA=1:1) yielded 17 g of yellow oil, which solidified (150 mg) after standing.
MS m/z(ESI):502[M+1]
+
MS m/z(ESI):502[M+1] +
第六步 N-(5-(3,5-二氟苯甲基)-1-((2-(三甲基甲硅烷基)乙氧基)甲基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺17h的合成The sixth step N-(5-(3,5-difluorobenzyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3, Synthesis of 4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 17h
将化合物17g(130mg，0.26mmol，1.0eq)，4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺9a(83mg，0.26mmol，1.0eq)，Cu(17mg，0.26mmol，1.0eq)，CuI(50mg，0.26mmol，1.0eq)，N,N’-二甲基乙二胺(23mg，0.26mmol，1.0eq)和K3PO4(0.17g，0.78mmol，3.0eq)投于甲苯(10ml)中，N2气换气5次，回流反应2h。将反应液直接拌样，flash柱纯化(DCM～DCM/MeOH＝10：1)得黄色固体17h(120mg)。Compound 17g (130mg, 0.26mmol, 1.0eq), 4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 9a( 83mg, 0.26mmol, 1.0eq), Cu (17mg, 0.26mmol, 1.0eq), CuI (50mg, 0.26mmol, 1.0eq), N,N'-dimethylethylenediamine (23mg, 0.26mmol, 1.0eq) ) And K3PO4 (0.17g, 0.78mmol, 3.0eq) were thrown into toluene (10ml), N2 gas was ventilated 5 times, and the reaction was refluxed for 2h. The reaction solution was mixed directly, and purified by flash column (DCM～DCM/MeOH=10:1) to obtain a yellow solid 17h (120mg).
MS m/z(ESI):692[M+1]
+
MS m/z(ESI):692[M+1] +
第七步 N-(5-(3,5-二氟苯甲基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺17的合成。The seventh step N-(5-(3,5-difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazine-1 Synthesis of -yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 17.
将化合物17h(84mg，0.12mmol，1.0eq)溶于DCM(8ml)中，加入TFA(2ml)，室温反应2h，减压蒸干，加入THF(8ml)和H2O(2ml)，加入LiOH(0.48g，20.0mmol)，室温反应过夜。加入H2O(20ml)，用EA(50ml×2)萃取，合并有机层，用brine洗涤，经无水硫酸钠干燥，浓缩，制备纯化(DCM/MeOH＝15：1)得近白色固体17(5mg)。Compound 17h (84mg, 0.12mmol, 1.0eq) was dissolved in DCM (8ml), added TFA (2ml), reacted at room temperature for 2h, evaporated to dryness under reduced pressure, added THF (8ml) and H2O (2ml), added LiOH (0.48) g, 20.0 mmol), react at room temperature overnight. Add H2O (20ml), extract with EA (50ml×2), combine the organic layers, wash with brine, dry over anhydrous sodium sulfate, concentrate, prepare and purify (DCM/MeOH=15:1) to obtain nearly white solid 17 (5mg ).
MS m/z(ESI):562[M+1]
+
MS m/z(ESI):562[M+1] +
1H NMR(400MHz,Chloroform-d)δ8.87(s,1H),8.55(s,1H),8.11(d,J＝7.4Hz,1H),7.77(s,1H),7.52(d,J＝9.0Hz,1H),6.86–6.74(m,2H),6.61(tt,J＝9.1,2.5Hz,1H),6.21(dd,J＝9.0,2.2Hz,1H),6.09(d,J＝2.3Hz,1H),4.23(s,2H),3.99(dt,J＝11.9,3.9Hz,2H),3.65–3.50(m,3H),3.38(t,J＝5.1Hz,4H),2.68(t,J＝5.0Hz,4H),2.44(s,3H),2.02(d,J＝5.1Hz,2H),1.65-1.57(m,2H).
1 H NMR (400MHz, Chloroform-d) δ 8.87 (s, 1H), 8.55 (s, 1H), 8.11 (d, J = 7.4 Hz, 1H), 7.77 (s, 1H), 7.52 (d, J ＝9.0Hz,1H), 6.86–6.74(m,2H), 6.61(tt,J＝9.1,2.5Hz,1H), 6.21(dd,J＝9.0,2.2Hz,1H), 6.09(d,J＝ 2.3Hz, 1H), 4.23 (s, 2H), 3.99 (dt, J = 11.9, 3.9 Hz, 2H), 3.65-3.50 (m, 3H), 3.38 (t, J = 5.1 Hz, 4H), 2.68 ( t,J=5.0Hz,4H),2.44(s,3H),2.02(d,J=5.1Hz,2H),1.65-1.57(m,2H).
实施例18Example 18
N-(6-(3,5-二氟苄基)咪唑并[1,2-b]哒嗪-3-基)-4-(4-甲基哌嗪-1基)-2-((四氢吡喃-4-基)胺基)-苯甲酰胺18的制备N-(6-(3,5-Difluorobenzyl)imidazo[1,2-b]pyridazin-3-yl)-4-(4-methylpiperazin-1 yl)-2-(( Preparation of tetrahydropyran-4-yl)amino)-benzamide 18
采用与实施例9相同的制备方法，以6-氯-咪唑并[1,2-b]哒嗪为原料，可得到化合物18。Using the same preparation method as in Example 9 and using 6-chloro-imidazo[1,2-b]pyridazine as a raw material, compound 18 can be obtained.
MS m/z(ESI):562[M+H]
+。
MS m/z (ESI): 562 [M+H] + .
1H NMR(400MHz,三氟乙酸盐，Methanol-d4)δ7.91(d,J＝9.4Hz,1H),7.85–7.71(m,2H),7.13(d,J＝9.4Hz,1H),7.01–6.92(m,2H),6.85–6.75(m,1H),6.38(dd,J＝9.0,2.4Hz,1H),6.29(d,J＝2.4Hz,1H),4.23(s,2H),3.92(dt,J＝11.9,4.0Hz,2H),3.74-3.65(m,1H),3.62-3.57(m,2H),3.42-3.24(m,8H),2.94(s,3H),2.01(d,J＝4.3Hz,2H),1.59–1.49(m,2H).
1 H NMR (400MHz, trifluoroacetate, Methanol-d4) δ7.91(d,J=9.4Hz,1H), 7.85–7.71(m,2H), 7.13(d,J=9.4Hz,1H) ,7.01–6.92(m,2H), 6.85–6.75(m,1H), 6.38(dd,J=9.0,2.4Hz,1H), 6.29(d,J=2.4Hz,1H), 4.23(s,2H ), 3.92(dt,J=11.9,4.0Hz,2H),3.74-3.65(m,1H),3.62-3.57(m,2H),3.42-3.24(m,8H),2.94(s,3H), 2.01(d,J=4.3Hz,2H), 1.59–1.49(m,2H).
实施例19Example 19
N-(5-((4-(2-羟基丙烷-2-基)哌啶-1-基)甲基)-1H-吲唑-3-基)-4-(4-甲基哌啶-1-基)-2-((四氢吡喃-4-基)胺基)苯甲酰胺19的制备N-(5-((4-(2-hydroxypropan-2-yl)piperidin-1-yl)methyl)-1H-indazol-3-yl)-4-(4-methylpiperidine- Preparation of 1-yl)-2-((tetrahydropyran-4-yl)amino)benzamide 19
第一步 2-氟-5-羟基苯氰19a的制备The first step is the preparation of 2-fluoro-5-hydroxybenzonitrile 19a
向2-氟-5-甲酰基苯氰I-1a(1g，6.7mmol)的二氯甲烷溶液(30mL)中加入三乙酰基硼氢化钠(4g，18.9mmol)，滴入两滴乙酸后室温搅拌反应过夜。加水(30mL*3)洗涤，无水Na2SO4干燥，过滤，浓缩，得到标题化合物19a(930mg，6.2mmol)，收率91.8％。Add sodium triacetyl borohydride (4g, 18.9 mmol) to the dichloromethane solution (30mL) of 2-fluoro-5-formyl benzocyanide I-1a (1g, 6.7mmol), add two drops of acetic acid and room temperature The reaction was stirred overnight. It was washed with water (30 mL*3), dried with anhydrous Na2SO4, filtered, and concentrated to obtain the title compound 19a (930 mg, 6.2 mmol) with a yield of 91.8%.
MS m/z(ESI):152[M+H]+。MS m/z(ESI): 152[M+H]+.
第二步 2-氟-5-溴甲基苯氰19b的制备The second step is the preparation of 2-fluoro-5-bromomethylbenzonitrile 19b
将19a(700mg，4.6mmol)溶于二氯甲烷(20mL)中，冰浴下加入三溴化磷(800mg，2.96mmol)，室温搅拌2小时后冰浴，加入水(20mL)，继续搅拌30分钟，静置分层，有机相加入无水Na2SO4干燥，过滤，浓缩，得到粗品标题化合物19b直接用于下一步。Dissolve 19a (700mg, 4.6mmol) in dichloromethane (20mL), add phosphorus tribromide (800mg, 2.96mmol) under ice bath, stir for 2 hours at room temperature and then ice bath, add water (20mL), continue stirring for 30 After 5 minutes, let stand for layering, add anhydrous Na2SO4 to the organic phase, dry, filter, and concentrate to obtain the crude title compound 19b and use it directly in the next step.
第三步 2-氟-5-((4-(2-羟基丙烷-2-基)哌啶-1-基)甲基)苯氰19d的制备The third step is the preparation of 2-fluoro-5-((4-(2-hydroxypropane-2-yl)piperidin-1-yl)methyl)benzonitrile 19d
化合物19b(985mg，4.6mmol)溶于二氯甲烷(20mL)中，加入二异丙基乙胺(2g，15.5mmol)和2-(4-哌啶基)-2-丙醇19c(700mg，4.89mmol)，室温搅拌反应过夜。浓缩，Flash柱层析纯化(DCM/MeOH,0～10％),得到标题化合物19d(510mg，1.85mmol)，收率39.9％。Compound 19b (985mg, 4.6mmol) was dissolved in dichloromethane (20mL), and diisopropylethylamine (2g, 15.5mmol) and 2-(4-piperidinyl)-2-propanol 19c (700mg, 4.89mmol), the reaction was stirred overnight at room temperature. Concentrated and purified by Flash column chromatography (DCM/MeOH, 0-10%) to obtain the title compound 19d (510 mg, 1.85 mmol) with a yield of 39.9%.
MS m/z(ESI):277[M+H]+。MS m/z(ESI): 277[M+H]+.
第四步 2-(1-((3-胺基-1-氢吲唑-5-基)甲基)哌啶-4-基)-2-丙醇19e的制备The fourth step is the preparation of 2-(1-((3-amino-1-hydroindazol-5-yl)methyl)piperidin-4-yl)-2-propanol 19e
化合物19d(510mg，1.85mmol)溶于乙醇(10mL)中，加入水合肼(10mL)，微波加热至100℃反应2小时后浓缩，Flash柱层析纯化(DCM/MeOH，0～25％)，得标题化合物19e(320mg，1.11mmol)，收率60.1％。Compound 19d (510mg, 1.85mmol) was dissolved in ethanol (10mL), hydrazine hydrate (10mL) was added, the reaction was heated to 100°C in microwave for 2 hours and then concentrated, and purified by Flash column chromatography (DCM/MeOH, 0-25%). The title compound 19e (320 mg, 1.11 mmol) was obtained with a yield of 60.1%.
MS m/z(ESI):289[M+H]+。MS m/z(ESI): 289[M+H]+.
第五步 N-(5-((4-(2-羟基丙烷-2-基)哌啶-1-基)甲基)-1H-吲唑-3-基)-4-(4-甲基哌啶-1-基)-2-(2,2,2-三氟-N-(四氢吡喃-4-基)乙酰胺基)苯甲酰胺19f的制备Step 5 N-(5-((4-(2-hydroxypropan-2-yl)piperidin-1-yl)methyl)-1H-indazol-3-yl)-4-(4-methyl Preparation of piperidin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydropyran-4-yl)acetamido)benzamide 19f
化合物19e(836mg，1.93mmol)溶于二氯甲烷(20mL)中，冰浴下加入1f(320mg，1.11mmol)和二异丙基乙胺(500mg，3.88mmol)，室温反应2小时后浓缩，薄板层析纯化(DCM/MeOH，25％)，得标题化合物19f(120mg，0.17mmol)，收率15.8％。Compound 19e (836mg, 1.93mmol) was dissolved in dichloromethane (20mL), 1f (320mg, 1.11mmol) and diisopropylethylamine (500mg, 3.88mmol) were added under ice bath, reacted at room temperature for 2 hours and then concentrated. Purification by thin-plate chromatography (DCM/MeOH, 25%) gave the title compound 19f (120 mg, 0.17 mmol) with a yield of 15.8%.
MS m/z(ESI):686[M+H]+。MS m/z(ESI): 686[M+H]+.
第七步 N-(5-((4-(2-羟基丙烷-2-基)哌啶-1-基)甲基)-1H-吲唑-3-基)-4-(4-甲基哌啶-1-基)-2-((四氢吡喃-4-基)胺基)苯甲酰胺19的制备The seventh step N-(5-((4-(2-hydroxypropan-2-yl)piperidin-1-yl)methyl)-1H-indazol-3-yl)-4-(4-methyl Preparation of piperidin-1-yl)-2-((tetrahydropyran-4-yl)amino)benzamide 19
化合物19f(120mg，0.17mmol)溶于甲醇(10mL)中，加入碳酸钾(1g，7.2mmol)，室温搅拌过夜，浓缩，薄板层析纯化(DCM/MeOH，25％)，得标题化合物19(10mg，0.017mmol)，收率9.7％。Compound 19f (120mg, 0.17mmol) was dissolved in methanol (10mL), potassium carbonate (1g, 7.2mmol) was added, stirred overnight at room temperature, concentrated, and purified by thin-plate chromatography (DCM/MeOH, 25%) to obtain the title compound 19 ( 10mg, 0.017mmol), yield 9.7%.
MS m/z(ESI):590[M+H]+。MS m/z(ESI): 590[M+H]+.
1H NMR(400MHz,DMSO-d6)δ12.62(s,1H),10.08(s,1H),8.25(d,J＝7.7Hz,1H),7.76(d,J＝9.1Hz,1H),7.49–7.32(m,2H),7.27(d,J＝8.6Hz,1H),6.25–6.16(m,1H),6.10(d,J＝2.3Hz,1H),4.00(s,1H),3.75(m,2H),3.64-3.45(m,5H),3.23(t,J＝5.1Hz,4H),2.87(d,J＝10.3Hz,2H),2.41(t,J＝5.0Hz,4H),2.19(s,3H),1.87-1.75(m,3H),1.59(d,J＝11.7Hz,2H),1.30-1.19(m,4H),0.96(s,6H).
1 H NMR (400MHz, DMSO-d6) δ 12.62 (s, 1H), 10.08 (s, 1H), 8.25 (d, J = 7.7 Hz, 1H), 7.76 (d, J = 9.1 Hz, 1H), 7.49–7.32(m,2H), 7.27(d,J=8.6Hz,1H), 6.25–6.16(m,1H), 6.10(d,J=2.3Hz,1H), 4.00(s,1H), 3.75 (m, 2H), 3.64-3.45 (m, 5H), 3.23 (t, J = 5.1 Hz, 4H), 2.87 (d, J = 10.3 Hz, 2H), 2.41 (t, J = 5.0 Hz, 4H) ,2.19(s,3H),1.87-1.75(m,3H),1.59(d,J=11.7Hz,2H),1.30-1.19(m,4H),0.96(s,6H).
实施例20Example 20
N-(5-(3,5-二氟苯基)-1H-吲唑-3-基)-4-(5-(二甲基胺基)六氢环戊烷[c]并吡咯-2(1H)-基)-2((四氢-2H-吡喃-4-基)胺基)苯甲酰胺20的制备N-(5-(3,5-Difluorophenyl)-1H-indazol-3-yl)-4-(5-(dimethylamino)hexahydrocyclopentane[c]pyrrole-2 Preparation of (1H)-yl)-2((tetrahydro-2H-pyran-4-yl)amino)benzamide 20
采用与实施例12相同的制备方法，以N,N-二甲基八氢环五烷[c]并吡咯-5-氨为原料，可得到化合物20。Using the same preparation method as in Example 12, using N,N-dimethyloctahydrocyclopenta[c]pyrrole-5-ammonia as the raw material, compound 20 can be obtained.
MS m/z(ESI):615[M+1]
+。
MS m/z(ESI): 615[M+1] + .
1H NMR(400MHz,DMSO-d6)δ12.58(s,1H),9.96(s,1H),8.32(d,J＝7.5Hz,1H),7.73(d,J＝9.0Hz,1H),7.44(s,1H),7.36(d,J＝8.6Hz,1H),7.20(dd,J＝8.6,1.6Hz,1H),7.04–6.88(m,3H),5.88(dd,J＝8.9,2.1Hz,1H),5.74(d,J＝2.2Hz,1H),4.00(s,2H),3.77(dt,J＝11.6,3.9Hz,2H),3.65-3.55(m,1H),3.50–3.40(m,2H),3.39–3.35(m,2H),3.19(dd,J＝10.2,3.0Hz,2H),2.65(d,J＝11.8Hz,2H),2.48-2.37(m,1H),2.14-2.03(s,8H),1.91(dd,J＝13.3,3.4Hz,2H),1.36-1.16(m,6H).
1 H NMR (400MHz, DMSO-d6) δ12.58 (s, 1H), 9.96 (s, 1H), 8.32 (d, J = 7.5 Hz, 1H), 7.73 (d, J = 9.0 Hz, 1H), 7.44 (s, 1H), 7.36 (d, J = 8.6 Hz, 1H), 7.20 (dd, J = 8.6, 1.6 Hz, 1H), 7.04-6.88 (m, 3H), 5.88 (dd, J = 8.9, 2.1Hz, 1H), 5.74 (d, J = 2.2 Hz, 1H), 4.00 (s, 2H), 3.77 (dt, J = 11.6, 3.9 Hz, 2H), 3.65-3.55 (m, 1H), 3.50- 3.40 (m, 2H), 3.39–3.35 (m, 2H), 3.19 (dd, J = 10.2, 3.0 Hz, 2H), 2.65 (d, J = 11.8 Hz, 2H), 2.48-2.37 (m, 1H) ,2.14-2.03(s,8H),1.91(dd,J=13.3,3.4Hz,2H),1.36-1.16(m,6H).
实施例21Example 21
N-(5-(1-(3,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺21的制备N-(5-(1-(3,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl) Preparation of -2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 21
第一步 5-(1-(3,5-二氟苯基)-2-羟基乙基)-2-氟苯乙腈21a的制备The first step is the preparation of 5-(1-(3,5-difluorophenyl)-2-hydroxyethyl)-2-fluorobenzeneacetonitrile 21a
化合物10b(2g，6mmol)，氰化锌(702mg,6mmol)和四三苯基磷钯(693mg,0.6mmol)混合在DMF(8mL)中,用氮气置换两次，反应液在微波上110℃反应1小时，冷却后将反应液倒入水(30mL)中，用乙酸乙酯(50×2mL)萃取，有机相干燥后旋干得到粗品，经过柱层析(PE/EA＝30：1)纯化后得到化合物21a(1.5g，5.4mmol)，收率90％。Compound 10b (2g, 6mmol), zinc cyanide (702mg, 6mmol) and palladium tetrakistriphenylphosphorus (693mg, 0.6mmol) were mixed in DMF (8mL), replaced with nitrogen twice, and the reaction solution was microwaved at 110°C Reacted for 1 hour. After cooling, the reaction solution was poured into water (30mL), extracted with ethyl acetate (50×2mL), the organic phase was dried and spin-dried to obtain the crude product, which was subjected to column chromatography (PE/EA=30:1) After purification, compound 21a (1.5 g, 5.4 mmol) was obtained with a yield of 90%.
MS m/z(ESI):278[M+1]
+。
MS m/z (ESI): 278 [M+1] + .
第二步 5-(1-(3,5-二氟苯基)-2-氟乙基)-2-氟苯乙腈21b的制备The second step is the preparation of 5-(1-(3,5-difluorophenyl)-2-fluoroethyl)-2-fluorobenzeneacetonitrile 21b
化合物21a(1.5g,5.41mmol)溶解在二氯甲烷(30mL)中，溶液冷却至0℃，向溶液中缓慢滴加DAST(4.36g,27.0mmol)，反应液在常温下搅拌过夜，然后将反应液缓慢滴加到冰水(30mL)中，用二氯甲烷(50mL)萃取，有机相干燥后旋干得到粗品，经过硅胶柱层析纯化后得到化合物21b(245mg)。Compound 21a (1.5g, 5.41mmol) was dissolved in dichloromethane (30mL), the solution was cooled to 0°C, DAST (4.36g, 27.0mmol) was slowly added dropwise to the solution, the reaction solution was stirred at room temperature overnight, and then The reaction solution was slowly added dropwise to ice water (30 mL), extracted with dichloromethane (50 mL), the organic phase was dried and then spin-dried to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 21b (245 mg).
MS m/z(ESI):280[M+H]
+。
MS m/z (ESI): 280 [M+H] + .
第三步 5-(1-(3,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-胺21c的制备The third step is the preparation of 5-(1-(3,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-amine 21c
化合物21b(245mg,0.88mmol)和水合肼(2.2g,44mmol)溶解在乙醇(6mL) 中，反应液在微波反应器上90℃反应4小时，旋干得到粗品，经过硅胶柱层析纯化(DCM/MeOH＝15:1)后得到化合物21c(94mg)，收率37％。Compound 21b (245mg, 0.88mmol) and hydrazine hydrate (2.2g, 44mmol) were dissolved in ethanol (6mL). The reaction solution was reacted on a microwave reactor at 90°C for 4 hours and spin-dried to obtain the crude product, which was purified by silica gel column chromatography ( After DCM/MeOH=15:1), compound 21c (94mg) was obtained with a yield of 37%.
MS m/z(ESI):292[M+H]
+。
MS m/z (ESI): 292 [M+H] + .
第四步 N-(5-(1-(3,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯甲酰胺21d的制备The fourth step N-(5-(1-(3,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methylpiperazine-1 -Yl)-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido) benzamide 21d
化合物1f(70mg,0.16mmol)溶于在二氯甲烷(10mL)中，滴加到冷却至-20℃的化合物21c(48mg,0.16mmol)和DIPEA(63mg,0.48mmol)的四氢呋喃(10mL)溶液中，反应在-20℃搅拌2小时，然后在常温下继续搅拌2小时，反应液用二氯甲烷(50mL)稀释，用饱和食盐水洗涤，干燥旋干得到粗品，经过柱层析(DCM/MeOH＝15:1)纯化后得到化合物21d(370mg,0.48mmol)，收率45％。Compound 1f (70mg, 0.16mmol) was dissolved in dichloromethane (10mL) and added dropwise to a solution of compound 21c (48mg, 0.16mmol) and DIPEA (63mg, 0.48mmol) in tetrahydrofuran (10mL) cooled to -20°C The reaction was stirred at -20°C for 2 hours, and then continued stirring at room temperature for 2 hours. The reaction solution was diluted with dichloromethane (50mL), washed with saturated brine, dried and spin-dried to obtain the crude product, which was subjected to column chromatography (DCM/ MeOH=15:1) After purification, compound 21d (370 mg, 0.48 mmol) was obtained with a yield of 45%.
MS m/z(ESI):689[M+H]
+。
MS m/z (ESI): 689 [M+H] + .
第五步 N-(5-(1-(3,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺21的制备Step 5 N-(5-(1-(3,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methylpiperazine-1 -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 21
化合物21d(77mg,0.11mmol)溶于甲醇(8mL)和水(8mL)的混合溶剂中，加入碳酸钾(77mg,0.55mmol)，反应液在常温下搅拌3小时，加入水(10ml)，用二氯甲烷(50ml)萃取。有机相用无水硫酸钠干燥，过滤浓缩得到粗品，经过硅胶柱层析纯化(DCM/MOH＝15:1)后得到化合物21(25mg,0.042mmol)，收率38％。Compound 21d (77mg, 0.11mmol) was dissolved in a mixed solvent of methanol (8mL) and water (8mL), potassium carbonate (77mg, 0.55mmol) was added, the reaction solution was stirred at room temperature for 3 hours, and water (10ml) was added. Extract with dichloromethane (50ml). The organic phase was dried with anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by silica gel column chromatography (DCM/MOH=15:1) to obtain compound 21 (25 mg, 0.042 mmol) with a yield of 38%.
MS m/z(ESI):593[M+1]+。MS m/z(ESI): 593[M+1]+.
1H NMR(400MHz,DMSO-d6)δ12.70(s,1H),10.12(s,1H),8.31(d,J＝7.7Hz,1H),7.77(d,J＝9.1Hz,1H),7.52(s,1H),7.45-7.23(m,3H),7.07(t,J＝8.6Hz,2H),6.21(dd,J＝9.0,2.3Hz,1H),6.11(d,J＝2.3Hz,1H),5.10(d,J＝7.9Hz,1H),4.98(d,J＝7.9Hz,1H),4.89-4.79(m,1H),3.84-3.60(m,3H),3.60-3.40(m,2H),3.34(s,4H),2.59(s,4H),2.32(s,3H),2.00-1.82(m,2H),1.24-1.19(m,2H).
1 H NMR(400MHz,DMSO-d6)δ12.70(s,1H), 10.12(s,1H), 8.31(d,J=7.7Hz,1H), 7.77(d,J=9.1Hz,1H), 7.52(s,1H),7.45-7.23(m,3H), 7.07(t,J=8.6Hz,2H), 6.21(dd,J=9.0,2.3Hz,1H),6.11(d,J=2.3Hz ,1H), 5.10(d,J=7.9Hz,1H), 4.98(d,J=7.9Hz,1H), 4.89-4.79(m,1H),3.84-3.60(m,3H), 3.60-3.40( m, 2H), 3.34 (s, 4H), 2.59 (s, 4H), 2.32 (s, 3H), 2.00-1.82 (m, 2H), 1.24-1.19 (m, 2H).
实施例22Example 22
5-(3,5-二氟苯基)-N-(2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)-1H-吲唑-3-甲酰胺22的制备5-(3,5-Difluorophenyl)-N-(2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)-1H-indazole-3-carboxamide 22 Preparation
第一步 1-(3-甲氧基-4-硝基苯基)-4-甲基哌嗪22b的制备The first step is the preparation of 1-(3-methoxy-4-nitrophenyl)-4-methylpiperazine 22b
化合物22a(5.1g，30mmol)和N-甲基哌嗪(6g,60mmol)溶解在N-甲基吡咯烷酮(40ml)中，溶液在100℃下搅拌2小时，冷却后倒入冰水(200mL)，溶液用乙酸乙酯(150mL)萃取，有机相用饱和食盐水洗涤，干燥后旋干得到的粗品经过柱层析纯化后得到化合物22b(6g)。Compound 22a (5.1g, 30mmol) and N-methylpiperazine (6g, 60mmol) were dissolved in N-methylpyrrolidone (40ml). The solution was stirred at 100°C for 2 hours. After cooling, poured into ice water (200mL) The solution was extracted with ethyl acetate (150 mL), the organic phase was washed with saturated brine, dried and then spin-dried to obtain the crude product after purification by column chromatography to obtain compound 22b (6g).
MS m/z(ESI):252[M+1]
+。
MS m/z (ESI): 252[M+1] + .
第二步 2-甲氧基-4-(4-甲基哌嗪-1-基)苯胺22c的制备The second step Preparation of 2-methoxy-4-(4-methylpiperazin-1-yl)aniline 22c
化合物22b(6g，23.9mmol)溶于甲醇(100mL)中，加入钯碳(0.6g)，反应液在氢气氛围下搅拌2小时。反应液过滤旋干得到化合物22c(4.5g,20mmol)，收率85％。Compound 22b (6 g, 23.9 mmol) was dissolved in methanol (100 mL), palladium carbon (0.6 g) was added, and the reaction solution was stirred under a hydrogen atmosphere for 2 hours. The reaction solution was filtered and spin-dried to obtain compound 22c (4.5 g, 20 mmol) with a yield of 85%.
MS m/z(ESI):222[M+1]
+。
MS m/z(ESI): 222[M+1] + .
第三步 二叔丁基5-溴-1H-吲唑-1,3-二羧酸酯22e的制备The third step Preparation of di-tert-butyl 5-bromo-1H-indazole-1,3-dicarboxylate 22e
向化合物22d(1g，4.1mmol)和Boc
2O(2.7g，12.4mmol)的二氯甲烷(10mL)溶液中加入叔丁醇(10mL)和DMAP(0.5g，4mmol)，体系在室温搅拌过夜。反应液旋干，加硅胶拌样，过柱纯化后得到化合物22e(1.5g,3.73mmol)，收率91％。
To the dichloromethane (10mL) solution of compound 22d (1g, 4.1mmol) and Boc 2 O (2.7g, 12.4mmol) was added tert-butanol (10mL) and DMAP (0.5g, 4mmol), and the system was stirred at room temperature overnight . The reaction solution was spin-dried, mixed with silica gel, and purified by column to obtain compound 22e (1.5 g, 3.73 mmol) with a yield of 91%.
MS m/z(ESI):397[M+1]
+。
MS m/z (ESI): 397 [M+1] + .
第四步 叔丁基5-溴-1H-吲唑-3-羧酸酯22f的制备The fourth step is the preparation of tert-butyl 5-bromo-1H-indazole-3-carboxylate 22f
向化合物22e(1.1g，2.8mmol)的甲醇(30mL)溶液中K
2CO
3(2g，14.5mmol)，体系在室温搅拌过夜。向反应液加入水(200mL)继续搅拌15分钟，过滤水洗干燥得到化合物22f(0.8g,2.7mmol)，收率97％。
To a solution of compound 22e (1.1 g, 2.8 mmol) in methanol (30 mL) with K 2 CO 3 (2 g, 14.5 mmol), the system was stirred at room temperature overnight. Water (200 mL) was added to the reaction solution to continue stirring for 15 minutes, filtered, washed with water and dried to obtain compound 22f (0.8 g, 2.7 mmol) with a yield of 97%.
MS m/z(ESI):297[M+1]
+。
MS m/z (ESI): 297 [M+1] + .
第五步 叔丁基5-溴-1-三苯甲基-1H-吲唑-3-羧酸酯22g的制备Step 5 Preparation of 22g tert-butyl 5-bromo-1-trityl-1H-indazole-3-carboxylate
向化合物22f(0.8g，2.7mmol)的二氯甲烷(20mL)溶液中三乙胺(0.55g，5.4mmol)和三氯苯甲烷(0.9g,3.2mmol)，体系在室温搅拌过夜。反应液旋干，加硅胶拌样，过柱纯化得到化合物22g(1.2g,2.2mmol)，收率83％。To a solution of compound 22f (0.8 g, 2.7 mmol) in dichloromethane (20 mL), triethylamine (0.55 g, 5.4 mmol) and trichlorophenylmethane (0.9 g, 3.2 mmol) were added, and the system was stirred at room temperature overnight. The reaction solution was spin-dried, mixed with silica gel, and purified by column to obtain 22 g (1.2 g, 2.2 mmol) of compound with a yield of 83%.
MS m/z(ESI):539[M+1]
+。
MS m/z (ESI): 539 [M+1] + .
第六步 叔丁基5-(4,4,5,5-四甲基-1，3，2-二氧杂硼烷-2-基)-1-三苯甲基-1H-吲唑-3-羧酸酯22h的制备The sixth step: tert-butyl 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-trityl-1H-indazole- Preparation of 3-carboxylate 22h
将化合物22g(100mg，0.18mmol)，频哪醇硼酸酯(100mg，0.39mmol)，Pd(dppf)Cl2(30mg,0.04mmol)和醋酸钾(50mg，0.5mmol)的DMF(5mL)溶液加热到100℃反应3小时后，冷却到室温，过滤，乙酸乙酯(50mL)稀释，饱和食盐水洗涤，干燥过滤浓缩得到化合物22h(100mg,0.17mmol)，收率92％。Heat the solution of compound 22g (100mg, 0.18mmol), pinacol borate (100mg, 0.39mmol), Pd(dppf)Cl2 (30mg, 0.04mmol) and potassium acetate (50mg, 0.5mmol) in DMF (5mL) After reacting at 100°C for 3 hours, it was cooled to room temperature, filtered, diluted with ethyl acetate (50 mL), washed with saturated brine, dried, filtered and concentrated to obtain compound 22h (100 mg, 0.17 mmol) with a yield of 92%.
MS m/z(ESI):587[M+1]
+。
MS m/z (ESI): 587 [M+1] + .
第七步 叔丁基5-(3,5-二氟苯基)-1-三苯甲基-1H-吲唑-3-羧酸酯22i的制备The seventh step is the preparation of tert-butyl 5-(3,5-difluorophenyl)-1-trityl-1H-indazole-3-carboxylate 22i
将化合物22h(500mg，0.85mmol)，1-(溴甲基)-3,5-二氟苯(500mg，2.41mmol)，Pd(PPh
3)
4(200mg，0.17mmol)和磷酸钾(500mg，2.36mmol)的甲苯(10 mL)溶液在氮气保护下加热到100℃反应3小时后，冷却到室温，过滤，乙酸乙酯(50mL)稀释，饱和食盐水洗涤，干燥过滤浓缩得到粗品，经过硅胶柱层析纯化后得到化合物22i(200mg,0.34mmol)，收率40％。
Compound 22h (500mg, 0.85mmol), 1-(bromomethyl)-3,5-difluorobenzene (500mg, 2.41mmol), Pd(PPh 3 ) 4 (200mg, 0.17mmol) and potassium phosphate (500mg, A solution of 2.36mmol) in toluene (10 mL) was heated to 100°C for 3 hours under the protection of nitrogen, then cooled to room temperature, filtered, diluted with ethyl acetate (50mL), washed with saturated brine, dried, filtered and concentrated to obtain a crude product. After purification by column chromatography, compound 22i (200 mg, 0.34 mmol) was obtained with a yield of 40%.
MS m/z(ESI):587[M+1]
+。
MS m/z (ESI): 587 [M+1] + .
第八步 叔丁基5-(3,5-二氟苯基)-1-三苯甲基-1H-吲唑-3-羧酸酯22j的制备Step 8: Preparation of tert-butyl 5-(3,5-difluorophenyl)-1-trityl-1H-indazole-3-carboxylate 22j
向化合物22i(200mg，0.34mmol)的二氯甲烷(8mL)溶液加入三氟乙酸(2mL)，反应液在室温下搅拌2小时，反应液浓缩旋干得到化合物22j(60mg,0.20mmol)，收率61％。Trifluoroacetic acid (2mL) was added to the dichloromethane (8mL) solution of compound 22i (200mg, 0.34mmol). The reaction solution was stirred at room temperature for 2 hours. The reaction solution was concentrated and spin-dried to obtain compound 22j (60mg, 0.20mmol). The rate is 61%.
MS m/z(ESI):307[M+1]
+。
MS m/z (ESI): 307[M+1] + .
第九步 叔丁基5-(3,5-二氟苯基)-N-(2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)-1H-吲唑-3-甲酰胺22的制备Step 9 tert-butyl 5-(3,5-difluorophenyl)-N-(2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)-1H-indazole Preparation of -3-carboxamide 22
将化合物22c(203mg,0.92mmol)和化合物22j(220mg,0.76mmol)溶解在二氯甲烷(15mL)中，加入HATU(433mg,1.14mmol)和DIPEA(44mg,1.52mmol)，反应液在常温下搅拌2小时，反应液用二氯甲烷(30mL)稀释，用饱和食盐水洗涤，有机相干燥旋干得到粗品，经过柱层析纯化再经过反相柱纯化后得到化合物22(55mg,0.11mmol)，收率15％。Dissolve compound 22c (203mg, 0.92mmol) and compound 22j (220mg, 0.76mmol) in dichloromethane (15mL), add HATU (433mg, 1.14mmol) and DIPEA (44mg, 1.52mmol), the reaction solution is at room temperature After stirring for 2 hours, the reaction solution was diluted with dichloromethane (30 mL), washed with saturated brine, and the organic phase was dried and spin-dried to obtain a crude product, which was purified by column chromatography and then purified by reverse phase column to obtain compound 22 (55 mg, 0.11 mmol) , The yield is 15%.
MS m/z(ESI):492[M+1]
+。
MS m/z (ESI): 492 [M+1] + .
1H NMR(400MHz,DMSO-d
6)δ13.66(s,1H),9.26(s,1H),8.14(d,J＝8.8Hz,1H),8.06(s,1H),7.63–7.52(m,1H),7.34(dd,J＝8.7,1.6Hz,1H),7.09–6.90(m,3H),6.68(d,J＝2.5Hz,1H),6.50(dd,J＝8.9,2.5Hz,1H),4.10(s,2H),3.88(s,3H),3.17(s,4H),2.62(s,4H),2.33(s,3H).
1 H NMR (400MHz, DMSO-d 6 ) δ 13.66 (s, 1H), 9.26 (s, 1H), 8.14 (d, J = 8.8 Hz, 1H), 8.06 (s, 1H), 7.63-7.52 ( m, 1H), 7.34 (dd, J = 8.7, 1.6 Hz, 1H), 7.09-6.90 (m, 3H), 6.68 (d, J = 2.5 Hz, 1H), 6.50 (dd, J = 8.9, 2.5 Hz ,1H), 4.10 (s, 2H), 3.88 (s, 3H), 3.17 (s, 4H), 2.62 (s, 4H), 2.33 (s, 3H).
实施例23Example 23
N-(5-(1-(3,5-二氟苯基)-2-甲氧基乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺23的制备N-(5-(1-(3,5-Difluorophenyl)-2-methoxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazine-1- Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 23
第一步 2-溴-4-(1-(3,5-二氟苯基)-2-甲氧基乙基)-1-氟苯23a的制备The first step is the preparation of 2-bromo-4-(1-(3,5-difluorophenyl)-2-methoxyethyl)-1-fluorobenzene 23a
将化合物10b(400mg,1.2mmol)溶解在四氢呋喃(15mL)中，冷却至0℃，加入钠氢(58mg,1.44mmol),反应液在0℃搅拌1小时，再向反应液中加入碘甲烷(198.8mg,1.44mmol)，反应液在室温下搅拌过夜，反应液用饱和氯化铵溶液淬灭，用乙酸乙酯(30mL)萃取，干燥旋干得到粗品，经过柱层析纯化后得到化合物23a(330mg,0.96mmol)，收率80％。Compound 10b (400mg, 1.2mmol) was dissolved in tetrahydrofuran (15mL), cooled to 0°C, sodium hydrogen (58mg, 1.44mmol) was added, the reaction solution was stirred at 0°C for 1 hour, and then methyl iodide ( 198.8mg, 1.44mmol), the reaction solution was stirred overnight at room temperature, the reaction solution was quenched with saturated ammonium chloride solution, extracted with ethyl acetate (30mL), dried and spin-dried to obtain the crude product, which was purified by column chromatography to obtain compound 23a (330mg, 0.96mmol), the yield is 80%.
MS m/z(ESI):345[M+H]
+。
MS m/z (ESI): 345 [M+H] + .
第二步 5-(1-(3,5-二氟苯基)-2-甲氧基乙基)-2-氟苯腈23b的制备The second step is the preparation of 5-(1-(3,5-difluorophenyl)-2-methoxyethyl)-2-fluorobenzonitrile 23b
化合物23a(330mg，0.96mmol)，氰化锌(112mg,0.96mmol)和四三苯基磷钯(115mg,0.1mmol)混合在DMF(4mL)中,用氮气置换两次，反应液在微波上110℃反应1小时，冷却后将反应液倒入水(30mL)中，用乙酸乙酯(50mL)萃取，有机相干燥后旋干得到粗品，经过柱层析(PE/EA＝30：1)纯化后得到化合物23b(200mg，0.69mmol)，收率72％。Compound 23a (330mg, 0.96mmol), zinc cyanide (112mg, 0.96mmol) and palladium tetrakistriphenylphosphorus (115mg, 0.1mmol) were mixed in DMF (4mL), replaced with nitrogen twice, and the reaction solution was microwaved React at 110°C for 1 hour. After cooling, the reaction solution is poured into water (30mL), extracted with ethyl acetate (50mL), the organic phase is dried and spin-dried to obtain the crude product, which is subjected to column chromatography (PE/EA=30:1) After purification, compound 23b (200 mg, 0.69 mmol) was obtained with a yield of 72%.
MS m/z(ESI):292[M+H]
+。
MS m/z (ESI): 292 [M+H] + .
第三步 5-(1-(3,5-二氟苯基)-2-甲氧基乙基)-1H-吲唑-3-胺23c的制备The third step is the preparation of 5-(1-(3,5-difluorophenyl)-2-methoxyethyl)-1H-indazol-3-amine 23c
化合物23b(200mg,0.69mmol)和水合肼(345mg,6.9mmol)混合在N-甲基吡咯烷酮(6mL)中，反应液在微波反应器上90℃反应4小时，冷却后加入到(30mL)中，用乙酸乙酯(50mL)萃取，有机相干燥后旋干得到粗品，经过硅胶柱层析纯化后得到化合物23c(194mg，0.64mmol)。Compound 23b (200mg, 0.69mmol) and hydrazine hydrate (345mg, 6.9mmol) were mixed in N-methylpyrrolidone (6mL), the reaction solution was reacted on a microwave reactor at 90°C for 4 hours, and then added to (30mL) after cooling After extraction with ethyl acetate (50 mL), the organic phase was dried and then spin-dried to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 23c (194 mg, 0.64 mmol).
MS m/z(ESI):304[M+H]
+。
MS m/z (ESI): 304 [M+H] + .
第四步 N-(5-(1-(3,5-二氟苯基)-2-甲氧基)乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯甲酰胺23d的制备The fourth step N-(5-(1-(3,5-difluorophenyl)-2-methoxy)ethyl)-1H-indazol-3-yl)-4-(4-methylpiperidine (Azin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 23d
化合物1f(100mg,0.33mmol)溶于四氢呋喃(5mL)中，加到化合物23c(174.6mg,0.33mmol)和DIPEA(85mg,0.66mmol)的四氢呋喃(15mL)溶液中，反应在-20℃搅拌2小时，然后在常温下继续搅拌2小时，反应液用二氯甲烷(50mL)稀释，用饱和食盐水洗涤，干燥旋干得到粗品，经过柱层析(DCM/MeOH＝10:1)纯化后得到化合物23d(140mg,0.2mmol)，收率60％。Compound 1f (100mg, 0.33mmol) was dissolved in tetrahydrofuran (5mL), added to compound 23c (174.6mg, 0.33mmol) and DIPEA (85mg, 0.66mmol) in tetrahydrofuran (15mL) solution, the reaction was stirred at -20℃2 Hours, then continue to stir at room temperature for 2 hours, the reaction solution was diluted with dichloromethane (50mL), washed with saturated brine, dried and spin-dried to obtain a crude product, which was purified by column chromatography (DCM/MeOH=10:1) Compound 23d (140 mg, 0.2 mmol), yield 60%.
MS m/z(ESI):701[M+H]
+。
MS m/z (ESI): 701 [M+H] + .
第五步 N-(5-(1-(3,5-二氟苯基)-2-甲氧基乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺23的制备Step 5 N-(5-(1-(3,5-Difluorophenyl)-2-methoxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazine -1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 23
化合物23d(140mg,0.2mmol)溶于甲醇(10mL)和水(5mL)的混合溶剂中，加入碳酸钾(82.8mg,0.6mmol)，反应液在常温下搅拌3小时，加入水(20ml)，用二氯甲烷(30ml)萃取。有机相用无水硫酸钠干燥，过滤浓缩得到粗品，经过硅胶柱层析纯化后得到化合物23(20mg,0.033mmol)，收率17％。Compound 23d (140mg, 0.2mmol) was dissolved in a mixed solvent of methanol (10mL) and water (5mL), potassium carbonate (82.8mg, 0.6mmol) was added, the reaction solution was stirred at room temperature for 3 hours, and water (20ml) was added. Extract with dichloromethane (30ml). The organic phase was dried with anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 23 (20 mg, 0.033 mmol) with a yield of 17%.
MS m/z(ESI):605[M+H]
+。
MS m/z (ESI): 605 [M+H] + .
1H NMR(400MHz,DMSO-d6)δ12.63(s,1H),10.09(s,1H),8.32(d,J＝7.7Hz,1H),7.76(d,J＝9.0Hz,1H),7.49(s,1H),7.37(d,J＝8.7Hz,1H),7.34–7.19(m,2H),7.01(t,J＝8.8Hz,2H),6.20(dd,J＝9.0,2.3Hz,1H),6.10(d,J＝2.4Hz,1H),4.67(t,J＝8.0Hz,1H),3.99(dt,J＝28.0,8.9Hz,2H),3.86–3.73(m,2H), 3.73–3.59(m,1H),3.54–3.41(m,2H),3.24(d,J＝2.6Hz,7H),2.40(t,J＝5.0Hz,4H),2.19(s,3H),1.92(d,J＝13.2Hz,2H),1.36-1.28(m,2H).
1 H NMR (400MHz, DMSO-d6) δ 12.63 (s, 1H), 10.09 (s, 1H), 8.32 (d, J = 7.7 Hz, 1H), 7.76 (d, J = 9.0 Hz, 1H), 7.49(s,1H),7.37(d,J=8.7Hz,1H),7.34-7.19(m,2H),7.01(t,J=8.8Hz,2H), 6.20(dd,J=9.0,2.3Hz ,1H),6.10(d,J=2.4Hz,1H),4.67(t,J=8.0Hz,1H),3.99(dt,J=28.0,8.9Hz,2H),3.86-3.73(m,2H) , 3.73–3.59(m,1H),3.54–3.41(m,2H), 3.24(d,J=2.6Hz,7H), 2.40(t,J=5.0Hz,4H), 2.19(s,3H), 1.92(d,J=13.2Hz,2H),1.36-1.28(m,2H).
实施例24Example 24
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-2-甲基-7-((四氢-2H-吡喃-4-基)氨基)-1,2,3,4-四氢异喹啉-6-甲酰胺24的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-2-methyl-7-((tetrahydro-2H-pyran-4-yl)amino) -Preparation of 1,2,3,4-tetrahydroisoquinoline-6-carboxamide 24
第一步 1-(6-溴-1,2,3,4-四氢异喹啉-2-基)-2,2,2-三氟乙烷-1-酮24bThe first step 1-(6-bromo-1,2,3,4-tetrahydroisoquinolin-2-yl)-2,2,2-trifluoroethane-1-one 24b
向干燥的反应瓶中加入24a(2.12g,10mmol)和10mL三氟乙酸酐，加完后室温搅拌1小时，TLC显示原料反应完。减压浓缩反应液，乙酸乙酯稀释，冷的碳酸氢钠溶液洗涤3次，有机相分出后用无水硫酸钠干燥，然后加压浓缩，得到的油状物24b直接用于下一步反应。Add 24a (2.12 g, 10 mmol) and 10 mL of trifluoroacetic anhydride to the dry reaction flask, and stir at room temperature for 1 hour after the addition. TLC shows that the reaction of the raw materials is complete. The reaction solution was concentrated under reduced pressure, diluted with ethyl acetate, washed with cold sodium bicarbonate solution 3 times, the organic phase was separated and dried with anhydrous sodium sulfate, and then concentrated under pressure, the obtained oily substance 24b was directly used in the next reaction.
MS m/z(ESI):308[M+H]+MS m/z(ESI):308[M+H]+
第二步 1-(6-溴-7-硝基-1,2,3,4-四氢异喹啉-2-基)-2,2,2-三氟乙烷-1-酮24c的制备The second step 1-(6-bromo-7-nitro-1,2,3,4-tetrahydroisoquinolin-2-yl)-2,2,2-trifluoroethane-1-one 24c preparation
向反应瓶中加入50mL浓硫酸，冷却至0～5℃后加入化合物24b(3.1g,10mmol)，加完后分批次加硝酸钾(1.2g,12.0mmol)，保持内温不高于10℃。加完后搅拌过夜。TLC显示原料反应完。反应液倒入冰水中，析出固体，过滤，得到的粗品24c纯度足够用于下一步反应。Add 50mL concentrated sulfuric acid to the reaction flask, cool to 0～5℃, add compound 24b (3.1g, 10mmol), add potassium nitrate (1.2g, 12.0mmol) in batches after the addition, keep the internal temperature not higher than 10 ℃. Stir overnight after addition. TLC showed that the reaction of the starting materials was complete. The reaction solution was poured into ice water, and solids were separated out and filtered. The crude product 24c obtained was pure enough to be used in the next reaction.
MS m/z(ESI):353[M+H]+MS m/z(ESI):353[M+H]+
第三步 7-硝基-2-(2,2,2-三氟乙酰基)-1,2,3,4-四氢异喹啉-6-甲腈24d的制备The third step is the preparation of 7-nitro-2-(2,2,2-trifluoroacetyl)-1,2,3,4-tetrahydroisoquinoline-6-carbonitrile 24d
向微波反应瓶加入24c(353mg,1.0mmol)，CuCN(180mg,2.0mmol)，和DMF(5mL).混合物在130℃下微波加热1小时，TLC显示原料基本反应完。反应液用大量乙酸乙酯稀释，过滤，滤液用氯化铵溶液洗涤。有机相用无水硫酸钠干燥后减压浓缩至较小体积，加入适量石油醚打浆，过滤得到的粗品24d直接用于下一步反应。24c (353 mg, 1.0 mmol), CuCN (180 mg, 2.0 mmol), and DMF (5 mL) were added to the microwave reaction flask. The mixture was heated in microwave at 130° C. for 1 hour. TLC showed that the raw materials were basically reacted. The reaction solution was diluted with a large amount of ethyl acetate, filtered, and the filtrate was washed with ammonium chloride solution. The organic phase was dried with anhydrous sodium sulfate and concentrated under reduced pressure to a smaller volume, and an appropriate amount of petroleum ether was added to make a slurry, and the crude product obtained by filtration for 24 d was directly used in the next reaction.
MS m/z(ESI):300[M+H]+MS m/z(ESI): 300[M+H]+
第四步 7-硝基-1,2,3,4-四氢异喹啉-6-甲腈24e的制备Step 4 Preparation of 7-nitro-1,2,3,4-tetrahydroisoquinoline-6-carbonitrile 24e
将24d(300mg,1.0mmol)溶于20ml甲醇中，加入10ml饱和碳酸钾溶液，搅拌半小时TLC显示原料反应完。减压浓缩出去反应液中的大部分甲醇，加入乙酸乙酯和水分液，有机相用无水硫酸钠干燥，flash纯化(二氯甲烷:甲醇＝20：1)得24e(142mg,0.7mmol,70％)。Dissolve 24d (300mg, 1.0mmol) in 20ml of methanol, add 10ml of saturated potassium carbonate solution, and stir for half an hour. TLC shows that the reaction of the raw materials is complete. Concentrate under reduced pressure to remove most of the methanol in the reaction solution, add ethyl acetate and water solution, dry the organic phase with anhydrous sodium sulfate, flash purification (dichloromethane: methanol = 20:1) to obtain 24e (142mg, 0.7mmol, 70%).
MS m/z(ESI):204[M+H]+MS m/z(ESI):204[M+H]+
第五步 2-甲基-7-硝基-1,2,3,4-四氢异喹啉-6-甲腈24f的制备The fifth step is the preparation of 2-methyl-7-nitro-1,2,3,4-tetrahydroisoquinoline-6-carbonitrile 24f
化合物24e(1.0g,5mmol)溶于50ml二氯甲烷中,加入多聚甲醛(1.5g，50mmol)和5ml醋酸，再加入NaBH(OAc)3(2.12g,10mmol).室温搅拌过夜，TLC显示大部分原料反应完。加入碳酸钾水溶液淬灭反应，分出有机相，加入硅胶拌样，flash纯化(二氯甲烷:甲醇＝20：1)得24f(650mg,3mmol,60％)。Compound 24e (1.0g, 5mmol) was dissolved in 50ml of dichloromethane, paraformaldehyde (1.5g, 50mmol) and 5ml of acetic acid were added, and NaBH(OAc)3 (2.12g, 10mmol) was added. Stir at room temperature overnight, TLC showed Most of the raw materials have been reacted. The reaction was quenched by adding potassium carbonate aqueous solution, the organic phase was separated, and silica gel was added to mix the sample, and flash purification (dichloromethane:methanol=20:1) to obtain 24f (650mg, 3mmol, 60%).
MS m/z(ESI):218[M+H]+MS m/z(ESI):218[M+H]+
第六步 2-甲基-7-硝基-1,2,3,4-四氢异喹啉-6-羧酸24g的制备Step 6 Preparation of 24g of 2-methyl-7-nitro-1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid
向微波反应瓶中加入24f(650mg,3mmol)，10ml溴化氢溶液(50％wt)，5ml醋酸，然后120℃下微波加热1小时。TLC显示大部分原料反应完。反应液室温过夜后有大量固体析出，过滤所得固体即是24g(350mg,1.5mmol,50％)。Add 24f (650mg, 3mmol), 10ml hydrogen bromide solution (50%wt), 5ml acetic acid to the microwave reaction flask, and then microwave heating at 120°C for 1 hour. TLC showed that most of the raw materials had reacted. After the reaction solution was at room temperature overnight, a large amount of solid precipitated out, and the solid obtained by filtration was 24 g (350 mg, 1.5 mmol, 50%).
MS m/z(ESI):237[M+H]+MS m/z(ESI):237[M+H]+
第七步 N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-2-甲基-7-硝基-1,2,3,4-四氢异喹啉-6-甲酰胺24h的制备Step 7 N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-2-methyl-7-nitro-1,2,3,4-tetrahydro Preparation of isoquinoline-6-carboxamide 24h
向干燥的反应瓶中加入化合物24g(100mg,0.42mmol)，二氯甲烷和1ml草酰氯。室温搅拌4小时后减压浓缩，用二氯甲烷带干，加入干燥的四氢呋喃和0.2ml DIPEA,氮气保护下冷却至-40～-50℃。然后缓慢加入5-(3,5-二氟苯甲基)-1H-吲唑-3-胺(109mg,0.42mmol)的四氢呋喃溶液。加完后自然升至室温。TLC显示大部分原料反应完。加入硅胶拌样，flash纯化(二氯甲烷:甲醇＝15：1)得24h(60mg,0.12mmol,28％).To the dry reaction flask was added 24 g (100 mg, 0.42 mmol) of compound, dichloromethane and 1 ml of oxalyl chloride. Stir at room temperature for 4 hours, then concentrate under reduced pressure, dry with dichloromethane, add dry tetrahydrofuran and 0.2ml DIPEA, and cool to -40～-50℃ under nitrogen protection. Then a solution of 5-(3,5-difluorobenzyl)-1H-indazol-3-amine (109 mg, 0.42 mmol) in tetrahydrofuran was slowly added. After the addition, it will naturally rise to room temperature. TLC showed that most of the raw materials had reacted. Add silica gel to mix the sample, flash purification (dichloromethane: methanol = 15:1) to obtain 24h (60mg, 0.12mmol, 28%).
MS m/z(ESI):478[M+H]+MS m/z(ESI):478[M+H]+
第八步 7-氨基-N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-2-甲基-1,2,3,4-四氢异喹啉-6-甲酰胺24i的制备Step 8 7-Amino-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-2-methyl-1,2,3,4-tetrahydroiso Preparation of quinoline-6-carboxamide 24i
向反应瓶中加入24h(60mg,0.12mmol),5ml四氢呋喃和0.5ml水。然后加入65mg还原性铁粉。加完后60℃下回流2小时。TLC显示原料反应完。加入大量乙酸乙酯稀释反应液，过滤所得滤液用flash纯化(二氯甲烷:甲醇＝15：1)得24i(30mg,0.067mmol,50％).Add 24h (60mg, 0.12mmol), 5ml tetrahydrofuran and 0.5ml water to the reaction flask. Then add 65mg of reducing iron powder. After the addition, reflux at 60°C for 2 hours. TLC showed that the reaction of the starting materials was complete. A large amount of ethyl acetate was added to dilute the reaction solution, and the filtrate obtained by filtration was purified with flash (dichloromethane: methanol = 15:1) to obtain 24i (30mg, 0.067mmol, 50%).
MS m/z(ESI):448[M+H]+MS m/z(ESI):448[M+H]+
第九步 N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-2-甲基-7-((四氢-2H-吡喃-4-基)氨基)-1,2,3,4-四氢异喹啉-6-甲酰胺24的制备Step 9 N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-2-methyl-7-((tetrahydro-2H-pyran-4-yl )Amino)-1,2,3,4-tetrahydroisoquinoline-6-carboxamide 24 preparation
将化合物24i(30mg,0.067mmol)溶于二氯甲烷中，加入I-2(100mg,1.0mmol)和0.2ml醋酸。然后加入NaBH(OAc)
3(21.2mg,0.1mmol).室温搅拌过夜。加入碳酸钾溶液，分出有机相，减压浓缩后用制备薄层色谱纯化(二氯甲烷:甲醇＝10：1)得24(10mg,0.0188mmol,30％).
Compound 24i (30 mg, 0.067 mmol) was dissolved in dichloromethane, and I-2 (100 mg, 1.0 mmol) and 0.2 ml of acetic acid were added. Then NaBH(OAc) 3 (21.2 mg, 0.1 mmol) was added. Stir at room temperature overnight. Potassium carbonate solution was added to separate the organic phase, concentrated under reduced pressure and purified by preparative thin-layer chromatography (dichloromethane: methanol = 10:1) to give 24 (10mg, 0.0188mmol, 30%).
MS m/z(ESI):532[M+H]+MS m/z(ESI): 532[M+H]+
1H NMR(400MHz,DMSO-d6)δ12.70(s,1H),10.39(s,1H),7.67(s,1H),7.59(d,J＝7.6Hz,1H),7.51(s,1H),7.42(d,J＝8.6Hz,1H),7.26(dd,J＝8.5,1.6Hz,1H),7.06–6.91(m,3H),6.52(s,1H),4.04(s,2H),3.86-3.77(m,2H),3.62-3.39(m,5H),2.78-2.70(m,2H),2.68-2.56(m,2H),2.35(s,3H),1.97-1.86(m,2H),1.38-1.27(m,2H).
1 H NMR(400MHz,DMSO-d6)δ12.70(s,1H), 10.39(s,1H), 7.67(s,1H), 7.59(d,J=7.6Hz,1H), 7.51(s,1H) ),7.42(d,J=8.6Hz,1H),7.26(dd,J=8.5,1.6Hz,1H),7.06-6.91(m,3H),6.52(s,1H),4.04(s,2H) ,3.86-3.77(m,2H),3.62-3.39(m,5H),2.78-2.70(m,2H),2.68-2.56(m,2H),2.35(s,3H),1.97-1.86(m, 2H), 1.38-1.27 (m, 2H).
实施例25Example 25
4-((1R,3S,5s,7s)-5-氨基金刚烷-2-基)-N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺盐酸盐25的制备4-((1R,3S,5s,7s)-5-aminoadamantan-2-yl)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl) Preparation of -2-((tetrahydro-2H-pyran-4-yl)amino)benzamide hydrochloride 25
第一步 乙基2-溴-4-碘苯酸酯25b的制备The first step: Preparation of ethyl 2-bromo-4-iodobenzoate 25b
向干燥的反应瓶中加入25a(3.28g,10mmol和50ml DCM，加完后加入10ml草酰氯，加完后室温搅拌1小时，TLC显示原料反应完。减压浓缩反应液，乙 酸乙酯稀释，碳酸氢钠溶液洗涤3次，有机相分出后用无水硫酸钠干燥，然后加压浓缩，得到的油状物25b直接用于下一步反应。Add 25a (3.28g, 10mmol and 50ml DCM to the dry reaction flask, add 10ml oxalyl chloride after the addition is complete, stir at room temperature for 1 hour after the addition, TLC shows that the reaction of the raw materials is complete. The reaction solution is concentrated under reduced pressure, diluted with ethyl acetate, Washed with sodium bicarbonate solution for 3 times, the organic phase was separated and dried with anhydrous sodium sulfate, and then concentrated under pressure. The obtained oily substance 25b was directly used in the next reaction.
MS m/z(ESI):357[M+H]
+
MS m/z(ESI): 357[M+H] +
第二步 苯甲基(1s,3R,5S,7s)-4-(3-溴-4-(乙氧羰基)苯基)-4-羟基金刚烷-1-羧酸酯25d的制备The second step is the preparation of benzyl (1s, 3R, 5S, 7s)-4-(3-bromo-4-(ethoxycarbonyl)phenyl)-4-hydroxyadamantane-1-carboxylate 25d
向反应瓶中加入25b(350mg,1.0mmol),和10ml四氢呋喃，冷却至-20℃然后加入i-PrMgCl(1.0ml,1M)，此温度下搅拌0.5小时，加入25c(284mg,1.0mmol)，自然升温过夜。TLC显示有产物生成。加入氯化铵溶液淬灭反应。加入乙酸乙酯萃取，食盐水洗涤有机相。有机相分出后加入硅胶拌样，flash纯化(PE:EA＝10：1)得25d(250mg,0.5mmol,50％)。Add 25b (350mg, 1.0mmol), and 10ml of tetrahydrofuran to the reaction flask, cool to -20°C and then add i-PrMgCl (1.0ml, 1M), stir at this temperature for 0.5 hours, add 25c (284mg, 1.0mmol), Heat up naturally overnight. TLC showed product formation. The reaction was quenched by adding ammonium chloride solution. Ethyl acetate was added for extraction, and the organic phase was washed with brine. After the organic phase was separated, silica gel was added to mix the sample, and the flash purified (PE:EA=10:1) to obtain 25d (250mg, 0.5mmol, 50%).
MS m/z(ESI):497[M-OH]
+
MS m/z(ESI):497[M-OH] +
第三步 苯甲基(1s,3R,5S,7s)-4-(3-溴-4-(乙氧羰基)苯基)金刚烷-1-羧酸酯25e的制备The third step is the preparation of benzyl (1s, 3R, 5S, 7s)-4-(3-bromo-4-(ethoxycarbonyl)phenyl)adamantane-1-carboxylate 25e
向单口瓶中加入25d(500mg,1.0mmol),5ml三氟乙酸，1ml Et
3SiH.室温搅拌2小时，TLC显示原料反应完。减压浓缩掉大部分三氟乙酸，加入DCM硅胶拌样，flash纯化(PE:EA＝10：1)得25e(495mg,1.0mmol,100％).
Add 25d (500mg, 1.0mmol), 5ml trifluoroacetic acid, 1ml Et 3 SiH to the single-neck flask. Stir at room temperature for 2 hours, TLC shows that the raw materials have reacted completely. Concentrate most of the trifluoroacetic acid under reduced pressure, add DCM silica gel to mix the sample, flash purification (PE:EA=10:1) to obtain 25e (495mg, 1.0mmol, 100%).
MS m/z(ESI):497[M+H]
+
MS m/z(ESI):497[M+H] +
第四步 苯甲基(1s,3R,5S,7s)-4-(3-((叔-丁氧基羰基)氨基)-4-(乙氧羰基)苯基)金刚烷-1-羧酸酯25f的制备Step 4 Benzyl(1s,3R,5S,7s)-4-(3-((tert-butoxycarbonyl)amino)-4-(ethoxycarbonyl)phenyl)adamantane-1-carboxylic acid Preparation of ester 25f
将25e(495mg,1.0mmol),碳酸铯(646mg,2.0mmol),醋酸钯(2.2mg,0.1mmol),XantPhos(5.78mg,0.1mmol),氨基甲酸叔丁酯(117mg,1.0mmol)混合，加入10ml二氧六环。100℃下搅拌1小时，TLC显示原料反应完。加入乙酸乙酯稀释反应液并过滤，得到的滤液用flash纯化(PE:EA＝10：1)得25f(265mg,0.5mmol,50％).Mix 25e (495mg, 1.0mmol), cesium carbonate (646mg, 2.0mmol), palladium acetate (2.2mg, 0.1mmol), XantPhos (5.78mg, 0.1mmol), t-butyl carbamate (117mg, 1.0mmol), Add 10ml of dioxane. After stirring at 100°C for 1 hour, TLC showed that the reaction of the starting materials was complete. Add ethyl acetate to dilute the reaction solution and filter, and the resulting filtrate was flash purified (PE:EA=10:1) to give 25f (265mg, 0.5mmol, 50%).
MS m/z(ESI):534[M+H]
+
MS m/z(ESI): 534[M+H] +
第五步 (1s,3R,5S,7s)-4-(3-((叔-丁氧基羰基)氨基)-4-(乙氧羰基)苯基)金刚烷-1-羧酸25g的制备Step 5 Preparation of (1s, 3R, 5S, 7s)-4-(3-((tert-butoxycarbonyl)amino)-4-(ethoxycarbonyl)phenyl)adamantane-1-carboxylic acid 25g
化合物25f(1.06g,2.0mmol)溶于四氢呋喃中，加入湿钯炭(500mg)，在氢气氛围中(氢气球)搅拌过夜。TLC显示原料反应完，硅藻土过滤，滤液浓缩即是25g(600mg,68％)。Compound 25f (1.06 g, 2.0 mmol) was dissolved in tetrahydrofuran, wet palladium carbon (500 mg) was added, and the mixture was stirred overnight in a hydrogen atmosphere (hydrogen balloon). TLC showed that the reaction of the raw materials was complete, filtered through Celite, and the filtrate was concentrated to be 25 g (600 mg, 68%).
MS m/z(ESI):444[M+H]
+
MS m/z(ESI):444[M+H] +
第六步 乙基4-((1R,3S,5s,7s)-5-(((苄氧基)羰基)氨基)金刚烷-2-基)-2-((叔-丁氧基羰基)氨基)苯酸酯25h的制备Step 6 Ethyl 4-((1R,3S,5s,7s)-5-(((benzyloxy)carbonyl)amino)adamantan-2-yl)-2-((tert-butoxycarbonyl) Preparation of amino) benzoate 25h
向干燥的反应瓶中加入25g(444mg,1.0mmol),10ml甲苯，DDPA(275mg,1.0mmol)，苯甲醇(216mg,2.0mmol)和三乙胺(200mg,2.0mmol)。混合物在120℃下反应4小时，TLC显示原料基本反应完。加入乙酸乙酯稀释反应液，用碳酸氢钠洗涤，有机相分出后用flash纯化(PE:EA＝15：1)得25h(400mg,73％).Add 25g (444mg, 1.0mmol), 10ml of toluene, DDPA (275mg, 1.0mmol), benzyl alcohol (216mg, 2.0mmol) and triethylamine (200mg, 2.0mmol) to the dry reaction flask. The mixture was reacted at 120°C for 4 hours, and TLC showed that the raw materials were basically reacted. Add ethyl acetate to dilute the reaction solution, wash with sodium bicarbonate, separate the organic phase and purify it with flash (PE:EA=15:1) to give 25h (400mg, 73%).
MS m/z(ESI):549[M+H]
+
MS m/z(ESI):549[M+H] +
第七步 乙基2-氨基-4-((1R,3S,5s,7s)-5-(((苄氧基)羰基)氨基)金刚烷-2-基)苯酸酯25i的制备The seventh step is the preparation of ethyl 2-amino-4-((1R,3S,5s,7s)-5-(((benzyloxy)carbonyl)amino)adamantan-2-yl)benzoate 25i
向干燥的反应瓶中加入化合物25h(548mg,1.0mmol)，20ml二氯甲烷，然后加入2ml三氟乙酸。加完后室温搅拌过夜。TLC显示原料反应完。减压浓缩反应液除去大部分三氟乙酸和二氯甲烷，然后加入乙酸乙酯稀释，碳酸氢钠溶液洗涤，无水硫酸钠干燥，减压浓缩得25i(450mg,100％).Add compound 25h (548 mg, 1.0 mmol), 20 ml of dichloromethane, and then 2 ml of trifluoroacetic acid to the dry reaction flask. After the addition, stir at room temperature overnight. TLC showed that the reaction of the starting materials was complete. The reaction solution was concentrated under reduced pressure to remove most of the trifluoroacetic acid and dichloromethane, then diluted with ethyl acetate, washed with sodium bicarbonate solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 25i (450mg, 100%).
MS m/z(ESI):449[M+H]
+
MS m/z(ESI):449[M+H] +
第八步 乙基4-((1R,3S,5s,7s)-5-(((苄氧基)羰基)氨基)金刚烷-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酸酯25j的制备Step 8 Ethyl 4-((1R,3S,5s,7s)-5-(((benzyloxy)carbonyl)amino)adamantan-2-yl)-2-((tetrahydro-2H-pyran) Preparation of -4-yl)amino)benzoate 25j
向反应瓶中加入四氢-4H-吡喃-4-酮(200mg,2.0mmol),25i(450mg,1.0mmol),二氯甲烷，醋酸(1.0ml)和NaBH(OAc)
3(636mg,3.0mmol)。室温下搅拌过夜，然后加入碳酸氢钠淬灭反应，有机相分出后加入硅胶拌样，flash纯化(PE/EA＝10：1)25j(370mg,0.7mmol,70％)。
Add tetrahydro-4H-pyran-4-one (200mg, 2.0mmol), 25i (450mg, 1.0mmol), dichloromethane, acetic acid (1.0ml) and NaBH(OAc) 3 (636mg, 3.0mmol) to the reaction flask. mmol). After stirring overnight at room temperature, sodium bicarbonate was added to quench the reaction. After the organic phase was separated, silica gel was added for sample mixing, and flash purification (PE/EA=10:1) 25j (370 mg, 0.7 mmol, 70%).
MS m/z(ESI):533[M+H]
+
MS m/z(ESI):533[M+H] +
第九步 4-((1R,3S,5s,7s)-5-(((苄氧基)羰基)氨基)金刚烷-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酸25k的制备Step 9 4-((1R,3S,5s,7s)-5-(((benzyloxy)carbonyl)amino)adamantan-2-yl)-2-((tetrahydro-2H-pyran-4 -(Yl)amino)benzoic acid 25k preparation
将化合物25j(532mg,1.0mmol)溶于10ml四氢呋喃中，加入2ml氢氧化锂溶液(10％)，室温搅拌2小时，TLC显示原料反应完。减压除去大部分四氢呋喃，加适量的水稀释残留物，加入适量柠檬酸中和反应液，加入乙酸乙酯萃取3次以上。合并有机相，饱和食盐水洗涤，无水硫酸钠干燥后减压浓缩得到25k(400mg,0.8mmol,80％)。Compound 25j (532 mg, 1.0 mmol) was dissolved in 10 ml of tetrahydrofuran, 2 ml of lithium hydroxide solution (10%) was added, and the mixture was stirred at room temperature for 2 hours. TLC indicated that the reaction of the raw materials was complete. Remove most of the tetrahydrofuran under reduced pressure, add an appropriate amount of water to dilute the residue, add an appropriate amount of citric acid to neutralize the reaction solution, and add ethyl acetate to extract more than 3 times. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 25k (400 mg, 0.8 mmol, 80%).
MS m/z(ESI):505[M+H]
+
MS m/z(ESI):505[M+H] +
第十步 4-((1R,3S,5s,7s)-5-(((苄氧基)羰基)氨基)金刚烷-2-基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯甲酸25l的制备Step 10 4-((1R,3S,5s,7s)-5-(((benzyloxy)carbonyl)amino)adamantan-2-yl)-2-(2,2,2-trifluoro-N -(Tetrahydro-2H-pyran-4-yl)acetamido)benzoic acid 25l
向干燥的反应瓶中加入25k(252mg,0.5mmol),0.5ml三氟乙酸酐和二氯甲烷。室温搅拌1小时后TLC显示原料反应完。加入水搅拌半小时，分出有机相用无水硫酸钠干燥，减压浓缩后即得25l(300mg,0.5mmol,100％).Add 25k (252mg, 0.5mmol), 0.5ml trifluoroacetic anhydride and dichloromethane to the dry reaction flask. After stirring at room temperature for 1 hour, TLC showed that the reaction of the starting materials was complete. Add water and stir for half an hour. Separate the organic phase and dry with anhydrous sodium sulfate. After concentration under reduced pressure, 25l (300mg, 0.5mmol, 100%) is obtained.
MS m/z(ESI):601[M+H]
+
MS m/z(ESI):601[M+H] +
第十一步 苯甲基((1s,3R,5S,7s)-4-(4-((5-(3,5-二氟苄基)-1H-吲唑-3-基)甲酰胺)-3-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)-乙酰氨基)苯基)金刚烷-1-基)氨基甲酸酯25m的制备Eleventh step Benzyl ((1s,3R,5S,7s)-4-(4-((5-(3,5-difluorobenzyl)-1H-indazol-3-yl)carboxamide) Preparation of -3-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)-acetylamino)phenyl)adamantan-1-yl)carbamate 25m
向干燥的反应瓶中加入25l(60mg,0.1mmol),5ml二氯甲烷和1ml草酰氯。室温下搅拌4-6小时，减压浓缩该反应液至干，加入四氢呋喃稀释，冷却至-40℃，氮气保护。然后加入0.2ml DIPEA，再缓慢加入I-2(26mg,0.1mmol)的THF溶液。加完后自然升温至室温。TLC显示大部分原料反应完。浓缩反应液至适当体积，然后用制备薄层色谱纯化(DCM:MeOH＝10:1)得25m(42mg,0.05mmol,50％)。Add 25l (60mg, 0.1mmol), 5ml dichloromethane and 1ml oxalyl chloride to the dry reaction flask. Stir at room temperature for 4-6 hours, concentrate the reaction solution under reduced pressure to dryness, add tetrahydrofuran to dilute, and cool to -40°C under nitrogen protection. Then add 0.2ml DIPEA, and then slowly add the THF solution of I-2 (26mg, 0.1mmol). After the addition, the temperature was naturally raised to room temperature. TLC showed that most of the raw materials had reacted. The reaction solution was concentrated to an appropriate volume, and then purified by preparative thin layer chromatography (DCM:MeOH=10:1) to obtain 25m (42mg, 0.05mmol, 50%).
MS m/z(ESI):842.9[M+H]
+
MS m/z(ESI): 842.9[M+H] +
第十二步 苯甲基((1s,3R,5S,7s)-4-(4-((5-(3,5-二氟苄基)-1H-吲唑-3-基)甲酰胺)-3-((四氢-2H-吡喃-4-基)胺)苯基)金刚烷-1-基)氨基甲酸酯25n的制备Step 12 Benzyl ((1s,3R,5S,7s)-4-(4-((5-(3,5-difluorobenzyl)-1H-indazol-3-yl)carboxamide) Preparation of -3-((tetrahydro-2H-pyran-4-yl)amine)phenyl)adamantan-1-yl)carbamate 25n
向反应瓶中加入25m(42mg,0.05mmol)和3ml甲醇，溶解后加入0.5ml碳酸钾溶液，10min后TLC显示原料反应完。减压浓缩出去甲醇，加入乙酸乙酯萃取，无水硫酸钠干燥，减压浓缩即得25n(30mg,0.04mmol,80％)。Add 25m (42mg, 0.05mmol) and 3ml of methanol to the reaction flask, add 0.5ml of potassium carbonate solution after dissolution, and TLC after 10min shows that the raw materials have reacted. Concentrate under reduced pressure to remove methanol, add ethyl acetate for extraction, dry with anhydrous sodium sulfate, and concentrate under reduced pressure to obtain 25n (30 mg, 0.04 mmol, 80%).
MS m/z(ESI):746.9[M+H]
+
MS m/z(ESI): 746.9[M+H] +
第十三步 4-((1R,3S,5s,7s)-5-氨基金刚烷-2-基)-N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺盐酸盐25The thirteenth step 4-((1R,3S,5s,7s)-5-aminoadamantan-2-yl)-N-(5-(3,5-difluorobenzyl)-1H-indazole- 3-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide hydrochloride 25
向反应瓶中加入25n(30mg,0.04mmol)，5ml甲醇和30mg Pd/C，然后在氢气氛围中反应2小时，TLC显示原料反应完。反应液用硅藻土过滤，滤液减压浓缩后用少量乙酸乙酯稀释，加入氯化氢的乙酸乙酯溶液(2M)，有固体析出，过滤后得纯的25(10mg,0.015mmol,38％)。Add 25n (30mg, 0.04mmol), 5ml methanol and 30mg Pd/C to the reaction flask, and then react in a hydrogen atmosphere for 2 hours. TLC shows that the raw materials have been reacted. The reaction solution was filtered with Celite. The filtrate was concentrated under reduced pressure and diluted with a small amount of ethyl acetate. After adding hydrogen chloride in ethyl acetate (2M), a solid precipitated out. After filtration, pure 25 (10mg, 0.015mmol, 38%) was obtained. .
MS m/z(ESI):612[M+H]
+
MS m/z(ESI):612[M+H] +
1H NMR(400MHz,Methanol-d
4)δ8.17(d,J＝8.3Hz,1H),7.76–7.69(m,1H),7.53–7.45(m,2H),7.42(s,1H),7.35(dd,J＝8.7,1.5Hz,1H),6.83(h,J＝4.1Hz,2H),6.74(tt,J＝9.2,2.4Hz,1H),4.10(s,2H),4.02-3.95(m,2H),3.90-3.78(m,1H),3.50-3.39(m,2H),3.20-3.11(m,1H),2.92-2.83(m,2H),2.25-2.08(m,4H),2.04-1.90(m,4H),1.88-1.69(m,4H),1.68-1.55(m,2H),1.35-1.26(m,1H).
1 H NMR(400MHz,Methanol-d 4 )δ8.17(d,J=8.3Hz,1H), 7.76–7.69(m,1H), 7.53–7.45(m,2H), 7.42(s,1H), 7.35(dd,J=8.7,1.5Hz,1H), 6.83(h,J=4.1Hz,2H), 6.74(tt,J=9.2,2.4Hz,1H), 4.10(s,2H),4.02-3.95 (m,2H),3.90-3.78(m,1H),3.50-3.39(m,2H),3.20-3.11(m,1H),2.92-2.83(m,2H),2.25-2.08(m,4H) , 2.04-1.90 (m, 4H), 1.88-1.69 (m, 4H), 1.68-1.55 (m, 2H), 1.35-1.26 (m, 1H).
实施例26Example 26
4-(5-氨基-金刚烷-2-基)-N-(5-(3,5-二氟苄基)-1H-吲哚-3-基)-2-(哌啶-4-基氨基)苯甲酰胺26的制备4-(5-Amino-adamantan-2-yl)-N-(5-(3,5-difluorobenzyl)-1H-indol-3-yl)-2-(piperidin-4-yl) (Amino) benzamide 26 preparation
第一步 4-((5-苄氧羰基氨基)金刚烷)-2-基)-2-((N-苄氧羰基哌啶-4-基)氨基)-苯甲酸乙酯26a的制备The first step is the preparation of 4-((5-benzyloxycarbonylamino)adamantane)-2-yl)-2-((N-benzyloxycarbonylpiperidin-4-yl)amino)-benzoic acid ethyl ester 26a
将25i(310mg，0.69mmol)，4-Cbz-哌啶酮(cas：19099-93-5，806mg，3.46mmol)混溶于10mL醋酸中，然后加入醋酸硼氢化钠(907mg，3.46mmol)。室温搅拌过夜，LCMS监测反应。旋干溶剂，残余物柱分离，石油醚/乙酸乙酯＝5/1(V/V)洗脱，得目标化合物26a(340mg)，白色固体，收率74％。25i (310 mg, 0.69 mmol), 4-Cbz-piperidone (cas: 19099-93-5, 806 mg, 3.46 mmol) was mixed in 10 mL of acetic acid, and then sodium acetate borohydride (907 mg, 3.46 mmol) was added. Stir at room temperature overnight, and monitor the reaction by LCMS. The solvent was spin-dried, the residue was separated by column, and eluted with petroleum ether/ethyl acetate=5/1 (V/V) to obtain target compound 26a (340 mg) as a white solid, with a yield of 74%.
LCMS:m/z＝666.5[M+H]+第三步4-((5-苄氧羰基氨基)金刚烷)-2-基)-2-((N-苄氧羰基哌啶-4-基)氨基)-苯甲酸26b的制备LCMS: m/z=666.5[M+H]+The third step 4-((5-benzyloxycarbonylamino)adamantane)-2-yl)-2-((N-benzyloxycarbonylpiperidine-4- (Yl)amino)-benzoic acid 26b preparation
将26a(900mg，1.35mmol)溶于15mL乙醇中，加入4M的氢氧化钠水溶液8mL。该体系回流16小时。LCMS监测反应。反应完毕，旋去溶剂大部分乙醇，加入10mL水后，调pH～4，有固体析出。滤出该固体，并用水洗，即得目标化合物26b(780mg)，土灰色固体，收率91％。未进一步纯化，直接用于下一步。26a (900 mg, 1.35 mmol) was dissolved in 15 mL of ethanol, and 8 mL of 4M sodium hydroxide aqueous solution was added. The system was refluxed for 16 hours. LCMS monitors the reaction. After the reaction was completed, the solvent was rotated to remove most of the ethanol, and after adding 10 mL of water, the pH was adjusted to 4, and a solid was precipitated. The solid was filtered out and washed with water to obtain target compound 26b (780 mg) as a grayish solid with a yield of 91%. It was used directly in the next step without further purification.
LCMS:m/z＝638.2[M+H]+LCMS: m/z=638.2[M+H]+
第四步 4-((5-苄氧羰基氨基)金刚烷)-2-基)-2-(N-(1-苄氧羰基哌啶-4-基)-2,2,2-三氟乙酰氨基)-苯甲酸26c的制备Step 4 4-((5-Benzyloxycarbonylamino)adamantane)-2-yl)-2-(N-(1-benzyloxycarbonylpiperidin-4-yl)-2,2,2-trifluoro Preparation of acetamido)-benzoic acid 26c
将26b(780mg，1.22mmol)溶于20mL二氯甲烷中，0℃下加入三氟乙酸酐(1.2eq)和三乙胺(1.5eq)。加毕，室温搅拌三小时。旋干溶剂，得到的粗品即为26c。未纯化，直接用于下一步。26b (780mg, 1.22mmol) was dissolved in 20mL of dichloromethane, and trifluoroacetic anhydride (1.2eq) and triethylamine (1.5eq) were added at 0°C. After the addition, stir at room temperature for three hours. Rotate the solvent to dryness, and the crude product obtained is 26c. It was not purified and used directly in the next step.
LCMS:m/z＝734.2[M+H]+LCMS: m/z=734.2[M+H]+
第五步 N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((5-苄氧羰基氨基)金刚烷)-2-基)-2-(N-(1-苄氧羰基哌啶-4-基)-2,2,2-三氟乙酰氨基)苯甲酰胺26e的制备Step 5 N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((5-benzyloxycarbonylamino)adamantane)-2-yl)- Preparation of 2-(N-(1-benzyloxycarbonylpiperidin-4-yl)-2,2,2-trifluoroacetamido)benzamide 26e
将粗品26c(～1.22mmol)溶于20mL二氯甲烷中，加入1mL草酰氯及1滴DMF，室温搅拌3小时，反应完毕，旋干溶剂及过量的草酰氯，得到酰氯26d，溶于10mL无水四氢呋喃中备用。将5-(3,5-二氟苯甲基)-1H-吲唑-3-胺I-2(316mg，1.22mmol)溶于10mL无水四氢呋喃，冰水浴冷却下，滴入上述酰氯溶液，然后加入三乙胺(0.33mL，2.44mmol)。加毕，室温搅拌过夜。滤除析出的固体，旋干滤液，残留物柱分离，石油醚/乙酸乙酯＝3/1(V/V)洗脱，得目标化合物26e(410mg)，黄色固体，三步合并收率34％。Dissolve the crude product 26c (~1.22mmol) in 20mL of dichloromethane, add 1mL of oxalyl chloride and 1 drop of DMF, stir at room temperature for 3 hours, after the reaction is complete, spin off the solvent and excess oxalyl chloride to obtain acid chloride 26d, which is dissolved in 10mL Reserve in water tetrahydrofuran. Dissolve 5-(3,5-difluorobenzyl)-1H-indazol-3-amine 1-2 (316 mg, 1.22 mmol) in 10 mL of anhydrous tetrahydrofuran, cool in an ice-water bath, drop the above acid chloride solution, Then triethylamine (0.33 mL, 2.44 mmol) was added. After the addition, stir overnight at room temperature. The precipitated solid was filtered off, the filtrate was spin-dried, and the residue was separated by column, and eluted with petroleum ether/ethyl acetate=3/1 (V/V) to obtain the target compound 26e (410mg) as a yellow solid. The combined yield of the three steps was 34. %.
LCMS:m/z＝975.2[M+H]+LCMS: m/z=975.2[M+H]+
第六步 N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((5-苄氧羰基氨基)金刚烷)-2-基)-2-((苄氧羰基)哌啶-4-基氨基)苯甲酰胺26f的制备The sixth step N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((5-benzyloxycarbonylamino)adamantane)-2-yl)- Preparation of 2-((benzyloxycarbonyl)piperidin-4-ylamino)benzamide 26f
于20mL封管中加入26e(410mg，0.42mmol)，用5mL甲醇溶解，然后加入1.12mL三乙胺(20eq)。封管，60℃加热搅拌反应1小时。LCMS监测反应。旋干反应体系，TLC制备分离，二氯甲烷/甲醇＝20/1(V/V)展开。得目标化合物26f(240mg)，收率65％。26e (410mg, 0.42mmol) was added to a 20mL sealed tube, dissolved with 5mL methanol, and then 1.12mL triethylamine (20eq) was added. The tube was sealed and heated and stirred at 60°C for 1 hour. LCMS monitors the reaction. The reaction system was spin-dried, TLC prepared and separated, and developed with dichloromethane/methanol=20/1 (V/V). The target compound 26f (240 mg) was obtained with a yield of 65%.
LCMS:m/z＝879.2[M+H]+LCMS: m/z=879.2[M+H]+
第七步 N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((5-氨基)金刚烷)-2-基)-2-(哌啶-4-基氨基)苯甲酰胺26的制备The seventh step N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((5-amino)adamantane)-2-yl)-2-( Preparation of piperidin-4-ylamino)benzamide 26
于50mL单口瓶中加入26f(110mg，0.13mmol)，用20mL甲醇溶解，加入50mg湿Pd/C(10％)。氢气球置换空气，氢气氛围下，室温搅拌20小时。反应完毕，滤除Pd/C，滤液旋干，即得目标化合物26(40mg)，白色固体，收率50％。26f (110 mg, 0.13 mmol) was added to a 50 mL single-necked flask, dissolved in 20 mL methanol, and 50 mg wet Pd/C (10%) was added. The hydrogen balloon replaces the air, and the mixture is stirred at room temperature for 20 hours under a hydrogen atmosphere. After the reaction was completed, the Pd/C was filtered off, and the filtrate was spin-dried to obtain the target compound 26 (40 mg) as a white solid with a yield of 50%.
LCMS:m/z＝611.2[M+H]+LCMS: m/z=611.2[M+H]+
1HNMR(400MHz,MeOD)δ＝8.03–8.05(d,J＝8.0Hz,1H),7.88(s,1H),7.44–7.53(m,2H),7.16–7.22(m,2H),6.73–6.85(m,3H),4.11(s,1H),3.97(brs,1H),3.57–3.59(d,J＝6.8Hz,1H),3.42–3.46(m,1H),3.11–3.19(m,3H),2.83(s,2H),2.25–2.29(m,2H),2.12(s,4H),1.82–1.99(m,6H),1.57–1.62(m,2H),1.14–1.26(m,1H).
1 HNMR(400MHz,MeOD)δ=8.03–8.05(d,J=8.0Hz,1H), 7.88(s,1H), 7.44–7.53(m,2H), 7.16–7.22(m,2H), 6.73– 6.85(m,3H),4.11(s,1H),3.97(brs,1H),3.57–3.59(d,J=6.8Hz,1H),3.42–3.46(m,1H),3.11–3.19(m, 3H), 2.83(s, 2H), 2.25–2.29(m, 2H), 2.12(s, 4H), 1.82–1.99(m, 6H), 1.57–1.62(m, 2H), 1.14–1.26(m, 1H).
实施例27Example 27
(R)-N-(5-(2-(2,5-二氟苯基)吡咯烷-1-基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺27的制备(R)-N-(5-(2-(2,5-Difluorophenyl)pyrrolidin-1-yl)-1H-indazol-3-yl)-4-(4-methylpiperazine- Preparation of 1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 27
采用与实施例3相同的制备方法，以(R)-2-(2,5-二氟苯基)吡咯烷为原料，可得到化合物27。Using the same preparation method as in Example 3, using (R)-2-(2,5-difluorophenyl)pyrrolidine as a raw material, compound 27 can be obtained.
MS m/z(ESI):616[M+1]
+
MS m/z(ESI):616[M+1] +
1H NMR(400MHz,DMSO-d
6)δ12.32(s,1H),9.94(s,1H),8.28(d,J＝7.7Hz,1H),7.76(d,J＝9.0Hz,1H),7.31–7.19(m,2H),7.14–7.04(m,1H),6.87(m,1H),6.66(dd,J＝9.1,2.3Hz,1H),6.49(d,J＝2.2Hz,1H),6.22(dd,J＝9.0,2.3Hz,1H),6.14(d,J＝2.3Hz,1H),4.91(d,J＝8.3Hz,1H),3.83(m,2H),3.71(m,2H),3.56–3.45(m,2H),3.27(m,5H),2.48–2.36(m,4H),2.24(s,3H),2.05–1.80(m,5H),1.42–1.25(m,3H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.32 (s, 1H), 9.94 (s, 1H), 8.28 (d, J = 7.7 Hz, 1H), 7.76 (d, J = 9.0 Hz, 1H) ,7.31–7.19(m,2H),7.14–7.04(m,1H),6.87(m,1H),6.66(dd,J=9.1,2.3Hz,1H),6.49(d,J=2.2Hz,1H ), 6.22 (dd, J = 9.0, 2.3 Hz, 1H), 6.14 (d, J = 2.3 Hz, 1H), 4.91 (d, J = 8.3 Hz, 1H), 3.83 (m, 2H), 3.71 (m , 2H), 3.56–3.45(m, 2H), 3.27(m, 5H), 2.48–2.36(m, 4H), 2.24(s, 3H), 2.05–1.80(m, 5H), 1.42–1.25(m ,3H).
实施例28Example 28
N-(5-(1-(2,5-二氟苯基)-2-羟基乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺28的制备N-(5-(1-(2,5-Difluorophenyl)-2-hydroxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl) Preparation of -2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 28
采用与实施例10相同的制备方法，以2,5-二氟溴苯为原料，可得到化合物28。Using the same preparation method as in Example 10, using 2,5-difluorobromobenzene as a raw material, compound 28 can be obtained.
MS m/z(ESI):591[M+H]
+。
MS m/z (ESI): 591 [M+H] + .
1H NMR(400MHz,DMSO-d
6)δ12.61(s,1H),10.07(s,1H),8.28(d,J＝7.7Hz,1H),7.75(d,J＝9.1Hz,1H),7.42(s,1H),7.35(d,J＝8.6Hz,1H),7.32–7.21(m,2H),7.13(td,J＝9.3,4.6Hz,1H),7.04(td,J＝8.7,8.2,4.2Hz,1H),6.20(dd,J＝9.0,2.3Hz,1H),6.10(d,J＝2.4Hz,1H),4.90(t,J＝5.2Hz,1H),4.38(t,J＝7.4Hz,1H),4.03–3.84(m,2H),3.78(d,J＝11.8Hz,2H),3.71–3.58(m,1H),3.46(t,J＝10.9Hz,2H),3.23(s,4H),2.43(s,4H),2.21(s,3H),1.90(d,J＝12.7Hz,2H),1.41–1.25(m,2H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.61 (s, 1H), 10.07 (s, 1H), 8.28 (d, J = 7.7 Hz, 1H), 7.75 (d, J = 9.1 Hz, 1H) ,7.42(s,1H),7.35(d,J=8.6Hz,1H),7.32–7.21(m,2H),7.13(td,J=9.3,4.6Hz,1H),7.04(td,J=8.7 , 8.2, 4.2 Hz, 1H), 6.20 (dd, J = 9.0, 2.3 Hz, 1H), 6.10 (d, J = 2.4 Hz, 1H), 4.90 (t, J = 5.2 Hz, 1H), 4.38 (t ,J=7.4Hz,1H),4.03-3.84(m,2H),3.78(d,J=11.8Hz,2H),3.71-3.58(m,1H),3.46(t,J=10.9Hz,2H) , 3.23(s,4H),2.43(s,4H),2.21(s,3H),1.90(d,J=12.7Hz,2H),1.41-1.25(m,2H).
实施例29Example 29
2-(3,5-二氟苯基)-2-(3-(4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰氨基)-1H-吲唑-5-基)乙酸乙酯29的制备2-(3,5-Difluorophenyl)-2-(3-(4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl) (Amino) benzoylamino) -1H-indazol-5-yl) ethyl acetate 29 preparation
第一步 5-(2-(苄氧基)-1-(3,5-二氟苯基)乙基)-3-(4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4)叔丁基基)氨基)苯甲酰氨基)-1H-吲唑-1-甲酸叔丁酯29aThe first step 5-(2-(benzyloxy)-1-(3,5-difluorophenyl)ethyl)-3-(4-(4-methylpiperazin-1-yl)-2- ((Tetrahydro-2H-pyran-4)tert-butyl)amino)benzamido)-1H-indazole-1-carboxylic acid tert-butyl ester 29a
化合物10g(1g，1.47mmol)的DCM(50mL)溶液在0℃滴加Boc
2O(320mg,56.8mmol)中，反应液在常温下搅拌2小时，然后小心向反应液中加入冰水(150mL),反应液在常温下继续搅拌3小时，将反应液浓缩后用DCM(200mL)萃取，有机相分别用饱和碳酸氢钠溶液和食盐水洗涤，干燥后旋干得到的粗品经过柱层析纯化后得到化合物29a的粗品(1.0g，1.28mmol)。
Compound 10g (1g, 1.47mmol) in DCM (50mL) was added dropwise to Boc 2 O (320mg, 56.8mmol) at 0℃, the reaction solution was stirred at room temperature for 2 hours, and then ice water (150mL) was carefully added to the reaction solution ), the reaction solution was stirred at room temperature for 3 hours, the reaction solution was concentrated and then extracted with DCM (200mL), the organic phase was washed with saturated sodium bicarbonate solution and brine, dried and then spin-dried to obtain the crude product after purification by column chromatography A crude product (1.0 g, 1.28 mmol) of compound 29a was obtained.
MS m/z(ESI):781[M+H]
+。
MS m/z (ESI): 781 [M+H] + .
第二步 5-(1-(3,5-二氟苯基)-2-羟基乙基)-3-(4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基))叔丁基氨基)苯甲酰氨基)-1H-吲唑-1-甲酸叔丁酯29bThe second step 5-(1-(3,5-difluorophenyl)-2-hydroxyethyl)-3-(4-(4-methylpiperazin-1-yl)-2-((tetrahydro -2H-pyran-4-yl)) tert-butylamino)benzamido)-1H-indazole-1-carboxylic acid tert-butyl ester 29b
化合物29a(1.0g，1.28mmol)溶于甲醇(60mL)中，加入甲酸铵(400mg，6.4mmol)和钯碳(100mg)，反应液加热到90℃℃，回流2h。过滤旋干得到初产品，经过硅胶柱层析纯化后的到化合物29b(500mg，0.72mmol)。Compound 29a (1.0 g, 1.28 mmol) was dissolved in methanol (60 mL), ammonium formate (400 mg, 6.4 mmol) and palladium carbon (100 mg) were added, and the reaction solution was heated to 90° C. and refluxed for 2 h. The initial product was filtered and spin-dried to obtain the initial product, which was purified by silica gel column chromatography to obtain compound 29b (500 mg, 0.72 mmol).
MS m/z(ESI):691[M+H]
+。
MS m/z (ESI): 691 [M+H] + .
第三步 5-(2-乙酰氧基-1-(3,5-二氟苯基)乙基)-3-(4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)叔丁基基)氨基)苯甲酰氨基)-1H-吲唑-1-甲酸叔丁酯29cThe third step 5-(2-acetoxy-1-(3,5-difluorophenyl)ethyl)-3-(4-(4-methylpiperazin-1-yl)-2-(( Tetrahydro-2H-pyran-4-yl)tert-butyl)amino)benzamido)-1H-indazole-1-carboxylate 29c
将化合物29b(100mg，0.14mmol)溶解在DCM(15mL)中，冷却至0℃，加入醋酸酐(14mg,0.14mmol),反应液在0℃搅拌1小时，再向反应液中加入 水，用DCM(50mL)萃取，有机相用饱和食盐水洗涤，干燥旋干得到粗品，经过柱层析纯化后得到化合物036c(60mg，0.082mmol)。Compound 29b (100mg, 0.14mmol) was dissolved in DCM (15mL), cooled to 0°C, acetic anhydride (14mg, 0.14mmol) was added, the reaction solution was stirred at 0°C for 1 hour, and then water was added to the reaction solution. It was extracted with DCM (50 mL), the organic phase was washed with saturated brine, dried and spin-dried to obtain a crude product, which was purified by column chromatography to obtain compound 036c (60 mg, 0.082 mmol).
MS m/z(ESI):733[M+H]
+。
MS m/z (ESI): 733 [M+H] + .
第四步 2-(3,5-二氟苯基)-2-(3-(4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰氨基)-1H-吲唑-5-基)乙酸乙酯29The fourth step 2-(3,5-difluorophenyl)-2-(3-(4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4 -Yl)amino)benzamido)-1H-indazol-5-yl)ethyl acetate 29
化合物29c(60mg，0.082mmol)，溶于DCM(10mL)中,加入TFA(2mL)，室温下搅拌1h，直接浓缩，经过柱层析纯化后得到化合物29三氟乙酸盐(40mg，0.063mmol)。Compound 29c (60mg, 0.082mmol) was dissolved in DCM (10mL), added TFA (2mL), stirred at room temperature for 1h, directly concentrated, and purified by column chromatography to obtain compound 29 trifluoroacetate (40mg, 0.063mmol) ).
MS m/z(ESI):633[M+H]
+。
MS m/z (ESI): 633 [M+H] + .
1H NMR(400MHz,DMSO-d
6)δ12.70(s,1H),10.20(s,1H),8.32(dd,J＝4.0,2.4Hz,1H),7.81(d,J＝8.8Hz,1H),7.50(s,1H),7.41(d,J＝8.9Hz,1H),7.37–7.31(m,1H),7.24(d,J＝7.2Hz,1H),7.10–7.01(m,2H),6.26(d,J＝8.7Hz,1H),6.17(s,1H),4.72(s,1H),4.65–4.59(m,1H),4.07–3.95(m,2H),3.84–3.75(m,2H),3.73-3.68(m,1H),3.60–3.56(m,4H),3.50–3.45(m,2H),3.12–3.09(m,4H),2.82(s,3H),1.99–1.81(m,5H),1.37–1.29(m,2H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.70 (s, 1H), 10.20 (s, 1H), 8.32 (dd, J = 4.0, 2.4 Hz, 1H), 7.81 (d, J = 8.8 Hz, 1H),7.50(s,1H),7.41(d,J=8.9Hz,1H),7.37–7.31(m,1H),7.24(d,J=7.2Hz,1H),7.10–7.01(m,2H ), 6.26(d,J=8.7Hz,1H), 6.17(s,1H), 4.72(s,1H), 4.65–4.59(m,1H), 4.07–3.95(m,2H), 3.84–3.75( m, 2H), 3.73-3.68 (m, 1H), 3.60--3.56 (m, 4H), 3.50--3.45 (m, 2H), 3.12--3.09 (m, 4H), 2.82 (s, 3H), 1.99-- 1.81 (m, 5H), 1.37-1.29 (m, 2H).
实施例30Example 30
N-(5-(1-(3,5-二氟苯基)乙基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺30的制备N-(5-(1-(3,5-Difluorophenyl)ethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazine -1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 30
采用与实施例17相同的制备方法，以1-(3,5-二氟苯基)-1-溴乙烷为原料，可得到化合物30。Using the same preparation method as in Example 17, using 1-(3,5-difluorophenyl)-1-bromoethane as a raw material, compound 30 can be obtained.
MS m/z(ESI):576[M+1]
+
MS m/z(ESI):576[M+1] +
1H NMR(400MHz,Chloroform-d)δ8.85(s,1H),8.63(s,1H),8.13(d,J＝7.3Hz,1H),7.78(s,1H),7.53(d,J＝8.9Hz,1H),6.88–6.77(m,2H),6.58(tt,J＝8.8,2.3Hz,1H),6.22(dd,J＝9.0,2.2Hz,1H),6.12–6.05(m,1H),4.37(q,J＝7.1Hz,1H),3.99(d,J＝11.7Hz,2H),3.66–3.45(m,3H),3.35(t,J＝5.2Hz,4H),2.63(t,J＝5.0Hz,4H),2.40(s,3H),2.03(d,J＝13.2Hz,2H),1.75–1.56(m,5H).
1 H NMR (400MHz, Chloroform-d) δ 8.85 (s, 1H), 8.63 (s, 1H), 8.13 (d, J = 7.3 Hz, 1H), 7.78 (s, 1H), 7.53 (d, J ＝8.9Hz,1H),6.88–6.77(m,2H),6.58(tt,J＝8.8,2.3Hz,1H), 6.22(dd,J＝9.0,2.2Hz,1H),6.12–6.05(m, 1H), 4.37 (q, J = 7.1 Hz, 1H), 3.99 (d, J = 11.7 Hz, 2H), 3.66–3.45 (m, 3H), 3.35 (t, J = 5.2 Hz, 4H), 2.63 ( t,J=5.0Hz,4H),2.40(s,3H),2.03(d,J=13.2Hz,2H),1.75-1.56(m,5H).
实施例31Example 31
N-(5-(3-氟苄基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺31的制备N-(5-(3-Fluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-( Preparation of (tetrahydro-2H-pyran-4-yl)amino)benzamide 31
采用与实施例17相同的制备方法，以3-氟溴苄为原料，可得到化合物31。Using the same preparation method as in Example 17, using 3-fluorobenzyl bromide as a raw material, compound 31 can be obtained.
MS m/z(ESI):544[M+1]
+
MS m/z(ESI):544[M+1] +
1H NMR(400MHz,DMSO-d
6)δ13.17(s,1H),10.30(s,1H),8.90(s,1H),8.24(d,J＝7.6Hz,1H),7.79(d,J＝9.1Hz,1H),7.50(s,1H),7.30(td,J＝8.0,6.3Hz,1H),7.16–7.06(m,2H),6.99(td,J＝8.7,2.8Hz,1H),6.24(dd,J＝9.0,2.3Hz,1H),6.13(d,J＝2.4Hz,1H),4.18(s,2H),3.82(dt,J＝11.7,4.0Hz,2H),3.67(t,J＝8.9Hz,1H),3.54–3.43(m,2H),3.28(t,J＝5.0Hz,4H),2.46(d,J＝5.2Hz,4H),2.24(s,3H),1.97–1.89(m,2H),1.36(ddd,J＝13.4,9.9,3.8Hz,2H).
1 H NMR (400MHz, DMSO-d 6 ) δ 13.17 (s, 1H), 10.30 (s, 1H), 8.90 (s, 1H), 8.24 (d, J = 7.6 Hz, 1H), 7.79 (d, J = 9.1Hz, 1H), 7.50 (s, 1H), 7.30 (td, J = 8.0, 6.3 Hz, 1H), 7.16-7.06 (m, 2H), 6.99 (td, J = 8.7, 2.8 Hz, 1H ), 6.24 (dd, J = 9.0, 2.3 Hz, 1H), 6.13 (d, J = 2.4 Hz, 1H), 4.18 (s, 2H), 3.82 (dt, J = 11.7, 4.0 Hz, 2H), 3.67 (t,J=8.9Hz,1H),3.54–3.43(m,2H), 3.28(t,J=5.0Hz,4H), 2.46(d,J=5.2Hz,4H), 2.24(s,3H) ,1.97–1.89(m,2H),1.36(ddd,J=13.4,9.9,3.8Hz,2H).
实施例32Example 32
N-(5-(1-(3-氟苯基)-2-羟基乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢 -2H-吡喃-4-基)氨基)苯甲酰胺32的制备N-(5-(1-(3-Fluorophenyl)-2-hydroxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2- Preparation of ((Tetrahydro-2H-pyran-4-yl)amino)benzamide 32
采用与实施例10相同的制备方法，以3-氟溴苯为原料，可得到化合物32。Using the same preparation method as in Example 10, using 3-fluorobromobenzene as a raw material, compound 32 can be obtained.
MS m/z(ESI):573[M+H]
+。
MS m/z (ESI): 573 [M+H] + .
1H NMR(400MHz,DMSO-d
6)δ12.59(s,1H),10.05(s,1H),8.28(d,J＝7.7Hz,1H),7.76(d,J＝9.1Hz,1H),7.45(d,J＝1.4Hz,1H),7.35(d,J＝8.6Hz,1H),7.31–7.19(m,2H),7.15–7.03(m,2H),7.00–6.89(m,1H),6.21(dd,J＝9.0,2.3Hz,1H),6.11(d,J＝2.3Hz,1H),4.81(t,J＝5.1Hz,1H),4.16(t,J＝7.3Hz,1H),3.94(td,J＝7.7,7.2,4.8Hz,2H),3.78(dt,J＝11.6,4.1Hz,2H),3.66(dq,J＝13.3,5.0Hz,1H),3.46(td,J＝11.0,2.5Hz,2H),3.24(d,J＝5.5Hz,4H),2.47(d,J＝3.1Hz,4H),2.23(s,3H),1.98–1.82(m,2H),1.32(dt,J＝12.8,3.2Hz,2H).
1 H NMR(400MHz,DMSO-d 6 )δ12.59(s,1H),10.05(s,1H), 8.28(d,J=7.7Hz,1H), 7.76(d,J=9.1Hz,1H) ,7.45(d,J=1.4Hz,1H),7.35(d,J=8.6Hz,1H),7.31–7.19(m,2H),7.15–7.03(m,2H),7.00–6.89(m,1H) ), 6.21 (dd, J = 9.0, 2.3 Hz, 1H), 6.11 (d, J = 2.3 Hz, 1H), 4.81 (t, J = 5.1 Hz, 1H), 4.16 (t, J = 7.3 Hz, 1H ), 3.94 (td, J = 7.7, 7.2, 4.8 Hz, 2H), 3.78 (dt, J = 11.6, 4.1 Hz, 2H), 3.66 (dq, J = 13.3, 5.0 Hz, 1H), 3.46 (td, J = 11.0, 2.5 Hz, 2H), 3.24 (d, J = 5.5 Hz, 4H), 2.47 (d, J = 3.1 Hz, 4H), 2.23 (s, 3H), 1.98-1.82 (m, 2H), 1.32(dt,J=12.8,3.2Hz,2H).
实施例33Example 33
N-(5-(1-(3-氟苯基)乙基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺33的制备N-(5-(1-(3-Fluorophenyl)ethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazine-1- Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 33
采用与实施例17相同的制备方法，以3,4-二氢-2H-吡喃、1-(3-氟苯基)-1-溴乙烷为原料，可得到化合物33。Using the same preparation method as in Example 17, using 3,4-dihydro-2H-pyran and 1-(3-fluorophenyl)-1-bromoethane as raw materials, compound 33 can be obtained.
MS m/z(ESI):558[M+1]
+
MS m/z(ESI):558[M+1] +
1H NMR(400MHz,Chloroform-d)δ8.87–8.81(m,1H),8.62(s,1H),8.12(d,J＝16.7Hz,1H),7.76(d,J＝1.2Hz,1H),7.53(d,J＝8.9Hz,1H),7.21(td,J＝8.0,6.0Hz,1H),7.11–7.06(m,1H),7.02(dt,J＝10.3,2.2Hz,1H),6.83(tdd,J＝8.3,2.6,1.1Hz,1H),6.21(dd,J＝9.1,2.2Hz,1H),6.09(d,J＝2.3Hz,1H),4.41(q,J＝7.1Hz,1H),3.99(d,J＝11.7Hz,2H),3.58(q,J＝13.0,10.8Hz,3H),3.35(t,J＝5.1Hz,4H),2.66(t,J＝5.0Hz,4H),2.42(s,3H),2.03(d,J＝14.5Hz,2H),1.73(d,J＝7.2Hz,3H),1.62(d,J＝11.9Hz,2H).
1 H NMR (400MHz, Chloroform-d) δ 8.87–8.81 (m, 1H), 8.62 (s, 1H), 8.12 (d, J = 16.7 Hz, 1H), 7.76 (d, J = 1.2 Hz, 1H ), 7.53 (d, J = 8.9 Hz, 1H), 7.21 (td, J = 8.0, 6.0 Hz, 1H), 7.11-7.06 (m, 1H), 7.02 (dt, J = 10.3, 2.2 Hz, 1H) ,6.83(tdd,J=8.3,2.6,1.1Hz,1H), 6.21(dd,J=9.1,2.2Hz,1H), 6.09(d,J=2.3Hz,1H), 4.41(q,J=7.1 Hz, 1H), 3.99 (d, J = 11.7 Hz, 2H), 3.58 (q, J = 13.0, 10.8 Hz, 3H), 3.35 (t, J = 5.1 Hz, 4H), 2.66 (t, J = 5.0 Hz, 4H), 2.42 (s, 3H), 2.03 (d, J = 14.5 Hz, 2H), 1.73 (d, J = 7.2 Hz, 3H), 1.62 (d, J = 11.9 Hz, 2H).
实施例34Example 34
N-(5-(1-(2,5-二氟苯基)乙基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺34的制备N-(5-(1-(2,5-Difluorophenyl)ethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazine -1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 34
采用与实施例33相同的制备方法，以3,4-二氢-2H-吡喃、1-(3-氟苯基)-1-溴乙烷为原料，可得到化合物34。Using the same preparation method as in Example 33, using 3,4-dihydro-2H-pyran and 1-(3-fluorophenyl)-1-bromoethane as raw materials, compound 34 can be obtained.
MS m/z(ESI):576[M+1]
+
MS m/z(ESI):576[M+1] +
1H NMR(400MHz,Chloroform-d)δ8.85(s,1H),8.64(s,1H),8.17(d,J＝7.3Hz,1H),7.79(s,1H),7.53(d,J＝8.9Hz,1H),7.04(ddd,J＝9.2,5.8,3.2Hz,1H),6.91(td,J＝9.1,4.6Hz,1H),6.80(ddd,J＝8.4,6.2,3.5Hz,1H),6.22–6.16(m,1H),6.08(d,J＝2.3Hz,1H),4.70(q,J＝7.2Hz,1H),3.98(d,J＝11.7Hz,2H),3.67–3.50(m,3H),3.43–3.30(m,4H),2.67(t,J＝5.0Hz,4H),2.43(s,3H),2.07–1.98(m,2H),1.76–1.56(m,5H).
1 H NMR (400MHz, Chloroform-d) δ 8.85 (s, 1H), 8.64 (s, 1H), 8.17 (d, J = 7.3 Hz, 1H), 7.79 (s, 1H), 7.53 (d, J ＝8.9Hz,1H), 7.04(ddd,J＝9.2,5.8,3.2Hz,1H), 6.91(td,J＝9.1,4.6Hz,1H), 6.80(ddd,J＝8.4,6.2,3.5Hz, 1H), 6.22–6.16 (m, 1H), 6.08 (d, J = 2.3 Hz, 1H), 4.70 (q, J = 7.2 Hz, 1H), 3.98 (d, J = 11.7 Hz, 2H), 3.67– 3.50(m,3H),3.43–3.30(m,4H), 2.67(t,J=5.0Hz,4H),2.43(s,3H),2.07–1.98(m,2H),1.76–1.56(m, 5H).
实施例35Example 35
N-(5-(1-(2,5-二氟苯基)-2-甲氧基乙基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺35的制备N-(5-(1-(2,5-Difluorophenyl)-2-methoxyethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-( Preparation of 4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 35
第一步 5-溴-1-(四氢-2H-吡喃-2-基)-1H-吡唑并[3,4-c]吡啶35bThe first step 5-bromo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridine 35b
化合物35a(10g，50.5mol)，DHP(7.6g,90.91mmol),甲磺酸(484mg,5.05mmol)溶于二氯甲烷(200mL)室温下搅拌过夜后，反应完毕。加入饱和碳酸氢钠用二氯甲烷萃取，有机相用饱和食盐水洗，无水硫酸钠干燥，过滤，浓缩后柱层析纯化得到标题化合物2(7g，24.822mmol)。Compound 35a (10g, 50.5mol), DHP (7.6g, 90.91mmol), methanesulfonic acid (484mg, 5.05mmol) were dissolved in dichloromethane (200mL) and stirred overnight at room temperature, the reaction was completed. Saturated sodium bicarbonate was added and extracted with dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography to obtain the title compound 2 (7 g, 24.822 mmol).
MS m/z(ESI):282.1[M+1]
+
MS m/z(ESI): 282.1[M+1] +
第二步 1-(四氢-2H-吡喃-2-基)-5-乙烯基-1H-吡唑并[3,4-c]吡啶35cStep 2 1-(Tetrahydro-2H-pyran-2-yl)-5-vinyl-1H-pyrazolo[3,4-c]pyridine 35c
化合物35b(5.62g，20mmol)和4,4,5,5-四甲基-2-乙烯基-1,3,2-二氧杂硼杂环戊烷(6.16g，40mmol)溶于1,4-二氧六环(200mL)和水(40mL)，加入Pd(PPh3)4(2.308g，2mmol)和碳酸钠(4.24g，40mmol),在氩气的保护下加热100℃搅拌6h后，反应完毕。反应液加水用乙酸乙酯萃取三次，用饱和食盐水洗，无水硫酸钠干燥，过滤，浓缩后柱层析纯化得到标题化合物35c(3.4g，14.847mmol)。Compound 35b (5.62g, 20mmol) and 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (6.16g, 40mmol) were dissolved in 1, 4-Dioxane (200mL) and water (40mL), add Pd(PPh3)4 (2.308g, 2mmol) and sodium carbonate (4.24g, 40mmol), heat and stir at 100℃ for 6h under the protection of argon, The reaction is complete. The reaction solution was added with water and extracted three times with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography to obtain the title compound 35c (3.4 g, 14.847 mmol).
MS m/z(ESI):230.1[M+H]
+
MS m/z(ESI): 230.1[M+H] +
第三步 5-(1,2-二溴乙基)-1-(四氢-2H-吡喃-2-基)-1H-吡唑并[3,4-c]吡啶35dThe third step 5-(1,2-dibromoethyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridine 35d
化合物35c(3.4g，14.847mmol)溶于乙酸(30mL)，加入溴化锂(1.547g，17.816mmol)室温下搅拌五分钟后加入高碘酸钠(794mg，3.712mmol)，室温下搅拌4h后，反应完毕。反应液加入水和乙酸乙酯萃取，有机相用饱和碳酸钠和食盐水洗，无水硫酸钠干燥，过滤，浓缩后得到标题化合物35d(4g，10.28mmol)，收率69％Compound 35c (3.4g, 14.847mmol) was dissolved in acetic acid (30mL). Lithium bromide (1.547g, 17.816mmol) was added and stirred at room temperature for five minutes. Sodium periodate (794mg, 3.712mmol) was added. After stirring for 4 hours at room temperature, the reaction complete. The reaction solution was extracted with water and ethyl acetate, the organic phase was washed with saturated sodium carbonate and brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the title compound 35d (4g, 10.28mmol) with a yield of 69%
MS m/z(ESI)390.1[M+1]
+
MS m/z(ESI)390.1[M+1] +
第四步 5-(1-溴乙烯基)-1-(四氢-2H-吡喃-2-基)-1H-吡唑并[3,4-c]吡啶35eThe fourth step 5-(1-bromovinyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridine 35e
化合物35d(4g，10.28mmol)溶于甲醇(20mL)和四氢呋喃(20mL)，然后加碳酸钾(2.838g，20.56mmol)，室温下搅拌过夜后，反应完毕。反应液旋干后加水，用乙酸乙酯萃取三次，用饱和食盐水洗，无水硫酸钠干燥，过滤，浓缩后柱层析纯化得到标题化合物35e(1.4g，4.545mmol)。Compound 35d (4g, 10.28mmol) was dissolved in methanol (20mL) and tetrahydrofuran (20mL), then potassium carbonate (2.838g, 20.56mmol) was added, and the reaction was completed after stirring overnight at room temperature. The reaction solution was spin-dried and added with water, extracted three times with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography to obtain the title compound 35e (1.4 g, 4.545 mmol).
MS m/z(ESI):310.1[M+H]
+
MS m/z(ESI): 310.1[M+H] +
第五步 5-(1-(2,5-二氟苯基)乙烯基)-1-(四氢-2H-吡喃-2-基)-1H-吡唑并[3,4-c]吡啶35fThe fifth step 5-(1-(2,5-difluorophenyl)vinyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c] Pyridine 35f
化合物35e(1.4g，4.54mmol)和(2,5-二氟苯基)硼酸(1.436mg，9.09mmol)溶于1，4-二氧六环(40mL)和水(10mL)中，加入[1,1'-双(二苯基膦基)二茂铁]二氯化钯(322mg，0.454mmol)和碳酸钠(963mg，9.09mmol)，在氩气的保护下加热 100℃搅拌5h后，反应完毕。反应液加入水和乙酸乙酯萃取，有机相用饱和食盐水洗，无水硫酸钠干燥，过滤，浓缩后柱层析纯化得到标题化合物35f(1g，2.932mmol)。Compound 35e (1.4g, 4.54mmol) and (2,5-difluorophenyl)boronic acid (1.436mg, 9.09mmol) were dissolved in 1,4-dioxane (40mL) and water (10mL), and added [ 1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (322mg, 0.454mmol) and sodium carbonate (963mg, 9.09mmol), heated at 100°C under the protection of argon and stirred for 5h, The reaction is complete. The reaction solution was extracted with water and ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography to obtain the title compound 35f (1 g, 2.932 mmol).
MS m/z(ESI):342.2[M+1]
+
MS m/z(ESI): 342.2[M+1] +
第六步(2-(2,5-二氟苯基)-2-(1-(四氢-2H-吡喃-2-基)-1H-吡唑并[3,4-c]吡啶-5-基)乙基)硼酸35gThe sixth step (2-(2,5-difluorophenyl)-2-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridine- 5-yl)ethyl)boronic acid 35g
化合物35f(720mg，2.11mmol)和4,4,4'，4'，5,5,5'，5'-八甲基-2,2'-二(1,3,2-二氧杂硼烷)(1072mg，4.22mmol)溶于甲醇(30mL)，加入氧化亚铜(151mg，1.055mmol)，KH2PO4(918mg，5.2775mmol),三苯基磷(274mg，1.055mmol)加热回流4h后，反应完毕。缩后柱层析纯化得到标题化合物35g(320mg，0.826mmol)，收率39％。Compound 35f (720mg, 2.11mmol) and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaboron Alkyl) (1072mg, 4.22mmol) was dissolved in methanol (30mL), and cuprous oxide (151mg, 1.055mmol), KH2PO4 (918mg, 5.2775mmol), triphenylphosphonium (274mg, 1.055mmol) were heated and refluxed for 4h after adding cuprous oxide (151mg, 1.055mmol). complete. After purification by column chromatography, 35 g (320 mg, 0.826 mmol) of the title compound was obtained with a yield of 39%.
MS m/z(ESI):388.3[M+H]
+
MS m/z(ESI): 388.3[M+H] +
第七步 2-(2,5-二氟苯基)-2-(1-(四氢-2H-吡喃-2-基)-1H-吡唑并[3,4-c]吡啶-5-基)乙-1-醇35hThe seventh step 2-(2,5-difluorophenyl)-2-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridine-5 -Base) Ethan-1-ol 35h
化合物35g(320mg，0.826mmol)溶于四氢呋喃(6mL)中，加入1M氢氧化钠水溶液(6mL),反应液冷却到0℃，缓慢滴加30％的双氧水(3mL)。反应液在常温下搅拌3小时，然后用饱和硫代硫酸钠溶液淬灭。用乙酸乙酯(150mL)萃取，有机相用饱和碳酸氢钠溶液和食盐水洗涤，干燥后用无水硫酸钠干燥，过滤旋干得到初产品，经过硅胶柱层析纯化后的到化合物35h(260mg,0.724mmol)收率87％。Compound 35g (320mg, 0.826mmol) was dissolved in tetrahydrofuran (6mL), 1M aqueous sodium hydroxide solution (6mL) was added, the reaction solution was cooled to 0°C, and 30% hydrogen peroxide (3mL) was slowly added dropwise. The reaction solution was stirred at room temperature for 3 hours, and then quenched with saturated sodium thiosulfate solution. It was extracted with ethyl acetate (150 mL), the organic phase was washed with saturated sodium bicarbonate solution and brine, dried and dried over anhydrous sodium sulfate, filtered and spin-dried to obtain the initial product, which was purified by silica gel column chromatography to obtain compound 35h (260mg , 0.724mmol), the yield was 87%.
MS m/z(ESI):360.2[M+H]
+
MS m/z(ESI): 360.2[M+H] +
第八步 5-(1-(2,5-二氟苯基)-2-甲氧基乙基)-1-(四氢-2H-吡喃-2-基)-1H-吡唑并[3,4-c]吡啶35iStep 8 5-(1-(2,5-Difluorophenyl)-2-methoxyethyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[ 3,4-c]pyridine 35i
化合物35h(260mg，0.724mmol)溶于四氢呋喃(10mL)，冰浴氩气保护下加入氢化钠(87mg，2.1727mmol,搅拌半小时后加入碘甲烷(308mg，2.1727mol)缓慢升到室温后搅拌过夜，反应完毕。反应液用稀盐酸淬灭后加饱和碳酸氢钠，用 乙酸乙酯萃取三次，用饱和食盐水洗，无水硫酸钠干燥，过滤，浓缩后柱层析纯化得到标题化合物35i(210mg，0.7266mmol)。Compound 35h (260mg, 0.724mmol) was dissolved in tetrahydrofuran (10mL), sodium hydride (87mg, 2.1727mmol) was added under the protection of argon in an ice bath, and after stirring for half an hour, methyl iodide (308mg, 2.1727mol) was added slowly to room temperature and stirred overnight After the reaction was completed, the reaction solution was quenched with dilute hydrochloric acid, then saturated sodium bicarbonate was added, extracted three times with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain the title compound 35i (210mg , 0.7266 mmol).
MS m/z(ESI):374.2[M+H]
+
MS m/z(ESI): 374.2[M+H] +
第九步 5-(1-(2,5-二氟苯基)-2-甲氧基乙基)-1H-吡唑并[3,4-c]吡啶35jStep 9 5-(1-(2,5-Difluorophenyl)-2-methoxyethyl)-1H-pyrazolo[3,4-c]pyridine 35j
化合物35i(260mg，0.697mmol)溶于二氯甲烷(4mL)，加入三氟乙酸(4mL),室温反应2h后，反应完毕。浓缩后板层析纯化得到标题化合物35j(210mg)，未纯，直接做下一步。Compound 35i (260 mg, 0.697 mmol) was dissolved in dichloromethane (4 mL), trifluoroacetic acid (4 mL) was added, and the reaction was completed at room temperature for 2 hours. After concentration and purification by plate chromatography, the title compound 35j (210mg) was obtained, which was not pure, so proceed directly to the next step.
MS m/z(ESI):290.1[M+H]
+
MS m/z(ESI): 290.1[M+H] +
第十步 5-(1-(2,5-二氟苯基)-2-甲氧基乙基)-3-碘-1H-吡唑并[3,4-c]吡啶35kThe tenth step 5-(1-(2,5-difluorophenyl)-2-methoxyethyl)-3-iodo-1H-pyrazolo[3,4-c]pyridine 35k
化合物35j(210mg，0.7266mmol)溶于DMF(5ml)中，依次加入K
2CO
3(401mg，2.905mmol，)和9a(367.6mg，1.4532mmol，)，室温反应3h。加入EA(50ml)稀释，依次用H2O(25ml)和brine(15ml×2)洗涤，经无水硫酸钠干燥，浓缩，柱纯化得标题化合物35k(250mg，0.6mmol)，收率83％
Compound 35j (210 mg, 0.7266 mmol) was dissolved in DMF (5 ml), K 2 CO 3 (401 mg, 2.905 mmol,) and 9a (367.6 mg, 1.4532 mmol,) were sequentially added, and reacted at room temperature for 3 hours. Dilute with EA (50ml), wash with H2O (25ml) and brine (15ml×2) successively, dry over anhydrous sodium sulfate, concentrate, and purify by column to obtain the title compound 35k (250mg, 0.6mmol) with a yield of 83%
MS m/z(ESI):416.1[M+1]
+
MS m/z(ESI): 416.1[M+1] +
第十一步 5-(1-(2,5-二氟苯基)-2-甲氧基乙基)-3-碘-1-(四氢-2H-吡喃-2-基)-1H-吡唑并[3,4-c]吡啶35lThe eleventh step 5-(1-(2,5-difluorophenyl)-2-methoxyethyl)-3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H -Pyrazolo[3,4-c]pyridine 35l
化化合物35k(250mg，0.602mol)，DHP(101mg,1.204mmol),溶于二氯甲烷(20mL)，冰浴下滴加甲磺酸(116mg,1.204mmol)，室温下搅拌过夜后，反应完毕。加入饱和碳酸氢钠后用乙酸乙酯萃取，有机相用饱和食盐水洗，无水硫酸钠干燥，过滤，浓缩后柱层析纯化得到标题化合物35l(160m g，0.32mmol)，收率53％Compound 35k (250mg, 0.602mol), DHP (101mg, 1.204mmol), dissolved in dichloromethane (20mL), methanesulfonic acid (116mg, 1.204mmol) was added dropwise under ice bath, after stirring overnight at room temperature, the reaction was complete . After adding saturated sodium bicarbonate and extracting with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography to obtain the title compound 35l (160 mg, 0.32 mmol), the yield was 53%
MS m/z(ESI):500.2[M+1]
+
MS m/z(ESI): 500.2[M+1] +
第十二步 5-(1-(2,5-二氟苯基)-2-甲氧基乙基)-3-碘-1-(四氢-2H-吡喃-2-基)-1H-吡唑并[3,4-c]吡啶35mStep 12 5-(1-(2,5-Difluorophenyl)-2-methoxyethyl)-3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H -Pyrazolo[3,4-c]pyridine 35m
化合物35l(80mg，0.16mmol)，4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基) 氨基)苯酰胺(76mg，0.24mmol)，Cu(10mg，0.16mmol)，CuI(31mg，0.16mmol)，N,N’-二甲基乙二胺(85mg，0.96mmol，)和K
3PO
4(102mg，0.48mmol)投于甲苯(15ml)中氩气换气5次，回流反应6h。将反应液直接拌样，柱纯化得化合物35m(48m g，0.069mmol)，收率43.6％
Compound 35l (80mg, 0.16mmol), 4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide (76mg, 0.24mmol) , Cu (10mg, 0.16mmol), CuI (31mg, 0.16mmol), N,N'-dimethylethylenediamine (85mg, 0.96mmol,) and K 3 PO 4 (102mg, 0.48mmol) were cast in toluene ( In 15ml) argon gas was exchanged 5 times, and the reaction was refluxed for 6h. The reaction solution was mixed directly and purified by column to obtain compound 35m (48mg, 0.069mmol) with a yield of 43.6%
MS m/z(ESI):690.6[M+1]
+
MS m/z(ESI): 690.6[M+1] +
第十三步 N-(5-(1-(2,5-二氟苯基)-2-甲氧基乙基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺HHT13056The thirteenth step N-(5-(1-(2,5-difluorophenyl)-2-methoxyethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl) -4-(4-Methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide HHT13056
化合物13(48mg，0.069mmol)溶于二氯甲烷(4mL)，加入三氟乙酸(4mL),室温反应2h后，反应完毕。旋干后加入饱和碳酸氢钠后用乙酸乙酯萃取，有机相用饱和食盐水洗，无水硫酸钠干燥，过滤，浓缩后制备TLC纯化得到标题化合物HHT13056(31m g，0.0512mmol)，收率73％Compound 13 (48 mg, 0.069 mmol) was dissolved in dichloromethane (4 mL), trifluoroacetic acid (4 mL) was added, and the reaction was completed at room temperature for 2 hours. After spin-drying, saturated sodium bicarbonate was added and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated and purified by preparative TLC to obtain the title compound HHT13056 (31 mg, 0.0512 mmol), with a yield of 73 %
MS m/z(ESI):606.5[M+1]
+
MS m/z(ESI):606.5[M+1] +
1H NMR(400MHz,DMSO-d
6)δ13.17(s,1H),10.29(s,1H),8.90(d,J＝1.2Hz,1H),8.23(d,J＝7.7Hz,1H),7.75(d,J＝9.1Hz,1H),7.50(d,J＝1.2Hz,1H),7.31(s,1H),7.15(dd,J＝9.3,4.7Hz,1H),7.06(d,J＝4.1Hz,1H),6.24–6.19(m,1H),6.10(d,J＝2.4Hz,1H),4.70(t,J＝7.3Hz,1H),4.02–3.91(m,2H),3.83–3.75(m,2H),3.71–3.63(m,1H),3.47(t,J＝10.5Hz,2H),3.23(d,J＝10.2Hz,7H),2.42(s,4H),2.20(s,3H),1.90(d,J＝12.6Hz,2H),1.31(d,J＝9.6Hz,2H).
1 H NMR (400MHz, DMSO-d 6 ) δ 13.17 (s, 1H), 10.29 (s, 1H), 8.90 (d, J = 1.2 Hz, 1H), 8.23 (d, J = 7.7 Hz, 1H) ,7.75(d,J=9.1Hz,1H),7.50(d,J=1.2Hz,1H),7.31(s,1H),7.15(dd,J=9.3,4.7Hz,1H),7.06(d, J = 4.1Hz, 1H), 6.24-6.19 (m, 1H), 6.10 (d, J = 2.4 Hz, 1H), 4.70 (t, J = 7.3 Hz, 1H), 4.02-3.91 (m, 2H), 3.83–3.75(m,2H), 3.71–3.63(m,1H), 3.47(t,J=10.5Hz,2H), 3.23(d,J=10.2Hz,7H),2.42(s,4H), 2.20 (s, 3H), 1.90 (d, J = 12.6 Hz, 2H), 1.31 (d, J = 9.6 Hz, 2H).
实施例36Example 36
2-(3-氟苯基)-2-(3-(4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)-1氢-吲唑-5-基)乙基异丁酯36的制备2-(3-Fluorophenyl)-2-(3-(4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)- Preparation of 1-Hydro-indazol-5-yl) ethyl isobutyl ester 36
采用与实施例29相同的制备方法，以3-氟溴苯、异丁酸酐为原料，可得到化合物36。Using the same preparation method as in Example 29, using 3-fluorobromobenzene and isobutyric anhydride as raw materials, compound 36 can be obtained.
MS m/z(ESI):643[M+1]
+。
MS m/z (ESI): 643 [M+1] + .
1H NMR(400MHz,DMSO-d
6)δ12.62(s,1H),10.06(s,1H),8.26(d,J＝7.7Hz,1H),7.75(d,J＝9.1Hz,1H),7.52(s,1H),7.42–7.23(m,3H),7.20–7.08(m,2H),6.99(td,J＝8.7,2.2Hz,1H),6.20(dd,J＝9.0,2.3Hz,1H),6.10(d,J＝2.3Hz,1H),4.58(qd,J＝10.9,7.5Hz,2H),4.46(t,J＝7.5Hz,1H),3.78(d,J＝11.6Hz,2H),3.71–3.60(m,1H),3.46(t,J＝10.9Hz,2H),3.23(t,J＝5.0Hz,4H),2.44–2.30(m,5H),2.20(s,3H),1.90(d,J＝13.0Hz,2H),1.31(q,J＝12.0,10.2Hz,2H),0.88(dd,J＝7.0,2.0Hz,6H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.62 (s, 1H), 10.06 (s, 1H), 8.26 (d, J = 7.7 Hz, 1H), 7.75 (d, J = 9.1 Hz, 1H) ,7.52(s,1H),7.42–7.23(m,3H),7.20–7.08(m,2H),6.99(td,J＝8.7,2.2Hz,1H),6.20(dd,J＝9.0,2.3Hz ,1H),6.10(d,J＝2.3Hz,1H),4.58(qd,J＝10.9,7.5Hz,2H),4.46(t,J＝7.5Hz,1H),3.78(d,J＝11.6Hz ,2H),3.71–3.60(m,1H), 3.46(t,J=10.9Hz,2H), 3.23(t,J=5.0Hz,4H), 2.44-2.30(m,5H), 2.20(s, 3H), 1.90 (d, J = 13.0Hz, 2H), 1.31 (q, J = 12.0, 10.2Hz, 2H), 0.88 (dd, J = 7.0, 2.0Hz, 6H).
实施例37Example 37
N-(5-(3-氟苄基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(5-甲基-2,5-二氮杂双环[2.2.1]庚-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺37的制备N-(5-(3-Fluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(5-methyl-2,5-diazabicyclo[2.2 .1]Heptan-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 37
采用与实施例17相同的制备方法，以3-氟溴苯、2-甲基-2,5-二氮杂二环[2.2.1]庚烷为原料，可得到化合物37。Using the same preparation method as in Example 17, using 3-fluorobromobenzene and 2-methyl-2,5-diazabicyclo[2.2.1]heptane as raw materials, compound 37 can be obtained.
MS m/z(ESI):556[M+1]
+
MS m/z(ESI):556[M+1] +
1H NMR(400MHz,DMSO-d
6)δ13.15(s,1H),10.21(s,1H),8.89(d,J＝1.3Hz,1H),8.31(d,J＝7.6Hz,1H),7.76(d,J＝9.0Hz,1H),7.49(d,J＝1.3Hz,1H),7.30(td,J＝7.9,6.3Hz,1H),7.16–7.07(m,2H),6.99(td,J＝8.3,2.5Hz,1H),5.94(dd,J＝8.8,2.2Hz,1H),5.77(d,J＝2.2Hz,1H),4.50(s,1H),4.18(s,2H),3.88–3.76(m,2H),3.66(s,2H),3.55–3.44(m,3H),3.35-3.28(m,2H),2.91(d,J＝9.8Hz,1H),2.43(s,3H),1.92(q,J＝14.2,13.1Hz,4H),1.42–1.29(m,2H).
1 H NMR (400MHz, DMSO-d 6 ) δ 13.15 (s, 1H), 10.21 (s, 1H), 8.89 (d, J = 1.3 Hz, 1H), 8.31 (d, J = 7.6 Hz, 1H) ,7.76(d,J=9.0Hz,1H),7.49(d,J=1.3Hz,1H),7.30(td,J=7.9,6.3Hz,1H),7.16-7.07(m,2H),6.99( td,J=8.3,2.5Hz,1H),5.94(dd,J=8.8,2.2Hz,1H),5.77(d,J=2.2Hz,1H),4.50(s,1H),4.18(s,2H ), 3.88–3.76(m, 2H), 3.66(s, 2H), 3.55–3.44(m, 3H), 3.35–3.28(m, 2H), 2.91(d, J=9.8Hz, 1H), 2.43( s,3H),1.92(q,J=14.2,13.1Hz,4H),1.42-1.29(m,2H).
实施例38Example 38
N-(5-(1-(3-氟苯基)-2-甲氧基乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺38的制备N-(5-(1-(3-Fluorophenyl)-2-methoxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)- Preparation of 2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 38
采用与实施例23相同的制备方法，以3-氟溴苯为原料，可得到化合物38。Using the same preparation method as in Example 23 and using 3-fluorobromobenzene as a raw material, compound 38 can be obtained.
MS m/z(ESI):587[M+1]
+。
MS m/z (ESI): 587 [M+1] + .
1H NMR(400MHz,DMSO-d
6)δ12.59(s,1H),10.05(s,1H),8.30(d,J＝7.7Hz,1H),7.76(d,J＝9.1Hz,1H),7.47(s,1H),7.35(d,J＝8.7Hz,1H),7.30–7.23(m,2H),7.18–7.03(m,2H),7.00–6.88(m,1H),6.21(dd,J＝9.0,2.3Hz,1H),6.10(d,J＝2.3Hz,1H),4.36(t,J＝7.4Hz,1H),3.97–3.72(m,4H),3.72–3.59(m,1H),3.52–3.42(m,2H),3.23(d,J＝10.0Hz,7H),2.48(s,4H),2.22(s,3H),1.90(dt,J＝13.0,4.2Hz,2H),1.42–1.24(m,2H).
1 H NMR(400MHz,DMSO-d 6 )δ12.59(s,1H),10.05(s,1H),8.30(d,J=7.7Hz,1H),7.76(d,J=9.1Hz,1H) ,7.47(s,1H),7.35(d,J=8.7Hz,1H),7.30–7.23(m,2H),7.18–7.03(m,2H),7.00–6.88(m,1H),6.21(dd ,J=9.0,2.3Hz,1H), 6.10(d,J=2.3Hz,1H), 4.36(t,J=7.4Hz,1H), 3.97–3.72(m,4H), 3.72–3.59(m, 1H),3.52–3.42(m,2H), 3.23(d,J=10.0Hz,7H), 2.48(s,4H),2.22(s,3H),1.90(dt,J=13.0,4.2Hz,2H ),1.42-1.24(m,2H).
实施例39Example 39
N-(5-(3,5-二氟苄基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(5-甲基-2,5-二氮杂双环[2.2.1]庚-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺39的合成N-(5-(3,5-Difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(5-methyl-2,5-diazepine Synthesis of bicyclo[2.2.1]hept-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 39
采用与实施例17相同的制备方法，以2-甲基-2,5-二氮杂二环[2.2.1]庚烷为原料，可得到化合物39。Using the same preparation method as in Example 17, using 2-methyl-2,5-diazabicyclo[2.2.1]heptane as a raw material, compound 39 can be obtained.
MS m/z(ESI):574[M+1]
+
MS m/z(ESI):574[M+1] +
1H NMR(400MHz,DMSO-d
6)δ13.14(s,1H),10.19(s,1H),8.90(d,J＝1.2Hz,1H),8.31(d,J＝7.5Hz,1H),7.75(d,J＝9.0Hz,1H),7.53(s,1H),7.07–6.95(m,3H),5.93(d,J＝8.7Hz,1H),5.76(s,1H),4.42(s,1H),4.19(s,2H),3.82(d,J＝11.4Hz,2H),3.64(s,1H),3.48(t,J＝10.8Hz,4H),3.32-3.28(m,2H),2.83(d,J＝9.3Hz,1H),2.30(s,3H),1.92(q,J＝12.5,12.0Hz,4H),1.35(d,J＝11.5Hz,2H).
1 H NMR (400MHz, DMSO-d 6 ) δ 13.14 (s, 1H), 10.19 (s, 1H), 8.90 (d, J = 1.2 Hz, 1H), 8.31 (d, J = 7.5 Hz, 1H) ,7.75(d,J=9.0Hz,1H),7.53(s,1H),7.07–6.95(m,3H),5.93(d,J=8.7Hz,1H),5.76(s,1H),4.42( s, 1H), 4.19 (s, 2H), 3.82 (d, J = 11.4 Hz, 2H), 3.64 (s, 1H), 3.48 (t, J = 10.8 Hz, 4H), 3.32-3.28 (m, 2H) ), 2.83 (d, J = 9.3 Hz, 1H), 2.30 (s, 3H), 1.92 (q, J = 12.5, 12.0 Hz, 4H), 1.35 (d, J = 11.5 Hz, 2H).
实施例40Example 40
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(3-(二甲基氨基)氮杂环丁烷-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺40的合成N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(3-(dimethylamino)azetidin-1-yl)-2- Synthesis of ((Tetrahydro-2H-pyran-4-yl)amino)benzamide 40
采用与实施例12相同的制备方法，以N，N-二甲基氮杂环-3-胺为原料，可得到化合物40。Using the same preparation method as in Example 12, using N,N-dimethylazacyclo-3-amine as a raw material, compound 40 can be obtained.
MS m/z(ESI):561[M+1]
+
MS m/z(ESI): 561[M+1] +
1H NMR(400MHz,DMSO-d
6)δ12.64(s,1H),10.05(s,1H),8.38(d,J＝7.5Hz,1H),7.78(d,J＝8.8Hz,1H),7.48(s,1H),7.40(d,J＝8.5Hz,1H),7.25(dd,J＝8.6,1.6Hz,1H),7.06–6.92(m,3H),5.78–5.68(m,1H),5.63(d,J＝2.2Hz,1H),4.03(s,2H),3.97(t,J＝7.4Hz,2H),3.81(dt,J＝11.6,3.9Hz,2H),3.72–3.53(m,3H),3.47(td,J＝11.5,10.9,2.4Hz,2H),3.27-3.19(m,1H),2.17(s,6H),1.98–1.87(m,2H),1.34(tdd,J＝13.4,8.6,3.6Hz,2H).
1 H NMR(400MHz,DMSO-d 6 )δ12.64(s,1H),10.05(s,1H),8.38(d,J=7.5Hz,1H),7.78(d,J=8.8Hz,1H) ,7.48(s,1H),7.40(d,J=8.5Hz,1H),7.25(dd,J=8.6,1.6Hz,1H),7.06–6.92(m,3H),5.78–5.68(m,1H ), 5.63 (d, J = 2.2 Hz, 1H), 4.03 (s, 2H), 3.97 (t, J = 7.4 Hz, 2H), 3.81 (dt, J = 11.6, 3.9 Hz, 2H), 3.72-3.53 (m, 3H), 3.47 (td, J = 11.5, 10.9, 2.4 Hz, 2H), 3.27-3.19 (m, 1H), 2.17 (s, 6H), 1.98-1.87 (m, 2H), 1.34 (tdd , J = 13.4, 8.6, 3.6 Hz, 2H).
实施例41Example 41
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(3-((2-甲氧基乙基)(甲基)氨基)氮杂环丁烷-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺41的合成N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(3-((2-methoxyethyl)(methyl)amino)azacyclo Synthesis of Butane-1-yl)-2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide 41
采用与实施例12相同的制备方法，以N-(2-甲氧基乙基)-N-甲基氮杂-3-胺为原料，可得到化合物41。Using the same preparation method as in Example 12, using N-(2-methoxyethyl)-N-methylaza-3-amine as a raw material, compound 41 can be obtained.
MS m/z(ESI):605[M+1]
+
MS m/z(ESI):605[M+1] +
1H NMR(400MHz,DMSO-d
6)δ12.64(s,1H),10.04(s,1H),8.38(d,J＝7.6Hz,1H),7.78(d,J＝8.8Hz,1H),7.48(s,1H),7.43–7.35(m,1H),7.25(dd,J＝8.6,1.6Hz,1H),7.01(ddd,J＝11.8,8.0,1.5Hz,3H),5.72(dd,J＝8.7,2.1Hz,1H),5.63(d,J＝2.1Hz,1H),4.03(s,2H),3.96(t,J＝7.5Hz,2H),3.80(dd,J＝9.8,5.9Hz,2H),3.66-3.55(m,3H),3.52–3.37(m,5H),3.24(s,3H),2.46(t,2H),2.17(s,3H),1.93(d,J＝12.8Hz,2H),1.38-1.28(m,2H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.64 (s, 1H), 10.04 (s, 1H), 8.38 (d, J = 7.6 Hz, 1H), 7.78 (d, J = 8.8 Hz, 1H) ,7.48(s,1H),7.43-7.35(m,1H),7.25(dd,J=8.6,1.6Hz,1H),7.01(ddd,J=11.8,8.0,1.5Hz,3H),5.72(dd ,J=8.7,2.1Hz,1H),5.63(d,J=2.1Hz,1H),4.03(s,2H),3.96(t,J=7.5Hz,2H), 3.80(dd,J=9.8, 5.9Hz, 2H), 3.66-3.55 (m, 3H), 3.52-3.37 (m, 5H), 3.24 (s, 3H), 2.46 (t, 2H), 2.17 (s, 3H), 1.93 (d, J =12.8Hz, 2H), 1.38-1.28 (m, 2H).
实施例42Example 42
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((2-(吡咯烷-1-基)乙基)氨基)-2-((四 氢-2H-吡喃-4-基)氨基)苯酰胺42的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((2-(pyrrolidin-1-yl)ethyl)amino)-2-( Preparation of (Tetrahydro-2H-pyran-4-yl)amino)benzamide 42
采用与实施例12相同的制备方法，以2-(吡咯烷-1-基)乙基胺为原料，可得到化合物42。Using the same preparation method as in Example 12, using 2-(pyrrolidin-1-yl)ethylamine as a raw material, compound 42 can be obtained.
MS m/z(ESI):575[M+H]+。MS m/z(ESI): 575[M+H]+.
1H NMR(400MHz,Methanol-d
4)δ7.68(d,J＝8.8Hz,1H),7.56(s,1H),7.40(d,J＝8.6Hz,1H),7.25–7.19(m,1H),6.85–6.76(m,2H),6.75–6.66(m,1H),6.07–5.95(m,2H),4.05(s,2H),3.91(d,J＝11.7Hz,2H),3.69–3.44(m,6H),3.15(d,J＝13.6Hz,5H),2.07–1.92(m,6H),1.54–1.45(m,2H).
1 H NMR(400MHz,Methanol-d 4 )δ7.68(d,J=8.8Hz,1H), 7.56(s,1H), 7.40(d,J=8.6Hz,1H), 7.25–7.19(m, 1H), 6.85–6.76(m,2H), 6.75–6.66(m,1H), 6.07–5.95(m,2H),4.05(s,2H), 3.91(d,J=11.7Hz,2H), 3.69 –3.44(m,6H), 3.15(d,J=13.6Hz,5H), 2.07–1.92(m,6H), 1.54–1.45(m,2H).
实施例43Example 43
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(乙基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺43的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2 -((Tetrahydro-2H-pyran-4-yl)amino)benzamide 43
第一步 叔-丁基4-(乙基(2-(吡咯烷-1-基)乙基)氨基)-2-硝基苯酸酯43a的合成The first step is the synthesis of tert-butyl 4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-nitrobenzoate 43a
将I-1b(500mg,2.1mmol),2N-乙基-2-(吡咯烷-1-基)乙烷-1-胺(587mg,4.1mmol)和DIPEA(1060mg,8.3mmol)溶于NMP中,在120℃搅拌2小时。冷却至室温，加水淬灭，用乙酸乙酯萃取，有机相合并后用饱和氯化钠洗，用无水硫酸钠干燥，过滤，浓缩，得到粗品，粗品用柱层析纯化(DCM：MeOH＝15:1)得到化合43a(500mg)呈黄色油状物。Dissolve I-1b (500mg, 2.1mmol), 2N-ethyl-2-(pyrrolidin-1-yl)ethane-1-amine (587mg, 4.1mmol) and DIPEA (1060mg, 8.3mmol) in NMP , Stir at 120°C for 2 hours. Cooled to room temperature, quenched with water, extracted with ethyl acetate, combined the organic phases, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was purified by column chromatography (DCM: MeOH = 15:1) Compound 43a (500mg) was obtained as a yellow oil.
MS m/z(ESI):364(M+H)
+
MS m/z(ESI):364(M+H) +
第二步 叔-丁基2-氨基-4-(乙基(2-(吡咯烷-1-基)乙基)氨基)苯酸酯43b的合成The second step is the synthesis of tert-butyl 2-amino-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)benzoate 43b
将43a(500mg,1.37mmol)溶于甲醇(14mL)中，加入5mL的饱和氯化铵水溶液和锌粉(447mg,6.88mmol)。该混合物在室温搅拌2小时，过滤，滤液用DCM萃取，有机相用水洗，无水硫酸钠干燥，浓缩，粗品43b(360mg)呈白色固体。该粗品直接用于下一步反应。Dissolve 43a (500mg, 1.37mmol) in methanol (14mL), add 5mL of saturated aqueous ammonium chloride solution and zinc powder (447mg, 6.88mmol). The mixture was stirred at room temperature for 2 hours, filtered, and the filtrate was extracted with DCM. The organic phase was washed with water, dried over anhydrous sodium sulfate, and concentrated. The crude product 43b (360 mg) was a white solid. The crude product was directly used in the next reaction.
MS m/z(ESI):334(M+H)
+
MS m/z(ESI):334(M+H) +
第三步 叔-丁基4-(乙基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酸酯43c的合成The third step tert-butyl 4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzoate Synthesis of 43c
将四甲基三乙酰氧基硼氢化铵(1130mg,4.32mmol)缓慢加入到化合物 43b(360mg,1.08mmol)的DCM(10mL)和三氟乙酸(1mL)溶液中，该混合物在室温搅拌2小时。加入饱和NaHCO
3水溶液淬灭反应。体系用DCM萃取，有机相用水洗，无水硫酸钠干燥，浓缩，粗品用柱层析纯化(DCM：MeOH＝10:1)得到化合物43c(300mg)黄色固体，收率66％。
Tetramethyltriacetoxyammonium borohydride (1130mg, 4.32mmol) was slowly added to a solution of compound 43b (360mg, 1.08mmol) in DCM (10mL) and trifluoroacetic acid (1mL), and the mixture was stirred at room temperature for 2 hours . The reaction was quenched by adding saturated aqueous NaHCO 3 solution. The system was extracted with DCM, the organic phase was washed with water, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by column chromatography (DCM:MeOH=10:1) to obtain compound 43c (300mg) as a yellow solid with a yield of 66%.
MS m/z(ESI):418(M+H)
+
MS m/z(ESI):418(M+H) +
第四步 叔-丁基4-(乙基(2-(吡咯烷-1-基)乙基)氨基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯酸酯43d的合成The fourth step: tert-butyl 4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyridine) Synthesis of pyran-4-yl)acetamido)benzoate 43d
将化合物43c(300mg,0.71mmol)和三乙胺(286mg,2.84mmol)分散在DCM中，在室温下滴加三氟乙酸酐(302mg,1.43mmol)，该混合物在室温搅拌2小时。加入水淬灭反应。体系用DCM萃取，有机相用水洗，无水硫酸钠干燥，浓缩，粗品用柱层析纯化(DCM：MeOH＝15:1)得到化合物43d(250mg)黄色油状物，收率68％。Compound 43c (300 mg, 0.71 mmol) and triethylamine (286 mg, 2.84 mmol) were dispersed in DCM, trifluoroacetic anhydride (302 mg, 1.43 mmol) was added dropwise at room temperature, and the mixture was stirred at room temperature for 2 hours. The reaction was quenched by adding water. The system was extracted with DCM, the organic phase was washed with water, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by column chromatography (DCM:MeOH=15:1) to obtain compound 43d (250mg) as a yellow oil with a yield of 68%.
MS m/z(ESI):514(M+H)
+
MS m/z(ESI):514(M+H) +
第五步 4-(乙基(2-(吡咯烷-1-基)乙基)氨基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯甲酸43e的合成The fifth step 4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4- (4) Acetylamino) benzoic acid 43e synthesis
将化合物43d(250mg,0.48mmol)分散在DCM(5mL)中，在室温下滴加三氟乙酸(5mL)，该混合物在室温搅拌2小时，浓缩，粗品化合物43e直接用于下一步反应。Compound 43d (250 mg, 0.48 mmol) was dispersed in DCM (5 mL), trifluoroacetic acid (5 mL) was added dropwise at room temperature, the mixture was stirred at room temperature for 2 hours, concentrated, and the crude compound 43e was directly used in the next reaction.
MS m/z(ESI):458(M+H)
+
MS m/z(ESI): 458(M+H) +
第六步 4-(乙基(2-(吡咯烷-1-基)乙基)氨基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯甲酰氯43f的合成Step 6 4-(Ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4- (4) Acetylamino) benzoyl chloride 43f synthesis
将化合物43e(200mg,0.43mmol)分散在DCM(5mL)中，在室温下滴加草酰氯(273mg,2.15mmol)和一滴DMF,该混合物在室温搅拌2小时，浓缩，粗品化合物43f直接用于下一步反应。Compound 43e (200mg, 0.43mmol) was dispersed in DCM (5mL), oxalyl chloride (273mg, 2.15mmol) and a drop of DMF were added dropwise at room temperature, the mixture was stirred at room temperature for 2 hours, concentrated, and the crude compound 43f was used directly Next reaction.
第七步 N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(乙基(2-(吡咯烷-1-基)乙基)氨基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)苯酰胺43g的合成The seventh step N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino Synthesis of )-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 43g
将化合物I-2(98mg,0.37mmol)和三乙胺(143mg,1.41mmol)溶于DCM中，在冰浴下滴加43f(180mg,0.37)的DCM溶液，滴加完毕，升至室温并继续搅拌2小时，加入水淬灭，用DCM萃取，有机相用水洗，无水硫酸钠干燥，浓缩，粗品化合物43g直接用于下一步反应。Compound I-2 (98mg, 0.37mmol) and triethylamine (143mg, 1.41mmol) were dissolved in DCM, and 43f (180mg, 0.37) in DCM was added dropwise under an ice bath. After the addition was completed, warmed to room temperature and Stirring was continued for 2 hours, quenched by adding water, extracted with DCM, the organic phase was washed with water, dried over anhydrous sodium sulfate, concentrated, and 43 g of the crude compound was directly used in the next reaction.
MS m/z(ESI):699(M+H)
+
MS m/z(ESI):699(M+H) +
第八步 N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(乙基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺43的合成Step 8 N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino )-2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide 43
将化合物43g(80mg,0.11mmol)和三乙胺(151mg,1.5mmol)溶于甲醇(2mL)中，在60℃搅拌2小时，浓缩，粗品柱层析纯化(DCM：MeOH＝10:1)得到化合物43(16.5mg)呈白色固体。Compound 43g (80mg, 0.11mmol) and triethylamine (151mg, 1.5mmol) were dissolved in methanol (2mL), stirred at 60°C for 2 hours, concentrated, and the crude product was purified by column chromatography (DCM:MeOH=10:1) Compound 43 (16.5 mg) was obtained as a white solid.
MS m/z(ESI):603(M+H)
+
MS m/z(ESI):603(M+H) +
1H NMR(400MHz,Methanol-d4)δ7.70(d,J＝9.0Hz,1H),7.57(dd,J＝1.7,0.8Hz,1H),7.40(dd,J＝8.6,0.8Hz,1H),7.22(dd,J＝8.7,1.6Hz,1H),6.85–6.77(m,2H),6.70(tt,J＝9.0,2.3Hz,1H),6.12(dd,J＝9.0,2.5Hz,1H),5.95(d,J＝2.4Hz,1H),4.05(s,2H),3.92(dt,J＝11.8,4.1Hz,2H),3.66(dt,J＝9.4,5.1Hz,1H),3.60–3.50(m,4H),3.45(q,J＝7.1Hz,2H),2.80–2.64(m,6H),2.03(d,J＝13.2Hz,2H),1.89–1.81(m,4H),1.59–1.46(m,2H),1.19(t,J＝7.0Hz,3H).1H NMR(400MHz,Methanol-d4)δ7.70(d,J＝9.0Hz,1H), 7.57(dd,J＝1.7,0.8Hz,1H),7.40(dd,J＝8.6,0.8Hz,1H) ,7.22(dd,J=8.7,1.6Hz,1H),6.85-6.77(m,2H),6.70(tt,J=9.0,2.3Hz,1H),6.12(dd,J=9.0,2.5Hz,1H) ), 5.95 (d, J = 2.4 Hz, 1H), 4.05 (s, 2H), 3.92 (dt, J = 11.8, 4.1 Hz, 2H), 3.66 (dt, J = 9.4, 5.1 Hz, 1H), 3.60 –3.50(m,4H),3.45(q,J=7.1Hz,2H),2.80–2.64(m,6H),2.03(d,J=13.2Hz,2H),1.89–1.81(m,4H), 1.59-1.46(m,2H),1.19(t,J=7.0Hz,3H).
实施例44Example 44
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((2-(二甲氨基)乙基)硫代)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺44的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((2-(dimethylamino)ethyl)thio)-2-((tetra Preparation of hydrogen-2H-pyran-4-yl)amino)benzamide 44
采用与实施例43相同的制备方法，以2-(二甲氨基)乙硫醇为原料，可得到化合物44。Using the same preparation method as in Example 43, using 2-(dimethylamino)ethanethiol as a raw material, compound 44 can be obtained.
MS m/z(ESI):562(M+H)
+
MS m/z(ESI):562(M+H) +
1H NMR(400MHz,DMSO-d6)δ12.68(s,1H),10.37(s,1H),8.08(d,J＝7.6Hz,1H),7.79(d,J＝8.5Hz,1H),7.46(s,1H),7.38(d,J＝8.5Hz,1H),7.23(dd,J＝8.6,1.5Hz,1H),7.01–6.90(m,3H),6.61(d,J＝1.7Hz,1H),6.48(dd,J＝8.4,1.7Hz,1H),4.01(s,2H),3.84–3.73(m,2H),3.64(d,J＝7.5Hz,1H),3.48–3.38(m,2H),3.15–3.05(m,2H),2.54–2.49(m,2H),2.17(s,6H),1.89(d,J＝12.5Hz,2H),1.41–1.26(m,2H).
1 H NMR(400MHz,DMSO-d6)δ12.68(s,1H), 10.37(s,1H), 8.08(d,J=7.6Hz,1H), 7.79(d,J=8.5Hz,1H), 7.46(s,1H),7.38(d,J=8.5Hz,1H),7.23(dd,J=8.6,1.5Hz,1H),7.01-6.90(m,3H),6.61(d,J=1.7Hz ,1H), 6.48(dd,J=8.4,1.7Hz,1H),4.01(s,2H),3.84–3.73(m,2H), 3.64(d,J=7.5Hz,1H),3.48–3.38( m,2H),3.15–3.05(m,2H),2.54–2.49(m,2H),2.17(s,6H),1.89(d,J=12.5Hz,2H),1.41–1.26(m,2H) .
实施例45Example 45
N-(5-(3-氟苯甲基)-1H-吲唑-3-基)-4-(乙基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺45的制备N-(5-(3-fluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(( Preparation of tetrahydro-2H-pyran-4-yl)amino)benzamide 45
采用与实施例43相同的制备方法，以3-氟溴苯为原料，可得到化合物45。Using the same preparation method as in Example 43 and using 3-fluorobromobenzene as a raw material, compound 45 can be obtained.
LCMS(ESI-MS)m/z:585(M+H
+).
LCMS (ESI-MS) m/z: 585 (M+H + ).
1H NMR(400MHz,Methanol-d
4)δ7.70(d,J＝9.0Hz,1H),7.55(dd,J＝1.7,0.9Hz,1H),7.38(dd,J＝8.7,0.9Hz,1H),7.28–7.19(m,2H),7.03(d,J＝7.6Hz,1H),6.96–6.82(m,2H),6.12(dd,J＝9.0,2.4Hz,1H),5.94(d,J＝2.4Hz,1H),4.05(s,2H),3.92(dt,J＝11.9,4.1Hz,2H),3.67(dt,J＝9.4,5.1Hz,1H),3.61–3.50(m,4H),3.45(q,J＝7.0Hz,2H),2.83–2.69(m,6H),2.03(d,J＝13.1Hz,2H),1.87(q,J＝4.9,3.7Hz,4H),1.52(ddt,J＝17.7,9.8,4.1Hz,2H),1.19(t,J＝7.0Hz,3H).
1 H NMR(400MHz,Methanol-d 4 )δ7.70(d,J＝9.0Hz,1H), 7.55(dd,J＝1.7,0.9Hz,1H), 7.38(dd,J＝8.7,0.9Hz, 1H), 7.28–7.19(m,2H), 7.03(d,J=7.6Hz,1H), 6.96–6.82(m,2H), 6.12(dd,J=9.0,2.4Hz,1H), 5.94(d ,J=2.4Hz,1H),4.05(s,2H),3.92(dt,J=11.9,4.1Hz,2H), 3.67(dt,J=9.4,5.1Hz,1H),3.61–3.50(m, 4H), 3.45(q,J=7.0Hz,2H), 2.83–2.69(m,6H), 2.03(d,J=13.1Hz,2H), 1.87(q,J=4.9,3.7Hz,4H), 1.52 (ddt, J = 17.7, 9.8, 4.1 Hz, 2H), 1.19 (t, J = 7.0 Hz, 3H).
实施例46Example 46
N-(5-(3-氟苯甲基)-1H-吲唑-3-基)-4-(甲基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺46的制备N-(5-(3-Fluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(( Preparation of tetrahydro-2H-pyran-4-yl)amino)benzamide 46
采用与实施例45相同的制备方法，以N-甲基-2-(吡咯烷-1-基)乙基胺为原料，可得到化合物46。Using the same preparation method as in Example 45 and using N-methyl-2-(pyrrolidin-1-yl)ethylamine as a raw material, compound 46 can be obtained.
LCMS(ESI-MS)m/z:571(M+H
+).
LCMS (ESI-MS) m/z: 571 (M+H + ).
1H NMR(400MHz,Methanol-d
4)δ7.71(d,J＝9.0Hz,1H),7.55(q,J＝0.9Hz,1H),7.38(dd,J＝8.7,0.9Hz,1H),7.27–7.20(m,2H),7.03(d,J＝7.8Hz,1H),6.95–6.83(m,2H),6.14(dd,J＝9.1,2.4Hz,1H),5.97(d,J＝2.4Hz,1H),4.05(s,2H),3.92(dt,J＝11.9,4.0Hz,2H),3.68(dt,J＝9.5,5.2Hz,1H),3.62–3.53(m,4H),3.03(s,3H),2.84–2.71(m,6H),2.03(d,J＝13.8Hz,2H),1.89–1.84(m,4H),1.57–1.47(m,2H).
1 H NMR(400MHz,Methanol-d 4 )δ7.71(d,J＝9.0Hz,1H), 7.55(q,J＝0.9Hz,1H), 7.38(dd,J＝8.7,0.9Hz,1H) ,7.27–7.20(m,2H),7.03(d,J=7.8Hz,1H), 6.95–6.83(m,2H), 6.14(dd,J=9.1,2.4Hz,1H), 5.97(d,J ＝2.4Hz,1H),4.05(s,2H),3.92(dt,J＝11.9,4.0Hz,2H), 3.68(dt,J＝9.5,5.2Hz,1H),3.62–3.53(m,4H) ,3.03(s,3H),2.84-2.71(m,6H),2.03(d,J=13.8Hz,2H),1.89-1.84(m,4H),1.57-1.47(m,2H).
实施例47Example 47
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((氧杂环丁烷-3-基甲基)氨基)苯甲酰胺47的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((oxetane Preparation of -3-ylmethyl)amino)benzamide 47
采用与实施例11相同的制备方法，以氧杂环丁烷-3-基甲醛为原料，可得到化合物47。Using the same preparation method as in Example 11, using oxetane-3-ylcarboxaldehyde as a raw material, compound 47 can be obtained.
LCMS(ESI-MS)m/z:547.4(M+H
+).
LCMS (ESI-MS) m/z: 547.4 (M+H + ).
1H NMR(400MHz,DMSO-d
6)δ12.64(s,1H),10.10(s,1H),8.21(t,J＝5.2Hz,1H),7.79(d,J＝9.0Hz,1H),7.47(s,1H),7.44–7.36(m,1H),7.24(dd,J＝8.6,1.6Hz,1H),7.08–6.89(m,3H),6.25(dd,J＝9.0,2.3Hz,1H),6.10(d,J＝2.3Hz,1H),4.66(dd,J＝7.7,6.0Hz,2H),4.32(t,J＝5.9Hz,2H),4.02(s,2H),3.43(dd,J＝7.2,5.2Hz,2H),3.33–3.10(m,5H),2.49–2.36(m,4H),2.22(s,3H).
1 H NMR(400MHz,DMSO-d 6 )δ12.64(s,1H), 10.10(s,1H), 8.21(t,J=5.2Hz,1H), 7.79(d,J=9.0Hz,1H) ,7.47(s,1H),7.44–7.36(m,1H),7.24(dd,J=8.6,1.6Hz,1H),7.08–6.89(m,3H),6.25(dd,J=9.0,2.3Hz ,1H),6.10(d,J=2.3Hz,1H),4.66(dd,J=7.7,6.0Hz,2H), 4.32(t,J=5.9Hz,2H),4.02(s,2H),3.43 (dd,J=7.2,5.2Hz,2H), 3.33–3.10(m,5H), 2.49–2.36(m,4H), 2.22(s,3H).
实施例48Example 48
N-(5-(3-氟苄基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((氧杂环丁烷-3-基甲基)氨基)苯甲酰胺48的制备N-(5-(3-Fluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((oxetane-3- Preparation of (methyl)amino)benzamide 48
采用与实施例47相同的制备方法，以3-氟溴苯为原料，可得到化合物48。Using the same preparation method as in Example 47 and using 3-fluorobromobenzene as a raw material, compound 48 can be obtained.
MS m/z(ESI):529[M+H]
+
MS m/z(ESI):529[M+H] +
1H NMR(400MHz,DMSO-d
6)δ12.62(s,1H),10.09(s,1H),8.21(t,J＝5.3Hz,1H),7.79(d,J＝9.0Hz,1H),7.45(s,1H),7.39(dd,J＝8.6,0.8Hz,1H),7.34–7.25(m,1H),7.22(dd,J＝8.6,1.6Hz,1H),7.11–7.02(m,2H),7.01–6.95(m,1H),6.26(dd,J＝9.0,2.3Hz,1H),6.10(d,J＝2.4Hz,1H),4.67(dd,J＝7.7,6.0Hz,2H), 4.32(t,J＝5.9Hz,2H),4.02(s,2H),3.44(dd,J＝7.2,5.2Hz,2H),3.31-3.25(m,4H),3.21(m,1H),2.45(m,4H),2.24(s,3H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.62 (s, 1H), 10.09 (s, 1H), 8.21 (t, J = 5.3 Hz, 1H), 7.79 (d, J = 9.0 Hz, 1H) ,7.45(s,1H),7.39(dd,J=8.6,0.8Hz,1H),7.34-7.25(m,1H),7.22(dd,J=8.6,1.6Hz,1H),7.11-7.02(m ,2H),7.01–6.95(m,1H), 6.26(dd,J=9.0,2.3Hz,1H), 6.10(d,J=2.4Hz,1H), 4.67(dd,J=7.7,6.0Hz, 2H), 4.32(t,J=5.9Hz,2H),4.02(s,2H),3.44(dd,J=7.2,5.2Hz,2H),3.31-3.25(m,4H),3.21(m,1H ), 2.45(m, 4H), 2.24(s, 3H).
实施例49Example 49
N-(5-(2,5-二氟苄基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((氧杂环丁烷-3-基甲基)氨基)苯甲酰胺49的制备N-(5-(2,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((oxetane Preparation of -3-ylmethyl)amino)benzamide 49
采用与实施例47相同的制备方法，以3-氟溴苯为原料，可得到化合物49。Using the same preparation method as in Example 47 and using 3-fluorobromobenzene as a raw material, compound 49 can be obtained.
MS m/z(ESI):547[M+H]
+
MS m/z(ESI):547[M+H] +
1H NMR(400MHz,DMSO-d
6)δ12.64(s,1H),10.10(s,1H),8.21(t,J＝5.2Hz,1H),7.79(d,J＝9.0Hz,1H),7.53–7.35(m,2H),7.28–7.12(m,3H),7.12–7.01(m,1H),6.26(dd,J＝9.0,2.3Hz,1H),6.10(d,J＝2.4Hz,1H),4.67(dd,J＝7.7,5.9Hz,2H),4.32(t,J＝5.9Hz,2H),4.02(s,2H),3.44(dd,J＝7.2,5.2Hz,2H),3.29(t,J＝5.0Hz,4H),3.25–3.13(m,1H),2.44(t,J＝5.0Hz,4H),2.23(s,3H).
1 H NMR(400MHz,DMSO-d 6 )δ12.64(s,1H), 10.10(s,1H), 8.21(t,J=5.2Hz,1H), 7.79(d,J=9.0Hz,1H) ,7.53–7.35(m,2H),7.28–7.12(m,3H),7.12–7.01(m,1H), 6.26(dd,J=9.0,2.3Hz,1H),6.10(d,J=2.4Hz ,1H), 4.67(dd,J=7.7,5.9Hz,2H), 4.32(t,J=5.9Hz,2H),4.02(s,2H),3.44(dd,J=7.2,5.2Hz,2H) , 3.29(t,J=5.0Hz,4H), 3.25–3.13(m,1H), 2.44(t,J=5.0Hz,4H), 2.23(s,3H).
实施例50Example 50
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺50的制备N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl) Preparation of -2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 50
采用与实施例21相同的制备方法，以2,5-二氟溴苯为原料，可得到化合物 50。Using the same preparation method as in Example 21, using 2,5-difluorobromobenzene as a raw material, compound 50 can be obtained.
MS m/z(ESI):593[M+H]
+。
MS m/z (ESI): 593 [M+H] + .
1H NMR(400MHz,DMSO-d
6)δ12.67(s,1H),10.07(s,1H),8.28(d,J＝7.7Hz,1H),7.76(d,J＝9.0Hz,1H),7.50(d,J＝1.5Hz,1H),7.43–7.34(m,2H),7.29(dd,J＝8.7,1.6Hz,1H),7.19(td,J＝9.3,4.7Hz,1H),7.15–7.04(m,1H),6.21(dd,J＝9.0,2.3Hz,1H),6.10(d,J＝2.3Hz,1H),5.10-4.86m,2H),4.72(dt,J＝14.8,7.3Hz,1H),3.78(dt,J＝11.6,4.1Hz,2H),3.65(q,J＝8.7Hz,1H),3.46(tt,J＝11.8,2.3Hz,2H),3.26(s,4H),2.46(s,4H),2.25(s,3H),1.96–1.81(m,2H),1.35-1.22(m,2H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.67 (s, 1H), 10.07 (s, 1H), 8.28 (d, J = 7.7 Hz, 1H), 7.76 (d, J = 9.0 Hz, 1H) ,7.50(d,J=1.5Hz,1H),7.43-7.34(m,2H),7.29(dd,J=8.7,1.6Hz,1H),7.19(td,J=9.3,4.7Hz,1H), 7.15–7.04 (m, 1H), 6.21 (dd, J = 9.0, 2.3 Hz, 1H), 6.10 (d, J = 2.3 Hz, 1H), 5.10-4.86m, 2H), 4.72 (dt, J = 14.8 ,7.3Hz,1H), 3.78(dt,J=11.6,4.1Hz,2H), 3.65(q,J=8.7Hz,1H), 3.46(tt,J=11.8,2.3Hz,2H), 3.26(s ,4H), 2.46(s,4H), 2.25(s,3H), 1.96-1.81(m,2H), 1.35-1.22(m,2H).
实施例51Example 51
N-(5-(1-(3-氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺51的制备N-(5-(1-(3-fluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2- Preparation of ((Tetrahydro-2H-pyran-4-yl)amino)benzamide 51
采用与实施例21相同的制备方法，以3-氟溴苯为原料，可得到化合物51。Using the same preparation method as in Example 21, using 3-fluorobromobenzene as a raw material, compound 51 can be obtained.
MS m/z(ESI):575[M+1]
+。
MS m/z (ESI): 575[M+1] + .
1H NMR(400MHz,DMSO-d
6)δ12.65(s,1H),10.06(s,1H),8.27(d,J＝7.7Hz,1H),7.76(d,J＝9.1Hz,1H),7.53(d,J＝1.6Hz,1H),7.39(d,J＝8.7Hz,1H),7.35–7.26(m,2H),7.22–7.14(m,2H),7.01(td,J＝8.7,2.3Hz,1H),6.21(dd,J＝9.0,2.3Hz,1H),6.10(d,J＝2.3Hz,1H),5.10-4.75(m,2H),4.55(dt,J＝14.8,7.1Hz,1H),3.85–3.56(m,3H),3.52–3.41(m,2H),3.23(d,J＝5.6Hz,4H),2.43(d,J＝5.8Hz,4H),2.22(s,3H),1.89(dd,J＝10.8,6.8Hz,2H),1.50–1.24(m,2H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.65 (s, 1H), 10.06 (s, 1H), 8.27 (d, J = 7.7 Hz, 1H), 7.76 (d, J = 9.1 Hz, 1H) ,7.53(d,J=1.6Hz,1H),7.39(d,J=8.7Hz,1H),7.35-7.26(m,2H),7.22-7.14(m,2H),7.01(td,J=8.7 ,2.3Hz,1H),6.21(dd,J=9.0,2.3Hz,1H),6.10(d,J=2.3Hz,1H),5.10-4.75(m,2H),4.55(dt,J=14.8, 7.1Hz, 1H), 3.85–3.56 (m, 3H), 3.52–3.41 (m, 2H), 3.23 (d, J = 5.6 Hz, 4H), 2.43 (d, J = 5.8 Hz, 4H), 2.22 ( s, 3H), 1.89 (dd, J = 10.8, 6.8 Hz, 2H), 1.50-1.24 (m, 2H).
实施例52Example 52
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-乙基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺52的制备N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-ethylpiperazin-1-yl) Preparation of -2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 52
采用与实施例50相同的制备方法，以N-乙基哌嗪为原料，可得到化合物52。Using the same preparation method as in Example 50 and using N-ethylpiperazine as a raw material, compound 52 can be obtained.
MS m/z(ESI):607[M+1]
+
MS m/z(ESI):607[M+1] +
1H NMR(400MHz,DMSO-d
6)δ12.67(s,1H),10.08(s,1H),8.27(d,J＝7.6Hz,1H),7.76(d,J＝9.0Hz,1H),7.50(s,1H),7.44–7.37(m,2H),7.29(dd,J＝8.7,1.7Hz,1H),7.19(td,J＝9.3,4.7Hz,1H),7.10(ddd,J＝8.9,7.5,3.5Hz,1H),6.20(dd,J＝9.0,2.3Hz,1H),6.09(d,J＝2.3Hz,1H),5.10–4.86(m,2H),4.72(dt,J＝14.8,7.3Hz,1H),3.87–3.72(m,2H),3.64(dd,J＝10.6,6.5Hz,1H),3.46(ddt,J＝13.0,11.2,2.2Hz,2H),3.25(s,4H),2.48(s,4H),2.34(q,2H),1.96–1.83(m,2H),1.30(dtd,J＝13.5,9.8,4.0Hz,2H),1.01(t,J＝7.1Hz,3H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.67 (s, 1H), 10.08 (s, 1H), 8.27 (d, J = 7.6 Hz, 1H), 7.76 (d, J = 9.0 Hz, 1H) ,7.50(s,1H),7.44-7.37(m,2H),7.29(dd,J=8.7,1.7Hz,1H),7.19(td,J=9.3,4.7Hz,1H),7.10(ddd,J ＝8.9,7.5,3.5Hz,1H), 6.20(dd,J＝9.0,2.3Hz,1H), 6.09(d,J＝2.3Hz,1H), 5.10–4.86(m,2H), 4.72(dt, J = 14.8, 7.3 Hz, 1H), 3.87–3.72 (m, 2H), 3.64 (dd, J = 10.6, 6.5 Hz, 1H), 3.46 (ddt, J = 13.0, 11.2, 2.2 Hz, 2H), 3.25 (s,4H),2.48(s,4H),2.34(q,2H),1.96-1.83(m,2H),1.30(dtd,J=13.5,9.8,4.0Hz,2H),1.01(t,J =7.1Hz, 3H).
实施例53Example 53
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-***啉哌啶-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺53的制备N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-morpholinepiperidin-1-yl) Preparation of -2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 53
采用与实施例50相同的制备方法，以4-(吗啉-4-基)哌啶为原料，可得到化 合物53。Using the same preparation method as in Example 50 and using 4-(morpholin-4-yl)piperidine as a raw material, compound 53 can be obtained.
MS m/z(ESI):663[M+H]
+。
MS m/z (ESI): 663 [M+H] + .
1H NMR(400MHz,DMSO-d
6)δ12.68–12.62(m,1H),10.04(s,1H),8.28(d,J＝7.6Hz,1H),7.74(d,J＝9.1Hz,1H),7.50(d,J＝1.5Hz,1H),7.45–7.34(m,2H),7.29(dd,J＝8.7,1.6Hz,1H),7.19(td,J＝9.3,4.7Hz,1H),7.10(ddd,J＝12.1,7.8,3.5Hz,1H),6.20(dd,J＝9.1,2.3Hz,1H),6.09(d,J＝2.3Hz,1H),5.10-4.80(m,2H),4.72(dt,J＝14.8,7.2Hz,1H),3.81(dd,J＝29.3,12.1Hz,4H),3.69–3.39(m,7H),3.30-3.25(m,2H),2.75(t,J＝12.3Hz,2H),2.48-2.28(m,3H),2.01-1.78(m,4H),1.47–1.11(m,4H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.68-12.62 (m, 1H), 10.04 (s, 1H), 8.28 (d, J = 7.6 Hz, 1H), 7.74 (d, J = 9.1 Hz, 1H), 7.50 (d, J = 1.5 Hz, 1H), 7.45–7.34 (m, 2H), 7.29 (dd, J = 8.7, 1.6 Hz, 1H), 7.19 (td, J = 9.3, 4.7 Hz, 1H ), 7.10 (ddd, J = 12.1, 7.8, 3.5 Hz, 1H), 6.20 (dd, J = 9.1, 2.3 Hz, 1H), 6.09 (d, J = 2.3 Hz, 1H), 5.10-4.80 (m, 2H), 4.72 (dt, J = 14.8, 7.2 Hz, 1H), 3.81 (dd, J = 29.3, 12.1 Hz, 4H), 3.69–3.39 (m, 7H), 3.30-3.25 (m, 2H), 2.75 (t,J=12.3Hz,2H), 2.48-2.28(m,3H), 2.01-1.78(m,4H), 1.47-1.11(m,4H).
实施例54Example 54
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-(四氢-2H-吡喃-4-基)哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺54的制备N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-(tetrahydro-2H-pyran- Preparation of 4-yl)piperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 54
采用与实施例50相同的制备方法，以1-(四氢-2H-吡喃-4-基)哌嗪为原料，可得到化合物54。Using the same preparation method as in Example 50, using 1-(tetrahydro-2H-pyran-4-yl)piperazine as a raw material, compound 54 can be obtained.
MS m/z(ESI):663[M+H]
+。
MS m/z (ESI): 663 [M+H] + .
1H NMR(400MHz,DMSO-d
6)δ12.67(s,1H),10.08(s,1H),8.27(d,J＝7.6Hz,1H),7.76(d,J＝9.0Hz,1H),7.50(s,1H),7.45–7.35(m,2H),7.29(dd,J＝8.7,1.6Hz,1H),7.19(td,J＝9.3,4.7Hz,1H),7.11(tt,J＝8.9,3.5Hz,1H),6.20(dd,J＝8.9,2.2Hz,1H),6.09(d,J＝2.4Hz,1H),5.10-4.80(m,2H),4.72(dt,J＝14.9,7.3Hz,1H),3.87(dd,J＝10.7,4.1Hz,2H),3.78(d,J＝11.9Hz,2H),3.64(d,J＝8.4Hz, 1H),3.52–3.39(m,2H),3.30–3.14(m,6H),2.58(s,4H),2.38(s,1H),1.96–1.84(m,2H),1.71(d,J＝12.4Hz,2H),1.46–1.21(m,4H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.67 (s, 1H), 10.08 (s, 1H), 8.27 (d, J = 7.6 Hz, 1H), 7.76 (d, J = 9.0 Hz, 1H) ,7.50(s,1H),7.45-7.35(m,2H),7.29(dd,J=8.7,1.6Hz,1H),7.19(td,J=9.3,4.7Hz,1H),7.11(tt,J ＝8.9,3.5Hz,1H), 6.20(dd,J＝8.9,2.2Hz,1H), 6.09(d,J＝2.4Hz,1H), 5.10-4.80(m,2H), 4.72(dt,J＝ 14.9, 7.3 Hz, 1H), 3.87 (dd, J = 10.7, 4.1 Hz, 2H), 3.78 (d, J = 11.9 Hz, 2H), 3.64 (d, J = 8.4 Hz, 1H), 3.52–3.39 ( m,2H),3.30–3.14(m,6H),2.58(s,4H),2.38(s,1H),1.96–1.84(m,2H),1.71(d,J=12.4Hz,2H),1.46 –1.21(m,4H).
实施例55Example 55
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-环丙基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺55的制备N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-cyclopropylpiperazin-1-yl )-2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide 55
采用与实施例50相同的制备方法，以4-环丙基哌嗪为原料，可得到化合物55。Using the same preparation method as in Example 50 and using 4-cyclopropylpiperazine as a raw material, compound 55 can be obtained.
MS m/z(ESI):619[M+1]
+
MS m/z(ESI):619[M+1] +
1H NMR(400MHz,DMSO-d
6)δ12.70(s,1H),10.11(s,1H),8.31(d,J＝7.6Hz,1H),7.79(d,J＝9.1Hz,1H),7.53(s,1H),7.47–7.39(m,2H),7.32(dd,J＝8.7,1.6Hz,1H),7.29–7.10(m,2H),6.23(dd,J＝9.1,2.2Hz,1H),6.12(d,J＝2.3Hz,1H),5.13–4.86(m,2H),4.75(dt,J＝14.8,7.3Hz,1H),3.81(d,J＝11.7Hz,2H),3.75–3.62(m,1H),3.49(t,J＝11.0Hz,2H),3.22(t,J＝5.0Hz,4H),2.65(t,J＝4.9Hz,4H),1.93(d,J＝12.5Hz,2H),1.66(tt,J＝6.7,3.7Hz,1H),1.40–1.27(m,2H),0.48–0.32(m,4H).
1 H NMR(400MHz,DMSO-d 6 )δ12.70(s,1H), 10.11(s,1H), 8.31(d,J=7.6Hz,1H), 7.79(d,J=9.1Hz,1H) ,7.53(s,1H),7.47–7.39(m,2H),7.32(dd,J=8.7,1.6Hz,1H),7.29–7.10(m,2H),6.23(dd,J=9.1,2.2Hz ,1H),6.12(d,J＝2.3Hz,1H),5.13–4.86(m,2H),4.75(dt,J＝14.8,7.3Hz,1H),3.81(d,J＝11.7Hz,2H) ,3.75–3.62(m,1H),3.49(t,J=11.0Hz,2H), 3.22(t,J=5.0Hz,4H), 2.65(t,J=4.9Hz,4H),1.93(d, J = 12.5Hz, 2H), 1.66 (tt, J = 6.7, 3.7Hz, 1H), 1.40–1.27 (m, 2H), 0.48–0.32 (m, 4H).
实施例56Example 56
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-三氟甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺56的制备N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-trifluoromethylpiperazine-1- Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 56
采用与实施例50相同的制备方法，以4-三氟甲基哌嗪为原料，可得到化合物56。Using the same preparation method as in Example 50 and using 4-trifluoromethylpiperazine as a raw material, compound 56 can be obtained.
MS m/z(ESI):647[M+1]
+
MS m/z(ESI):647[M+1] +
1H NMR(400MHz,DMSO-d
6)δ12.68(s,1H),10.12(s,1H),8.28(d,J＝7.7Hz,1H),7.79(d,J＝9.0Hz,1H),7.50(d,J＝1.6Hz,1H),7.44–7.36(m,2H),7.29(dd,J＝8.8,1.6Hz,1H),7.22–7.17(m,1H),7.10(td,J＝8.2,3.7Hz,1H),6.26–6.21(m,1H),6.15(d,J＝2.3Hz,1H),5.14–4.83(m,2H),4.72(dt,J＝14.7,7.1Hz,1H),3.79(d,J＝11.6Hz,2H),3.65(s,1H),3.46(t,J＝10.9Hz,2H),3.33(t,J＝5.0Hz,4H),2.98(t,J＝5.0Hz,4H),1.92(t,J＝13.1Hz,2H),1.36–1.26(m,2H).
1 H NMR(400MHz,DMSO-d 6 )δ12.68(s,1H), 10.12(s,1H), 8.28(d,J=7.7Hz,1H), 7.79(d,J=9.0Hz,1H) ,7.50(d,J=1.6Hz,1H),7.44-7.36(m,2H),7.29(dd,J=8.8,1.6Hz,1H),7.22-7.17(m,1H),7.10(td,J =8.2,3.7Hz,1H), 6.26–6.21(m,1H), 6.15(d,J=2.3Hz,1H), 5.14–4.83(m,2H), 4.72(dt,J=14.7,7.1Hz, 1H), 3.79 (d, J = 11.6 Hz, 2H), 3.65 (s, 1H), 3.46 (t, J = 10.9 Hz, 2H), 3.33 (t, J = 5.0 Hz, 4H), 2.98 (t, J = 5.0Hz, 4H), 1.92 (t, J = 13.1Hz, 2H), 1.36-1.26 (m, 2H).
实施例57Example 57
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(甲基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺57的制备N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidine-1- (Pyryl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 57
采用与实施例50相同的制备方法，以N-甲基-2-(吡咯烷-1-基)乙基胺为原料，可得到化合物57。Using the same preparation method as in Example 50, using N-methyl-2-(pyrrolidin-1-yl)ethylamine as a raw material, compound 57 can be obtained.
MS m/z(ESI):621(M+H)
+
MS m/z(ESI):621(M+H) +
1H NMR(400MHz,Methanol-d4)δ7.72–7.59(m,2H),7.42(dd,J＝8.8,0.8Hz,1H),7.30(dd,J＝8.9,1.6Hz,1H),7.18(ddd,J＝9.1,5.7,3.1Hz,1H),7.07(td,J＝9.3,4.5Hz,1H),6.98(ddd,J＝12.2,8.3,3.6Hz,1H),6.14(dd,J＝9.2,2.4Hz,1H),5.96(d,J＝2.3Hz,1H),5.09–4.88(m,2H),4.84–4.71(m,1H),3.93(dt,J＝11.7,4.1Hz,2H),3.68(dt,J＝9.4,5.1Hz,1H),3.57(td,J＝9.7,9.2,3.0Hz,4H),3.02(s,3H),2.80–2.58(m,6H),2.04(d,J＝13.3Hz,2H),1.85(t,J＝4.5Hz,4H),1.59-1.48(m,2H).
1 H NMR(400MHz,Methanol-d4)δ7.72–7.59(m,2H), 7.42(dd,J＝8.8,0.8Hz,1H), 7.30(dd,J＝8.9,1.6Hz,1H), 7.18 (ddd,J=9.1,5.7,3.1Hz,1H), 7.07(td,J=9.3,4.5Hz,1H), 6.98(ddd,J=12.2,8.3,3.6Hz,1H), 6.14(dd,J ＝9.2,2.4Hz,1H),5.96(d,J＝2.3Hz,1H),5.09–4.88(m,2H),4.84–4.71(m,1H),3.93(dt,J＝11.7,4.1Hz, 2H), 3.68(dt,J=9.4,5.1Hz,1H),3.57(td,J=9.7,9.2,3.0Hz,4H),3.02(s,3H),2.80–2.58(m,6H),2.04 (d, J = 13.3Hz, 2H), 1.85 (t, J = 4.5 Hz, 4H), 1.59-1.48 (m, 2H).
实施例58Example 58
N-(5-(3,5-二氟苄基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-((1R，4R)-5-甲基-2,5-二氮杂双环[2.2.10.1]庚烷-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺58的制备N-(5-(3,5-Difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-((1R,4R)-5-methyl-2 Preparation of ,5-diazabicyclo[2.2.10.1]heptan-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 58
采用与实施例39相同的制备方法，以(1R，4R)-2-甲基-2,5-二氮杂二环[2.2.1]庚烷为原料，可得到化合物58。Using the same preparation method as in Example 39, using (1R,4R)-2-methyl-2,5-diazabicyclo[2.2.1]heptane as a raw material, compound 58 can be obtained.
MS m/z(ESI):574[M+1]
+
MS m/z(ESI):574[M+1] +
1H NMR(400MHz,DMSO-d
6)δ13.14(s,1H),10.19(s,1H),8.90(d,J＝1.2Hz,1H),8.31(d,J＝7.5Hz,1H),7.75(d,J＝9.0Hz,1H),7.53(s,1H),7.07–6.95(m,3H),5.93(d,J＝8.7Hz,1H),5.76(s,1H),4.42(s,1H),4.19(s,2H),3.82(d,J＝11.4Hz,2H),3.64(s,1H),3.48(t,J＝10.8Hz,4H),3.32-3.28(m,2H),2.83(d,J＝9.3Hz,1H),2.30(s,3H),1.92(q,J＝12.5,12.0Hz,4H),1.35(d,J＝11.5Hz,2H).
1 H NMR (400MHz, DMSO-d 6 ) δ 13.14 (s, 1H), 10.19 (s, 1H), 8.90 (d, J = 1.2 Hz, 1H), 8.31 (d, J = 7.5 Hz, 1H) ,7.75(d,J=9.0Hz,1H),7.53(s,1H),7.07–6.95(m,3H),5.93(d,J=8.7Hz,1H),5.76(s,1H),4.42( s, 1H), 4.19 (s, 2H), 3.82 (d, J = 11.4 Hz, 2H), 3.64 (s, 1H), 3.48 (t, J = 10.8 Hz, 4H), 3.32-3.28 (m, 2H) ), 2.83 (d, J = 9.3 Hz, 1H), 2.30 (s, 3H), 1.92 (q, J = 12.5, 12.0 Hz, 4H), 1.35 (d, J = 11.5 Hz, 2H).
实施例59Example 59
N-(5-(3,5-二氟苄基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-((1s，4s)-5-甲基-2,5-二氮杂双环[2.2.10.1]庚烷-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺59的制备N-(5-(3,5-Difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-((1s,4s)-5-methyl-2 Preparation of ,5-diazabicyclo[2.2.10.1]heptan-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 59
采用与实施例39相同的制备方法，以(1S，4S)-2-甲基-2,5-二氮杂二环[2.2.1]庚烷为原料，可得到化合物59。Using the same preparation method as in Example 39, using (1S,4S)-2-methyl-2,5-diazabicyclo[2.2.1]heptane as a raw material, compound 59 can be obtained.
MS m/z(ESI):574[M+1]
+
MS m/z(ESI):574[M+1] +
1H NMR(400MHz,DMSO-d
6)δ13.14(s,1H),10.19(s,1H),8.90(d,J＝1.2Hz,1H),8.31(d,J＝7.5Hz,1H),7.75(d,J＝9.0Hz,1H),7.53(s,1H),7.07–6.95(m,3H),5.93(d,J＝8.7Hz,1H),5.76(s,1H),4.42(s,1H),4.19(s,2H),3.82(d,J＝11.4Hz,2H),3.64(s,1H),3.48(t,J＝10.8Hz,4H),3.32-3.28(m,2H),2.83(d,J＝9.3Hz,1H),2.30(s,3H),1.92(q,J＝12.5,12.0Hz,4H),1.35(d,J＝11.5Hz,2H).
1 H NMR (400MHz, DMSO-d 6 ) δ 13.14 (s, 1H), 10.19 (s, 1H), 8.90 (d, J = 1.2 Hz, 1H), 8.31 (d, J = 7.5 Hz, 1H) ,7.75(d,J=9.0Hz,1H),7.53(s,1H),7.07–6.95(m,3H),5.93(d,J=8.7Hz,1H),5.76(s,1H),4.42( s, 1H), 4.19 (s, 2H), 3.82 (d, J = 11.4 Hz, 2H), 3.64 (s, 1H), 3.48 (t, J = 10.8 Hz, 4H), 3.32-3.28 (m, 2H) ), 2.83 (d, J = 9.3 Hz, 1H), 2.30 (s, 3H), 1.92 (q, J = 12.5, 12.0 Hz, 4H), 1.35 (d, J = 11.5 Hz, 2H).
实施例60Example 60
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-2-(2-(二甲氨基)乙基)-1-羰基-6-((四氢-2H-吡喃-4-基)氨基)异二氢吲哚-5-甲酰胺60的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-2-(2-(dimethylamino)ethyl)-1-carbonyl-6-((tetra Preparation of Hydro-2H-pyran-4-yl)amino)isoindole-5-carboxamide 60
第一步 二甲基2-((四氢-2H-吡喃-4-基)氨基)对苯二甲酸酯60bThe first step Dimethyl 2-((tetrahydro-2H-pyran-4-yl)amino)terephthalate 60b
向干燥的反应瓶中加入60a(2.09g,10mmol和50ml DCM，加入醋酸硼氢化钠(4.24g，20mmol)，4-四氢吡喃酮(2.2g,22mmol)和5mL醋酸。反应室温搅拌过夜，TLC显示反应完毕。加入碳酸钾溶液搅拌30分钟，分出有机相，无水硫酸钠干燥后减压浓缩得到粗品60b直接用于下一步。Add 60a (2.09g, 10mmol and 50ml DCM) to the dry reaction flask, add sodium acetate borohydride (4.24g, 20mmol), 4-tetrahydropyrone (2.2g, 22mmol) and 5mL acetic acid. The reaction is stirred at room temperature overnight , TLC showed that the reaction was complete. Potassium carbonate solution was added and stirred for 30 minutes, the organic phase was separated, dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain crude product 60b, which was used directly in the next step.
MS m/z(ESI):294[M+H]+MS m/z(ESI):294[M+H]+
第二步 二甲基2-溴-5-((四氢-2H-吡喃-4-基)氨基)对苯二甲酸酯60cThe second step Dimethyl 2-bromo-5-((tetrahydro-2H-pyran-4-yl)amino)terephthalate 60c
向反应瓶中加入60b(2.93g,10.0mmol)和100ml乙腈，然后加入NBS(1.78g,10.0mmol)，加完后室温搅拌过夜。TLC显示原料反应完。减压浓缩反应液，倒入水中，搅拌0.5小时，过滤并干燥所得固体即是60c(3.2g,8.6mmol)。Add 60b (2.93g, 10.0mmol) and 100ml of acetonitrile to the reaction flask, then add NBS (1.78g, 10.0mmol), and stir at room temperature overnight after addition. TLC showed that the reaction of the starting materials was complete. The reaction solution was concentrated under reduced pressure, poured into water, stirred for 0.5 hours, filtered and dried, and the resulting solid was 60c (3.2g, 8.6mmol).
MS m/z(ESI):373[M+2]+MS m/z(ESI):373[M+2]+
1H NMR(400MHz,DMSO-d6)δ7.97(s,1H),7.77–7.69(d,J＝8.0,1H),7.17(s,1H),3.85(s,3H),3.84(s,3H),3.78-3.66(m,1H),3.53-3.41(m,2H),1.96-1.85(m,2H),1.50-1.33(m,2H).1H NMR(400MHz,DMSO-d6)δ7.97(s,1H), 7.77–7.69(d,J=8.0,1H), 7.17(s,1H), 3.85(s,3H), 3.84(s,3H) ), 3.78-3.66 (m, 1H), 3.53-3.41 (m, 2H), 1.96-1.85 (m, 2H), 1.50-1.33 (m, 2H).
第三步 二甲基2-氰基-5-((四氢-2H-吡喃-4-基)氨基)对苯二甲酸酯60dThe third step Dimethyl 2-cyano-5-((tetrahydro-2H-pyran-4-yl)amino)terephthalate 60d
向单口瓶中加入60c(1.86g,5mmol),CuCN(890mg,10mmol)和DMF(30ml),加 热至150℃,在此温度下维持1小时，冷却室温，倒入乙酸乙酯-氨水体系中并搅拌，澄清后分液。有机相用食盐水洗三次，无水硫酸钠干燥，减压浓缩后得60d(1.27g,4.0mmol).Add 60c (1.86g, 5mmol), CuCN (890mg, 10mmol) and DMF (30ml) to the single-mouth flask, heat to 150℃, maintain at this temperature for 1 hour, cool to room temperature, and pour into the ethyl acetate-ammonia system Stir and separate after clarification. The organic phase was washed three times with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 60d (1.27g, 4.0mmol).
MS m/z(ESI):319[M+H]+MS m/z(ESI):319[M+H]+
第四步 甲基1-羰基-6-((四氢-2H-吡喃-4-基)氨基)异二氢吲哚-5-羧酸酯60eStep 4 Methyl 1-carbonyl-6-((tetrahydro-2H-pyran-4-yl)amino)isoindoline-5-carboxylate 60e
将60d(319mg,1.0mmol)溶于THF-MeOH体系中，加入100mg Raney-Ni，在氢气氛围下搅拌过夜，TLC显示反应完成，过滤并减压浓缩滤液，所得固体用EtOAc-PE体系打浆，得60e(230mg,0.8mmol).60d (319mg, 1.0mmol) was dissolved in the THF-MeOH system, 100mg Raney-Ni was added, and the mixture was stirred overnight under a hydrogen atmosphere. TLC showed that the reaction was complete. The filtrate was filtered and concentrated under reduced pressure. The obtained solid was slurried with the EtOAc-PE system. Get 60e (230mg, 0.8mmol).
MS m/z(ESI):291[M+H]+MS m/z(ESI): 291[M+H]+
第五步 甲基2-(2-溴乙基)-1-羰基-6-((四氢-2H-吡喃-4-基)氨基)异二氢吲哚-5-羧酸酯60fStep 5 Methyl 2-(2-bromoethyl)-1-carbonyl-6-((tetrahydro-2H-pyran-4-yl)amino)isoindoline-5-carboxylate 60f
化合物60e(1.45g,5.0mmol)溶于DMF中，加入NaH(0.8g,60％,20mmol,4eq)并在室温下搅拌0.5小时，然后加入二溴乙烷(9.25g,50mmol)并在室温下搅拌4小时，倒入氯化铵水溶液中，加入乙酸乙酯萃取3次，有机相用无水硫酸钠干燥，加入硅胶拌样，flash纯化(DCM:MeOH＝20:1)得60f(0.79g,2.0mmol).Compound 60e (1.45g, 5.0mmol) was dissolved in DMF, NaH (0.8g, 60%, 20mmol, 4eq) was added and stirred at room temperature for 0.5 hours, then dibromoethane (9.25g, 50mmol) was added and kept at room temperature After stirring for 4 hours, pour into aqueous ammonium chloride solution, add ethyl acetate to extract 3 times, dry the organic phase with anhydrous sodium sulfate, add silica gel to mix the sample, flash purification (DCM:MeOH=20:1) to obtain 60f(0.79) g, 2.0mmol).
MS m/z(ESI):433[M+H]+MS m/z(ESI):433[M+H]+
第六步 甲基2-(2-(二甲氨基)乙基)-1-羰基-6-((四氢-2H-吡喃-4-基)氨基)异二氢吲哚-5-羧酸酯60gStep 6 Methyl 2-(2-(dimethylamino)ethyl)-1-carbonyl-6-((tetrahydro-2H-pyran-4-yl)amino)isoindoline-5-carboxyl Acid ester 60g
向干燥的反应瓶中加入60f(433mg,1.0mmol),二甲胺盐酸盐(400mg,5.0mmol),碳酸钾(1.38g,10.0mmol)和DMF(20ml)。搅拌过夜后加入二氯甲烷稀释反应液，过滤所得滤液减压浓缩除去高沸点的DMF，flash纯化(DCM:MeOH＝20:1)得60g(180mg,0.5mmol)。To the dry reaction flask was added 60f (433mg, 1.0mmol), dimethylamine hydrochloride (400mg, 5.0mmol), potassium carbonate (1.38g, 10.0mmol) and DMF (20ml). After stirring overnight, dichloromethane was added to dilute the reaction solution, the filtrate obtained by filtration was concentrated under reduced pressure to remove high boiling DMF, and flash purification (DCM:MeOH=20:1) gave 60g (180mg, 0.5mmol).
MS m/z(ESI):362[M+H]+MS m/z(ESI):362[M+H]+
第七步 2-(2-(二甲氨基)乙基)-1-羰基-6-((四氢-2H-吡喃-4-基)氨基)异二氢吲哚-5-羧酸60hStep 7 2-(2-(Dimethylamino)ethyl)-1-carbonyl-6-((tetrahydro-2H-pyran-4-yl)amino)isoindoline-5-carboxylic acid 60h
向干燥的反应瓶中加入化合物60g(361mg,1.0mmol)，10ml四氢呋喃，然后加入5ml的氢氧化锂溶液(2M)，加完后80℃下反应2小时。TLC显示原料反应完。减压浓缩反应液除去大部分四氢呋喃，加入盐酸中和反应液，用制备HPLC得60h(300mg,0.86mmol).Add 60 g (361 mg, 1.0 mmol) of compound, 10 ml of tetrahydrofuran, and then add 5 ml of lithium hydroxide solution (2M) to the dry reaction flask, and react at 80° C. for 2 hours after the addition is complete. TLC showed that the reaction of the starting materials was complete. The reaction solution was concentrated under reduced pressure to remove most of the tetrahydrofuran, and hydrochloric acid was added to neutralize the reaction solution. Preparative HPLC was used to obtain 60h (300mg, 0.86mmol).
MS m/z(ESI):348[M+H]+MS m/z(ESI):348[M+H]+
第八步 2-(2-(二甲氨基)乙基)-1-羰基-6-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)异二氢吲哚-5-羧酸60iStep 8 2-(2-(Dimethylamino)ethyl)-1-carbonyl-6-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido )Isoindole-5-carboxylic acid 60i
向反应瓶中加入60h(200mg,0.57mmol)，二氯甲烷和0.5ml三氟乙酸酐，几分钟后反应液由浑浊变澄清，反应完成，减压浓缩反应液，残留物再加入少量水，用高真空泵继续浓缩至干，得固体状的60i(252mg,0.57mmol)。Add 60h (200mg, 0.57mmol), dichloromethane and 0.5ml trifluoroacetic anhydride to the reaction flask. After a few minutes, the reaction solution turns from turbidity to clear, and the reaction is complete. The reaction solution is concentrated under reduced pressure, and a small amount of water is added to the residue. Concentrate to dryness with a high vacuum pump to obtain a solid 60i (252 mg, 0.57 mmol).
MS m/z(ESI):444[M+H]+MS m/z(ESI):444[M+H]+
第九步 N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-2-(2-(二甲氨基)乙基)-1-羰基-6-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰氨基)异二氢吲哚-5-甲酰胺60jStep 9 N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-2-(2-(dimethylamino)ethyl)-1-carbonyl-6- (2,2,2-Trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)isoindole-5-carboxamide 60j
向干燥的反应瓶中加入60i(221mg,0.5mmol),25ml二氯甲烷和1ml草酰氯。室温下搅拌4-6小时，减压浓缩该反应液至干，加入四氢呋喃稀释，冷却至-40℃，氮气保护。然后加入1ml DIPEA，再缓慢加入I-2(130mg,0.5mmol)的THF溶液。加完后自然升温至室温。TLC显示大部分原料反应完。浓缩反应液至适当体积，然后用制备薄层色谱纯化(DCM:MeOH＝10:1)得60j(171mg,0.25mmol,50％).Add 60i (221mg, 0.5mmol), 25ml of dichloromethane and 1ml of oxalyl chloride to the dry reaction flask. Stir at room temperature for 4-6 hours, concentrate the reaction solution under reduced pressure to dryness, add tetrahydrofuran to dilute, and cool to -40°C under nitrogen protection. Then add 1ml DIPEA, and then slowly add the THF solution of I-2 (130mg, 0.5mmol). After the addition, the temperature was naturally raised to room temperature. TLC showed that most of the raw materials had reacted. The reaction solution was concentrated to an appropriate volume, and then purified by preparative thin layer chromatography (DCM:MeOH=10:1) to obtain 60j (171mg, 0.25mmol, 50%).
MS m/z(ESI):685[M+H]+MS m/z(ESI):685[M+H]+
第十步 N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-2-(2-(二甲氨基)乙基)-1-羰基-6-((四氢-2H-吡喃-4-基)氨基)异二氢吲哚-5-甲酰胺60Step 10 N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-2-(2-(dimethylamino)ethyl)-1-carbonyl-6- ((Tetrahydro-2H-pyran-4-yl)amino)isoindole-5-carboxamide 60
向反应瓶中加入13051l(120mg,0.175mmol)和10ml甲醇，溶解后加入0.5ml碳酸钾溶液，10min后TLC显示原料反应完。减压浓缩出去甲醇，加入乙酸乙酯萃取，无水硫酸钠干燥，减压浓缩并用制备薄层色谱纯化(DCM:MeOH＝10:1)的60(70mg,0.12mmol)。13051l (120mg, 0.175mmol) and 10ml methanol were added to the reaction flask, 0.5ml potassium carbonate solution was added after dissolution, and TLC showed that the raw materials had reacted after 10 minutes. Concentrate under reduced pressure to remove methanol, add ethyl acetate for extraction, dry over anhydrous sodium sulfate, concentrate under reduced pressure and purify 60 (70 mg, 0.12 mmol) by preparative thin layer chromatography (DCM:MeOH=10:1).
MS m/z(ESI):589[M+H]+MS m/z(ESI):589[M+H]+
1H NMR(400MHz,Chloroform-d)δ10.86(s,1H),9.85(s,1H),7.77(s,1H),7.63(s,1H),7.57(d,J＝7.3Hz,1H),7.32(d,J＝8.7Hz,1H),7.17(dd,J＝8.6,1.6Hz,1H),7.01(s,1H),6.76–6.67(m,2H),6.65–6.56(m,1H),4.29(s,2H),4.04(s,2H),3.95(m,2H),3.72(t,J＝6.2Hz,2H),3.49(m,3H),2.63(t,J＝6.2Hz,2H),2.33(s,6H),1.98(d,J＝13.3Hz,2H),1.52(m,2H).
1 H NMR (400MHz, Chloroform-d) δ 10.86 (s, 1H), 9.85 (s, 1H), 7.77 (s, 1H), 7.63 (s, 1H), 7.57 (d, J = 7.3 Hz, 1H) ), 7.32(d,J=8.7Hz,1H),7.17(dd,J=8.6,1.6Hz,1H),7.01(s,1H),6.76-6.67(m,2H),6.65-6.56(m, 1H), 4.29 (s, 2H), 4.04 (s, 2H), 3.95 (m, 2H), 3.72 (t, J = 6.2 Hz, 2H), 3.49 (m, 3H), 2.63 (t, J = 6.2 Hz, 2H), 2.33 (s, 6H), 1.98 (d, J = 13.3 Hz, 2H), 1.52 (m, 2H).
实施例61Example 61
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(N-甲基-2-吗啉代乙酰氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺61的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(N-methyl-2-morpholinoacetamido)-2-((tetrahydro -2H-pyran-4-yl)amino)benzamide 61
第一步 N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(甲基氨基)-2-硝基苯酰胺61aThe first step N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(methylamino)-2-nitrobenzamide 61a
化合物11b(500mg，1.17mmol),甲胺盐酸盐(157mg,2.3mmol)和DIPEA(603mg,4.68mmol)溶于DMSO(10mL)。该混合物在130℃下搅拌3小时，冷却，加入水淬灭，用乙酸乙酯萃取，有机相合并，用饱和氯化钠水溶液洗2次，用无水硫酸钠干燥，过滤，浓缩，得到的粗品用柱层析纯化(DCM:MeOH＝15:1)得到61a 400mg呈黄色固体。Compound 11b (500 mg, 1.17 mmol), methylamine hydrochloride (157 mg, 2.3 mmol) and DIPEA (603 mg, 4.68 mmol) were dissolved in DMSO (10 mL). The mixture was stirred at 130°C for 3 hours, cooled, quenched by adding water, extracted with ethyl acetate, the organic phases were combined, washed twice with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain The crude product was purified by column chromatography (DCM:MeOH=15:1) to obtain 61a 400mg as a yellow solid.
MS m/z(ESI):438[M+H]
+
MS m/z(ESI):438[M+H] +
第二步 N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(N-甲基-2-吗啉代乙酰氨基)-2-硝基苯酰胺61bThe second step N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(N-methyl-2-morpholinoacetamido)-2-nitro Benzamide 61b
将化合物61a(150mg,0.34mmol),2-吗啉代乙酸(50mg,0.34mmol),HATU(130mg,0.34mmol)和三乙胺(68mg,0.68mmol)溶于DMF(10mL)中。该混合物在室温下搅拌2小时，加入水淬灭，用乙酸乙酯萃取，有机相合并，用饱和氯化钠水溶液洗2次，用无水硫酸钠干燥，过滤，浓缩，得到的粗品用柱层析纯化(DCM:MeOH＝10:1)得到61b(120mg)呈黄色固体。Compound 61a (150 mg, 0.34 mmol), 2-morpholinoacetic acid (50 mg, 0.34 mmol), HATU (130 mg, 0.34 mmol) and triethylamine (68 mg, 0.68 mmol) were dissolved in DMF (10 mL). The mixture was stirred at room temperature for 2 hours, quenched by adding water, extracted with ethyl acetate, the organic phases were combined, washed twice with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product obtained was columnar Purification by chromatography (DCM:MeOH=10:1) gave 61b (120mg) as a yellow solid.
MS m/z(ESI):565[M+H]
+。
MS m/z (ESI): 565 [M+H] + .
第三步 2-氨基-N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(N-甲基-2-吗啉代乙酰氨基)苯酰胺61cThe third step 2-amino-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(N-methyl-2-morpholinoacetamido) Benzamide 61c
将61b(120mg，0.21mmol)N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(N-甲基-2-吗啉代乙酰氨基)-2-硝基苯酰胺分散在乙酸乙酯(10mL)中，加入10％Pd/C(15mg),该混合物在氢气条件下室温搅拌2小时，过滤，浓缩，粗品61c直接用于下一步61b (120mg, 0.21mmol) N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(N-methyl-2-morpholinoacetamido )-2-Nitrobenzamide was dispersed in ethyl acetate (10mL), 10% Pd/C (15mg) was added, the mixture was stirred at room temperature under hydrogen for 2 hours, filtered, concentrated, and the crude 61c was used directly in the next step
MS m/z(ESI):535[M+H]
+。
MS m/z (ESI): 535 [M+H] + .
第四步 N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(N-甲基-2-吗啉代乙酰氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺61The fourth step N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(N-methyl-2-morpholinoacetamido)-2-( (Tetrahydro-2H-pyran-4-yl)amino)benzamide 61
将四甲基三乙酰氧基硼氢化铵(86mg,0.73mmol)慢慢加入到化合物61c(60mg,0.11mmol),四氢-4H-吡喃-4-酮(16mg,0.16mmol)和三氟乙酸(1mL)的DCM(3mL)溶液中，该混合物在室温继续搅拌2小时。加入饱和NaHCO
3水溶液淬灭反应，DCM萃取，有机相合并，用饱和氯化钠水溶液洗2次，用无水硫酸钠干燥，过滤，浓缩，得到的粗品用柱层析纯化得到61(4mg)呈白色固体。
Tetramethyltriacetoxyammonium borohydride (86mg, 0.73mmol) was slowly added to compound 61c (60mg, 0.11mmol), tetrahydro-4H-pyran-4-one (16mg, 0.16mmol) and trifluoro In a solution of acetic acid (1 mL) in DCM (3 mL), the mixture was continuously stirred at room temperature for 2 hours. The reaction was quenched by adding saturated aqueous NaHCO 3 solution, extracted with DCM, and the organic phases were combined, washed twice with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The obtained crude product was purified by column chromatography to obtain 61 (4mg) It is a white solid.
MS m/z(ESI):619[M+H]
+。
MS m/z (ESI): 619 [M+H] + .
1H NMR(400MHz,Chloroform-d)δ8.34(d,J＝8.7Hz,1H),8.17(d,J＝7.4Hz,1H),8.12(s,1H),7.81(s,1H),7.44(d,J＝8.8Hz,1H),7.35(d,J＝8.7Hz,1H), 6.65(dd,J＝33.6,8.1Hz,3H),5.92(d,J＝8.8Hz,1H),5.79(s,1H),4.09–3.91(m,6H),3.81(s,4H),3.62-3.50(m,3H),2.89(s,3H),2.74(s,4H),2.03(d,J＝13.1Hz,2H),1.62-1.51(m,2H).
1 H NMR(400MHz,Chloroform-d)δ8.34(d,J=8.7Hz,1H), 8.17(d,J=7.4Hz,1H), 8.12(s,1H), 7.81(s,1H), 7.44(d,J=8.8Hz,1H), 7.35(d,J=8.7Hz,1H), 6.65(dd,J=33.6,8.1Hz,3H), 5.92(d,J=8.8Hz,1H), 5.79(s,1H),4.09--3.91(m,6H),3.81(s,4H),3.62-3.50(m,3H),2.89(s,3H),2.74(s,4H),2.03(d, J = 13.1Hz, 2H), 1.62-1.51 (m, 2H).
实施例62Example 62
N-(5-(1-(3,5-二氟苯基)-2,2,2-三氟-乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺62的制备N-(5-(1-(3,5-Difluorophenyl)-2,2,2-trifluoro-ethyl)-1H-indazol-3-yl)-4-(4-methylpiper Preparation of azin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 62
第一步 1-(3,5-二氟苯基)-2,2,2-三氟乙基-1-醇62bThe first step 1-(3,5-difluorophenyl)-2,2,2-trifluoroethyl-1-ol 62b
化合物62a(5.0g，35.2mmol)的DMF(50mL)溶液，加入CF
3-TMS(6.0g，42.3mmol)，和碳酸钾(9.7g，70.4mmol)，反应液在常温下继续搅拌过夜，将反应液加水后，用乙酸乙酯(200mL)萃取，有机相用饱和食盐水洗涤三次，干燥后旋干得到的粗品经过柱层析纯化后得到化合物62b的(7.0g，33.0mmol)。
Compound 62a (5.0g, 35.2mmol) in DMF (50mL) solution was added CF 3 -TMS (6.0g, 42.3mmol), and potassium carbonate (9.7g, 70.4mmol), the reaction solution was stirred overnight at room temperature, the After adding water to the reaction solution, it was extracted with ethyl acetate (200 mL), the organic phase was washed three times with saturated brine, dried and then spin-dried to obtain the crude product after purification by column chromatography to obtain compound 62b (7.0 g, 33.0 mmol).
第二步 1-(3,5-二氟苯基)-2,2,2-三氟乙基4-甲基苯磺酸盐62cThe second step 1-(3,5-difluorophenyl)-2,2,2-trifluoroethyl 4-methylbenzenesulfonate 62c
化合物62b(7g，33.0mmol)溶于四氢呋喃(100mL)中，于冰浴下加入NaH(1.6g，39.6mmol),搅拌0.5h，缓慢加入Tos-Cl(8.1g，39.6mmol)。反应液在常 温下搅拌2小时，反应液倒入冰水中，用乙酸乙酯(200mL)萃取，有机相用饱和碳酸氢钠溶液和食盐水洗涤，干燥后用无水硫酸钠干燥，过滤旋干得到初产品，经过硅胶柱层析纯化后的到化合物62c(7.0g，19.1mmol)。Compound 62b (7g, 33.0mmol) was dissolved in tetrahydrofuran (100mL), NaH (1.6g, 39.6mmol) was added under ice bath, stirred for 0.5h, and Tos-Cl (8.1g, 39.6mmol) was slowly added. The reaction solution was stirred at room temperature for 2 hours. The reaction solution was poured into ice water and extracted with ethyl acetate (200 mL). The organic phase was washed with saturated sodium bicarbonate solution and brine, dried, dried over anhydrous sodium sulfate, filtered and spin-dried to obtain The initial product was purified by silica gel column chromatography to obtain compound 62c (7.0 g, 19.1 mmol).
第三步 5-(1-(3,5-二氟苯基)-2,2,2-三氟乙基)-2-氟苄腈62eThe third step 5-(1-(3,5-difluorophenyl)-2,2,2-trifluoroethyl)-2-fluorobenzonitrile 62e
将化合物62c(1g,2.73mmol)溶解在1,4-二氧六环(15mL)中，加入化合物62d(900mg，5.46mmol)，Xphos-Pd-G2(110mg，0.14mmol)和磷酸钾(926mg，4.37mmol)，反应液在100℃微波下反应6小时，N
2保护。冷却后反应液倒入水(50mL)中，用乙酸乙酯(50mL)萃取，有机相用饱和食盐水洗涤，干燥旋干得到粗品，经过柱层析纯化后得到化合物62e(200mg,0.63mmol)。
Compound 62c (1g, 2.73mmol) was dissolved in 1,4-dioxane (15mL), compound 62d (900mg, 5.46mmol), Xphos-Pd-G2 (110mg, 0.14mmol) and potassium phosphate (926mg) were added , 4.37mmol), the reaction solution was reacted at 100°C in a microwave for 6 hours, protected by N 2. After cooling, the reaction solution was poured into water (50mL), extracted with ethyl acetate (50mL), the organic phase was washed with saturated brine, dried and spin-dried to obtain a crude product, which was purified by column chromatography to obtain compound 62e (200mg, 0.63mmol) .
第四步 5-(1-(3,5-二氟苯基)-2,2,2-三氟-乙基)-1H-吲唑-3-胺62fThe fourth step 5-(1-(3,5-difluorophenyl)-2,2,2-trifluoro-ethyl)-1H-indazol-3-amine 62f
化合物62e(400mg，1.26mmol)和水合肼(313mg,6.3mmol)混合在N-甲基吡咯烷酮(6mL)中，反应液在微波反应器上120℃反应1小时，冷却后加入到(30mL)中，用乙酸乙酯(50mL)萃取，有机相干燥后旋干得到粗品，经过硅胶柱层析纯化后得到化合物62f(250mg，0.76mmol)。Compound 62e (400mg, 1.26mmol) and hydrazine hydrate (313mg, 6.3mmol) were mixed in N-methylpyrrolidone (6mL), the reaction solution was reacted on a microwave reactor at 120°C for 1 hour, and then added to (30mL) after cooling After extraction with ethyl acetate (50 mL), the organic phase was dried and then spin-dried to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 62f (250 mg, 0.76 mmol).
MS m/z(ESI):328[M+H]
+。
MS m/z (ESI): 328 [M+H] + .
第五步 N-(5-(1-(3,5-二氟苯基)-2,2,2-三氟-乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-(2-，2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰胺基)苯甲酰胺62gStep 5 N-(5-(1-(3,5-Difluorophenyl)-2,2,2-trifluoro-ethyl)-1H-indazol-3-yl)-4-(4- Methylpiperazin-1-yl)-2-(2-,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 62g
化合物62f(500mg,1.2mmol)悬浮在二氯甲烷(30mL)中，冷却至0℃，向溶液中滴加草酰氯(254mg,2.0mmol)，反应液在常温下搅拌过夜，旋干再加入二氯甲烷(10mL)后再旋干，将产物溶解在四氢呋喃(5mL)中，冷却至-20℃，将化合物1f(250mg,0.76mmol)和DIPEA(196mg,1.52mmol)的四氢呋喃(15mL)溶液滴加到上述冷却液中，反应在-20℃搅拌2小时，然后在常温下继续搅拌2小时，反应液用二氯甲烷(50mL)稀释，用拌合食盐水洗涤，干燥旋干得到粗品，经过柱层析(DCM/MeOH＝10:1)纯化后得到化合物62g(50mg,0.069mmol)。Compound 62f (500mg, 1.2mmol) was suspended in dichloromethane (30mL), cooled to 0°C, oxalyl chloride (254mg, 2.0mmol) was added dropwise to the solution, and the reaction solution was stirred overnight at room temperature, then spin-dried and then added to the solution. After chloromethane (10mL), spin dry, dissolve the product in tetrahydrofuran (5mL), cool to -20°C, drop compound 1f (250mg, 0.76mmol) and DIPEA (196mg, 1.52mmol) in tetrahydrofuran (15mL) solution Add to the above cooling liquid, the reaction was stirred at -20 ℃ for 2 hours, and then continue to stir at room temperature for 2 hours, the reaction solution was diluted with dichloromethane (50 mL), washed with mixed brine, dried and spin-dried to obtain the crude product. After purification by column chromatography (DCM/MeOH=10:1), compound 62g (50mg, 0.069mmol) was obtained.
MS m/z(ESI):725[M+H]
+。
MS m/z (ESI): 725 [M+H] + .
第六步 N-(5-(1-(3,5-二氟苯基)-2,2,2-三氟-乙基)-1H-吲唑-3-基)-4-(4-甲基 哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺62Step 6 N-(5-(1-(3,5-Difluorophenyl)-2,2,2-trifluoro-ethyl)-1H-indazol-3-yl)-4-(4- Methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 62
化合物62g(50mg,0.069mmol)溶于甲醇(10mL)和水(5mL)的混合溶剂中，加入碳酸钾(90.7mg,0.66mmol)，反应液在常温下搅拌3小时，加入水(20ml)，用二氯甲烷(50ml)萃取。有机相用无水硫酸钠干燥，过滤浓缩得到粗品，经过硅胶柱层析纯化后得到化合物62(10mg,0.016mmol)。Compound 62g (50mg, 0.069mmol) was dissolved in a mixed solvent of methanol (10mL) and water (5mL), potassium carbonate (90.7mg, 0.66mmol) was added, the reaction solution was stirred at room temperature for 3 hours, and water (20ml) was added. Extract with dichloromethane (50ml). The organic phase was dried with anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 62 (10 mg, 0.016 mmol).
MS m/z(ESI):629[M+H]+。MS m/z(ESI):629[M+H]+.
1H NMR(400MHz,DMSO-d
6)δ12.78(s,1H),10.17(s,1H),8.28(d,J＝7.9Hz,1H),7.76–7.74(m,1H),7.48–7.45(m,1H),7.41–7.37(m,1H),7.27–7.11(m,4H),6.23–6.19(m,1H),6.12–6.08(m,1H),5.50–5.44(m,1H),3.81–3.73(m,2H),3.69–3.63(m,1H),3.50–3.42(m,2H),3.25–3.20(m,4H),2.42–2.36(m,4H),2.19(s,3H),1.93–1.87(m,2H),1.32–1.26(m,2H).
1 H NMR(400MHz,DMSO-d 6 )δ12.78(s,1H), 10.17(s,1H), 8.28(d,J=7.9Hz,1H), 7.76–7.74(m,1H), 7.48– 7.45(m,1H),7.41--7.37(m,1H),7.27-7.11(m,4H),6.23--6.19(m,1H),6.12-6.08(m,1H),5.50-5.44(m,1H) ), 3.81–3.73(m,2H), 3.69–3.63(m,1H), 3.50–3.42(m,2H), 3.25–3.20(m,4H), 2.42–2.36(m,4H), 2.19(s ,3H),1.93-1.87(m,2H),1.32-1.26(m,2H).
实施例63Example 63
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(乙基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺63的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2 -((Tetrahydro-2H-pyran-4-yl)amino)benzamide 63
采用与实施例43相同的制备方法，以3,5-二氟溴苯为原料，可得到化合物63。Using the same preparation method as in Example 43, using 3,5-difluorobromobenzene as a raw material, compound 63 can be obtained.
MS m/z(ESI):603[M+H]+。MS m/z(ESI): 603[M+H]+.
1H NMR(400MHz,Methanol-d
4)δ7.70(d,J＝9.0Hz,1H),7.57(d,J＝1.4Hz,1H),7.39(dd,J＝8.7,0.8Hz,1H),7.25(dd,J＝8.5,1.6Hz,1H),7.04(td,J＝9.1,4.6Hz,1H),6.98–6.86(m,2H),6.12(dd,J＝9.1,2.4Hz,1H),5.95(d,J＝2.4Hz,1H),4.06(s,2H),3.93(dt,J＝11.9,4.1Hz,2H),3.67(dd,J＝9.0,4.7Hz,1H),3.62–3.51(m,4H),3.45(q,J＝7.0Hz,2H),2.86–2.69(m,6H),2.03(d,J＝13.5Hz, 2H),1.87(t,J＝5.4Hz,4H),1.60–1.45(m,3H),1.19(t,J＝7.0Hz,3H).
1 H NMR(400MHz,Methanol-d 4 )δ7.70(d,J＝9.0Hz,1H), 7.57(d,J＝1.4Hz,1H), 7.39(dd,J＝8.7,0.8Hz,1H) ,7.25(dd,J=8.5,1.6Hz,1H),7.04(td,J=9.1,4.6Hz,1H),6.98–6.86(m,2H),6.12(dd,J=9.1,2.4Hz,1H) ), 5.95 (d, J = 2.4 Hz, 1H), 4.06 (s, 2H), 3.93 (dt, J = 11.9, 4.1 Hz, 2H), 3.67 (dd, J = 9.0, 4.7 Hz, 1H), 3.62 –3.51(m,4H), 3.45(q,J=7.0Hz,2H), 2.86–2.69(m,6H), 2.03(d,J=13.5Hz, 2H), 1.87(t,J=5.4Hz, 4H), 1.60-1.45 (m, 3H), 1.19 (t, J = 7.0Hz, 3H).
实施例64Example 64
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(乙基(2-(3-氟吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺64的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(3-fluoropyrrolidin-1-yl)ethyl)amino) Preparation of -2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 64
第一步 4-(乙基(2-羟乙基)氨基)-2-硝基苯甲酸叔丁酯64bThe first step 4-(ethyl(2-hydroxyethyl)amino)-2-nitrobenzoic acid tert-butyl ester 64b
将化合物I-1b(3.0g，12.4mmol，1.0eq)和2-(乙基氨基)乙烷-1-醇(64a)(3.3g，37.2mmol，3.0eq)投于NMP(10ml)混合溶剂中，加入DIPEA(8.6g，49.6mmol，4.0eq),105℃反应8h。Compound I-1b (3.0g, 12.4mmol, 1.0eq) and 2-(ethylamino)ethane-1-ol (64a) (3.3g, 37.2mmol, 3.0eq) were put into NMP (10ml) mixed solvent DIPEA (8.6g, 49.6mmol, 4.0eq) was added to the mixture and reacted at 105°C for 8h.
冷至室温，加入乙酸乙酯(80ml)和H
2O(20ml)萃取，有机层用brine(20ml)洗涤，经无水硫酸钠干燥，浓缩，flash柱纯化得黄色油状物64b(4.4g).
Cool to room temperature, add ethyl acetate (80ml) and H 2 O (20ml) for extraction, the organic layer was washed with brine (20ml), dried over anhydrous sodium sulfate, concentrated, and purified by flash column to obtain a yellow oil 64b (4.4g) .
MS m/z(ESI):311[M+1]
+
MS m/z(ESI):311[M+1] +
第二步 4-(乙基(2-氧乙基)氨基)-2-硝基苯甲酸叔丁酯64cThe second step 4-(ethyl(2-oxyethyl)amino)-2-nitrobenzoic acid tert-butyl ester 64c
将化合物64b(3.4g，11.0mmol，1.0eq)溶于DCM(80ml)中，加入dess-martin(5.6g,13.2mmol，1.2eq)，室温反应3h。加入碳酸氢钠和硫代硫酸钠水溶液淬灭，分出有机层，经无水硫酸钠干燥，浓缩后得64c直接用于下步反应。Compound 64b (3.4g, 11.0mmol, 1.0eq) was dissolved in DCM (80ml), dess-martin (5.6g, 13.2mmol, 1.2eq) was added and reacted at room temperature for 3h. Add sodium bicarbonate and sodium thiosulfate aqueous solution for quenching, separate the organic layer, dry over anhydrous sodium sulfate, and concentrate to obtain 64c directly for the next reaction.
MS m/z(ESI):309[M+1]
+
MS m/z(ESI): 309[M+1] +
第三步 4-(2-(3-(3-氟吡咯烷-1-基)乙基)氨基)-2-硝基苯甲酸叔丁酯64dThe third step 4-(2-(3-(3-fluoropyrrolidin-1-yl)ethyl)amino)-2-nitrobenzoic acid tert-butyl ester 64d
将3-氟吡咯烷盐酸盐(1.1g，8.7mmol，1.5eq)投于于DCM(50ml)中，依次加入DIPEA(1.5ml，8.7mmol，1.5eq)和化合物64c(1.8g，5.8mmol，1.0eq)室温反应30min后，加入醋酸硼氢化钠(3.7g，17.4mmol，3.0eq)，继续反应1h。Put 3-fluoropyrrolidine hydrochloride (1.1g, 8.7mmol, 1.5eq) in DCM (50ml), add DIPEA (1.5ml, 8.7mmol, 1.5eq) and compound 64c (1.8g, 5.8mmol) , 1.0 eq) After reacting at room temperature for 30 min, sodium acetate borohydride (3.7 g, 17.4 mmol, 3.0 eq) was added, and the reaction was continued for 1 h.
加入碳酸氢钠水溶液中和后，DCM(50ml×2)萃取，合并有机层，经无水硫酸钠干燥，浓缩，flash柱纯化得黄色油状物64d(1.7g)。After adding sodium bicarbonate aqueous solution for neutralization, extraction with DCM (50ml×2), the organic layers were combined, dried over anhydrous sodium sulfate, concentrated, and purified by flash column to obtain a yellow oil 64d (1.7g).
MS m/z(ESI):382[M+1]
+
MS m/z(ESI): 382[M+1] +
第四步 N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(乙基(2-(3-氟吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺64The fourth step N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(3-fluoropyrrolidin-1-yl)ethyl )Amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 64
采用与实施例43相同的制备方法，以64d为原料替代43a，可得到化合物63。Using the same preparation method as in Example 43, using 64d as a raw material instead of 43a, compound 63 can be obtained.
MS m/z(ESI):621[M+1]
+
MS m/z(ESI):621[M+1] +
1H NMR(400MHz,DMSO-d
6)δ12.62(s,1H),9.97(s,1H),8.35(d,J＝7.3Hz,1H),7.77(d,J＝9.1Hz,1H),7.47(s,1H),7.43–7.36(m,1H),7.25(dd,J＝8.6,1.6Hz,1H),7.06–6.92(m,3H),5.99(dd,J＝9.1,2.3Hz,1H),5.83(d,J＝2.3Hz,1H),5.21(dt,J＝56.1,5.6Hz,1H),4.03(s,2H),3.83(dt,J＝11.7,3.9Hz,2H),3.65–3.53(m,1H),3.51–3.36(m,5H),2.99–2.82(m,2H),2.74–2.55(m,3H),2.36(q,J＝7.9Hz,1H),2.14(ddq,J＝27.6,14.1,6.8Hz,1H),2.01–1.78(m,3H),1.45–1.28(m,2H),1.12(t,J＝6.9Hz,3H).
1 H NMR(400MHz,DMSO-d 6 )δ12.62(s,1H),9.97(s,1H),8.35(d,J=7.3Hz,1H),7.77(d,J=9.1Hz,1H) ,7.47(s,1H),7.43-7.36(m,1H),7.25(dd,J=8.6,1.6Hz,1H),7.06-6.92(m,3H),5.99(dd,J=9.1,2.3Hz ,1H),5.83(d,J=2.3Hz,1H),5.21(dt,J=56.1,5.6Hz,1H),4.03(s,2H),3.83(dt,J=11.7,3.9Hz,2H) ,3.65–3.53(m,1H),3.51–3.36(m,5H),2.99–2.82(m,2H),2.74–2.55(m,3H),2.36(q,J=7.9Hz,1H),2.14 (ddq,J=27.6,14.1,6.8Hz,1H),2.01–1.78(m,3H),1.45–1.28(m,2H),1.12(t,J=6.9Hz,3H).
实施例65Example 65
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((2-(吡咯烷-1-基)乙基)硫代)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺65的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((2-(pyrrolidin-1-yl)ethyl)thio)-2- Preparation of ((Tetrahydro-2H-pyran-4-yl)amino)benzamide 65
采用与实施例44相同的制备方法，以2-(1-吡咯烷基)乙硫醇为原料，可得到化合物65。Using the same preparation method as in Example 44 and using 2-(1-pyrrolidinyl)ethanethiol as a raw material, compound 65 can be obtained.
MS m/z(ESI):592(M+H)
+
MS m/z(ESI): 592(M+H) +
1H NMR(400MHz,Methanol-d4)δ7.74(d,J＝8.3Hz,1H),7.57(dd,J＝1.6,0.8Hz,1H),7.41(dd,J＝8.6,0.8Hz,1H),7.24(dd,J＝8.6,1.6Hz,1H),6.84–6.78(m,2H),6.75–6.67(m,2H),6.61(dd,J＝8.3,1.8Hz,1H),4.06(s,2H),3.92(dt,J＝11.8,4.0Hz,2H),3.69(td,J＝9.4,4.6Hz,1H),3.56(ddd,J＝11.9,10.2,2.5Hz,2H),3.20–3.13(m,2H),2.86–2.79(m,2H),2.72–2.64(m,4H),2.05–1.96(m,2H),1.88–1.80(m,4H),1.50(dtd,J＝13.7,10.0,4.2Hz,2H).
1 H NMR(400MHz,Methanol-d4)δ7.74(d,J＝8.3Hz,1H), 7.57(dd,J＝1.6,0.8Hz,1H),7.41(dd,J＝8.6,0.8Hz,1H ), 7.24(dd,J=8.6,1.6Hz,1H), 6.84–6.78(m,2H), 6.75–6.67(m,2H), 6.61(dd,J=8.3,1.8Hz,1H),4.06( s, 2H), 3.92 (dt, J = 11.8, 4.0 Hz, 2H), 3.69 (td, J = 9.4, 4.6 Hz, 1H), 3.56 (ddd, J = 11.9, 10.2, 2.5 Hz, 2H), 3.20 –3.13(m,2H),2.86–2.79(m,2H),2.72–2.64(m,4H),2.05–1.96(m,2H),1.88–1.80(m,4H),1.50(dtd,J= 13.7, 10.0, 4.2Hz, 2H).
实施例66Example 66
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-((2-(3,3-二氟吡咯烷-1-基)乙基)(乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺66的制备N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((2-(3,3-difluoropyrrolidin-1-yl)ethyl)( (Ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 66
采用与实施例64相同的制备方法，以3,3-二氟吡咯烷盐酸盐为原料，可得到化合物66。Using the same preparation method as in Example 64, using 3,3-difluoropyrrolidine hydrochloride as a raw material, compound 66 can be obtained.
MS m/z(ESI):639[M+1]
+
MS m/z(ESI):639[M+1] +
1H NMR(400MHz,DMSO-d
6)δ12.59(s,1H),9.95(s,1H),8.33(d,J＝7.3Hz,1H),7.74(d,J＝9.1Hz,1H),7.44(d,J＝1.5Hz,1H),7.37(dd,J＝8.6,0.8Hz,1H),7.22(dd,J＝8.6,1.6Hz,1H),7.03–6.89(m,3H),5.96(dd,J＝9.0,2.3Hz,1H),5.79(d,J＝2.4Hz,1H),4.00(s,2H),3.80(dt,J＝11.6,4.0Hz,2H),3.63–3.51(m,1H),3.41(ddt,J＝15.6,10.2,5.1Hz,6H),2.94(t,J＝13.4Hz,2H),2.74(t,J＝6.9Hz,2H),2.59(t,J＝7.3Hz,2H),2.21(tq,J＝13.8,6.8Hz,2H),1.97–1.87(m,2H),1.34(tdd,J＝13.5,9.7,3.9Hz,2H),1.08(t,J＝6.9Hz,3H).
1 H NMR(400MHz,DMSO-d 6 )δ12.59(s,1H),9.95(s,1H),8.33(d,J=7.3Hz,1H),7.74(d,J=9.1Hz,1H) ,7.44(d,J=1.5Hz,1H),7.37(dd,J=8.6,0.8Hz,1H),7.22(dd,J=8.6,1.6Hz,1H),7.03-6.89(m,3H), 5.96 (dd, J = 9.0, 2.3 Hz, 1H), 5.79 (d, J = 2.4 Hz, 1H), 4.00 (s, 2H), 3.80 (dt, J = 11.6, 4.0 Hz, 2H), 3.63-3.51 (m, 1H), 3.41 (ddt, J = 15.6, 10.2, 5.1 Hz, 6H), 2.94 (t, J = 13.4 Hz, 2H), 2.74 (t, J = 6.9 Hz, 2H), 2.59 (t, J = 7.3 Hz, 2H), 2.21 (tq, J = 13.8, 6.8 Hz, 2H), 1.97–1.87 (m, 2H), 1.34 (tdd, J = 13.5, 9.7, 3.9 Hz, 2H), 1.08 (t ,J=6.9Hz,3H).
实施例67Example 67
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-((2-(3,4-二氟吡咯烷-1-基)乙基)(乙基) 氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺67的制备N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((2-(3,4-difluoropyrrolidin-1-yl)ethyl)( (Ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 67
采用与实施例64相同的制备方法，以3,4-二氟吡咯烷盐酸盐为原料，可得到化合物67。Using the same preparation method as in Example 64, using 3,4-difluoropyrrolidine hydrochloride as a raw material, compound 67 can be obtained.
MS m/z(ESI):639[M+1]
+
MS m/z(ESI):639[M+1] +
1H NMR(400MHz,DMSO-d
6)δ12.61(s,1H),9.97(s,1H),8.36(d,J＝7.3Hz,1H),7.77(d,J＝9.1Hz,1H),7.47(s,1H),7.40(dd,J＝8.5,0.8Hz,1H),7.25(dd,J＝8.6,1.6Hz,1H),7.05–6.92(m,3H),5.99(dd,J＝9.1,2.3Hz,1H),5.83(d,J＝2.4Hz,1H),5.27–5.03(m,2H),4.04(s,2H),3.83(dt,J＝11.6,4.0Hz,2H),3.67–3.54(m,1H),3.52–3.35(m,6H),3.07–2.82(m,4H),2.66(t,J＝7.2Hz,2H),2.01–1.90(m,2H),1.45–1.31(m,2H),1.11(t,J＝6.9Hz,3H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.61 (s, 1H), 9.97 (s, 1H), 8.36 (d, J = 7.3 Hz, 1H), 7.77 (d, J = 9.1 Hz, 1H) ,7.47(s,1H),7.40(dd,J=8.5,0.8Hz,1H),7.25(dd,J=8.6,1.6Hz,1H),7.05-6.92(m,3H),5.99(dd,J ＝9.1,2.3Hz,1H),5.83(d,J＝2.4Hz,1H), 5.27–5.03(m,2H),4.04(s,2H), 3.83(dt,J＝11.6,4.0Hz,2H) ,3.67–3.54(m,1H),3.52–3.35(m,6H),3.07–2.82(m,4H),2.66(t,J=7.2Hz,2H),2.01–1.90(m,2H),1.45 –1.31(m,2H),1.11(t,J=6.9Hz,3H).
实施例68Example 68
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(3,4-二甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺68的制备N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(3,4-dimethylpiperazine-1 -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 68
采用与实施例50相同的制备方法，以1,2-二甲基哌嗪为原料，可得到化合物68。Using the same preparation method as in Example 50 and using 1,2-dimethylpiperazine as a raw material, compound 68 can be obtained.
MS m/z(ESI):607[M+H]
+。
MS m/z (ESI): 607 [M+H] + .
1H NMR(400MHz,DMSO-d
6)δ12.67(s,1H),10.08(s,1H),8.30(d,J＝7.7Hz,1H),7.76(d,J＝9.1Hz,1H),7.50(d,J＝1.6Hz,1H),7.45–7.35(m,2H),7.30(dd,J＝8.7,1.7Hz,1H),7.20(td,J＝9.3,4.7Hz,1H),7.15–7.06(m,1H),6.22(dd,J＝9.1,2.3Hz,1H),6.09(d,J＝2.3Hz,1H),5.10(t,J＝8.7Hz,0H),5.03–4.93(m,1H),4.87(dd,J＝9.3,6.4Hz,0H),4.73(dt,J＝14.8,7.2Hz,1H),3.84–3.73(m,2H),3.69(d,J＝13.8Hz,3H),3.47(ddt,J＝11.8,9.8,2.3Hz,2H),2.82(s,2H),2.22(s,4H),1.90(d,J＝13.4Hz,2H),1.40–1.25(m,2H),1.05(d,J＝6.1Hz,3H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.67 (s, 1H), 10.08 (s, 1H), 8.30 (d, J = 7.7 Hz, 1H), 7.76 (d, J = 9.1 Hz, 1H) ,7.50(d,J=1.6Hz,1H),7.45-7.35(m,2H),7.30(dd,J=8.7,1.7Hz,1H), 7.20(td,J=9.3,4.7Hz,1H), 7.15–7.06(m,1H), 6.22(dd,J=9.1,2.3Hz,1H), 6.09(d,J=2.3Hz,1H), 5.10(t,J=8.7Hz,0H), 5.03-4.93 (m, 1H), 4.87 (dd, J = 9.3, 6.4 Hz, 0H), 4.73 (dt, J = 14.8, 7.2 Hz, 1H), 3.84–3.73 (m, 2H), 3.69 (d, J = 13.8 Hz, 3H), 3.47 (ddt, J = 11.8, 9.8, 2.3 Hz, 2H), 2.82 (s, 2H), 2.22 (s, 4H), 1.90 (d, J = 13.4 Hz, 2H), 1.40-1.25 (m,2H),1.05(d,J=6.1Hz,3H).
实施例69Example 69
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(3-甲基-4-乙基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺69的制备N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(3-methyl-4-ethylpiperazine -1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 69
采用与实施例50相同的制备方法，以1-乙基-2-甲基哌嗪为原料，可得到化合物69。Using the same preparation method as in Example 50 and using 1-ethyl-2-methylpiperazine as a raw material, compound 69 can be obtained.
MS m/z(ESI):621[M+H]
+。
MS m/z (ESI): 621 [M+H] + .
1H NMR(400MHz,DMSO-d
6)δ12.67(s,1H),10.07(s,1H),8.29(d,J＝7.7Hz,1H),7.76(d,J＝9.1Hz,1H),7.50(d,J＝1.5Hz,1H),7.45–7.34(m,2H),7.30(dd,J＝8.7,1.6Hz,1H),7.20(td,J＝9.3,4.7Hz,1H),7.15–7.07(m,1H),6.21(dd,J＝9.1,2.3Hz,1H),6.08(d,J＝2.4Hz,1H),5.10(t,J＝8.7Hz,1H),4.98(dd,J＝9.1,6.2Hz,1H),4.87(dd,J＝9.3,6.4Hz,1H),4.73(dt,J＝14.8,7.2Hz,1H),3.85–3.73(m,2H),3.72–3.53(m,3H),3.47(ddt,J＝11.8,9.7,2.3Hz,2H),2.95–2.68(m,3H),2.57(t,J＝10.5Hz,1H),2.43(s,1H),2.29(d,J＝12.2Hz,2H),2.00–1.79(m,2H),1.39–1.25(m,2H),1.11–0.87(m,6H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.67 (s, 1H), 10.07 (s, 1H), 8.29 (d, J = 7.7 Hz, 1H), 7.76 (d, J = 9.1 Hz, 1H) ,7.50(d,J=1.5Hz,1H),7.45-7.34(m,2H),7.30(dd,J=8.7,1.6Hz,1H), 7.20(td,J=9.3,4.7Hz,1H), 7.15–7.07(m,1H), 6.21(dd,J=9.1,2.3Hz,1H), 6.08(d,J=2.4Hz,1H), 5.10(t,J=8.7Hz,1H), 4.98(dd ,J=9.1,6.2Hz,1H), 4.87(dd,J=9.3,6.4Hz,1H), 4.73(dt,J=14.8,7.2Hz,1H), 3.85–3.73(m,2H), 3.72– 3.53 (m, 3H), 3.47 (ddt, J = 11.8, 9.7, 2.3 Hz, 2H), 2.95-2.68 (m, 3H), 2.57 (t, J = 10.5 Hz, 1H), 2.43 (s, 1H) , 2.29 (d, J = 12.2 Hz, 2H), 2.00-1.79 (m, 2H), 1.39-1.25 (m, 2H), 1.11-0.87 (m, 6H).
实施例70Example 70
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(2,4-二甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺70的制备N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(2,4-dimethylpiperazine-1 -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 70
采用与实施例50相同的制备方法，以1,3-二甲基哌嗪为原料，可得到化合物69。Using the same preparation method as in Example 50, using 1,3-dimethylpiperazine as a raw material, compound 69 can be obtained.
MS m/z(ESI):607[M+H]
+。
MS m/z (ESI): 607 [M+H] + .
1H NMR(400MHz,DMSO-d
6)δ12.68(s,1H),10.07(s,1H),8.32(d,J＝7.6Hz,1H),7.79(d,J＝9.1Hz,1H),7.54(d,J＝1.5Hz,1H),7.46–7.37(m,2H),7.32(dd,J＝8.7,1.7Hz,1H),7.23(td,J＝9.3,4.7Hz,1H),7.18–7.09(m,1H),6.20(dd,J＝9.2,2.3Hz,1H),6.06(d,J＝2.3Hz,1H),5.13(t,J＝8.7Hz,0H),5.01(dd,J＝9.4,6.9Hz,1H),4.90(dd,J＝9.3,6.4Hz,1H),4.76(dt,J＝14.7,7.2Hz,1H),4.15(s,1H),3.81(dd,J＝11.6,3.5Hz,2H),3.66(dd,J＝11.5,6.0Hz,1H),3.50(t,J＝11.2Hz,3H),3.00(td,J＝12.0,3.5Hz,1H),2.84(d,J＝10.9Hz,1H),2.69(d,J＝11.1Hz,1H),2.20(s,4H),2.07–1.84(m,1H),1.34(ddt,J＝12.1,8.8,4.7Hz,2H),1.10(d,J＝6.5Hz,3H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.68 (s, 1H), 10.07 (s, 1H), 8.32 (d, J = 7.6 Hz, 1H), 7.79 (d, J = 9.1 Hz, 1H) ,7.54(d,J=1.5Hz,1H),7.46-7.37(m,2H),7.32(dd,J=8.7,1.7Hz,1H),7.23(td,J=9.3,4.7Hz,1H), 7.18–7.09(m,1H), 6.20(dd,J=9.2,2.3Hz,1H), 6.06(d,J=2.3Hz,1H), 5.13(t,J=8.7Hz,0H),5.01(dd ,J=9.4,6.9Hz,1H), 4.90(dd,J=9.3,6.4Hz,1H), 4.76(dt,J=14.7,7.2Hz,1H), 4.15(s,1H), 3.81(dd, J = 11.6, 3.5 Hz, 2H), 3.66 (dd, J = 11.5, 6.0 Hz, 1H), 3.50 (t, J = 11.2 Hz, 3H), 3.00 (td, J = 12.0, 3.5 Hz, 1H), 2.84 (d, J = 10.9 Hz, 1H), 2.69 (d, J = 11.1 Hz, 1H), 2.20 (s, 4H), 2.07-1.84 (m, 1H), 1.34 (ddt, J = 12.1, 8.8, 4.7Hz, 2H), 1.10 (d, J=6.5Hz, 3H).
实施例71Example 71
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-((2-(3,3,4,4-四氟吡咯烷-1-基)乙基)(乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺71的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((2-(3,3,4,4-tetrafluoropyrrolidin-1-yl) (Ethyl)(ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 71
采用与实施例64相同的制备方法，以3,3-二氟吡咯烷盐酸盐为原料，可得到化合物66。Using the same preparation method as in Example 64, using 3,3-difluoropyrrolidine hydrochloride as a raw material, compound 66 can be obtained.
MS m/z(ESI):639[M+1]
+
MS m/z(ESI):639[M+1] +
1H NMR(400MHz,DMSO-d
6)δ12.61(s,1H),9.98(s,1H),8.36(d,J＝7.3Hz,1H),7.77(d,J＝9.1Hz,1H),7.47(s,1H),7.40(dd,J＝8.6,0.8Hz,1H),7.25(dd,J＝8.6,1.6Hz,1H),7.00(ddd,J＝12.1,8.0,2.0Hz,3H),6.00(dd,J＝9.0,2.3Hz,1H),5.82(d,J＝2.4Hz,1H),4.04(s,2H),3.83(dt,J＝11.6,4.0Hz,2H),3.51–3.37(m,6H),3.26(s,3H),2.70(t,J＝7.2Hz,2H),1.99–1.89(m,2H),1.43–1.31(m,2H),1.11(t,J＝6.9Hz,3H).
1 H NMR(400MHz,DMSO-d 6 )δ12.61(s,1H),9.98(s,1H),8.36(d,J=7.3Hz,1H),7.77(d,J=9.1Hz,1H) ,7.47(s,1H),7.40(dd,J＝8.6,0.8Hz,1H),7.25(dd,J＝8.6,1.6Hz,1H),7.00(ddd,J＝12.1,8.0,2.0Hz,3H ),6.00(dd,J=9.0,2.3Hz,1H),5.82(d,J=2.4Hz,1H),4.04(s,2H),3.83(dt,J=11.6,4.0Hz,2H),3.51 –3.37(m,6H), 3.26(s,3H), 2.70(t,J=7.2Hz,2H),1.99–1.89(m,2H),1.43–1.31(m,2H),1.11(t,J =6.9Hz, 3H).
实施例72Example 72
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-2-((四氢-2H-吡喃-4-基)氨基)-4-(4-(2,2,2-三氟乙基)哌嗪-1-基)苯甲酰胺72的制备N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-2-((tetrahydro-2H-pyran-4- (Yl)amino)-4-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)benzamide 72
采用与实施例50相同的制备方法，以1,3-二甲基哌嗪为原料，可得到化合物72。Using the same preparation method as in Example 50 and using 1,3-dimethylpiperazine as a raw material, compound 72 can be obtained.
MS m/z(ESI):661[M+H]
+。
MS m/z (ESI): 661 [M+H] + .
1H NMR(400MHz,DMSO-d
6)δ12.67(s,1H),10.08(s,1H),8.27(d,J＝7.7Hz,1H),7.77(d,J＝9.0Hz,1H),7.51(d,J＝1.5Hz,1H),7.45–7.34(m,2H),7.29(dd,J＝8.7,1.7Hz,1H),7.20(td,J＝9.3,4.7Hz,1H),7.15–7.07(m,1H),6.21(dd,J＝9.0,2.3Hz,1H),6.11(d,J＝2.3Hz,1H),5.10(t,J＝8.7Hz,1H),4.98(dd,J＝9.4,6.9Hz,1H),4.87(dd,J＝9.3,6.4Hz,1H),4.73(dt,J＝14.7,7.3Hz,1H),3.79(d,J＝11.6Hz,2H),3.66(dd,J＝10.9,6.6Hz,1H),3.47(ddt,J＝13.3,11.5,2.4Hz,2H),3.28–3.13(m,7H),2.72(t,J＝5.0Hz,4H),1.99–1.81(m,2H),1.32(qd,J＝9.7,4.9Hz,2H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.67 (s, 1H), 10.08 (s, 1H), 8.27 (d, J = 7.7 Hz, 1H), 7.77 (d, J = 9.0 Hz, 1H) ,7.51(d,J=1.5Hz,1H),7.45-7.34(m,2H),7.29(dd,J=8.7,1.7Hz,1H), 7.20(td,J=9.3,4.7Hz,1H), 7.15–7.07 (m, 1H), 6.21 (dd, J = 9.0, 2.3 Hz, 1H), 6.11 (d, J = 2.3 Hz, 1H), 5.10 (t, J = 8.7 Hz, 1H), 4.98 (dd ,J=9.4,6.9Hz,1H), 4.87(dd,J=9.3,6.4Hz,1H), 4.73(dt,J=14.7,7.3Hz,1H), 3.79(d,J=11.6Hz,2H) , 3.66 (dd, J = 10.9, 6.6 Hz, 1H), 3.47 (ddt, J = 13.3, 11.5, 2.4 Hz, 2H), 3.28-3.13 (m, 7H), 2.72 (t, J = 5.0 Hz, 4H ),1.99–1.81(m,2H),1.32(qd,J=9.7,4.9Hz,2H).
实施例73Example 73
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(2-甲基-4-乙基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺73的制备N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(2-methyl-4-ethylpiperazine -1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 73
采用与实施例50相同的制备方法，以1-乙基-3-甲基哌嗪为原料，可得到化合物73。Using the same preparation method as in Example 50 and using 1-ethyl-3-methylpiperazine as a raw material, compound 73 can be obtained.
MS m/z(ESI):621[M+H]
+。
MS m/z (ESI): 621 [M+H] + .
1H NMR(400MHz,DMSO-d
6)δ12.72–12.63(m,1H),10.07(d,J＝10.0Hz,1H),8.37–8.25(m,1H),7.84–7.70(m,1H),7.59–7.49(m,1H),7.48–7.36(m,2H),7.36–7.28(m,1H),7.26–7.07(m,2H),6.23–6.15(m,1H),6.09–6.00(m,1H),5.16–5.07(m,1H),4.99(dd,J＝9.8,6.9Hz,1H),4.87(dd,J＝9.4,6.4Hz,0H),4.73(dt,J＝14.6,7.4Hz,1H),4.15(s,1H),3.79(d,J＝11.7Hz,2H),3.65(d,J＝7.5Hz,1H),3.55–3.40(m,3H),2.95(q,J＝13.2,12.3Hz,2H),2.76(d,J＝11.1Hz,1H),2.33(s,2H),2.15(d,J＝10.9Hz,1H),1.95(d,J＝20.6Hz,3H),1.34(td,J＝8.8,8.3,4.7Hz,2H),1.05(tdd,J＝14.0,9.9,6.9Hz,6H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.72-12.63 (m, 1H), 10.07 (d, J = 10.0 Hz, 1H), 8.37-8.25 (m, 1H), 7.84-7.70 (m, 1H) ), 7.59--7.49(m,1H),7.48--7.36(m,2H), 7.36--7.28(m,1H), 7.26-7.07(m,2H), 6.23--6.15(m,1H), 6.09-6.00 (m, 1H), 5.16–5.07 (m, 1H), 4.99 (dd, J = 9.8, 6.9 Hz, 1H), 4.87 (dd, J = 9.4, 6.4 Hz, 0H), 4.73 (dt, J = 14.6 ,7.4Hz,1H),4.15(s,1H),3.79(d,J=11.7Hz,2H), 3.65(d,J=7.5Hz,1H),3.55-3.40(m,3H),2.95(q ,J＝13.2,12.3Hz,2H),2.76(d,J＝11.1Hz,1H),2.33(s,2H),2.15(d,J＝10.9Hz,1H),1.95(d,J＝20.6Hz , 3H), 1.34 (td, J = 8.8, 8.3, 4.7 Hz, 2H), 1.05 (tdd, J = 14.0, 9.9, 6.9 Hz, 6H).
实施例74Example 74
4-(环丙基(2-(吡咯烷-1-基)乙基)氨基)-N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺744-(Cyclopropyl(2-(pyrrolidin-1-yl)ethyl)amino)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-2 -((Tetrahydro-2H-pyran-4-yl)amino)benzamide 74
第一步 4-氟-2-硝基苯甲酸苄酯74a的合成The first step is the synthesis of benzyl 4-fluoro-2-nitrobenzoate 74a
4-氟-2-硝基苯甲酸(20g,108mmol)溶于DMF(150mL)，逐滴加入苄溴(20.3g,119mmol)后，加入碳酸钾(41.4g,300mmol)90℃反应3-5小时。加水淬灭，二氯甲烷萃取，依次用水(100mL)和饱和食盐水(100mL)洗涤，无水硫酸钠干燥，过滤，浓缩，所得粗品经柱层析纯化得标题化合物74a(23g，83.6mmol)收率77.4％。4-Fluoro-2-nitrobenzoic acid (20g, 108mmol) was dissolved in DMF (150mL), benzyl bromide (20.3g, 119mmol) was added dropwise, potassium carbonate (41.4g, 300mmol) was added at 90℃ to react 3-5 Hour. Quench with water, extract with dichloromethane, wash with water (100mL) and saturated brine (100mL) successively, dry with anhydrous sodium sulfate, filter, and concentrate. The resulting crude product is purified by column chromatography to obtain the title compound 74a (23g, 83.6mmol) The yield was 77.4%.
第二步 苄基2-硝基-4-((2-(吡咯烷-1-基)乙基)氨基)苯甲酸酯74b的合成The second step is the synthesis of benzyl 2-nitro-4-((2-(pyrrolidin-1-yl)ethyl)amino)benzoate 74b
化合物74a(14.3g，125.4mmol)溶于NMP(100mL)，加入50mL二异丙基乙基氨(DIPEA)搅拌0.5小时后，加入化合物13079-a(23g，83.6mmol)室温搅拌 3-5小时。加水淬灭，二氯甲烷萃取。依次用水(150mL)和饱和食盐水(150mL)洗涤，无水硫酸钠干燥，过滤，浓缩，所得粗品经柱层析纯化(DCM:MeOH＝98:2)得标题化合物74b(22g，59.6mmol)收率71.3％。Compound 74a (14.3g, 125.4mmol) was dissolved in NMP (100mL), 50mL of diisopropylethylammonium (DIPEA) was added and stirred for 0.5 hours, compound 13079-a (23g, 83.6mmol) was added and stirred at room temperature for 3-5 hours . Quench with water and extract with dichloromethane. Wash with water (150 mL) and saturated brine (150 mL) successively, dry with anhydrous sodium sulfate, filter, and concentrate. The resulting crude product is purified by column chromatography (DCM:MeOH=98:2) to obtain the title compound 74b (22g, 59.6mmol) The yield was 71.3%.
MS m/z(ESI):370.2[M+H]
+。
MS m/z (ESI): 370.2 [M+H] + .
第三步 苄基4-(环丙基(2-(吡咯烷-1-基)乙基)氨基)-2-硝基苯甲酸酯74c的合成The third step is the synthesis of benzyl 4-(cyclopropyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-nitrobenzoate 74c
化合物74b(20g，54.2mmol)溶于甲苯(100mL)，加入化合物环丙基硼酸(13.9g,162.6mmol)加入吡啶(12.8g，162.6mmol)，加入Cu(OAc)
2(10.84g,54.2mmol)，加入Cs
2CO
3(8.8g,27.1mmol)搅拌0.5小时后，充氧气球加热至140℃反应8小时。加水淬灭，二氯甲烷萃取。依次用水(150mL)和饱和食盐水(150mL)洗涤，无水硫酸钠干燥，过滤，浓缩，所得粗品经柱层析纯化(DCM:MeOH＝96:4)得标题化合物74c(6.0g，14.7mmol)收率27.1％。
Compound 74b (20g, 54.2mmol) was dissolved in toluene (100mL), the compound cyclopropylboronic acid (13.9g, 162.6mmol) was added, pyridine (12.8g, 162.6mmol) was added, and Cu(OAc) 2 (10.84g, 54.2mmol) was added ), Cs 2 CO 3 (8.8 g, 27.1 mmol) was added and stirred for 0.5 hour, and then the oxygen-filled balloon was heated to 140° C. for reaction for 8 hours. Quench with water and extract with dichloromethane. Wash with water (150 mL) and saturated brine (150 mL) successively, dry with anhydrous sodium sulfate, filter, and concentrate. The resulting crude product is purified by column chromatography (DCM:MeOH=96:4) to obtain the title compound 74c (6.0g, 14.7mmol) ) The yield is 27.1%.
MS m/z(ESI):410.3[M+H]
+。
MS m/z (ESI): 410.3 [M+H] + .
第四步 苄基2-氨基-4-(环丙基(2-(吡咯烷-1-基)乙基)氨基)苯甲酸酯74d的合成The fourth step is the synthesis of benzyl 2-amino-4-(cyclopropyl(2-(pyrrolidin-1-yl)ethyl)amino)benzoate 74d
化合物74c(4.6g，11.2mmol)溶于甲醇和水5:1混合液(25mL)，搅拌均匀后，加入锌粉(13.8g,211mmol)和氯化铵(13.8g,258mmol)室温条件下搅拌约3小时。抽滤，将滤液旋干，所得粗品经柱层析纯化(DCM:MeOH＝95:5)得标题化合物74d(3.0g，7.9mmol)收率70.6％。Compound 74c (4.6g, 11.2mmol) was dissolved in a 5:1 mixture of methanol and water (25mL). After stirring, zinc powder (13.8g, 211mmol) and ammonium chloride (13.8g, 258mmol) were added and stirred at room temperature. About 3 hours. Suction filtration, the filtrate was spin-dried, and the obtained crude product was purified by column chromatography (DCM:MeOH=95:5) to obtain the title compound 74d (3.0 g, 7.9 mmol) with a yield of 70.6%.
MS m/z(ESI):380.3[M+H]
+。
MS m/z (ESI): 380.3 [M+H] + .
第五步 4-(环丙基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酸苄酯74e的合成The fifth step 4-(cyclopropyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzyl benzoate 74e synthesis
化合物74d(2.9g，7.6mmol)溶于DCM(25mL)，搅拌条件下加入1mL三氟乙酸，搅拌均匀后加入四氢-2H-吡喃-4-酮室温下搅拌0.5小时后加入NaBH(OAc)
3搅拌5小时。逐滴加入饱和氯化铵溶液淬灭。二氯甲烷萃取，多次用水(100mL)和饱和食盐水(100mL)洗涤，无水硫酸钠干燥，过滤，浓缩，柱层析(DCM:MeOH＝97:3)得标题化合物74e(2.0g，4.3mmol)，收率56.8％。
Compound 74d (2.9g, 7.6mmol) was dissolved in DCM (25mL), 1mL of trifluoroacetic acid was added under stirring, and tetrahydro-2H-pyran-4-one was added after stirring evenly, and after stirring for 0.5 hours at room temperature, NaBH(OAc ) 3 Stir for 5 hours. It was quenched by adding saturated ammonium chloride solution dropwise. Extracted with dichloromethane, washed with water (100mL) and saturated brine (100mL) several times, dried over anhydrous sodium sulfate, filtered, concentrated, and column chromatography (DCM:MeOH=97:3) to obtain the title compound 74e (2.0g, 4.3 mmol), the yield is 56.8%.
MS m/z(ESI):464.4[M+H]
+。
MS m/z (ESI): 464.4 [M+H] + .
第六步 苄基4-(环丙基(2-(吡咯烷-1-基)乙基)氨基)-2-(2,2,2-三氟-N-(四氢 -2H-吡喃-4-基)乙酰胺)苯甲酸酯74f的合成Step 6 Benzyl 4-(cyclopropyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran) -4-yl)acetamide) benzoate 74f synthesis
化合物74e(2.0g，4.3mmol)溶于无水DCM(25mL)，加入二异丙基乙基氨(DIPEA)4mL搅拌均匀后，冰水浴冷却下逐滴加入三氟乙酸酐，随后缓慢升温至室温搅拌约3小时。逐滴加入饱和氯化铵溶液淬灭。二氯甲烷萃取，多次用水(100mL)和饱和食盐水(100mL)洗涤，无水硫酸钠干燥，过滤，浓缩，柱层析(DCM:MeOH＝98:2)得标题化合物74f(1.0g,1.8mmol)，收率41.6％。Compound 74e (2.0g, 4.3mmol) was dissolved in anhydrous DCM (25mL), 4mL of diisopropylethylammonium (DIPEA) was added and stirred uniformly, trifluoroacetic anhydride was added dropwise under cooling in an ice-water bath, and then slowly heated to Stir at room temperature for about 3 hours. It was quenched by adding saturated ammonium chloride solution dropwise. Extracted with dichloromethane, washed with water (100mL) and saturated brine (100mL) several times, dried over anhydrous sodium sulfate, filtered, concentrated, and column chromatography (DCM:MeOH=98:2) to obtain the title compound 74f (1.0g, 1.8mmol), the yield is 41.6%.
MS m/z(ESI):560.4[M+H]
+。
MS m/z (ESI): 560.4 [M+H] + .
第七步 4-(环丙基(2-(吡咯烷-1-基)乙基)氨基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰胺)苯甲酸74g的合成The seventh step 4-(cyclopropyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4) -(Yl)acetamide) benzoic acid 74g synthesis
化合物74f(1.0g,1.8mmol)溶于甲醇(20mL)中，加入Pd/C(1.0g,9.4mmol)，充氢气置换三次后，在氢气环境下常温反应1-3小时，反应完全后，抽滤，将反应液旋干备用得粗产品74g(0.6g，1.27mmol)，收率71％。Compound 74f (1.0g, 1.8mmol) was dissolved in methanol (20mL), added Pd/C (1.0g, 9.4mmol), filled with hydrogen for three times, and reacted at room temperature for 1-3 hours under hydrogen environment. After the reaction was complete, After suction filtration, the reaction solution was spin-dried for later use to obtain 74 g (0.6 g, 1.27 mmol) of the crude product, with a yield of 71%.
MS m/z(ESI):470.4[M+H]
+。
MS m/z (ESI): 470.4 [M+H] + .
第八步 4-(环丙基(2-(吡咯烷-1-基)乙基)氨基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰胺)苯甲酰氯74h的合成Step 8 4-(Cyclopropyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4) -Yl)acetamide) benzoyl chloride 74h synthesis
化合物74g(0.6g，1.27mmol)溶于二氯甲烷，在搅拌状态下加入一滴N,N-二甲基甲酰胺(DMF)，随后逐滴加入草酰氯(0.4g,1.84mmol)后室温反应1-2小时。反应完全后旋干备用，得标题化合物74hCompound 74g (0.6g, 1.27mmol) was dissolved in dichloromethane, a drop of N,N-dimethylformamide (DMF) was added under stirring, and then oxalyl chloride (0.4g, 1.84mmol) was added dropwise and reacted at room temperature 1-2 hours. After the reaction is complete, spin dry for later use to obtain the title compound 74h
第九步 4-(环丙基(2-(吡咯烷-1-基)乙基)氨基)-N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-2-(2,2,2-三氟-N-(四氢-2H-吡喃-4-基)乙酰胺)苯甲酰胺74i的合成Step 9 4-(Cyclopropyl(2-(pyrrolidin-1-yl)ethyl)amino)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl )-2-(2,2,2-Trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamide)benzamide 74i
化合物I-5(390mg，1.5mmol)溶于无水DCM(15mL)，加入二异丙基乙基氨(DIPEA)1mL搅拌均匀后，冰水浴冷却下逐滴加入化合物74h(600mg,1.2mmol)，随后缓慢升温至室温搅拌约1小时。逐滴加入饱和氯化铵溶液淬灭。二氯甲烷萃取，无水硫酸钠干燥，过滤，浓缩，柱层析(DCM:MeOH＝98:2)得标题化合物74i(240mg,0.33mmol)，收率22.5％。Compound I-5 (390mg, 1.5mmol) was dissolved in anhydrous DCM (15mL), 1mL of diisopropylethylammonium (DIPEA) was added and stirred uniformly, compound 74h (600mg, 1.2mmol) was added dropwise under cooling in an ice-water bath , Then slowly warmed to room temperature and stirred for about 1 hour. It was quenched by adding saturated ammonium chloride solution dropwise. Extracted with dichloromethane, dried with anhydrous sodium sulfate, filtered, concentrated, and column chromatography (DCM:MeOH=98:2) to obtain the title compound 74i (240 mg, 0.33 mmol) with a yield of 22.5%.
MS m/z(ESI):711.4[M+H]
+。
MS m/z (ESI): 711.4 [M+H] + .
第十步 4-(环丙基(2-(吡咯烷-1-基)乙基)氨基)-N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺74的合成Step 10 4-(Cyclopropyl(2-(pyrrolidin-1-yl)ethyl)amino)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl )-2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide 74
化合物74i(240mg,0.33mmol)溶于无水甲醇(10mL)，加入三乙胺1mL搅 拌均匀后，80℃下回流1-3小时。随后缓慢升温至室温搅拌约1小时。逐滴加入饱和氯化铵溶液淬灭。二氯甲烷萃取，无水硫酸钠干燥，过滤，浓缩，柱层析(DCM:MeOH＝95:5)得标题化合物74(61mg,0.10mmol)，收率30％Compound 74i (240 mg, 0.33 mmol) was dissolved in anhydrous methanol (10 mL), and 1 mL of triethylamine was added and stirred uniformly, and then refluxed at 80°C for 1-3 hours. Then slowly warm to room temperature and stir for about 1 hour. It was quenched by adding saturated ammonium chloride solution dropwise. Extraction with dichloromethane, drying with anhydrous sodium sulfate, filtration, concentration, column chromatography (DCM:MeOH=95:5) to obtain the title compound 74 (61mg, 0.10mmol), the yield is 30%
MS m/z(ESI):615.5[M+H]
+。
MS m/z (ESI): 615.5 [M+H] + .
实施例75Example 75
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-((2-(1,4-二氧-7-氮杂螺[4.4]壬烷-1-基)乙基)(乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺75的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((2-(1,4-diox-7-azaspiro[4.4]nonane -1-yl)ethyl)(ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 75
采用与实施例64相同的制备方法，以3-吡咯烷酮缩乙二醇为原料，可得到化合物75。Using the same preparation method as in Example 64 and using 3-pyrrolidone ethylene ketal as the raw material, compound 75 can be obtained.
MS m/z(ESI):661[M+1]
+
MS m/z(ESI):661[M+1] +
1H NMR(400MHz,DMSO-d
6)δ12.60(s,1H),9.96(s,1H),8.35(d,J＝7.3Hz,1H),7.76(d,J＝9.1Hz,1H),7.47(s,1H),7.40(d,J＝8.6Hz,1H),7.24(dd,J＝8.6,1.6Hz,1H),7.05–6.93(m,3H),5.99(dd,J＝9.0,2.3Hz,1H),5.82(d,J＝2.4Hz,1H),4.04(s,2H),3.83(tt,J＝6.3,2.7Hz,6H),3.59(s,1H),3.51–3.35(m,6H),2.71–2.53(m,6H),2.02–1.85(m,4H),1.43–1.32(m,2H),1.11(t,J＝6.9Hz,3H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.60 (s, 1H), 9.96 (s, 1H), 8.35 (d, J = 7.3 Hz, 1H), 7.76 (d, J = 9.1 Hz, 1H) ,7.47(s,1H),7.40(d,J=8.6Hz,1H),7.24(dd,J=8.6,1.6Hz,1H),7.05-6.93(m,3H),5.99(dd,J=9.0 ,2.3Hz,1H),5.82(d,J=2.4Hz,1H),4.04(s,2H),3.83(tt,J=6.3,2.7Hz,6H),3.59(s,1H),3.51–3.35 (m,6H),2.71-2.53(m,6H),2.02-1.85(m,4H),1.43-1.32(m,2H),1.11(t,J=6.9Hz,3H).
实施例76Example 76
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-((2-(3-氧代吡咯烷-1-基)乙基)(乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺76的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((2-(3-oxopyrrolidin-1-yl)ethyl)(ethyl )Amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 76
将化合物75(120mg，0.2mmol)投于2-丁酮(10ml)中，加入6M HCl(2.5ml)，回流反应3h，减压蒸除溶剂。加入碳酸氢钠中和，DCM(50ml×2)萃取，合并有机层，柱层析(DCM:MeOH＝98:2)得化合物76(40mg)。Compound 75 (120mg, 0.2mmol) was put into 2-butanone (10ml), 6M HCl (2.5ml) was added, the reaction was refluxed for 3h, and the solvent was evaporated under reduced pressure. Add sodium bicarbonate to neutralize, extract with DCM (50ml×2), combine the organic layers, and column chromatography (DCM:MeOH=98:2) to obtain compound 76 (40mg).
MS m/z(ESI):617[M+1]
+
MS m/z(ESI):617[M+1] +
1H NMR(400MHz,DMSO-d
6)δ12.58(s,1H),9.94(s,1H),8.33(d,J＝7.3Hz,1H),7.74(d,J＝9.1Hz,1H),7.44(s,1H),7.37(d,J＝8.6Hz,1H),7.22(dd,J＝8.5,1.6Hz,1H),7.03–6.89(m,3H),5.98(dd,J＝9.3,2.3Hz,1H),5.82(d,J＝2.7Hz,1H),4.01(s,2H),3.84–3.75(m,2H),3.57(d,J＝8.7Hz,1H),3.50–3.35(m,5H),2.99(s,2H),2.91(t,J＝6.9Hz,2H),2.68(t,J＝7.3Hz,2H),2.31(t,J＝6.9Hz,2H),1.92(d,J＝12.9Hz,2H),1.42–1.28(m,2H),1.10(t,J＝6.9Hz,3H).
1 H NMR(400MHz,DMSO-d 6 )δ12.58(s,1H),9.94(s,1H),8.33(d,J=7.3Hz,1H),7.74(d,J=9.1Hz,1H) ,7.44(s,1H),7.37(d,J=8.6Hz,1H),7.22(dd,J=8.5,1.6Hz,1H),7.03-6.89(m,3H),5.98(dd,J=9.3 ,2.3Hz,1H),5.82(d,J=2.7Hz,1H),4.01(s,2H),3.84–3.75(m,2H),3.57(d,J=8.7Hz,1H),3.50–3.35 (m, 5H), 2.99 (s, 2H), 2.91 (t, J = 6.9 Hz, 2H), 2.68 (t, J = 7.3 Hz, 2H), 2.31 (t, J = 6.9 Hz, 2H), 1.92 (d,J=12.9Hz,2H),1.42-1.28(m,2H),1.10(t,J=6.9Hz,3H).
实施例77Example 77
N-(5-(2,5-二氟苯甲基)-1H-吲唑-3-基)-4-(甲基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺77的制备N-(5-(2,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidin-1-yl)ethyl)amino)-2 -((Tetrahydro-2H-pyran-4-yl)amino)benzamide 77
采用与实施例46相同的制备方法，以2,5-二氟溴苯为原料，可得到化合物77。Using the same preparation method as in Example 46, using 2,5-difluorobromobenzene as a raw material, compound 77 can be obtained.
MS m/z(ESI):589[M+H]
+。
MS m/z (ESI): 589 [M+H] + .
1H NMR(400MHz,Methanol-d
4)δ7.71(d,J＝9.0Hz,1H),7.57(s,1H),7.40(dd,J＝8.6,0.8Hz,1H),7.25(dd,J＝8.8,1.6Hz,1H),7.05(td,J＝9.0,4.5Hz,1H),6.99–6.87(m,2H),6.14(dd,J＝9.1,2.4Hz,1H),5.97(d,J＝2.3Hz,1H),4.06(s,2H),3.93(dt,J＝11.7,4.0Hz,2H),3.69(dt,J＝9.4,5.1Hz,1H),3.63–3.52(m,4H),3.03(s,3H),2.80–2.63(m,6H),2.04(d,J＝12.7Hz,2H),1.86(s,4H),1.53(ddd,J＝18.7,9.3,4.5Hz,3H).
1 H NMR(400MHz,Methanol-d 4 )δ7.71(d,J＝9.0Hz,1H), 7.57(s,1H), 7.40(dd,J＝8.6,0.8Hz,1H), 7.25(dd, J = 8.8, 1.6 Hz, 1H), 7.05 (td, J = 9.0, 4.5 Hz, 1H), 6.99-6.87 (m, 2H), 6.14 (dd, J = 9.1, 2.4 Hz, 1H), 5.97 (d ,J=2.3Hz,1H),4.06(s,2H),3.93(dt,J=11.7,4.0Hz,2H),3.69(dt,J=9.4,5.1Hz,1H),3.63-3.52(m, 4H),3.03(s,3H),2.80–2.63(m,6H),2.04(d,J=12.7Hz,2H),1.86(s,4H),1.53(ddd,J=18.7,9.3,4.5Hz ,3H).
实施例78Example 78
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(乙基(2-(氮杂环丁烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺78的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(azetidin-1-yl)ethyl)amino) Preparation of -2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 78
采用与实施例64相同的制备方法，以氮杂环丁烷盐酸盐为原料，可得到化合物78。Using the same preparation method as in Example 64 and using azetidine hydrochloride as a raw material, compound 78 can be obtained.
MS m/z(ESI):589[M+1]
+
MS m/z(ESI):589[M+1] +
1H NMR(400MHz,DMSO-d
6)δ12.59(d,J＝7.7Hz,1H),9.98(s,1H),8.35(d,J＝7.4Hz,1H),7.75(d,J＝9.0Hz,1H),7.47–7.33(m,2H),7.22(dd,J＝8.6,1.6Hz,1H),6.97(tdd,J＝12.8,7.7,4.2Hz,3H),5.99(d,J＝9.3Hz,1H),5.82(d,J＝2.4Hz,1H),4.01(s,2H),3.85–3.74(m,2H),3.60(s,3H),3.50–3.32(m,8H),2.80–2.74(m,1H),2.14(s,2H),1.92(d,J＝13.1Hz,2H),1.42–1.29(m,2H),1.08(q,J＝7.6,7.0Hz,3H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.59 (d, J = 7.7 Hz, 1H), 9.98 (s, 1H), 8.35 (d, J = 7.4 Hz, 1H), 7.75 (d, J = 9.0Hz, 1H), 7.47–7.33 (m, 2H), 7.22 (dd, J = 8.6, 1.6 Hz, 1H), 6.97 (tdd, J = 12.8, 7.7, 4.2 Hz, 3H), 5.99 (d, J =9.3Hz,1H),5.82(d,J=2.4Hz,1H),4.01(s,2H), 3.85–3.74(m,2H), 3.60(s,3H), 3.50–3.32(m,8H) ,2.80–2.74(m,1H),2.14(s,2H),1.92(d,J=13.1Hz,2H),1.42–1.29(m,2H),1.08(q,J=7.6,7.0Hz,3H ).
实施例79Example 79
N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((2-(吡咯烷-1-基)乙基)亚硫酰基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺79的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((2-(pyrrolidin-1-yl)ethyl)sulfinyl)-2 -((Tetrahydro-2H-pyran-4-yl)amino)benzamide 79
将化合物65(100mg)溶于DCM(10ml)中，冰水浴冷却下，加入m-CPBA(29mg，0.17mmol，1.0eq)，继续反应4h。加入碳酸氢钠水溶液中和，DCM(15ml×2)萃取，合并有机层，浓缩，制备版纯化得79(HHT0159-101-1A,50mg)。Compound 65 (100 mg) was dissolved in DCM (10 ml), and m-CPBA (29 mg, 0.17 mmol, 1.0 eq) was added under ice-water bath cooling, and the reaction was continued for 4 h. Add sodium bicarbonate aqueous solution to neutralize, extract with DCM (15ml×2), combine the organic layers, concentrate, and purify the preparation plate to obtain 79 (HHT0159-101-1A, 50mg).
MS m/z(ESI):608[M+1]
+
MS m/z(ESI):608[M+1] +
1H NMR(400MHz,DMSO-d
6)δ12.98(s,1H),10.44(s,1H),8.09(d,J＝7.8Hz,1H),7.81(d,J＝8.5Hz,1H),7.50–7.34(m,2H),7.23(d,J＝8.6Hz,1H),7.03–6.87(m,2H),6.83(s,1H),6.55(d,J＝8.4Hz,1H),4.00(s,2H),3.84–3.52(m,5H),3.51–3.40(m,6H),3.24(d,J＝8.0Hz,2H),2.25–2.07(m,2H),1.86(q,J＝9.9,7.6Hz,3H),1.38–1.14(m,3H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.98 (s, 1H), 10.44 (s, 1H), 8.09 (d, J = 7.8 Hz, 1H), 7.81 (d, J = 8.5 Hz, 1H) ,7.50–7.34(m,2H), 7.23(d,J=8.6Hz,1H), 7.03–6.87(m,2H), 6.83(s,1H), 6.55(d,J=8.4Hz,1H), 4.00(s,2H), 3.84–3.52(m,5H), 3.51–3.40(m,6H), 3.24(d,J=8.0Hz,2H), 2.25–2.07(m,2H), 1.86(q, J = 9.9, 7.6 Hz, 3H), 1.38-1.14 (m, 3H).
实施例80Example 80
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-甲基-4,7-二氮杂螺[2.5]辛烷-7-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺80的制备N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methyl-4,7-diazepine Preparation of heterospiro[2.5]octane-7-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 80
采用与实施例50相同的制备方法，以4-甲基-4,7-二氮杂螺[2.5]辛烷为原料，可得到化合物80。Using the same preparation method as in Example 50, using 4-methyl-4,7-diazaspiro[2.5]octane as a raw material, compound 80 can be obtained.
MS m/z(ESI):619[M+H]
+。
MS m/z (ESI): 619 [M+H] + .
1H NMR(400MHz,DMSO-d
6)δ12.66(s,1H),10.06(s,1H),8.31(d,J＝7.7Hz,1H),7.75(d,J＝9.1Hz,1H),7.51(s,1H),7.44–7.34(m,2H),7.30(dd,J＝8.7,1.6Hz,1H),7.20(td,J＝9.3,4.7Hz,1H),7.11(tt,J＝8.0,3.5Hz,1H),6.19(dd,J＝ 9.2,2.2Hz,1H),6.06(d,J＝2.4Hz,1H),5.10(t,J＝8.7Hz,1H),4.99(t,J＝8.2Hz,1H),4.91–4.83(m,0H),4.73(dt,J＝14.8,7.2Hz,1H),3.78(d,J＝11.7Hz,3H),3.72–3.60(m,1H),3.47(t,J＝10.9Hz,2H),3.24(t,J＝5.4Hz,2H),3.05(s,2H),2.89(t,J＝5.2Hz,2H),2.27(s,3H),1.91(d,J＝13.3Hz,2H),1.39–1.24(m,2H),0.61(q,J＝4.4,3.8Hz,2H),0.54–0.43(m,2H).
1 H NMR(400MHz,DMSO-d 6 )δ12.66(s,1H), 10.06(s,1H), 8.31(d,J=7.7Hz,1H), 7.75(d,J=9.1Hz,1H) ,7.51(s,1H),7.44-7.34(m,2H),7.30(dd,J=8.7,1.6Hz,1H),7.20(td,J=9.3,4.7Hz,1H),7.11(tt,J ＝8.0,3.5Hz,1H),6.19(dd,J＝9.2,2.2Hz,1H),6.06(d,J＝2.4Hz,1H), 5.10(t,J＝8.7Hz,1H), 4.99(t ,J=8.2Hz,1H), 4.91–4.83(m,0H), 4.73(dt,J=14.8,7.2Hz,1H), 3.78(d,J=11.7Hz,3H), 3.72–3.60(m, 1H), 3.47 (t, J = 10.9 Hz, 2H), 3.24 (t, J = 5.4 Hz, 2H), 3.05 (s, 2H), 2.89 (t, J = 5.2 Hz, 2H), 2.27 (s, 3H), 1.91(d,J=13.3Hz,2H), 1.39-1.24(m,2H), 0.61(q,J=4.4,3.8Hz,2H), 0.54-0.43(m,2H).
实施例81Example 81
N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(3,5-二甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺81的制备N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(3,5-dimethylpiperazine-1 -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 81
采用与实施例50相同的制备方法，以4-甲基-4,7-二氮杂螺[2.5]辛烷为原料，可得到化合物81。Using the same preparation method as in Example 50 and using 4-methyl-4,7-diazaspiro[2.5]octane as a raw material, compound 81 can be obtained.
MS m/z(ESI):607[M+H]
+。
MS m/z (ESI): 607 [M+H] + .
1H NMR(400MHz,DMSO-d
6)δ12.67(s,1H),10.08(s,1H),8.30(d,J＝7.7Hz,1H),7.76(d,J＝9.0Hz,1H),7.50(s,1H),7.43–7.34(m,2H),7.29(dd,J＝8.7,1.6Hz,1H),7.19(td,J＝9.3,4.7Hz,1H),7.10(ddd,J＝8.9,7.5,3.5Hz,1H),6.22(dd,J＝9.0,2.3Hz,1H),6.08(d,J＝2.3Hz,1H),5.10(t,J＝8.7Hz,0H),4.98(t,J＝8.2Hz,1H),4.86(dd,J＝9.3,6.4Hz,0H),4.72(dt,J＝14.7,7.2Hz,1H),3.82–3.62(m,5H),3.48(tt,J＝11.7,2.3Hz,2H),2.29(t,J＝11.4Hz,2H),1.89(dd,J＝11.4,6.9Hz,2H),1.32(tq,J＝9.3,5.6,4.7Hz,2H),1.06(d,J＝6.3Hz,6H).
1 H NMR (400MHz, DMSO-d 6 ) δ 12.67 (s, 1H), 10.08 (s, 1H), 8.30 (d, J = 7.7 Hz, 1H), 7.76 (d, J = 9.0 Hz, 1H) ,7.50(s,1H),7.43-7.34(m,2H),7.29(dd,J=8.7,1.6Hz,1H),7.19(td,J=9.3,4.7Hz,1H),7.10(ddd,J ＝8.9,7.5,3.5Hz,1H), 6.22(dd,J＝9.0,2.3Hz,1H), 6.08(d,J＝2.3Hz,1H), 5.10(t,J＝8.7Hz,0H), 4.98 (t,J=8.2Hz,1H), 4.86(dd,J=9.3,6.4Hz,0H), 4.72(dt,J=14.7,7.2Hz,1H), 3.82-3.62(m,5H), 3.48( tt,J＝11.7,2.3Hz,2H),2.29(t,J＝11.4Hz,2H),1.89(dd,J＝11.4,6.9Hz,2H),1.32(tq,J＝9.3,5.6,4.7Hz ,2H),1.06(d,J=6.3Hz,6H).
实施例82Example 82
N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(甲基(2-(3-氰基吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺82的制备N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(3-cyanopyrrolidin-1-yl)ethyl)amino )-2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide 82
采用与实施例64相同的制备方法，以N-甲基-乙醇胺、3-氰基吡咯烷盐酸盐为原料，可得到化合物82。Using the same preparation method as in Example 64, using N-methyl-ethanolamine and 3-cyanopyrrolidine hydrochloride as raw materials, compound 82 can be obtained.
MS m/z(ESI):614[M+H]
+。
MS m/z (ESI): 614 [M+H] + .
1H NMR(400MHz,Methanol-d
4)δ7.70(d,J＝9.0Hz,1H),7.57(d,J＝1.5Hz,1H),7.42–7.37(m,1H),7.22(dd,J＝8.7,1.7Hz,1H),6.85–6.78(m,2H),6.74–6.67(m,1H),6.14(dd,J＝9.1,2.4Hz,1H),5.96(d,J＝2.4Hz,1H),4.06(s,2H),3.92(dt,J＝12.0,4.1Hz,2H),3.73–3.63(m,1H),3.63–3.50(m,4H),3.23–3.13(m,1H),3.03(s,3H),2.91–2.79(m,3H),2.77–2.69(m,2H),2.68–2.58(m,1H),2.26(ddd,J＝10.1,7.9,5.5Hz,2H),2.11–1.98(m,3H),1.58–1.46(m,2H).
1 H NMR(400MHz,Methanol-d 4 )δ7.70(d,J=9.0Hz,1H), 7.57(d,J=1.5Hz,1H), 7.42–7.37(m,1H), 7.22(dd, J=8.7,1.7Hz,1H), 6.85–6.78(m,2H), 6.74–6.67(m,1H), 6.14(dd,J=9.1,2.4Hz,1H), 5.96(d,J=2.4Hz ,1H),4.06(s,2H),3.92(dt,J=12.0,4.1Hz,2H),3.73-3.63(m,1H),3.63-3.50(m,4H),3.23-3.13(m,1H) ),3.03(s,3H),2.91–2.79(m,3H),2.77–2.69(m,2H),2.68–2.58(m,1H),2.26(ddd,J=10.1,7.9,5.5Hz,2H ), 2.11--1.98(m,3H), 1.58--1.46(m,2H).
实施例83Example 83
(R)-N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(甲基(2-(3-羟基吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺83的制备(R)-N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(3-hydroxypyrrolidin-1-yl)ethyl (Yl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 83
第一步 叔-丁基2-硝基-4-(乙基(2-(对甲苯磺酰氧基)乙基)氨基)苯甲酸酯83b的合成The first step is the synthesis of tert-butyl 2-nitro-4-(ethyl(2-(p-toluenesulfonyloxy)ethyl)amino)benzoate 83b
将化合物83a(500mg,1.69mmol)和TEA(338mg,3.38mmol)分散在DCM中，在室温下慢慢加入对甲苯磺酰氯(385mg,2.0mmol)的DCM溶液，该混合物在室温继续搅拌4小时，加入水淬灭，用DCM萃取，有机相用水洗，无水硫酸钠干燥，浓缩，粗品用flash纯化(PE：EA＝1:1)得到83b(700mg)淡黄色固体，收率91％。Compound 83a (500mg, 1.69mmol) and TEA (338mg, 3.38mmol) were dispersed in DCM, p-toluenesulfonyl chloride (385mg, 2.0mmol) in DCM was slowly added at room temperature, and the mixture was stirred at room temperature for 4 hours It was quenched by adding water, extracted with DCM, the organic phase was washed with water, dried over anhydrous sodium sulfate, concentrated, and the crude product was flash purified (PE:EA=1:1) to obtain 83b (700mg) as a pale yellow solid with a yield of 91%.
MS m/z(ESI):451[M+H]+。MS m/z(ESI): 451[M+H]+.
第二步(R)-叔-丁基2-硝基-4-(乙基(2-(3-羟基吡咯烷-1-基)乙基)氨基)苯甲酸酯83c的合成The second step is the synthesis of (R)-tert-butyl 2-nitro-4-(ethyl(2-(3-hydroxypyrrolidin-1-yl)ethyl)amino)benzoate 83c
将化合物83b(300mg,0.66mmol),(R)-吡咯烷-3-醇(114mg,1.32mmol)和碳酸铯(860mg,2.64mmol)溶于DMF(30mL)中，该混合物在室温搅拌12小时，加水淬灭，用DCM萃取2次，有机相用水洗，干燥，浓缩，粗品用flash纯化(DCM:MeOH＝15:1)得到83c(200mg)黄色固体。收率83％Compound 83b (300mg, 0.66mmol), (R)-pyrrolidin-3-ol (114mg, 1.32mmol) and cesium carbonate (860mg, 2.64mmol) were dissolved in DMF (30mL), and the mixture was stirred at room temperature for 12 hours , Quenched with water, extracted with DCM twice, the organic phase was washed with water, dried, concentrated, and the crude product was flash purified (DCM:MeOH=15:1) to obtain 83c (200mg) as a yellow solid. Yield 83%
MS m/z(ESI):366[M+H]+。MS m/z(ESI): 366[M+H]+.
第三步 (R)-叔-丁基2-硝基-4-(乙基(2-(3-乙酰氧基吡咯烷-1-基)乙基)氨基) 苯甲酸酯83d的合成The third step is the synthesis of (R)-tert-butyl 2-nitro-4-(ethyl(2-(3-acetoxypyrrolidin-1-yl)ethyl)amino) benzoate 83d
将化合物83c(150mg,0.41mmol)和TEA(123mg,1.23mmol)溶于DCM中，在冰浴下加入乙酰氯(48mg,0.61mmol),该混合物在室温搅拌2小时，加入水淬灭，用DCM萃取，有机相用水洗，干燥，浓缩，粗品用flash纯化(PE:EA＝1:1)得到160mg化合物83d呈黄色固体。收率95％。Compound 83c (150mg, 0.41mmol) and TEA (123mg, 1.23mmol) were dissolved in DCM, acetyl chloride (48mg, 0.61mmol) was added in an ice bath, the mixture was stirred at room temperature for 2 hours, and water was added for quenching. It was extracted with DCM, the organic phase was washed with water, dried, concentrated, and the crude product was flash purified (PE:EA=1:1) to obtain 160 mg of compound 83d as a yellow solid. The yield was 95%.
MS m/z(ESI):408[M+H]+。MS m/z(ESI): 408[M+H]+.
第四步 (R)-叔-丁基2-氨基-4-(乙基(2-(3-乙酰氧基吡咯烷-1-基)乙基)氨基)苯甲酸酯83e的合成The fourth step is the synthesis of (R)-tert-butyl 2-amino-4-(ethyl(2-(3-acetoxypyrrolidin-1-yl)ethyl)amino)benzoate 83e
将化合物83d(160mg,0.39mmol)溶于甲醇(10mL)中，加入锌粉(127mg，1.9mmol)和饱和氯化铵溶液(10mL)。该混合物在室温搅拌2小时，反应完全后过滤，滤液用DCM萃取，有机相用水洗，干燥，浓缩，得到粗品化合物83e直接用于下一步反应。Compound 83d (160 mg, 0.39 mmol) was dissolved in methanol (10 mL), and zinc powder (127 mg, 1.9 mmol) and saturated ammonium chloride solution (10 mL) were added. The mixture was stirred at room temperature for 2 hours. After the reaction was completed, it was filtered. The filtrate was extracted with DCM. The organic phase was washed with water, dried and concentrated to obtain the crude compound 83e, which was directly used in the next reaction.
MS m/z(ESI):378[M+H]+。MS m/z(ESI): 378[M+H]+.
第五步 (R)-叔-丁基2-((四氢-2H-吡喃-4-基)氨基)-4-(乙基(2-(3-乙酰氧基吡咯烷-1-基)乙基)氨基)苯甲酸酯83f的合成Step 5 (R)-tert-Butyl 2-((tetrahydro-2H-pyran-4-yl)amino)-4-(ethyl(2-(3-acetoxypyrrolidin-1-yl) ) Ethyl) amino) benzoate 83f synthesis
将四甲基三乙酰氧基硼氢化铵(512mg,1.95mmol)缓慢加入到化合物83e(150mg,0.39mmol)和四氢吡喃糖酮(116mg,1.16mmol)的DCM(10mL)和三氟乙酸(1mL)溶液中，该混合物在室温搅拌2小时。加入饱和NaHCO3水溶液淬灭反应。体系用DCM萃取，有机相用水洗，无水硫酸钠干燥，浓缩，粗品83f直接用于下一步。Tetramethyltriacetoxyammonium borohydride (512mg, 1.95mmol) was slowly added to compound 83e (150mg, 0.39mmol) and tetrahydropyranosone (116mg, 1.16mmol) in DCM (10mL) and trifluoroacetic acid (1 mL) In solution, the mixture was stirred at room temperature for 2 hours. The reaction was quenched by adding saturated aqueous NaHCO3 solution. The system was extracted with DCM, the organic phase was washed with water, dried over anhydrous sodium sulfate, concentrated, and the crude product 83f was used directly in the next step.
MS m/z(ESI):462[M+H]+。MS m/z(ESI): 462[M+H]+.
第六步 (R)-叔-丁基2-(N-(四氢-2H-吡喃-4-基)-三氟乙酰氨基)-4-(乙基(2-(3-乙酰氧基吡咯烷-1-基)乙基)氨基)苯甲酸酯83g的合成Step 6 (R)-tert-Butyl 2-(N-(tetrahydro-2H-pyran-4-yl)-trifluoroacetamido)-4-(ethyl(2-(3-acetoxy) Synthesis of pyrrolidin-1-yl)ethyl)amino)benzoate 83g
将化合物83f(150mg,0.32mmol)和TEA(128mg,1.28mmol)分散在DCM中，在室温下滴加三氟乙酸酐(136mg,0.64mmol)，该混合物在室温搅拌2小时。加入水淬灭反应。体系用DCM萃取，有机相用水洗，无水硫酸钠干燥，浓缩，粗品83g直接用于下一步反应。Compound 83f (150 mg, 0.32 mmol) and TEA (128 mg, 1.28 mmol) were dispersed in DCM, trifluoroacetic anhydride (136 mg, 0.64 mmol) was added dropwise at room temperature, and the mixture was stirred at room temperature for 2 hours. The reaction was quenched by adding water. The system was extracted with DCM, the organic phase was washed with water, dried over anhydrous sodium sulfate, concentrated, and 83 g of the crude product was directly used in the next reaction.
MS m/z(ESI):558[M+H]+。MS m/z(ESI): 558[M+H]+.
第七步 (R)-2-(N-(四氢-2H-吡喃-4-基)-三氟乙酰氨基)-4-(乙基(2-(3-乙酰氧基 吡咯烷-1-基)乙基)氨基)苯甲酸83h的合成The seventh step (R)-2-(N-(tetrahydro-2H-pyran-4-yl)-trifluoroacetamido)-4-(ethyl(2-(3-acetoxypyrrolidine-1) -(Yl) ethyl) amino) benzoic acid 83h synthesis
将化合物83g(130mg,0.23mmol)分散在DCM(5mL)中，在室温下滴加三氟乙酸(5mL)，该混合物在室温搅拌2小时，浓缩，粗品化合物83h直接用于下一步反应。Compound 83g (130mg, 0.23mmol) was dispersed in DCM (5mL), trifluoroacetic acid (5mL) was added dropwise at room temperature, the mixture was stirred at room temperature for 2 hours, concentrated, and the crude compound 83h was directly used in the next reaction.
MS m/z(ESI):502[M+H]+。MS m/z(ESI): 502[M+H]+.
第八步 (R)-2-(N-(四氢-2H-吡喃-4-基)-三氟乙酰氨基)-4-(乙基(2-(3-乙酰氧基吡咯烷-1-基)乙基)氨基)苯甲酰氯83i的合成Step 8 (R)-2-(N-(tetrahydro-2H-pyran-4-yl)-trifluoroacetamido)-4-(ethyl(2-(3-acetoxypyrrolidine-1) -(Yl)ethyl)amino)benzoyl chloride 83i synthesis
将化合物83h(100mg,0.19mmol)分散在DCM(5mL)中，在室温下滴加草酰氯(273mg,2.15mmol)和一滴DMF,该混合物在室温搅拌2小时，浓缩，粗品化合物83i直接用于下一步反应。Compound 83h (100mg, 0.19mmol) was dispersed in DCM (5mL), and oxalyl chloride (273mg, 2.15mmol) and a drop of DMF were added dropwise at room temperature. The mixture was stirred at room temperature for 2 hours and concentrated. The crude compound 83i was used directly Next reaction.
第九步 (R)-N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(甲基(2-(3-羟基吡咯烷-1-基)乙基)氨基)-2-(N-(四氢-2H-吡喃-4-基)-三氟乙酰氨基)苯甲酰胺83j的合成Step 9 (R)-N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(3-hydroxypyrrolidine-1- Synthesis of yl)ethyl)amino)-2-(N-(tetrahydro-2H-pyran-4-yl)-trifluoroacetamido)benzamide 83j
将化合物5-(3,5-二氟苯甲基)-1H-吲唑-3-胺(50mg,0.19mmol)和DIPEA(73mg,0.57mmol)溶于DCM中，在冰浴下滴加83i(100mg,0.19)的DCM溶液，滴加完毕，升至室温并继续搅拌2小时，加入水淬灭，用DCM萃取，有机相用水洗，无水硫酸钠干燥，浓缩，粗品用flash纯化(DCM：MeOH＝10：1)得到83j(40mg)呈淡黄色固体。收率28％。The compound 5-(3,5-difluorobenzyl)-1H-indazol-3-amine (50mg, 0.19mmol) and DIPEA (73mg, 0.57mmol) were dissolved in DCM, and 83i was added dropwise under an ice bath (100mg, 0.19) DCM solution, after the addition was completed, warmed to room temperature and continued stirring for 2 hours, quenched by adding water, extracted with DCM, the organic phase was washed with water, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified with flash (DCM : MeOH = 10:1) to obtain 83j (40 mg) as a pale yellow solid. The yield was 28%.
MS m/z(ESI):743[M+H]+。MS m/z(ESI): 743[M+H]+.
第十步(R)-N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(甲基(2-(3-羟基吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺83的合成The tenth step (R)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(3-hydroxypyrrolidine-1- Synthesis of phenyl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 83
将化合物83j(40mg,0.053mmol)溶于甲醇(2mL)中，加入LiOH溶液(5mL)，在室温搅拌2小时，反应完用DCM萃取，有机相干燥，浓缩，粗品柱层析纯化(DCM：MeOH＝10:1)得到化合物83(5mg)呈白色固体，收率15％。Compound 83j (40mg, 0.053mmol) was dissolved in methanol (2mL), LiOH solution (5mL) was added, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, it was extracted with DCM, the organic phase was dried, concentrated, and the crude product was purified by column chromatography (DCM: MeOH = 10:1) to obtain compound 83 (5 mg) as a white solid with a yield of 15%.
MS m/z(ESI):605[M+H]+。MS m/z(ESI): 605[M+H]+.
1H NMR(400MHz,Methanol-d4)δ7.71(d,J＝9.0Hz,1H),7.57(s,1H),7.40(d,J＝8.6Hz,1H),7.22(dd,J＝8.6,1.6Hz,1H),6.81(d,J＝7.3Hz,2H),6.75–6.66(m,1H),6.14(dd,J＝9.1,2.4Hz,1H),5.97(d,J＝2.2Hz,1H),4.37(t,J＝6.5Hz,1H),4.05(s,2H),3.92(dt,J＝11.8,4.1Hz,2H),3.68(dt,J＝9.5,5.2Hz,1H),3.61–3.51(m,3H),3.03(s,3H),2.95–2.83(m,2H),2.81–2.60(m,4H),2.21– 2.09(m,1H),2.04(d,J＝13.2Hz,2H),1.82–1.71(m,1H),1.53(ddt,J＝13.8,9.5,4.9Hz,2H).
1 H NMR(400MHz,Methanol-d4)δ7.71(d,J＝9.0Hz,1H), 7.57(s,1H), 7.40(d,J＝8.6Hz,1H), 7.22(dd,J＝8.6 ,1.6Hz,1H),6.81(d,J=7.3Hz,2H),6.75-6.66(m,1H),6.14(dd,J=9.1,2.4Hz,1H),5.97(d,J=2.2Hz ,1H), 4.37(t,J=6.5Hz,1H),4.05(s,2H),3.92(dt,J=11.8,4.1Hz,2H), 3.68(dt,J=9.5,5.2Hz,1H) ,3.61–3.51(m,3H),3.03(s,3H),2.95–2.83(m,2H), 2.81–2.60(m,4H),2.21–2.09(m,1H),2.04(d,J= 13.2Hz, 2H), 1.82-1.71 (m, 1H), 1.53 (ddt, J=13.8, 9.5, 4.9Hz, 2H).
实施例84Example 84
(S)-N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(甲基(2-(3-羟基吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺84的制备(S)-N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(3-hydroxypyrrolidin-1-yl)ethyl (Yl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 84
采用与实施例83相同的制备方法，以(S)-3-羟基吡咯烷为原料，可得到化合物84。Using the same preparation method as in Example 83 and using (S)-3-hydroxypyrrolidine as a raw material, compound 84 can be obtained.
MS m/z(ESI):605[M+H]
+。
MS m/z (ESI): 605 [M+H] + .
1H NMR(400MHz,Methanol-d
4)δ7.71(d,J＝9.0Hz,1H),7.57(s,1H),7.40(d,J＝8.6Hz,1H),7.22(dd,J＝8.6,1.6Hz,1H),6.81(d,J＝7.3Hz,2H),6.75–6.66(m,1H),6.14(dd,J＝9.1,2.4Hz,1H),5.97(d,J＝2.2Hz,1H),4.37(t,J＝6.5Hz,1H),4.05(s,2H),3.92(dt,J＝11.8,4.1Hz,2H),3.68(dt,J＝9.5,5.2Hz,1H),3.61–3.51(m,3H),3.03(s,3H),2.95–2.83(m,2H),2.81–2.60(m,4H),2.21–2.09(m,1H),2.04(d,J＝13.2Hz,2H),1.82–1.71(m,1H),1.53(ddt,J＝13.8,9.5,4.9Hz,2H).
1 H NMR(400MHz,Methanol-d 4 )δ7.71(d,J＝9.0Hz,1H), 7.57(s,1H), 7.40(d,J＝8.6Hz,1H), 7.22(dd,J＝ 8.6, 1.6 Hz, 1H), 6.81 (d, J = 7.3 Hz, 2H), 6.75-6.66 (m, 1H), 6.14 (dd, J = 9.1, 2.4 Hz, 1H), 5.97 (d, J = 2.2 Hz, 1H), 4.37 (t, J = 6.5 Hz, 1H), 4.05 (s, 2H), 3.92 (dt, J = 11.8, 4.1 Hz, 2H), 3.68 (dt, J = 9.5, 5.2 Hz, 1H ),3.61-3.51(m,3H),3.03(s,3H),2.95-2.83(m,2H),2.81-2.60(m,4H),2.21-2.09(m,1H),2.04(d,J = 13.2Hz, 2H), 1.82-1.71 (m, 1H), 1.53 (ddt, J = 13.8, 9.5, 4.9Hz, 2H).
生物学实验实施例：Examples of biological experiments:
实验一、化合物的体外酶学活性的测定Experiment 1. Determination of the in vitro enzymatic activity of the compound
1.实验目的和方法1. The purpose and method of the experiment
本实验目的是为了测试本发明化合物对原肌球蛋白受体激酶A(TRKA)、间变性淋巴瘤激酶(ALK)、原癌基因酪氨酸激酶(ROS1)的抑制作用，根据半数抑制浓度IC
50评价化合物的体外活性。
The purpose of this experiment is to test the inhibitory effect of the compound of the present invention on tropomyosin receptor kinase A (TRKA), anaplastic lymphoma kinase (ALK), and proto-oncogene tyrosine kinase (ROS1), according to the half inhibitory concentration IC 50 to evaluate the in vitro activity of the compound.
2.实验方案2. Experimental protocol
2.1化合物的配制与测定板的准备2.1 Compound preparation and preparation of assay plate
采用二甲基亚砜(DMSO)将待测化合物稀释至50微摩尔每升(μM)，转移到96孔板中并呈3倍梯度稀释成10个浓度。向96孔板的两个空孔中加入100微升(μL)二甲基亚砜(DMSO)，并将板标记为源板。然后将10微升(μL)化合物从源板转移到新96孔板中(中间板)。向中间板的每个孔中加入90微升(μL)1x激酶缓冲液。在振荡器上将化合物在中间板中混合10分钟。然后从中间板每孔吸取5微升(μL)转移到384孔板中，每个浓度2个复孔。Dimethyl sulfoxide (DMSO) was used to dilute the compound to be tested to 50 micromoles per liter (μM), transferred to a 96-well plate and diluted 3-fold to 10 concentrations. Add 100 microliters (μL) of dimethyl sulfoxide (DMSO) to the two empty wells of the 96-well plate, and mark the plate as the source plate. Then transfer 10 microliters (μL) of compound from the source plate to a new 96-well plate (middle plate). Add 90 microliters (μL) of 1x kinase buffer to each well of the middle plate. The compound was mixed in the middle plate on a shaker for 10 minutes. Then transfer 5 microliters (μL) from each well of the middle plate to a 384-well plate, with 2 replicate wells for each concentration.
2.2激酶反应2.2 Kinase reaction
向384孔测定板的每个孔中加入10微升(μL)2.5x激酶溶液(含原肌球蛋白受体激酶A(TRKA)、间变性淋巴瘤激酶(ALK)或原癌基因酪氨酸激酶(ROS1)蛋白终浓度分别为2.5nM，1.25nM，0.3nM)，在室温下孵育10分钟(min)。再向每孔中加入10微升(μL)的2.5x底物肽溶液(1x激酶碱性缓冲液中加入羟基荧光素(FAM)标记的底物肽和三磷酸腺苷(ATP)。同时设置对照组(最大值(Max)孔为只有DMSO无化合物组；最小值(Min)孔为不含原肌球蛋白受体激酶A(TRKA)、间变性淋巴瘤激酶(ALK)或原癌基因酪氨酸激酶(ROS1)蛋白组)，在28℃孵育一定时间后加入25微升(μL)终止缓冲液以停止激酶反应，利用Caliper仪器测定底物与产物的量，并计算出产物的转化率，进而计算出抑制率。Add 10 microliters (μL) of 2.5x kinase solution (containing tropomyosin receptor kinase A (TRKA), anaplastic lymphoma kinase (ALK) or proto-oncogene tyrosine to each well of the 384-well assay plate The final concentration of kinase (ROS1) protein was 2.5nM, 1.25nM, 0.3nM), and incubated at room temperature for 10 minutes (min). Then add 10 microliters (μL) of 2.5x substrate peptide solution (1x kinase alkaline buffer with hydroxyfluorescein (FAM) labeled substrate peptide and adenosine triphosphate (ATP) into each well. At the same time, set a control group ( The maximum (Max) hole is only DMSO without compound group; the minimum (Min) hole is without tropomyosin receptor kinase A (TRKA), anaplastic lymphoma kinase (ALK) or proto-oncogene tyrosine kinase (ROS1 protein group), after incubating at 28°C for a certain period of time, add 25 microliters (μL) of stop buffer to stop the kinase reaction, use the Caliper instrument to measure the amount of substrate and product, and calculate the conversion rate of the product, and then calculate Inhibition rate.
2.3数据处理及统计2.3 Data processing and statistics
将数据进行曲线拟合，计算半数抑制浓度(IC
50)值。抑制百分比＝(最大值(Max)孔转化率-每孔化合物转化率)/(最大值(Max)孔转化率-最小值(min)孔转化率)*100。“最大值(max)”代表二甲基亚砜(DMSO)不含化合物组；“最小值(min)”代表不含原肌球蛋白受体激酶A(TRKA)、间变性淋巴瘤激酶(ALK)或原癌基因酪氨酸激酶(ROS1)蛋白组。
The data was curve-fitted, and the median inhibitory concentration (IC 50 ) value was calculated. Inhibition percentage = (Maximum (Max) pore conversion rate-Per pore compound conversion rate) / (Maximum (Max) pore conversion rate-Minimum (min) pore conversion rate) * 100. "Maximum (max)" means that dimethyl sulfoxide (DMSO) does not contain the compound group; "minimum (min)" means that it does not contain tropomyosin receptor kinase A (TRKA), anaplastic lymphoma kinase (ALK) ) Or proto-oncogene tyrosine kinase (ROS1) proteome.
2.4测试结果和结论2.4 Test results and conclusions
本发明化合物对原肌球蛋白受体激酶A(TRKA)、间变性淋巴瘤激酶(ALK)、原癌基因酪氨酸激酶(ROS1)的活性测定的具体IC
50值见以下表1。由表1可知，本发明的化合物在体外生化实验水平可以有效抑制原肌球蛋白受体激酶A(TRKA)、间变性淋巴瘤激酶(ALK)、原癌基因酪氨酸激酶(ROS1)的活性。
The specific IC 50 values of the compounds of the present invention on the activity of tropomyosin receptor kinase A (TRKA), anaplastic lymphoma kinase (ALK), and proto-oncogene tyrosine kinase (ROS1) are shown in Table 1 below. It can be seen from Table 1 that the compounds of the present invention can effectively inhibit the activities of tropomyosin receptor kinase A (TRKA), anaplastic lymphoma kinase (ALK), and proto-oncogene tyrosine kinase (ROS1) at the level of in vitro biochemical experiments. .
表1.化合物对原肌球蛋白受体激酶A(TRKA)、间变性淋巴瘤激酶(ALK)、原 癌基因酪氨酸激酶(ROS1)活性结果Table 1. Activity results of compounds on tropomyosin receptor kinase A (TRKA), anaplastic lymphoma kinase (ALK), and proto-oncogene tyrosine kinase (ROS1)
实验二、本发明化合物对人间变性大细胞淋巴瘤细胞(Karpas299细胞)增殖抑制活性分析Experiment 2: Analysis of the compound of the present invention on the proliferation inhibitory activity of human anaplastic large cell lymphoma cells (Karpas299 cells)
1.实验目的和方法1. The purpose and method of the experiment
本实验用发光法细胞活力检测(
(CTG))法测试本发明化合物对人间变性大细胞淋巴瘤细胞(Karpas299细胞)的增殖抑制活性，根据半数抑制浓度IC
50评价化合物的细胞活性。
This experiment uses the luminescence method to detect cell viability ( (CTG)) compounds of present invention were tested lymphoma cell proliferation (Karpas299 cells) of human anaplastic large cell inhibitory activity, cellular activity IC 50 concentration of the compound according to the evaluation half inhibition.
2.实验方案2. Experimental protocol
2.1细胞培养2.1 Cell culture
人间变性大细胞淋巴瘤细胞(Karpas299)订购于南京科佰生物科技有限公司，使用洛斯维细胞培养基(RPMI 1640)(康宁，09919004)加10％的胎牛血清(Gibco，10817010)和1％青霉素/链霉素双抗(康宁，30002297)进行培养，显微镜下观察，确定细胞状态良好，将细胞转移到15毫升(mL)离心管中，1000转(rpm)离心5分钟，弃上清，加入完全培养基，吹打成单细胞悬液，置于37℃，5％二氧化碳(CO
2)培养箱(赛默飞，3111)中培养。
Human anaplastic large cell lymphoma cells (Karpas299) were ordered from Nanjing Kebai Biotechnology Co., Ltd., using Loswell Cell Medium (RPMI 1640) (Corning, 0919904) plus 10% fetal bovine serum (Gibco, 10817010) and 1% Penicillin/streptomycin double antibody (Kangning, 30002297) was cultured and observed under a microscope to confirm that the cells were in good condition. Transfer the cells to a 15 milliliter (mL) centrifuge tube, centrifuge at 1000 rpm for 5 minutes, and discard the supernatant. Add complete medium, pipette into a single cell suspension, and place it in a 37°C, 5% carbon dioxide (CO 2 ) incubator (Thermo Fisher, 3111).
2.2化合物的配制和化合物板的准备2.2 Compound preparation and compound board preparation
将待测化合物用二甲基亚砜(DMSO)配成10毫摩尔每升(mM)的工作液。测 试时的最高浓度为1微摩尔每升(μM)，:1：3进行稀释，共9个浓度梯度。The compound to be tested was prepared with dimethyl sulfoxide (DMSO) into a working solution of 10 millimoles per liter (mM). The highest concentration during the test is 1 micromole per liter (μM), with 1:3 dilution, a total of 9 concentration gradients.
2.3实验流程2.3 Experimental process
第一天：细胞铺板Day 1: Cell plating
取上述细胞悬液用细胞计数仪进行细胞计数，用完全培养基将细胞悬液调整到所需细胞密度然后接种至96孔板中，每孔90微升(μL)使细胞数为10000/孔，设置T0板，空白对照加入90微升(μL)完全培养基，置于37℃，5％二氧化碳(CO
2)培养箱(赛默飞，3111)中过夜。
Take the above cell suspension and use a cell counter to count the cells, adjust the cell suspension to the required cell density with complete medium and then inoculate it into a 96-well plate, 90 microliters (μL) per well to make the number of cells 10,000/well , Set the T0 plate, add 90 microliters (μL) of complete medium to the blank control, and place it in a 37°C, 5% carbon dioxide (CO 2 ) incubator (Invitrogen, 3111) overnight.
第二天：T0板读值Day 2: T0 board reading
每孔加入10微升(μL)含有溶媒的培养基后进行CTG分析，融化CTG试剂(普洛麦格，G7573)并平衡细胞板至室温30分钟，每孔加入50微升(μL)CTG溶液，在定轨摇床上振动2分钟使细胞裂解，把细胞板放置于室温10分钟以稳定冷光信号，用Envision(购买于珀金埃尔默(PerkinElmer))读取冷光值。Add 10 microliters (μL) of medium containing solvent to each well for CTG analysis, melt the CTG reagent (Promag, G7573) and equilibrate the cell plate to room temperature for 30 minutes, add 50 microliters (μL) of CTG solution to each well , Vibrate on an orbital shaker for 2 minutes to lyse the cells, place the cell plate at room temperature for 10 minutes to stabilize the luminescence signal, and read the luminescence value with Envision (purchased from PerkinElmer).
第二天：加入待测药Day 2: Add the drug to be tested
将，96孔细胞培养板中加入10微升(μL)稀释好的待测化合物溶液，每个浓度3个复孔并设置阴性对照组(不含化合物的溶剂对照组及不含细胞只有培养液的空白对照组)。在37℃、5％CO
2、95％湿度条件下进行细胞培养72小时(h)。
Add 10 microliters (μL) of the diluted test compound solution to a 96-well cell culture plate, 3 replicate wells for each concentration, and set a negative control group (solvent control without compound and only culture medium without cells Blank control group). Cell culture was performed for 72 hours (h) at 37°C, 5% CO 2, and 95% humidity.
第五天：试验板读值(药物处理72小时)Day 5: Reading the test board (72 hours of drug treatment)
融化CTG试剂(普洛麦格，G7573)并平衡细胞板至室温30分钟，每孔加入50微升(μL)CTG溶液，在定轨摇床上振动2分钟使细胞裂解，把细胞板放置于室温10分钟以稳定冷光信号，用Envision(购买于珀金埃尔默(PerkinElmer))读取冷光值。Melt CTG reagent (Promag, G7573) and equilibrate the cell plate to room temperature for 30 minutes, add 50 microliters (μL) of CTG solution to each well, shake on an orbital shaker for 2 minutes to lyse the cells, and place the cell plate at room temperature For 10 minutes to stabilize the luminescence signal, use Envision (purchased from PerkinElmer) to read the luminescence value.
2.4数据处理与统计2.4 Data processing and statistics
细胞增殖抑制率用公式：1-(V
sample-V
空白)/(V
control-V
空白)×100％计算。其中V
sample为药物处理组的读数值，V
空白为不含细胞只有培养液的空白对照的读数值，V
control为不含化合物的溶剂对照组的读数值。应用棱镜5(GraphPad Prism 5)软件，使用非线性回归模型绘制S型剂量-存活率曲线并计算化合物对细胞增殖的半抑制浓度(IC
50)值。
The cell proliferation inhibition rate is calculated with the formula: 1-(V sample -V blank )/(V control -V blank )×100%. Among them, V sample is the reading value of the drug treatment group, V blank is the reading value of the blank control without cells and only culture medium, and V control is the reading value of the solvent control group without the compound. Application Prism 5 (GraphPad Prism 5) software using a nonlinear regression model plotted dose-S - survival curves and calculate% inhibition of cell proliferation of the compound (IC 50) values.
2.5测试结果和结论2.5 Test results and conclusions
结果显示，实施例化合物对人间变性大细胞淋巴瘤细胞(Karpas299)增殖均有 较强的抑制作用，结果见表2。The results show that the compounds of the examples have a strong inhibitory effect on the proliferation of human anaplastic large cell lymphoma cells (Karpas299). The results are shown in Table 2.
实验三、本发明化合物对小鼠原B细胞稳转株BaF3 LMNA-NTRK1的增殖抑制活性分析Experiment 3: Analysis of the proliferation inhibitory activity of the compound of the present invention on the stable transgenic mouse B cell strain BaF3 LMNA-NTRK1
1.实验目的和方法1. The purpose and method of the experiment
本实验目的是为了测试本发明化合物对小鼠原B细胞稳转株BaF3LMNA-NTRK1的增殖抑制活性，根据半数抑制浓度IC
50评价化合物的细胞活性。
Purpose of this study was to test the compounds of the present invention, pro-B cell line stably transfected mouse BaF3LMNA-NTRK1 proliferation inhibitory activity, the activity of cell 50 according to the evaluation of compound half inhibition concentration IC.
2.实验方案2. Experimental protocol
2.1细胞培养2.1 Cell culture
小鼠原B细胞稳转株BaF3 LMNA-NTRK1订购于康源博创生物科技(北京)有限公司，使用洛斯维细胞培养基(RPMI 1640)(康宁，09919004)加10％的胎牛血清(Gibco，10817010)和1％青霉素/链霉素双抗(康宁，30002297)进行培养，显微镜下观察，确定细胞状态良好，将细胞转移到15毫升(mL)离心管中，1000转(rpm)离心5分钟，弃上清，加入完全培养基，吹打成单细胞悬液，置于37℃，5％二氧化碳(CO
2)培养箱(赛默飞，3111)中培养。
The mouse original B cell stable transgenic strain BaF3 LMNA-NTRK1 was ordered from Kangyuan Bochuang Biotechnology (Beijing) Co., Ltd., using Losvi Cell Medium (RPMI 1640) (Corning, 0919904) plus 10% fetal bovine serum (Gibco) , 10817010) and 1% penicillin/streptomycin double antibody (Kangning, 30002297) were cultured, observed under a microscope to confirm that the cells are in good condition, transfer the cells to a 15 milliliter (mL) centrifuge tube, and centrifuge at 1000 revolutions (rpm) for 5 Minutes, discard the supernatant, add complete medium, pipette to form a single cell suspension, and place it in a 37°C, 5% carbon dioxide (CO 2 ) incubator (Invitrogen, 3111).
2.2化合物的配制和化合物板的准备2.2 Compound preparation and compound board preparation
将待测化合物用二甲基亚砜(DMSO)配成10毫摩尔每升(mM)的工作液。测试时的最高浓度为1微摩尔每升(μM)，1：3进行稀释，共9个浓度梯度。The compound to be tested was prepared with dimethyl sulfoxide (DMSO) into a working solution of 10 millimoles per liter (mM). The highest concentration in the test is 1 micromole per liter (μM), and the dilution is 1:3, a total of 9 concentration gradients.
2.3实验流程2.3 Experimental process
第一天：细胞铺板Day 1: Cell plating
取上述细胞悬液用细胞计数仪进行细胞计数，用完全培养基将细胞悬液调整到所需细胞密度然后接种至96孔板中，每孔90微升(μL)使细胞数为10000/孔，设置T0板，空白对照加入90微升(μL)完全培养基，置于37℃，5％二氧化碳(CO
2)培养箱(赛默飞，3111)中过夜。
Take the above cell suspension and use a cell counter to count the cells, adjust the cell suspension to the required cell density with complete medium and then inoculate it into a 96-well plate, 90 microliters (μL) per well to make the number of cells 10,000/well , Set the T0 plate, add 90 microliters (μL) of complete medium to the blank control, and place it in a 37°C, 5% carbon dioxide (CO 2 ) incubator (Invitrogen, 3111) overnight.
第二天：T0板读值Day 2: T0 board reading
每孔加入10微升(μL)含有溶媒的培养基后进行CTG分析，融化CTG试剂(普洛麦格，G7573)并平衡细胞板至室温30分钟，每孔加入50微升(μL)CTG溶液，在定轨摇床上振动2分钟使细胞裂解，把细胞板放置于室温10分钟以稳定冷光信号，用Envision(购买于珀金埃尔默(PerkinElmer))读取冷光值。Add 10 microliters (μL) of medium containing solvent to each well for CTG analysis, melt the CTG reagent (Promag, G7573) and equilibrate the cell plate to room temperature for 30 minutes, add 50 microliters (μL) of CTG solution to each well , Vibrate on an orbital shaker for 2 minutes to lyse the cells, place the cell plate at room temperature for 10 minutes to stabilize the luminescence signal, and read the luminescence value with Envision (purchased from PerkinElmer).
第二天：加入待测药Day 2: Add the drug to be tested
96孔细胞培养板中加入10微升(μL)稀释好的待测化合物溶液，每个浓度3个复孔并设置阴性对照组(不含化合物的溶剂对照组及不含细胞只有培养液的空白对照组)。在37℃、5％CO
2、95％湿度条件下进行细胞培养72小时(h)。
Add 10 microliters (μL) of the diluted test compound solution to the 96-well cell culture plate, 3 replicate wells for each concentration, and set a negative control group (solvent control group without compound and blank without cell only culture medium Control group). Cell culture was performed for 72 hours (h) at 37°C, 5% CO 2, and 95% humidity.
第五天：试验板读值(药物处理72小时)Day 5: Reading the test board (72 hours of drug treatment)
融化CTG试剂(普洛麦格，G7573)并平衡细胞板至室温30分钟，每孔加入50微升(μL)CTG溶液，在定轨摇床上振动2分钟使细胞裂解，把细胞板放置于室温10分钟以稳定冷光信号，用Envision(购买于珀金埃尔默(PerkinElmer))读取冷光值。Melt CTG reagent (Promag, G7573) and equilibrate the cell plate to room temperature for 30 minutes, add 50 microliters (μL) of CTG solution to each well, shake on an orbital shaker for 2 minutes to lyse the cells, and place the cell plate at room temperature For 10 minutes to stabilize the luminescence signal, use Envision (purchased from PerkinElmer) to read the luminescence value.
2.4数据处理与统计2.4 Data processing and statistics
细胞增殖抑制率用公式：1-(V
sample-V
空白)/(V
control-V
空白)×100％计算。其中V
sample为药物处理组的读数值，V
空白为不含细胞只有培养液的空白对照的读数值，V
control为不含化合物的溶剂对照组的读数值。应用棱镜5(GraphPad Prism 5)软件，使用非线性回归模型绘制S型剂量-存活率曲线并计算化合物对细胞增殖的半抑制浓度(IC
50)值。
The cell proliferation inhibition rate is calculated with the formula: 1-(V sample -V blank )/(V control -V blank )×100%. Among them, V sample is the reading value of the drug treatment group, V blank is the reading value of the blank control without cells and only culture medium, and V control is the reading value of the solvent control group without the compound. Application Prism 5 (GraphPad Prism 5) software using a nonlinear regression model plotted dose-S - survival curves and calculate% inhibition of cell proliferation of the compound (IC 50) values.
2.5测试结果和结论2.5 Test results and conclusions
结果显示，实施例化合物对小鼠原B细胞稳转株BaF3 LMNA-NTRK1增殖均有较强的抑制作用，结果见表2。The results show that the compounds of the examples have a strong inhibitory effect on the proliferation of the stable mouse original B cell strain BaF3 LMNA-NTRK1. The results are shown in Table 2.
实验四、本发明化合物对小鼠原B细胞稳转株BaF3 SLC34A2-ROS1的增殖抑制活性分析Experiment 4. Analysis of the proliferation inhibitory activity of the compound of the present invention on the stable transgenic mouse B cell strain BaF3 SLC34A2-ROS1
1.实验目的和方法1. The purpose and method of the experiment
本实验目的是为了测试本发明化合物对小鼠原B细胞稳转株BaF3SLC34A2-ROS1的增殖抑制活性，根据半数抑制浓度IC
50评价化合物的细胞活性。
Purpose of this study was to test the compounds of the present invention, pro-B cell line stably transfected mouse BaF3SLC34A2-ROS1 proliferation inhibitory activity, the activity of cell 50 according to the evaluation of compound half inhibition concentration IC.
2.实验方案2. Experimental protocol
2.1细胞培养2.1 Cell culture
小鼠原B细胞稳转株BaF3 SLC34A2-ROS1稳转细胞株订购于康源博创生物科技(北京)有限公司，使用洛斯维细胞培养基(RPMI 1640)(康宁，09919004)加10％的胎牛血清(Gibco，10817010)和1％青霉素/链霉素双抗(康宁，30002297)进行培养，显微镜下观察，确定细胞状态良好，将细胞转移到15毫升(mL)离心管中， 1000转(rpm)离心5分钟，弃上清，加入完全培养基，吹打成单细胞悬液，置于37℃，5％二氧化碳(CO
2)培养箱(赛默飞，3111)中培养。
The stable transgenic mouse B cell strain BaF3 SLC34A2-ROS1 was ordered from Kangyuan Bochuang Biotechnology (Beijing) Co., Ltd., using Losvi Cell Medium (RPMI 1640) (Corning, 0919904) plus 10% fetus Bovine serum (Gibco, 10817010) and 1% penicillin/streptomycin double antibody (Corning, 30002297) were cultured and observed under a microscope to confirm that the cells were in good condition. The cells were transferred to a 15 milliliter (mL) centrifuge tube at 1000 revolutions ( rpm) centrifugation for 5 minutes, discard the supernatant, add complete medium, pipette to form a single cell suspension, and place it in a 37°C, 5% carbon dioxide (CO 2 ) incubator (Invitrogen, 3111) for culture.
2.2化合物的配制和化合物板的准备2.2 Compound preparation and compound board preparation
将待测化合物用二甲基亚砜(DMSO)配成10毫摩尔每升(mM)的工作液。测试时的最高浓度为1微摩尔每升(μM)，1：3进行稀释，共9个浓度梯度。The compound to be tested was prepared with dimethyl sulfoxide (DMSO) into a working solution of 10 millimoles per liter (mM). The highest concentration in the test is 1 micromole per liter (μM), and the dilution is 1:3, a total of 9 concentration gradients.
2.3实验流程2.3 Experimental process
第一天：细胞铺板Day 1: Cell plating
取上述细胞悬液用细胞计数仪进行细胞计数，用完全培养基将细胞悬液调整到所需细胞密度然后接种至96孔板中，每孔90微升(μL)使细胞数为10000/孔，设置T0板，空白对照加入90微升(μL)完全培养基，置于37℃，5％二氧化塔(CO
2)培养箱(赛默飞，3111)中过夜。
Take the above cell suspension and use a cell counter to count the cells, adjust the cell suspension to the required cell density with complete medium and then inoculate it into a 96-well plate, 90 microliters (μL) per well to make the number of cells 10,000/well , Set the T0 plate, add 90 microliters (μL) of complete medium to the blank control, and place it in a 37°C, 5% CO 2 incubator (Thermo Scientific, 3111) overnight.
第二天：T0板读值Day 2: T0 board reading
每孔加入10微升(μL)含有溶媒的培养基后进行CTG分析，融化CTG试剂(普洛麦格，G7573)并平衡细胞板至室温30分钟，每孔加入50微升(μL)CTG溶液，在定轨摇床上振动2分钟使细胞裂解，把细胞板放置于室温10分钟以稳定冷光信号，用Envision(购买于珀金埃尔默(PerkinElmer))读取冷光值。Add 10 microliters (μL) of medium containing solvent to each well for CTG analysis, melt the CTG reagent (Promag, G7573) and equilibrate the cell plate to room temperature for 30 minutes, add 50 microliters (μL) of CTG solution to each well , Vibrate on an orbital shaker for 2 minutes to lyse the cells, place the cell plate at room temperature for 10 minutes to stabilize the luminescence signal, and read the luminescence value with Envision (purchased from PerkinElmer).
第二天：加入待测药Day 2: Add the drug to be tested
96孔细胞培养板中加入10微升(μL)稀释好的待测化合物溶液，每个浓度3个复孔并设置阴性对照组(不含化合物的溶剂对照组及不含细胞只有培养液的空白对照组)。在37℃、5％CO
2、95％湿度条件下进行细胞培养72小时(h)。
Add 10 microliters (μL) of the diluted test compound solution to the 96-well cell culture plate, 3 replicate wells for each concentration, and set a negative control group (solvent control group without compound and blank without cell only culture medium Control group). Cell culture was performed for 72 hours (h) at 37°C, 5% CO 2, and 95% humidity.
第五天：试验板读值(药物处理72小时)Day 5: Reading the test board (72 hours of drug treatment)
融化CTG试剂(普洛麦格，G7573)并平衡细胞板至室温30分钟，每孔加入50微升(μL)CTG溶液，在定轨摇床上振动2分钟使细胞裂解，把细胞板放置于室温10分钟以稳定冷光信号，用Envision(购买于珀金埃尔默(PerkinElmer))读取冷光值。Melt CTG reagent (Promag, G7573) and equilibrate the cell plate to room temperature for 30 minutes, add 50 microliters (μL) of CTG solution to each well, shake on an orbital shaker for 2 minutes to lyse the cells, and place the cell plate at room temperature For 10 minutes to stabilize the luminescence signal, use Envision (purchased from PerkinElmer) to read the luminescence value.
2.4数据处理与统计2.4 Data processing and statistics
细胞增殖抑制率用公式：1-(V
sample-V
空白)/(V
control-V
空白)×100％计算。其中V
sample为药物处理组的读数值，V
空白为不含细胞只有培养液的空白对照的读数值，V
control为不含化合物的溶剂对照组的读数值。应用棱镜5(GraphPad Prism 5)软件， 使用非线性回归模型绘制S型剂量-存活率曲线并计算化合物对细胞增殖的半抑制浓度(IC
50)值。
The cell proliferation inhibition rate is calculated with the formula: 1-(V sample -V blank )/(V control -V blank )×100%. Among them, V sample is the reading value of the drug treatment group, V blank is the reading value of the blank control without cells and only culture medium, and V control is the reading value of the solvent control group without the compound. Application Prism 5 (GraphPad Prism 5) software using a nonlinear regression model plotted dose-S - survival curves and calculate% inhibition of cell proliferation of the compound (IC 50) values.
2.5测试结果和结论2.5 Test results and conclusions
结果显示，实施例化合物对小鼠原B细胞稳转株BaF3 SLC34A2-ROS1增殖均有较强的抑制作用，结果见表2。The results show that the compounds of the examples have a strong inhibitory effect on the proliferation of the stable mouse original B cell strain BaF3 SLC34A2-ROS1. The results are shown in Table 2.
表2.化合物对细胞增殖的抑制作用Table 2. Inhibitory effects of compounds on cell proliferation
实验五、本发明化合物在大鼠中药物代谢动力学实验Experiment 5. Pharmacokinetics experiment of the compound of the present invention in rats
1.摘要1. Summary
以体重在200-300g、8周龄雄性斯泼累格·多雷(Sprague Dawley，SD)大鼠为实验动物，应用液相色谱-质谱/质谱联用(LC/MS/MS)技术测定静脉、口服给予实施例2化合物、实施例4化合物、实施例6化合物、实施例9化合物、实施例10化合物、实施例14化合物、实施例16化合物、实施例17化合物、实施例18化合物、实施例20化合物、实施例21化合物、实施例23化合物、实施例24化合物、实施例29化合物、实施例31化合物、实施例32化合物、实施例33化合物、实施例35化合物、实施例37化合物、实施例38化合物、实施例39化合物、实施例40化合物、实施例42化合物、实施例44化合物、实施例45化合物、实 施例54化合物、实施例55化合物、实施例59化合物、实施例61化合物、实施例67化合物、实施例70化合物、实施例73化合物、实施例84化合物后，不同时刻血浆中的药物浓度并计算相关药代参数。研究本发明的化合物在大鼠体内的药代动力学行为，评价其药动学特征。Male Sprague Dawley (SD) rats weighing 200-300g and 8 weeks old were used as experimental animals, and the veins were measured using liquid chromatography-mass spectrometry/mass spectrometry (LC/MS/MS) technology , Oral administration of Example 2 compound, Example 4 compound, Example 6 compound, Example 9 compound, Example 10 compound, Example 14 compound, Example 16 compound, Example 17 compound, Example 18 compound, Example 20 compound, example 21 compound, example 23 compound, example 24 compound, example 29 compound, example 31 compound, example 32 compound, example 33 compound, example 35 compound, example 37 compound, example 38 compound, Example 39 compound, Example 40 compound, Example 42 compound, Example 44 compound, Example 45 compound, Example 54 compound, Example 55 compound, Example 59 compound, Example 61 compound, Example After 67 compound, Example 70 compound, Example 73 compound, and Example 84 compound, the concentration of the drug in plasma at different times was calculated and related pharmacokinetic parameters were calculated. Study the pharmacokinetic behavior of the compound of the present invention in rats and evaluate its pharmacokinetic characteristics.
2.实验方案2. Experimental protocol
2.1实验用化合物2.1 Experimental compounds
测定实施例2化合物、实施例4化合物、实施例6化合物、实施例9化合物、实施例10化合物、实施例14化合物、实施例16化合物、实施例17化合物、实施例18化合物、实施例20化合物、实施例21化合物、实施例23化合物、实施例24化合物、实施例29化合物、实施例31化合物、实施例32化合物、实施例33化合物、实施例35化合物、实施例37化合物、实施例38化合物、实施例39化合物、实施例40化合物、实施例42化合物、实施例44化合物、实施例45化合物、实施例54化合物、实施例55化合物、实施例59化合物、实施例61化合物、实施例67化合物、实施例70化合物、实施例73化合物、实施例84化合物。Determination of the compound of Example 2, the compound of Example 4, the compound of Example 6, the compound of Example 9, the compound of Example 10, the compound of Example 14, the compound of Example 16, the compound of Example 17, the compound of Example 18, the compound of Example 20 , Example 21 compound, Example 23 compound, Example 24 compound, Example 29 compound, Example 31 compound, Example 32 compound, Example 33 compound, Example 35 compound, Example 37 compound, Example 38 compound , Example 39 compound, Example 40 compound, Example 42 compound, Example 44 compound, Example 45 compound, Example 54 compound, Example 55 compound, Example 59 compound, Example 61 compound, Example 67 compound , Example 70 compound, Example 73 compound, and Example 84 compound.
2.2化合物的配制2.2 Compound preparation
称取一定量的化合物，静脉给药用10％吐温80于生理盐水溶解，口服给药用0.5％甲基纤维素于水中溶解，配制成均一的溶液。A certain amount of the compound is weighed, 10% Tween 80 is dissolved in physiological saline for intravenous administration, and 0.5% methylcellulose is dissolved in water for oral administration to prepare a uniform solution.
2.3采血时间及样品处理2.3 Blood collection time and sample processing
以10毫克每千克(mg/kg)的剂量对大鼠静脉和口服给予上述化合物，在给药后0.083、0.25、0.5、1、2、4、8、24小时通过眼眶采血，每次采血0.2毫升(mL)，至于乙二胺四乙酸二钾(EDTA-2K)抗凝管中，4℃，6000转/分钟离心10分钟分离血浆，-80℃保存。The above compounds were administered intravenously and orally to rats at a dose of 10 milligrams per kilogram (mg/kg). Blood was collected through the orbit at 0.083, 0.25, 0.5, 1, 2, 4, 8, 24 hours after administration, and blood was collected at 0.2 per time. Milliliters (mL), as for the dipotassium ethylenediaminetetraacetate (EDTA-2K) anticoagulation tube, centrifuge at 6000 rpm at 4°C for 10 minutes to separate the plasma, and store at -80°C.
2.4实验结果及结论2.4 Experimental results and conclusions
本发明的化合物给药后药代动力学参数见以下表3。由表3所示，本发明的化合物具有较好的代谢特征及较好的生物利用度。The pharmacokinetic parameters of the compound of the present invention after administration are shown in Table 3 below. As shown in Table 3, the compounds of the present invention have better metabolic characteristics and better bioavailability.
表3.SD大鼠经实施例化合物静脉、口服给药后药代动力学参数Table 3. Pharmacokinetic parameters of SD rats after intravenous and oral administration of the compounds of the examples
实验六、本发明实施例4、6、17、20、21、23、24、29、37、39、40、45、69、82化合物在人间变性大细胞淋巴瘤细胞(Karpas299细胞)小鼠移植瘤模型中的药效实验Experiment VI. Example 4, 6, 17, 20, 21, 23, 24, 29, 37, 39, 40, 45, 69, 82 compounds transplanted in mice of human anaplastic large cell lymphoma cells (Karpas299 cells) Pharmacodynamic experiment in tumor model
1.摘要1. Summary
以体重在16g-18g的8周龄雌性重症联合免疫缺陷(CB-17 SCID)小鼠为实验动物，测定实施例4化合物、实施例6化合物、实施例17化合物、实施例20化合物、实施例21化合物、实施例23化合物、实施例24化合物、实施例29化合物、实施例37化合物、实施例39化合物、实施例40化合物、实施例45化合物、实施例69化合物、实施例82化合物灌胃给予移植瘤小鼠后对小鼠肿瘤的药效结果。探讨发明化合物对肿瘤生长的影响。Using 8-week-old female severe combined immunodeficiency (CB-17 SCID) mice weighing 16g-18g as experimental animals, the compound of Example 4, the compound of Example 6, the compound of Example 17, and the compound of Example 20 were tested. 21 Compound, Example 23 Compound, Example 24 Compound, Example 29 Compound, Example 37 Compound, Example 39 Compound, Example 40 Compound, Example 45 Compound, Example 69 Compound, and Example 82 Compound The results of the drug effect on mouse tumors after transplantation of tumor mice. Explore the effect of the compound of the invention on tumor growth.
2.实验方案2. Experimental protocol
2.1实验用化合物2.1 Experimental compounds
实施例4化合物、实施例6化合物、实施例17化合物、实施例20化合物、实施例21化合物、实施例23化合物、实施例24化合物、实施例29化合物、实施例37化合物、实施例39化合物、实施例40化合物、实施例45化合物、实施例69化合物、实施例82化合物。Example 4 compound, Example 6 compound, Example 17 compound, Example 20 compound, Example 21 compound, Example 23 compound, Example 24 compound, Example 29 compound, Example 37 compound, Example 39 compound, The compound of Example 40, the compound of Example 45, the compound of Example 69, and the compound of Example 82.
2.2化合物的配制2.2 Compound preparation
称取一定量的化合物，溶于10％吐温80的生理盐水溶液，配制成均一的溶液。Weigh a certain amount of the compound and dissolve it in a 10% Tween 80 saline solution to prepare a uniform solution.
2.3细胞的培养2.3 Cell culture
人间变性大细胞淋巴瘤细胞(Karpas299细胞)订购于南京科佰生物科技有限公司，使用洛斯维细胞培养基(RPMI 1640)(康宁，09919004)加10％的胎牛血清 (Gibco，10817010)和1％青霉素/链霉素双抗(康宁，30002297)进行培养，显微镜下观察，确定细胞状态良好，将细胞转移到15毫升(mL)离心管中，1000转(rpm)离心5分钟，弃上清，加入完全培养基，吹打成单细胞悬液，置于37℃，5％二氧化碳(CO
2)培养箱(赛默飞，3111)中培养。
Human anaplastic large cell lymphoma cells (Karpas299 cells) were ordered from Nanjing Kebai Biotechnology Co., Ltd., using Losvi Cell Medium (RPMI 1640) (Corning, 0919904) plus 10% fetal bovine serum (Gibco, 10817010) and 1 % Penicillin/streptomycin double antibody (Kangning, 30002297) was cultured and observed under a microscope to confirm that the cells were in good condition. Transfer the cells to a 15 ml centrifuge tube, centrifuge at 1000 rpm for 5 minutes, and discard the supernatant , Add complete medium, pipette into a single cell suspension, and place it in a 37°C, 5% carbon dioxide (CO 2 ) incubator (Thermo Fisher, 3111).
2.4实验流程2.4 Experimental procedure
无菌条件下，将对数生长期的人间变性大细胞淋巴瘤细胞(Karpas299细胞)与基质胶(Matrigel)(康宁，8274014)混合后移植于雌性重症联合免疫缺陷(CB-17 SCID)小鼠背部右侧皮下，每只小鼠接种3×10
6个细胞，体积100微升(μL)，接种后，待肿瘤体积生长到150立方毫米(mm
3)时，将小鼠随机分成15组，每组6只进行体内药效实验，阴性对照组给等量的溶媒。具体设计见表4。
Under sterile conditions, the logarithmic growth phase of human anaplastic large cell lymphoma cells (Karpas299 cells) and Matrigel (Corning, 8274014) were mixed and transplanted into female severe combined immunodeficiency (CB-17 SCID) mice Subcutaneously on the right side of the back, each mouse was inoculated with 3×10 6 cells in a volume of 100 microliters (μL). After the inoculation, when the tumor volume grew to 150 cubic millimeters (mm 3 ), the mice were randomly divided into 15 groups. Six animals in each group were tested for in vivo efficacy, and the negative control group was given the same amount of solvent. The specific design is shown in Table 4.
表4.化合物体内药效试验方案Table 4. Compound in vivo efficacy test protocol
2.5实验结果及结论2.5 Experimental results and conclusions
根据公式计算小鼠肿瘤体积＝长×宽×宽/2(mm
3)，抑瘤率的公式＝1-(终点时给药组肿瘤体积-给药组分组时肿瘤体积)/(终点时对照组肿瘤体积-对照组分组时肿瘤体积)×100％，实施例化合物4、6、17、20、21、23、24、29、37、39、40、45、69、82在10毫克每千克(mg/kg)的给药浓度下对肿瘤生长抑制率分别达到96.25％、99.59％、96.94％、97.21％、98.49％、95.18％、95.35％、96.67％、96.49％、98.11％、95.66％、97.23％、96.41％、97.53％；表明本发明实施例化合物4、6、17、20、21、23、24、29、37、39、40、45、69、82在人间变性大细胞淋巴瘤细胞(Karpas299细胞)异种移植模型中具有较强的肿瘤生长抑制作用。
Calculate the mouse tumor volume according to the formula = length × width × width / 2 (mm 3 ), the formula of tumor inhibition rate = 1-(the tumor volume in the administration group at the end point-the tumor volume in the administration component group) / (the control at the end point Group tumor volume-tumor volume at the time of grouping in the control group)×100%. Example compounds 4, 6, 17, 20, 21, 23, 24, 29, 37, 39, 40, 45, 69, 82 are at 10 mg/kg (mg/kg) administration concentration of tumor growth inhibition rate reached 96.25%, 99.59%, 96.94%, 97.21%, 98.49%, 95.18%, 95.35%, 96.67%, 96.49%, 98.11%, 95.66%, 97.23%, 96.41%, 97.53%; indicating that the compounds 4, 6, 17, 20, 21, 23, 24, 29, 37, 39, 40, 45, 69, 82 of the examples of the present invention are in human anaplastic large cell lymphoma cells (Karpas299 cells) has a strong tumor growth inhibitory effect in the xenograft model.
实验七、本发明实施例2、9、10、14、16、18、29、33、42、45、55、59、72、80化合物在小鼠原B细胞稳转株BaF3 LMNA-NTRK1小鼠移植瘤模型中的药效实验Experiment VII. Example 2, 9, 10, 14, 16, 18, 29, 33, 42, 45, 55, 59, 72, and 80 of the compound were stably transformed into the mouse original B cell strain BaF3 LMNA-NTRK1 mouse Pharmacodynamic experiment in transplanted tumor model
1.摘要1. Summary
以体重在16g-18g的8周龄雌性重症联合免疫缺陷(CB-17 SCID)小鼠为实验动物，测定实施例2化合物、实施例9化合物、实施例10化合物、实施例14化合物、实施例16化合物、实施例18化合物、实施例29化合物、实施例33化合物、实施例42化合物、实施例45化合物、实施例55化合物、实施例59化合物、实施例72化合物、实施例80化合物灌胃给予移植瘤小鼠后对小鼠肿瘤的药效结果。探讨发明化合物对肿瘤生长的影响。Using 8-week-old female severe combined immunodeficiency (CB-17 SCID) mice weighing 16g-18g as experimental animals, the compounds of Example 2, Example 9, Compound 10, Compound 14 and Example Compound 16, Compound of Example 18, Compound of Example 29, Compound of Example 33, Compound of Example 42, Compound of Example 45, Compound of Example 55, Compound of Example 59, Compound of Example 72, Compound of Example 80 The results of the drug effect on mouse tumors after transplanting tumor mice. Explore the effect of the compound of the invention on tumor growth.
2.实验方案2. Experimental protocol
2.1实验用化合物2.1 Experimental compounds
实施例2化合物、实施例9化合物、实施例10化合物、实施例14化合物、实施例16化合物、实施例18化合物、实施例29化合物、实施例33化合物、实施例42化合物、实施例45化合物、实施例55化合物、实施例59化合物、实施例72化合物、实施例80化合物。Example 2 compound, Example 9 compound, Example 10 compound, Example 14 compound, Example 16 compound, Example 18 compound, Example 29 compound, Example 33 compound, Example 42 compound, Example 45 compound, The compound of Example 55, the compound of Example 59, the compound of Example 72, and the compound of Example 80.
2.2化合物的配制2.2 Compound preparation
称取一定量的化合物，溶于10％吐温80的生理盐水溶液，配制成均一的溶液。Weigh a certain amount of the compound and dissolve it in a 10% Tween 80 saline solution to prepare a uniform solution.
2.3细胞的培养2.3 Cell culture
小鼠原B细胞稳转株BaF3 LMNA-NTRK1订购于康源博创生物科技(北京)有限公司，使用洛斯维细胞培养基(RPMI 1640)(康宁，09919004)加10％的胎牛血清(Gibco，10817010)和1％青霉素/链霉素双抗(康宁，30002297)进行培养，显微镜下观察，确定细胞状态良好，将细胞转移到15毫升(mL)离心管中，1000转(rpm)离心5分钟，弃上清，加入完全培养基，吹打成单细胞悬液，置于37℃，5％二氧化碳(CO
2)培养箱(赛默飞，3111)中培养。
The mouse original B cell stable transgenic strain BaF3 LMNA-NTRK1 was ordered from Kangyuan Bochuang Biotechnology (Beijing) Co., Ltd., using Losvi Cell Medium (RPMI 1640) (Corning, 0919904) plus 10% fetal bovine serum (Gibco) , 10817010) and 1% penicillin/streptomycin double antibody (Kangning, 30002297) were cultured, observed under a microscope to confirm that the cells are in good condition, transfer the cells to a 15 milliliter (mL) centrifuge tube, and centrifuge at 1000 revolutions (rpm) for 5 Minutes, discard the supernatant, add complete medium, pipette to form a single cell suspension, and place it in a 37°C, 5% carbon dioxide (CO 2 ) incubator (Invitrogen, 3111).
2.4实验流程2.4 Experimental procedure
无菌条件下，将对数生长期的小鼠原B细胞稳转株BaF3 LMNA-NTRK1与基质胶(Matrigel)(康宁，8274014)混合后移植于雌性重症联合免疫缺陷(CB-17 SCID)小鼠背部右侧皮下，每只小鼠接种5×10
6个细胞，体积100微升(μL)，接种后，待肿瘤体积生长到150立方毫米(mm
3)时，将小鼠随机分成15组，每组6只进行体内药效实验，阴性对照组给等量的溶媒。具体设计见表5。
Under aseptic conditions, the logarithmic growth phase mouse original B cell stable transgenic strain BaF3 LMNA-NTRK1 was mixed with Matrigel (Kang Ning, 8274014) and then transplanted into female severe combined immunodeficiency (CB-17 SCID) small Subcutaneously on the right side of the back of the mouse, each mouse was inoculated with 5×10 6 cells in a volume of 100 microliters (μL). After the inoculation, when the tumor volume grew to 150 cubic millimeters (mm 3 ), the mice were randomly divided into 15 groups , 6 animals in each group were tested in vivo, and the negative control group was given the same amount of solvent. The specific design is shown in Table 5.
表5.化合物体内药效试验方案Table 5. Compound in vivo efficacy test protocol
2.5实验结果及结论2.5 Experimental results and conclusions
根据公式计算小鼠肿瘤体积＝长×宽×宽/2(mm
3)，抑瘤率公式＝1-(终点时给药组肿瘤体积-给药组分组时肿瘤体积)/(终点时对照组肿瘤体积-对照组分组时肿瘤体积)×100％，实施例化合物2、9、10、14、16、18、29、33、42、45、55、59、72、80在10毫克每千克(mg/kg)的给药浓度下对肿瘤生长抑制率分别达到96.25％、97.49％、96.34％、97.17％、98.23％、97.35％、95.25％、96.46％、96.20％、97.42％、97.23％、98.36％、96.34％、98.06％；表明本发明实施例化合物2、9、10、14、16、18、29、33、42、45、55、59、72、80在小鼠原B细胞稳转株BaF3LMNA-NTRK1异种移植模型中具有较强的肿瘤生长抑制作用。
Calculate the mouse tumor volume according to the formula = length × width × width / 2 (mm 3 ), tumor inhibition rate formula = 1-(the tumor volume at the end of the administration group-the tumor volume at the end of the administration group) / (the control group at the end Tumor volume-tumor volume at the time of grouping in the control group) × 100%, the example compounds 2, 9, 10, 14, 16, 18, 29, 33, 42, 45, 55, 59, 72, 80 are at 10 mg/kg ( mg/kg) the tumor growth inhibition rate reached 96.25%, 97.49%, 96.34%, 97.17%, 98.23%, 97.35%, 95.25%, 96.46%, 96.20%, 97.42%, 97.23%, 98.36 %, 96.34%, 98.06%; indicating that the compounds 2, 9, 10, 14, 16, 18, 29, 33, 42, 45, 55, 59, 72, 80 of the examples of the present invention are stable in mouse B cell strains BaF3LMNA-NTRK1 has a strong tumor growth inhibitory effect in the xenograft model.
实验八、本发明实施例4、9、14、17、18、21、29、33、42、45、59、61、76、83化合物在小鼠原B细胞稳转株Ba/F3 SLC34A2-ROS1小鼠移植瘤模型中的药效实验Experiment 8. The compounds of Examples 4, 9, 14, 17, 18, 21, 29, 33, 42, 45, 59, 61, 76, and 83 of the present invention were stably transformed into the mouse original B cell strain Ba/F3 SLC34A2-ROS1 Pharmacodynamic experiment in mouse transplanted tumor model
1.摘要1. Summary
以体重在16g-18g的8周龄雌性重症联合免疫缺陷(CB-17 SCID)小鼠为实验动物，测定实施例4化合物、实施例9化合物、实施例14化合物、实施例17化合物、实施例18化合物、实施例21化合物、实施例29化合物、实施例33化合物、实施例42化合物、实施例45化合物、实施例59化合物、实施例61化合物、实施例76化合物、实施例83化合物灌胃给予移植瘤小鼠后对小鼠肿瘤的药效结果。探讨发明化合物对肿瘤生长的影响。Using 8-week-old female severe combined immunodeficiency (CB-17 SCID) mice weighing 16g-18g as experimental animals, the compound of Example 4, the compound of Example 9, the compound of Example 14 and the compound of Example 17 were tested. 18 compound, Example 21 compound, Example 29 compound, Example 33 compound, Example 42 compound, Example 45 compound, Example 59 compound, Example 61 compound, Example 76 compound, and Example 83 compound were administered intragastrically The results of the drug effect on mouse tumors after transplanting tumor mice. Explore the effect of the compound of the invention on tumor growth.
2.实验方案2. Experimental protocol
2.1实验用化合物2.1 Experimental compounds
实施例4化合物、实施例9化合物、实施例14化合物、实施例17化合物、实施例18化合物、实施例21化合物、实施例29化合物、实施例33化合物、实施例42化合物、实施例45化合物、实施例59化合物、实施例61化合物、实施例76化合物、实施例83化合物。Example 4 compound, Example 9 compound, Example 14 compound, Example 17 compound, Example 18 compound, Example 21 compound, Example 29 compound, Example 33 compound, Example 42 compound, Example 45 compound, The compound of Example 59, the compound of Example 61, the compound of Example 76, and the compound of Example 83.
2.2化合物的配制2.2 Compound preparation
称取一定量的化合物，溶于10％吐温80的生理盐水溶液，配制成均一的溶液。Weigh a certain amount of the compound and dissolve it in a 10% Tween 80 saline solution to prepare a uniform solution.
2.3细胞的培养2.3 Cell culture
小鼠原B细胞稳转株Ba/F3 SLC34A2-ROS1订购于康源博创生物科技(北京)有限公司，使用洛斯维细胞培养基(RPMI 1640)(康宁，09919004)加10％的胎牛血清(Gibco，10817010)和1％青霉素/链霉素双抗(康宁，30002297)进行培养，显微镜下观察，确定细胞状态良好，将细胞转移到15毫升(mL)离心管中，1000转(rpm)离心5分钟，弃上清，加入完全培养基，吹打成单细胞悬液，置于37℃，5％二氧化碳(CO
2)培养箱(赛默飞，3111)中培养。
The mouse original B cell stable transgenic strain Ba/F3 SLC34A2-ROS1 was ordered from Kangyuan Bochuang Biotechnology (Beijing) Co., Ltd., using Losvi Cell Culture Medium (RPMI 1640) (Corning, 0919904) plus 10% fetal bovine serum (Gibco, 10817010) and 1% penicillin/streptomycin double antibody (Corning, 30002297) were cultured, observed under a microscope to confirm that the cells were in good condition, and transferred the cells to a 15 milliliter (mL) centrifuge tube at 1000 revolutions (rpm) Centrifuge for 5 minutes, discard the supernatant, add complete medium, pipette to form a single cell suspension, and place it in a 37°C, 5% carbon dioxide (CO 2 ) incubator (Invitrogen, 3111).
2.4实验流程2.4 Experimental procedure
无菌条件下，将对数生长期的小鼠原B细胞稳转株Ba/F3 SLC34A2-ROS1与基质胶(Matrigel)(康宁，8274014)混合后移植于雌性重症联合免疫缺陷(CB-17 SCID)小鼠背部右侧皮下，每只小鼠接种1*10
6个细胞，体积100微升(μL)，接种后，待肿瘤体积生长到150立方毫米(mm
3)时，将小鼠随机分成15组，每组6只进行体内药效实验，阴性对照组给等量的溶媒。具体设计见表6。
Under aseptic conditions, the logarithmic growth phase mouse original B cell stable transgenic strain Ba/F3 SLC34A2-ROS1 was mixed with Matrigel (Kangning, 8274014) and then transplanted into female severe combined immunodeficiency (CB-17 SCID) ) Subcutaneously on the right side of the back of the mouse, each mouse is inoculated with 1*10 6 cells with a volume of 100 microliters (μL). After the inoculation, when the tumor volume grows to 150 cubic millimeters (mm 3 ), the mice are randomly divided into In 15 groups, 6 animals in each group were subjected to in vivo efficacy experiments, and the negative control group was given the same amount of solvent. The specific design is shown in Table 6.
表6.化合物体内药效试验方案Table 6. Compound in vivo efficacy test protocol
2.5实验结果及结论2.5 Experimental results and conclusions
根据公式计算小鼠肿瘤体积＝长×宽×宽/2(mm
3)，抑瘤率公式＝1-(终点时给药组肿瘤体积-给药组分组时肿瘤体积)/(终点时对照组肿瘤体积-对照组分组时肿瘤体积)×100％，实施例化合物4、9、14、17、18、21、29、33、42、45、59、61、76、83在10毫克每千克(mg/kg)的给药浓度下对肿瘤生长抑制率分别达到92.13％、96.72％、94.36％、91.17％、97.28％、96.46％、94.03％、96.22％、95.37％、95.24％、97.80％、96.69％、97.32％、96.27％；表明本发明实施例化合物4、9、14、17、18、21、29、33、42、45、59、61、76、83在小鼠原B细胞稳转株Ba/F3SLC34A2-ROS1异种移植模型中具有较强的肿瘤生长抑制作用。
Calculate the mouse tumor volume according to the formula = length × width × width / 2 (mm 3 ), tumor inhibition rate formula = 1-(the tumor volume at the end of the administration group-the tumor volume at the end of the administration group) / (the control group at the end Tumor volume-tumor volume at the time of grouping in the control group) × 100%. Example compounds 4, 9, 14, 17, 18, 21, 29, 33, 42, 45, 59, 61, 76, 83 are at 10 mg/kg ( mg/kg), the tumor growth inhibition rate reached 92.13%, 96.72%, 94.36%, 91.17%, 97.28%, 96.46%, 94.03%, 96.22%, 95.37%, 95.24%, 97.80%, 96.69 %, 97.32%, 96.27%; indicating that the compounds 4, 9, 14, 17, 18, 21, 29, 33, 42, 45, 59, 61, 76, and 83 of the examples of the present invention are stable in mouse original B cell strains The Ba/F3SLC34A2-ROS1 xenograft model has a strong tumor growth inhibitory effect.
实验九、实施例11、实施例15、本发明实施例24、实施例45、实施例59、实施例62、实施例77、实施例81化合物的大鼠急毒实验Experiment 9, Example 11, Example 15, Example 24, Example 45, Example 59, Example 62, Example 77, and Example 81 Compound Rat Acute Toxicity Experiment
1.实验目的和方法1. The purpose and method of the experiment
本实验目的是为了测试化合物在大鼠上的毒性效果。The purpose of this experiment is to test the toxic effect of the compound on rats.
大鼠单次灌胃给予100毫克每公斤(mg/kg)、200毫克每公斤(mg/kg)以及500毫克每公斤(mg/kg)实施例11、实施例15、本发明实施例24、实施例45、实施例59、实施例62化合物、实施例77、实施例81化合物，观察14天，记录动物死亡情况，中毒反应，体重变化，饮食，外观，行为等。终点解剖动物，取脏器，进行组织病理学检查。Rats were given a single intragastric administration of 100 milligrams per kilogram (mg/kg), 200 milligrams per kilogram (mg/kg) and 500 milligrams per kilogram (mg/kg) Example 11, Example 15, Example 24 of the present invention, The compound of Example 45, Example 59, Example 62, Example 77, and Example 81 were observed for 14 days, and animal death, poisoning reaction, weight change, diet, appearance, behavior, etc. were recorded. The animals were dissected at the end point, and organs were taken for histopathological examination.
2.实验结果与结论2. Experimental results and conclusions
本发明实施例11、实施例15、本发明实施例24、实施例45、实施例59、实施例62、实施例77、实施例81化合物的半数致死剂量(LD50)均大于>500毫克每公斤(mg/kg)，安全性好。与对照组大鼠比较，给药组大鼠自给药日起14天内 未见体重及行为异常，本发明实施例11、实施例15、本发明实施例24、实施例45、实施例59、实施例64、实施例70、实施例82化合物并未显示出明显毒性。Example 11, Example 15, Example 24, Example 45, Example 59, Example 62, Example 77 and Example 81 (mg/kg), good safety. Compared with rats in the control group, the rats in the administration group showed no abnormal body weight and behavior within 14 days from the day of administration. Example 11, Example 15, Example 24, Example 45, Example 59 of the present invention The compounds of Example 64, Example 70, and Example 82 did not show significant toxicity.
以上所述仅为本发明的较佳实施例，并不用以限制本发明，凡在本发明的精神和原则之内，所做的任何修改、等同替换、改进等，均应包含在本发明保护的范围之内。The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection of the present invention. Within the range.
参考文献：references:
1.Menichincheri M,Ardini E,Magnaghi P,et al.Discovery of Entrectinib:a new 3-aminoindazole as a potent Anaplastic Lymphoma Kinase(ALK),c-ros Oncogene 1 Kinase(ROS1),and Pan-Tropomyosin Receptor Kinases(Pan-TRKs)inhibitor[J].Journal of Medicinal Chemistry,2016,59,3392-3408.1.Menichincheri M, Ardini E, Magnaghi P, et al. Discovery of Entrectinib: a new 3-aminoindazole as a potent Anaplastic Lymphoma Kinase (ALK), c-ros Oncogene 1 Kinase (ROS1), andomy Panses(ROS1), andomyPan-Troporinases(ROS1) Pan-TRKs)inhibitor[J].Journal of Medicinal Chemistry,2016,59,3392-3408.
Claims (11)
- 一种通式(II)所示的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、及其混合物形式、及其可药用盐、多晶型物、溶剂合物、前药、代谢物、同位素衍生物，A compound represented by general formula (II), or tautomers, mesosomes, racemates, enantiomers, diastereomers, and mixtures thereof, and Pharmaceutically acceptable salts, polymorphs, solvates, prodrugs, metabolites, isotope derivatives,
其中，Y 1为碳时，Y 3为氮、Y 2为碳、Y 4为碳、Y 5为碳； Wherein, when Y 1 is carbon, Y 3 is nitrogen, Y 2 is carbon, Y 4 is carbon, and Y 5 is carbon;Y 1为碳时，Y 2为碳、Y 3为氮、Y 4为碳、Y 5为氮； When Y 1 is carbon, Y 2 is carbon, Y 3 is nitrogen, Y 4 is carbon, and Y 5 is nitrogen;Y 1为氮时，Y 4为氮、Y 2为碳、Y 3为碳、Y 5为碳； When Y 1 is nitrogen, Y 4 is nitrogen, Y 2 is carbon, Y 3 is carbon, and Y 5 is carbon;Y 1为氮时，Y 2为氮、Y 3为碳、Y 4为碳、Y 5为碳； When Y 1 is nitrogen, Y 2 is nitrogen, Y 3 is carbon, Y 4 is carbon, and Y 5 is carbon;R 0为R 0 is
X为碳或氮；X is carbon or nitrogen;R 1为-CH 2-R 6、-CF 2-R 6、
R 1 is -CH 2 -R 6 ,
-CF 2 -R 6 、R 2为-NR aR b、-SO 2-NR aR b、-S(O)-NR aR b、-S(O)-(CH 2) 2-NR aR b、-S-(CH 2) 2-NR aR b、4-10元杂环基、-C 4-10环烷基，其中所述的4-10元杂环基或-C 4-10环烷基包括单环、双环、三环、桥环或螺环结构的化合物基团； R 2 is -NR a R b , -SO 2 -NR a R b , -S(O)-NR a R b , -S(O)-(CH 2 ) 2 -NR a R b , -S-( CH 2 ) 2 -NR a R b , 4-10 membered heterocyclic group, -C 4-10 cycloalkyl group, wherein the 4-10 membered heterocyclic group or -C 4-10 cycloalkyl group includes a single ring , Bicyclic, tricyclic, bridged ring or spiro ring structure compound group;所述的4-10元杂环基或-C 4-10环烷基可被以下基团单取代或多取代：-OH、-C 1-3烷基、-C 3-6环烷基、4-6元杂环基、5-10元杂芳基、-O-C 1-4烷基、-NH 2、卤素、-C 1-3卤代烷基、苯基或氰基； The 4-10 membered heterocyclic group or -C 4-10 cycloalkyl group may be mono- or multi-substituted by the following groups: -OH, -C 1-3 alkyl, -C 3-6 cycloalkyl, 4-6 membered heterocyclic group, 5-10 membered heteroaryl group, -OC 1-4 alkyl, -NH 2 , halogen, -C 1-3 haloalkyl, phenyl or cyano;R a和R b分别独立的为氢、-C 1-3烷基、-C 3-6环烷基、-C 1-4亚烷基-OH、-C 1-3亚烷基-C 3-9杂环烷基、-C 1-3亚烷基-C 4-6氧代杂环烷基、-C 1-3亚烷基-C 4-6环烷基、-C(O)-(CH 2) n-T，其中所述的-C 3-9杂环烷基包括单环、双环、三环、桥环或螺环 结构的化合物基团，其中所述的-C 3-9杂环烷基可被以下基团单取代或多取代：-C 1-3烷基、卤素、-C 1-3卤代烷基、-OH或氰基； R a and R b are independently hydrogen, -C 1-3 alkyl, -C 3-6 cycloalkyl, -C 1-4 alkylene-OH, -C 1-3 alkylene-C 3 -9 heterocycloalkyl, -C 1-3 alkylene-C 4-6 oxoheterocycloalkyl , -C 1-3 alkylene-C 4-6 cycloalkyl, -C(O)- (CH 2 ) n -T, wherein the -C 3-9 heterocycloalkyl group includes a compound group with a monocyclic, bicyclic, tricyclic, bridged ring or spiro ring structure, wherein the -C 3-9 The heterocycloalkyl group may be mono- or poly-substituted by the following groups: -C 1-3 alkyl, halogen, -C 1-3 haloalkyl, -OH or cyano;R a和R b或是连同它们所连接的氮原子一起形成未被取代或被T基团或-NR cR d取代的4-9元杂环烷基，T为-C 1-4烷基、被-C 1-3烷基取代的-C 2-4烷基、-C 3-6环烷基、4-6元杂环基、-C 1-3卤代烷基、-(CH 2) n-NR cR d；其中所述的4-9元杂环烷基包括单环、双环、三环、桥环或螺环结构的化合物基团； R a and R b or together with the nitrogen atom to which they are attached form a 4-9 membered heterocycloalkyl which is unsubstituted or substituted with a T group or -NR c R d , and T is -C 1-4 alkyl , -C 2-4 alkyl substituted by -C 1-3 alkyl, -C 3-6 cycloalkyl, 4-6 membered heterocyclic group, -C 1-3 haloalkyl, -(CH 2 ) n -NR c R d ; wherein the 4-9 membered heterocycloalkyl group includes a compound group of monocyclic, bicyclic, tricyclic, bridged ring or spiro ring structure;R c和R d分别独立的为氢、-C 1-3烷基、-C 1-4亚烷基-OH、-C 2-4亚烷基-OCH 3、-(CH 2) 2-OC 1-3烷基； R c and R d are independently hydrogen, -C 1-3 alkyl, -C 1-4 alkylene-OH, -C 2-4 alkylene -OCH 3 , -(CH 2 ) 2 -OC 1-3 alkyl;R 3为-NH-C 4-8环烷基、-NH-C 3-8杂环烷基、-NH-CH 2-C 3-8杂环烷基、-O-C 1-4烷基； R 3 is -NH-C 4-8 cycloalkyl, -NH-C 3-8 heterocycloalkyl, -NH-CH 2 -C 3-8 heterocycloalkyl, -OC 1-4 alkyl;R 4为苯基、杂芳基、-NH-苯基，其中所述的苯基可被R 2和/或R k取代，所述的杂芳基可被R 2取代；R k为卤素或-O-C 1-4烷基； R 4 is phenyl, heteroaryl, -NH-phenyl, wherein the phenyl group may be substituted by R 2 and/or R k , and the heteroaryl group may be substituted by R 2 ; R k is halogen or -OC 1-4 alkyl;R 6为苯基或4-6元杂环烷基，其中所述的苯基或4-6元杂环烷基被以下基团单取代或双取代：卤素、-OH、-C 1-3烷基、-C(CH 3) 2-OH或氰基； R 6 is a phenyl group or a 4-6 membered heterocycloalkyl group, wherein the phenyl group or a 4-6 membered heterocycloalkyl group is mono- or di-substituted by the following groups: halogen, -OH, -C 1-3 Alkyl, -C(CH 3 ) 2 -OH or cyano;R 7为氢、-C 1-3烷基、-(CH 2) n-OH、C 1-3卤代烷基、-(CH 2) n-OC 1-3烷基、-(CH 2) n-OC(O)-C 1-3烷基； R 7 is hydrogen, -C 1-3 alkyl, -(CH 2 ) n -OH, C 1-3 haloalkyl, -(CH 2 ) n -OC 1-3 alkyl, -(CH 2 ) n- OC(O)-C 1-3 alkyl;n为1或2。n is 1 or 2. - 根据权利要求1所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、及其混合物形式、及其可药用盐、多晶型物、溶剂合物、前药、代谢物、同位素衍生物，其特征在于，The compound according to claim 1, or its tautomers, mesosomes, racemates, enantiomers, diastereomers, and mixtures thereof, and pharmaceutically acceptable Salts, polymorphs, solvates, prodrugs, metabolites, isotopic derivatives, characterized in that:R 2为-NR aR b、-S(O)-NR aR b、-SO 2-NR aR b、-S(O)-(CH 2) 2-NR aR b、-S-(CH 2) 2-NR aR b、4-10元杂环基、-C 4-10环烷基，其中所述的4-10元杂环基或-C 4-10环烷基可被以下基团取代：-OH、-C 1-3烷基、-C 3-6环烷基、-O-C 1-4烷基、-NR cR d、-C 1-3氟代烷基； R 2 is -NR a R b , -S(O)-NR a R b , -SO 2 -NR a R b , -S(O)-(CH 2 ) 2 -NR a R b , -S-( CH 2 ) 2 -NR a R b , 4-10 membered heterocyclic group, -C 4-10 cycloalkyl group, wherein the 4-10 membered heterocyclic group or -C 4-10 cycloalkyl group may be Group substitution: -OH, -C 1-3 alkyl, -C 3-6 cycloalkyl, -OC 1-4 alkyl, -NR c R d , -C 1-3 fluoroalkyl;R 4为X 1为碳或氮；R k为-O-C 1-4烷基； R 4 is
X 1 is carbon or nitrogen; R k is -OC 1-4 alkyl;R 6为苯基或4-6元杂环烷基，其中所述的苯基被氟取代，所述的4-6元杂环烷基被-OH或-C(CH 3) 2-OH取代； R 6 is a phenyl group or a 4-6 membered heterocycloalkyl group, wherein the phenyl group is substituted by fluorine, and the 4-6 membered heterocycloalkyl group is substituted by -OH or -C(CH 3 ) 2 -OH ；R 7为氢、甲基、乙基、-CH 2-OH、-CH 2-OC 1-3烷基、-CH 2-OC(O)-C 1-3烷基、-CH 2-F、-CF 3、-CH 2-OCH 3； R 7 is hydrogen, methyl, ethyl, -CH 2 -OH, -CH 2 -OC 1-3 alkyl, -CH 2 -OC(O)-C 1-3 alkyl, -CH 2 -F, -CF 3 , -CH 2 -OCH 3 ;R a和R b分别独立的为氢、-C 1-3烷基、-C 3-6环烷基、-C 1-3亚烷基-C 3-9杂环烷基、-C 1-3亚烷基-C 4-6氧代杂环烷基、-C(O)-CH 2-C 4-6氮杂环烷基，其中所述的-C 3-9氮杂环烷基可被以下基团单取代或多取代：-C 1-3烷基、卤素、-C 1-3卤代烷基、-OH或氰基；其中所述的-C 3-9杂环烷基包括单环、双环、三环、桥环或螺环结构的化合物基团；R a和R b或是连同它们所连接的氮原子一起形成被T基团或-NR cR d取代的4-9元杂环烷基，T为-C 1-4烷基、-C 3-6环烷基、-CF 3、4-6元杂环基、-(CH 2) 2-N(CH 3) 2； R a and R b are each independently hydrogen, -C 1-3 alkyl, -C 3-6 cycloalkyl, -C 1-3 alkylene-C 3-9 heterocycloalkyl, -C 1- 3 alkylene-C 4-6 oxoheterocycloalkyl , -C(O)-CH 2 -C 4-6 azacycloalkyl, wherein the -C 3-9 azacycloalkyl can be Mono- or multi-substituted by the following groups: -C 1-3 alkyl, halogen, -C 1-3 haloalkyl, -OH or cyano; wherein the -C 3-9 heterocycloalkyl includes a single ring , Bicyclic, tricyclic, bridged ring or spiro ring structure; R a and R b or together with the nitrogen atom to which they are connected form a 4-9 membered heterocyclic group substituted by T group or -NR c R d Cycloalkyl, T is -C 1-4 alkyl, -C 3-6 cycloalkyl, -CF 3 , 4-6 membered heterocyclic group, -(CH 2 ) 2 -N(CH 3 ) 2 ;R c和R d分别独立的为氢、-C 1-3烷基、-(CH 2) 2-OC 1-3烷基。 R c and R d are each independently hydrogen, -C 1-3 alkyl, and -(CH 2 ) 2 -OC 1-3 alkyl. - 根据权利要求1或2所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、及其混合物形式、及其可药用盐、多晶型物、溶剂合物、前药、代谢物、同位素衍生物，其特征在于，The compound according to claim 1 or 2, or its tautomer, meso, racemate, enantiomer, diastereomer, and mixture forms thereof, and Pharmaceutical salts, polymorphs, solvates, prodrugs, metabolites, isotopic derivatives, characterized in that:R 2为
R 2 is
R 3为-O-C 1-4烷基、-NH-CH 2-C 4-5杂环烷基； R 3 is
-OC 1-4 alkyl, -NH-CH 2 -C 4-5 heterocycloalkyl;T 1为-O、-S、-NH； T 1 is -O, -S, -NH;R 6为R 6 is
- 根据权利要求1所述的化合物，其特征在于，所述化合物为：The compound of claim 1, wherein the compound is:N-(5-(2-(2,5-二氟苯基)吡咯烷-1-基)吡唑并[1,5-a]嘧啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(2-(2,5-Difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiper (Azin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(6-(2-(2,5-二氟苯基)吡咯烷-1-基)咪唑并[1,2-b]哒嗪-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(6-(2-(2,5-Difluorophenyl)pyrrolidin-1-yl)imidazo[1,2-b]pyridazin-3-yl)-4-(4-methylpiper (Azin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(2-(2,5-二氟苯基)吡咯烷-1-基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(2-(2,5-Difluorophenyl)pyrrolidin-1-yl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;5-(3,5-二氟苯基)-3-(2’-(4-甲基哌嗪-1-基)-[2,4’-联吡啶]-6-基)-1H-吲唑；5-(3,5-Difluorophenyl)-3-(2'-(4-methylpiperazin-1-yl)-[2,4'-bipyridine]-6-yl)-1H-indyl Azole;6-(5-(3,5-二氟苯基)-1H-吲唑-3-基)-N-(2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)吡啶-2胺；6-(5-(3,5-Difluorophenyl)-1H-indazol-3-yl)-N-(2-methoxy-4-(4-methylpiperazin-1-yl)benzene Base) pyridine-2 amine;N-(5-(1-(3,5-二氟苯基)环丙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯并酰胺；N-(5-(1-(3,5-Difluorophenyl)cyclopropyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2- ((Tetrahydro-2H-pyran-4-yl)amino)benzoamide;N-(5-((3,5-二氟苯基)二氟甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-((3,5-Difluorophenyl)difluoromethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-( (Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((4-甲基哌嗪-1-基)磺酰)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((4-methylpiperazin-1-yl)sulfonyl)-2-(( Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苯甲基)吡唑并[1,5-a]嘧啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiperazin-1-yl)-2- ((Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(1-(3,5-二氟苯基)-2-羟基乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(3,5-Difluorophenyl)-2-hydroxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-(噁丁环-3-基氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-(oxbutane-3 -Base amino) benzamide;N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(5-甲基六氢吡咯并[3,4-c]吡咯-2(1H)-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(5-methylhexahydropyrrolo[3,4-c]pyrrole-2(1H )-Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(甲基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidin-1-yl)ethyl)amino)-2 -((Tetrahydro-2H-pyran-4-yl)amino)benzamide;(R)-N-(5-(2-(2,5-二氟苯基)吡咯烷-1-基)吡唑并[1,5-a]嘧啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；(R)-N-(5-(2-(2,5-Difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4 -Methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(7-甲基-2,7-二氮杂螺[3.5]壬烷-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(7-methyl-2,7-diazaspiro[3.5]nonane-2 -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(6-甲基-2,6-二氮杂螺[3.3]庚烷-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(6-methyl-2,6-diazaspiro[3.3]heptane-2 -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苯甲基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(6-(3,5-二氟苄基)咪唑并[1,2-b]哒嗪-3-基)-4-(4-甲基哌嗪-1基)-2-((四氢吡喃-4-基)胺基)-苯甲酰胺；N-(6-(3,5-Difluorobenzyl)imidazo[1,2-b]pyridazin-3-yl)-4-(4-methylpiperazin-1 yl)-2-(( Tetrahydropyran-4-yl)amino)-benzamide;N-(5-((4-(2-羟基丙烷-2-基)哌啶-1-基)甲基)-1H-吲唑-3-基)-4-(4-甲基哌啶-1-基)-2-((四氢吡喃-4-基)胺基)苯甲酰胺；N-(5-((4-(2-hydroxypropan-2-yl)piperidin-1-yl)methyl)-1H-indazol-3-yl)-4-(4-methylpiperidine- 1-yl)-2-((tetrahydropyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苯基)-1H-吲唑-3-基)-4-(5-(二甲基胺基)六氢环戊烷[c]并吡咯-2(1H)-基)-2((四氢-2H-吡喃-4-基)胺基)苯甲酰胺；N-(5-(3,5-Difluorophenyl)-1H-indazol-3-yl)-4-(5-(dimethylamino)hexahydrocyclopentane[c]pyrrole-2 (1H)-yl)-2((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(1-(3,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(3,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;5-(3,5-二氟苯基)-N-(2-甲氧基-4-(4-甲基哌嗪-1-基)苯基)-1H-吲唑-3-甲酰胺；5-(3,5-Difluorophenyl)-N-(2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)-1H-indazole-3-carboxamide;N-(5-(1-(3,5-二氟苯基)-2-甲氧基乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(3,5-Difluorophenyl)-2-methoxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazine-1- Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-2-甲基-7-((四氢-2H-吡喃-4-基)氨基)-1,2,3,4-四氢异喹啉-6-甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-2-methyl-7-((tetrahydro-2H-pyran-4-yl)amino) -1,2,3,4-Tetrahydroisoquinoline-6-carboxamide;4-((1R,3S,5s,7s)-5-氨基金刚烷-2-基)-N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺盐酸盐；4-((1R,3S,5s,7s)-5-aminoadamantan-2-yl)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide hydrochloride;4-(5-氨基-金刚烷-2-基)-N-(5-(3,5-二氟苄基)-1H-吲哚-3-基)-2-(哌啶-4-基氨基)苯甲酰胺；4-(5-Amino-adamantan-2-yl)-N-(5-(3,5-difluorobenzyl)-1H-indol-3-yl)-2-(piperidin-4-yl) Amino) benzamide;(R)-N-(5-(2-(2,5-二氟苯基)吡咯烷-1-基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；(R)-N-(5-(2-(2,5-Difluorophenyl)pyrrolidin-1-yl)-1H-indazol-3-yl)-4-(4-methylpiperazine- 1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(1-(2,5-二氟苯基)-2-羟基乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-hydroxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;2-(3,5-二氟苯基)-2-(3-(4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰氨基)-1H-吲唑-5-基)乙酸乙酯；2-(3,5-Difluorophenyl)-2-(3-(4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl) Amino)benzoylamino)-1H-indazol-5-yl)ethyl acetate;N-(5-(1-(3,5-二氟苯基)乙基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(3,5-Difluorophenyl)ethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazine -1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3-氟苄基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3-Fluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-( (Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(1-(3-氟苯基)-2-羟基乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(3-Fluorophenyl)-2-hydroxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2- ((Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(1-(3-氟苯基)乙基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(3-Fluorophenyl)ethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazine-1- Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(1-(2,5-二氟苯基)乙基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)ethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazine -1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(1-(2,5-二氟苯基)-2-甲氧基乙基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-methoxyethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-( 4-Methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;2-(3-氟苯基)-2-(3-(4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)-1氢-吲唑-5-基)乙基异丁酯；2-(3-Fluorophenyl)-2-(3-(4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)- 1 Hydro-indazol-5-yl) ethyl isobutyl ester;N-(5-(3-氟苄基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(5-甲基-2,5-二氮杂双环[2.2.1]庚-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3-Fluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(5-methyl-2,5-diazabicyclo[2.2 .1]hept-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(1-(3-氟苯基)-2-甲氧基乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(3-Fluorophenyl)-2-methoxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)- 2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苄基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-(5-甲基-2,5-二氮杂双环[2.2.1]庚-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(5-methyl-2,5-diazepine Bicyclo[2.2.1]hept-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(3-(二甲基氨基)氮杂环丁烷-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(3-(dimethylamino)azetidin-1-yl)-2- ((Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(3-((2-甲氧基乙基)(甲基)氨基)氮杂环 丁烷-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(3-((2-methoxyethyl)(methyl)amino)azacyclo Butane-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((2-(pyrrolidin-1-yl)ethyl)amino)-2-( (Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(乙基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2 -((Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((2-(二甲氨基)乙基)硫代)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((2-(dimethylamino)ethyl)thio)-2-((tetra Hydrogen-2H-pyran-4-yl)amino)benzamide;N-(5-(3-氟苯甲基)-1H-吲唑-3-基)-4-(乙基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3-fluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(( Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3-氟苯甲基)-1H-吲唑-3-基)-4-(甲基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3-Fluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(( Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((氧杂环丁烷-3-基甲基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((oxetane -3-ylmethyl)amino)benzamide;N-(5-(3-氟苄基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((氧杂环丁烷-3-基甲基)氨基)苯甲酰胺；N-(5-(3-Fluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((oxetane-3- (Methyl)amino)benzamide;N-(5-(2,5-二氟苄基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((氧杂环丁烷-3-基甲基)氨基)苯甲酰胺；N-(5-(2,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((oxetane -3-ylmethyl)amino)benzamide;N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(1-(3-氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(3-fluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2- ((Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-乙基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-ethylpiperazin-1-yl) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-***啉哌啶-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-morpholinepiperidin-1-yl) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-(四氢-2H-吡喃-4-基)哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-(tetrahydro-2H-pyran- 4-yl)piperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-环丙基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-cyclopropylpiperazin-1-yl )-2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-三氟甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-trifluoromethylpiperazine-1- Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(甲基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidine-1- (Yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苄基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-((1R，4R)-5-甲基-2,5-二氮杂双环[2.2.10.1]庚烷-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-((1R,4R)-5-methyl-2 ,5-Diazabicyclo[2.2.10.1]heptan-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苄基)-1H-吡唑并[3,4-c]吡啶-3-基)-4-((1s，4s)-5-甲基-2,5-二氮杂双环[2.2.10.1]庚烷-2-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-((1s,4s)-5-methyl-2 ,5-Diazabicyclo[2.2.10.1]heptan-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-2-(2-(二甲氨基)乙基)-1-羰基-6-((四氢-2H-吡喃-4-基)氨基)异二氢吲哚-5-甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-2-(2-(dimethylamino)ethyl)-1-carbonyl-6-((tetra Hydrogen-2H-pyran-4-yl)amino)isoindole-5-carboxamide;N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(N-甲基-2-吗啉代乙酰氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(N-methyl-2-morpholinoacetamido)-2-((tetrahydro -2H-pyran-4-yl)amino)benzamide;N-(5-(1-(3,5-二氟苯基)-2,2,2-三氟-乙基)-1H-吲唑-3-基)-4-(4-甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(3,5-Difluorophenyl)-2,2,2-trifluoro-ethyl)-1H-indazol-3-yl)-4-(4-methylpiper (Azin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-(乙基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2 -((Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(乙基(2-(3-氟吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(3-fluoropyrrolidin-1-yl)ethyl)amino) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((2-(吡咯烷-1-基)乙基)硫代)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((2-(pyrrolidin-1-yl)ethyl)thio)-2- ((Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)--4-((2-(3,3-二氟吡咯烷-1-基)-乙基)-(乙 基)-氨基)--2-((四氢-2H-吡喃-4-基)氨基)-苯甲酰胺；N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)--4-((2-(3,3-difluoropyrrolidin-1-yl)-ethyl )-(Ethyl)-amino)-2-((tetrahydro-2H-pyran-4-yl)amino)-benzamide;N-(5-(3,5-二氟苄基)--1H-吲唑-3-基)--4-((2-(3,4-二氟吡咯烷-1-基)-乙基)(乙基)-氨基)--2-((四氢-2H-吡喃-4-基)-氨基)-苯甲酰胺；N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)--4-((2-(3,4-difluoropyrrolidin-1-yl)-ethyl Yl)(ethyl)-amino)-2-((tetrahydro-2H-pyran-4-yl)-amino)-benzamide;N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(3,4-二甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(3,4-dimethylpiperazine-1 -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(3-甲基-4-乙基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(3-methyl-4-ethylpiperazine -1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(2,4-二甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(2,4-dimethylpiperazine-1 -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苄基(-1H-吲唑-3-基)-4-((2-(3,3,4,4-四氟吡咯烷-1-基)-乙基)-(乙基)-氨基)--2-((四氢-2H-吡喃-4-基)-氨基)-苯甲酰胺；N-(5-(3,5-Difluorobenzyl(-1H-indazol-3-yl)-4-((2-(3,3,4,4-tetrafluoropyrrolidin-1-yl) -Ethyl)-(ethyl)-amino)-2-((tetrahydro-2H-pyran-4-yl)-amino)-benzamide;N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-2-((四氢-2H-吡喃-4-基)氨基)-4-(4-(2,2,2-三氟乙基)哌嗪-1-基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-2-((tetrahydro-2H-pyran-4- (Yl)amino)-4-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)benzamide;N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(2-甲基-4-乙基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(2-methyl-4-ethylpiperazine -1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;4-(环丙基(2-(吡咯烷-1-基)-乙基)-氨基)-N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)--2-((四氢-2H-吡喃-4-基)-氨基)-苯甲酰胺；4-(Cyclopropyl(2-(pyrrolidin-1-yl)-ethyl)-amino)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl) --2-((Tetrahydro-2H-pyran-4-yl)-amino)-benzamide;N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)--4-((2-(1,4-二氧-7-氮杂螺[4.4]壬烷-1-基)乙基)-(乙基)氨基)-2-((四氢-2H-吡喃-4-基)-氨基)-苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)--4-((2-(1,4-diox-7-azaspiro[4.4]non Alk-1-yl)ethyl)-(ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)-amino)-benzamide;N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-((2-(3-氧代吡咯烷-1-基)乙基)(乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((2-(3-oxopyrrolidin-1-yl)ethyl)(ethyl )Amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(2,5-二氟苯甲基)-1H-吲唑-3-基)-4-(甲基(2-(吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(2,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidin-1-yl)ethyl)amino)-2 -((Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(乙基(2-(氮杂环丁烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(azetidin-1-yl)ethyl)amino) -2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苯甲基)-1H-吲唑-3-基)-4-((2-(吡咯烷-1-基)乙基)亚硫酰基)-2-((四氢-2H-吡喃-4-基)氨基)苯酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-((2-(pyrrolidin-1-yl)ethyl)sulfinyl)-2 -((Tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(1-(2,5-二氟苯基)-2-氟乙基)-1H-吲唑-3-基)-4-(4-甲基-4,7-二氮杂螺[2.5]辛烷-7-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methyl-4,7-diazepine Heterosspiro[2.5]octane-7-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(1-(2,5-二氟苯基(-2-氟乙基)-1H-吲唑-3-基)-4-(3,5-二甲基哌嗪-1-基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(1-(2,5-Difluorophenyl(-2-fluoroethyl)-1H-indazol-3-yl)-4-(3,5-dimethylpiperazine-1 -Yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(甲基(2-(3-氰基吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(3-cyanopyrrolidin-1-yl)ethyl)amino )-2-((Tetrahydro-2H-pyran-4-yl)amino)benzamide;(R)-N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(甲基(2-(3-羟基吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺；(R)-N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(3-hydroxypyrrolidin-1-yl)ethyl (Yl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;(S)-N-(5-(3,5-二氟苄基)-1H-吲唑-3-基)-4-(甲基(2-(3-羟基吡咯烷-1-基)乙基)氨基)-2-((四氢-2H-吡喃-4-基)氨基)苯甲酰胺。(S)-N-(5-(3,5-Difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(3-hydroxypyrrolidin-1-yl)ethyl Yl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide.
- 制备如权利要求1中限定的通式II所示的化合物的方法，所述通式II所示的化合物通过以下方案制备，The method for preparing the compound represented by the general formula II as defined in claim 1, the compound represented by the general formula II is prepared by the following scheme,方案1，当其中，Y 1为碳时，Y 3为氮、Y 2为碳、Y 4为碳、Y 5为碳；R 1为-CF 2-R 6时，合成路线归纳如下： Scheme 1, when Y 1 is carbon, Y 3 is nitrogen, Y 2 is carbon, Y 4 is carbon, and Y 5 is carbon; when R 1 is -CF 2 -R 6 , the synthetic route is summarized as follows:
其中R 2、R 3、R 6如权利要求1中定义的，所述氧化试剂优选为Dess-Martin氧化剂、PCC、PDC等氧化剂，氟化试剂优选为BAST、DAST等氟化试剂，成酰胺方法优选为羧酸经酰氯于胺反应制成酰胺，或羧酸经1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐(EDCI)/1-羟基苯并***(HOBT)、环己基碳二亚胺(DCC)、2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(HATU)等缩合剂活化与胺成酰胺， Wherein R 2 , R 3 , R 6 are as defined in claim 1, the oxidizing reagent is preferably an oxidizing agent such as Dess-Martin oxidizing agent, PCC, PDC, etc., and the fluorinating reagent is preferably a fluorinating reagent such as BAST, DAST, etc., forming an amide method Preferably, the carboxylic acid is reacted with an amine to form an amide, or the carboxylic acid is reacted with 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI)/1-hydroxybenzotrimide Azole (HOBT), cyclohexylcarbodiimide (DCC), 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU), etc. Condensing agent activation and amine to form amide,方案2，当其中，Y 1为碳时，Y 3为氮、Y 2为碳、Y 4为碳、Y 5为碳；R 1为
合成路线归纳如下： Scheme 2, when Y 1 is carbon, Y 3 is nitrogen, Y 2 is carbon, Y 4 is carbon, and Y 5 is carbon; R 1 is
The synthetic route is summarized as follows:
其中R 2、R 3、R 6如权利要求1中定义的，所述氧化试剂优选为Dess-Martin氧化剂、PCC、PDC等氧化剂；成三元环方法优选为二乙基锌/二碘甲烷、三甲基碘化亚砜盐或三甲基溴化亚砜盐/钠氢；硼化试剂优选为硼烷、9-BBN等；氧化试剂优选为双氧水；氟化试剂优选为BAST、DAST等氟化试剂，成酰胺方法优选为羧酸经酰氯于胺反应制成酰胺，或羧酸经1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐(EDCI)/1-羟基苯并***(HOBT)、环己基碳二亚胺(DCC)、2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(HATU)等缩合剂活化与胺成酰胺， Wherein R 2 , R 3 , R 6 are as defined in claim 1, the oxidizing agent is preferably an oxidizing agent such as Dess-Martin oxidant, PCC, PDC, etc.; the method of forming a three-membered ring is preferably diethyl zinc/diiodomethane, Trimethyl sulfoxide iodide salt or trimethyl sulfoxide bromide salt/sodium hydrogen; boronating reagent is preferably borane, 9-BBN, etc.; oxidizing reagent is preferably hydrogen peroxide; fluorinating reagent is preferably fluorine such as BAST and DAST Chemical reagent, the method of amide formation is preferably the reaction of carboxylic acid with acid chloride and amine to make amide, or carboxylic acid with 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI)/ 1-Hydroxybenzotriazole (HOBT), cyclohexylcarbodiimide (DCC), 2-(7-oxybenzotriazole)-N,N,N',N'-tetramethylurea hexafluoro Phosphate (HATU) and other condensing agents are activated to form amides with amines,方案3，当其中，Y 1为碳时，Y 2为碳、Y 3为氮、Y 4为碳、Y 5为氮；或Y 1为氮时，Y 4为氮、Y 2为碳、Y 3为碳、Y 5为碳；或Y 1为氮时，Y 2为氮、Y 3为碳、Y 4为碳、Y 5为碳；R 1为-CH 2-R 6时；合成路线归纳如下： Scheme 3, when Y 1 is carbon, Y 2 is carbon, Y 3 is nitrogen, Y 4 is carbon, and Y 5 is nitrogen; or when Y 1 is nitrogen, Y 4 is nitrogen, Y 2 is carbon, and Y 3 is carbon and Y 5 is carbon; or when Y 1 is nitrogen, Y 2 is nitrogen, Y 3 is carbon, Y 4 is carbon, and Y 5 is carbon; when R 1 is -CH 2 -R 6 ; synthetic route is summarized as follows:
其中R 2、R 3、R 6如权利要求1中定义的，卤代实际优选为溴、碘、N-溴代丁二酰亚胺、N-碘代丁二酰亚胺等，偶联反应优选为Stille反应； Wherein R 2 , R 3 , R 6 are as defined in claim 1, and halogenation is actually preferably bromine, iodine, N-bromosuccinimide, N-iodosuccinimide, etc., coupling reaction Preferably it is Stille reaction;方案4，当其中，Y 1为碳时，Y 2为碳、Y 3为氮、Y 4为碳、Y 5为碳；或Y 1Scheme 4, when Y 1 is carbon, Y 2 is carbon, Y 3 is nitrogen, Y 4 is carbon, and Y 5 is carbon; or Y 1
其中R 2、R 3、R 6如权利要求1中定义的，硝化试剂优选为硝酸、硝酸钾、硝酸钠等；还原试剂优选为铁粉、锌粉、氯化亚锡、氢气/钯碳、氢气/镍等；成酰胺方法优选为羧酸经酰氯于胺反应制成酰胺，或羧酸经1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐(EDCI)/1-羟基苯并***(HOBT)、环己基碳二亚胺(DCC)、2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(HATU)等缩合剂活化与胺成酰胺， Wherein R 2 , R 3 , R 6 are as defined in claim 1, the nitrating agent is preferably nitric acid, potassium nitrate, sodium nitrate, etc.; the reducing agent is preferably iron powder, zinc powder, stannous chloride, hydrogen/palladium carbon, Hydrogen/nickel, etc.; the method of amide formation is preferably the reaction of carboxylic acid with acid chloride and amine to make amide, or carboxylic acid with 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI )/1-hydroxybenzotriazole (HOBT), cyclohexylcarbodiimide (DCC), 2-(7-oxybenzotriazole)-N,N,N',N'-tetramethylurea Hexafluorophosphate (HATU) and other condensing agents are activated to form amides with amines,方案5，当其中，R 0为R 1为-CH 2-R 6；合成路线归纳如下： Scheme 5, when R 0 is
R 1 is -CH 2 -R 6 ; the synthetic route is summarized as follows:
其中R 4、R 6如权利要求1中定义的。 Wherein R 4 and R 6 are as defined in claim 1. - 一种药物组合物，其特征在于，包括治疗有效量的权利要求1-4任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、及其混合物形式、及其可药用盐、多晶型物、溶剂合物、前药、代谢物、同位素衍生物，以及任选一种或多种医药上可接受的载剂和/或稀释剂。A pharmaceutical composition, characterized by comprising a therapeutically effective amount of the compound according to any one of claims 1 to 4, or its tautomer, mesosome, racemate, or enantiomer Isomers, diastereomers, and mixtures thereof, and pharmaceutically acceptable salts, polymorphs, solvates, prodrugs, metabolites, isotopic derivatives, and optionally one or more pharmaceutically acceptable Acceptable carriers and/or diluents.
- 根据权利要求1-4任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、及其混合物形式、及其可药用盐、多晶型物、溶剂合物、前药、代谢物、同位素衍生物，或者根据权利要求6所述的药物组合物在制备用于预防和/或治疗癌症的药物中的用途。The compound according to any one of claims 1 to 4, or its tautomer, meso, racemate, enantiomer, diastereomer, and mixtures thereof, And its pharmaceutically acceptable salts, polymorphs, solvates, prodrugs, metabolites, isotopic derivatives, or the pharmaceutical composition according to claim 6 in the preparation of drugs for the prevention and/or treatment of cancer the use of.
- 根据权利要求7所述的用途，其特征在于，所述的癌症为肺癌、骨髓癌、鳞状细胞癌、甲状腺毛囊癌、***癌、乳腺癌、膀胱癌、肺癌、胃癌、直肠癌、淋巴瘤或白血病。The use according to claim 7, wherein the cancer is lung cancer, bone marrow cancer, squamous cell carcinoma, thyroid hair follicle cancer, prostate cancer, breast cancer, bladder cancer, lung cancer, gastric cancer, rectal cancer, lymphoma Or leukemia.
- 一种用于预防和/或治疗肺癌、骨髓癌、鳞状细胞癌、甲状腺毛囊癌、***癌、乳腺癌、膀胱癌、肺癌、胃癌、直肠癌、淋巴瘤或白血病的方法，所述方法包括对有需要的对象施用治疗有效量的根据权利要求1-4任一项所述的化合物及其药学上可接受的盐或立体异构体或者根据权利要求6所述的药物组合物。A method for preventing and/or treating lung cancer, bone marrow cancer, squamous cell carcinoma, thyroid hair follicle cancer, prostate cancer, breast cancer, bladder cancer, lung cancer, gastric cancer, rectal cancer, lymphoma or leukemia, said method comprising A therapeutically effective amount of the compound according to any one of claims 1 to 4 and a pharmaceutically acceptable salt or stereoisomer thereof or the pharmaceutical composition according to claim 6 is administered to a subject in need.
- 根据权利要求1-4任一项所述的化合物，或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、及其混合物形式、及其可药用盐、多晶型物、溶剂合物、前药、代谢物、同位素衍生物，或者根据权利要求6所述的药物组合物用于预防和/或治疗癌症。The compound according to any one of claims 1 to 4, or its tautomer, meso, racemate, enantiomer, diastereomer, and mixtures thereof, And its pharmaceutically acceptable salts, polymorphs, solvates, prodrugs, metabolites, isotopic derivatives, or the pharmaceutical composition according to claim 6 for the prevention and/or treatment of cancer.
- 根据权利要求10所述之用途的化合物或药物组合物，其特征在于，所述的癌症为肺癌、骨髓癌、鳞状细胞癌、甲状腺毛囊癌、***癌、乳腺癌、膀胱癌、肺癌、胃癌、直肠癌、淋巴瘤或白血病。The compound or pharmaceutical composition for use according to claim 10, wherein the cancer is lung cancer, bone marrow cancer, squamous cell carcinoma, thyroid follicular carcinoma, prostate cancer, breast cancer, bladder cancer, lung cancer, gastric cancer , Rectal cancer, lymphoma or leukemia.
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