WO2023169453A1 - 一类含杂芳环炔基化合物及其制备方法和用途 - Google Patents

一类含杂芳环炔基化合物及其制备方法和用途 Download PDF

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WO2023169453A1
WO2023169453A1 PCT/CN2023/080231 CN2023080231W WO2023169453A1 WO 2023169453 A1 WO2023169453 A1 WO 2023169453A1 CN 2023080231 W CN2023080231 W CN 2023080231W WO 2023169453 A1 WO2023169453 A1 WO 2023169453A1
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substituted
alkyl
methyl
unsubstituted
membered
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French (fr)
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胡有洪
楼丽广
谢志铖
李琳
王蕾
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中国科学院上海药物研究所
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    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
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    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
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    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
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    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
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    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/472—Non-condensed isoquinolines, e.g. papaverine
    • A61K31/4725—Non-condensed isoquinolines, e.g. papaverine containing further heterocyclic rings
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    • A61K31/50—Pyridazines; Hydrogenated pyridazines
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    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
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    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
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    • C07D401/02—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 two hetero rings
    • C07D401/04—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 two hetero rings directly linked by a ring-member-to-ring-member bond
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    • 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
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Definitions

  • the present invention relates to KIT mutation inhibitors, in particular to a class of compounds containing heteroaromatic ring alkynyl groups and their preparation methods and uses. More specifically, the present invention relates to a compound that can inhibit the proliferation of KIT mutant cell lines, a preparation method of such compounds, and pharmaceutical uses for mutant KIT-related diseases.
  • KIT also known as CD117 or stem cell factor receptor
  • RTK Receptor Tyrosine Kinase
  • KIT belongs to the third type of RTK, which consists of 5 immunoglobulin-like domains (D1 ⁇ D5), 1 transmembrane domain, and 1 juxtamembrane domain (JMD) and tyrosine kinase (TK) domain. composed of cytoplasmic region.
  • PDGFR ⁇ platelet-derived growth factor receptor ⁇
  • PDGFR ⁇ platelet-derived growth factor receptor ⁇
  • FLT3 FLT3
  • CSF-1R colony-stimulating factor 1 receptor
  • the ligand stem cell factor binds to the extracellular domain to dimerize the receptor, leading to tyrosine autophosphorylation in the TK domain in the cytoplasmic region, further causing downstream signaling pathways (such as PI3K, JAK-STAT , Ras-Erk, Src family kinases and PLC signaling pathways), and trigger a variety of physiological processes, such as cell proliferation, division, and tissue growth and survival.
  • SCF ligand stem cell factor
  • KIT gain-of-function mutations are associated with various diseases such as tumors, inflammation, and autoimmune diseases.
  • GIST gastrointestinal stromal tumors
  • systemic mastocytosis a small amount of acute myeloid leukemia
  • glioma a small amount of acute myeloid leukemia
  • lung cancer a small amount of acute myeloid leukemia
  • KIT mutations Its role in GIST has been studied most extensively and deeply. About 85% of GISTs are caused by KIT mutations. GIST is the most common mesenchymal tumor of the gastrointestinal tract, with an incidence rate of approximately 1/100,000 to 2/100,000, accounting for 1 to 3% of all gastrointestinal tumors.
  • Imatinib As a first-line drug for the treatment of GIST, Imatinib is effective against most GISTs with primary KIT mutations, but 90% of patients will eventually develop drug resistance due to secondary mutations in KIT, leading to tumor recurrence.
  • These resistance mutations mainly occur in the ATP binding pocket of KIT (exon 13, such as K642E, V654A; exon 14, such as T670I) and activation loop (A-loop; exon 17, such as D816H, D816V, D820A, N822K, N822H, N822V ).
  • Sunitinib and Regorafenib as second- and third-line treatments for GIST, can only overcome a small number of Imatinib-resistant mutations (such as V654A and T670I mutations that occur in the ATP-binding pocket), and the clinical response rate is low. It is reported that among Imatinib-resistant cases, 50% of patients have secondary mutations in A-loop (Demetri GD, et al. Lancet. 2006, 368: 1329-1338; Nishida T, et al. Int.J. Clin.Oncol.2009,14:143-149). Therefore, the development of new KIT inhibitors that can reverse drug resistance caused by A-loop (exon 17) mutations has important clinical significance.
  • CN104211639A, CN108456163A and CN111662227A all relate to heteroaryl ring alkynyl compounds, which involve compounds whose connecting chain L is -C(O)NH-, -NHC(O)-, ether chain or amino chain.
  • the connecting chain of the heteroaryl ring alkynyl compound disclosed in WO2015089210A1 includes -C(O)NHCH 2 -, its pyridine ring must have a sulfone imine as the dominant structure in the meta position, and the middle benzene ring must be unsubstituted. situation; in addition, its focus is on VEGFR and PDGFR, and does not involve the inhibitory activity against KIT mutant cell lines.
  • the present invention provides a class of heteroaryl ring alkynyl small molecule kinase inhibitors with novel structures. These compounds have very strong proliferation inhibitory activity on KIT mutant cell lines (including various mutant forms including KIT D816V). At the same time, this type of compound has high selectivity, has no obvious cytotoxicity to non-tumor cells (32D cells) and KIT wild-type cells (NCI-H526, Moe7), and has weak inhibitory activity against EGFR and PDGFR-dependent cells, which reflects the Compounds with higher selectivity. In addition, representative compounds have significant anti-tumor effects in vivo and have low toxicity.
  • the invention provides a compound of formula (I), or its deuterated compound, pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug:
  • X is selected from -(C(R a )(R b )) q -;
  • R a and R b are each independently selected from hydrogen, deuterium, halogen, and C1-4 alkyl; or R a and R b connected to the same carbon atom together with the carbon atom form a 3-5-membered carbon ring;
  • q is selected from 1 and 2;
  • R 1 is selected from hydrogen, halogen, C1-6 alkyl
  • Ring M1 is selected from the following structures:
  • a 1 and A 2 are independent CR H4 or N atoms
  • a 3 is CR H2 or N atom
  • a 4 is CR H3 , NR H3 or N atom
  • R 2 and the atom connected thereto form a substituted or unsubstituted 3-7 membered carbocyclic ring or heterocyclic ring, and the substitution refers to being substituted by 1-5 R 3 ;
  • R H1 , R H2 and R H4 are each independently selected from hydrogen, halogen, amino, cyano, hydroxyl, -N(R 4 )(R 5 );
  • the R 4 and R 5 are each independently selected from hydrogen, C1-6 alkyl, C1-6 heteroalkyl, C3-6 cycloalkyl, C3-6 heterocycloalkyl, C1-6 alkyl acyl, C3- 6 cycloalkyl acyl;
  • the substitution means each is independently substituted by 1-5 R 6 ; in cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclyl, one or more
  • R 6 and the atoms to which they are connected together form a substituted or unsubstituted 5-7 membered carbocyclic ring or 5-7 membered heterocyclic ring, and the substitution refers to being substituted by 1 to 5 R 7 groups;
  • Ring M 2 is selected from unsubstituted or substituted C6-10 aryl, unsubstituted or substituted 5-7 membered heteroaryl, unsubstituted or substituted 5-7 heterocyclyl; the substitution means that each is independently replaced by Replaced by 1-5 R M ;
  • R M Two R M and the atoms connected to them together form a substituted or unsubstituted 3-7 membered carbocyclic ring or heterocyclic ring, and the substitution refers to being substituted by 1-5 R 8 groups;
  • the R 8 are each independently selected from hydroxyl, cyano, amino, halogen, -NH (C1-6 alkyl), C1-6 alkyl, C1-6 heteroalkyl, C1-6 haloalkyl, C1-6 hydroxyl Alkyl, C3-6 cycloalkyl, C3-6 heterocycloalkyl.
  • the compound has the structure of the following formula (II):
  • R 1 is selected from halogen, C1-3 alkyl; preferably selected from methyl, F, Cl;
  • R a and R b are each independently selected from hydrogen, deuterium, halogen, and methyl; or R a and R b and the carbon atoms connected to them together form a three-membered carbon ring;
  • Ring M1 is selected from the following structures:
  • a 1 , A 2 , A 3 , A 4 , rings W, Z and R H1 are each as defined above;
  • Ring M2 is defined as above.
  • ring M1 is selected from the following structures:
  • C,N on the ring atom indicates that it is CH or N.
  • R 2 , RH1 , RH2 and RH3 are each defined as described above.
  • Ring M 2 is selected from unsubstituted or substituted phenyl, unsubstituted or substituted thienyl, unsubstituted or substituted pyrazolyl, unsubstituted or substituted thiazolyl, unsubstituted or substituted pyridyl, unsubstituted or substituted oxazolyl, unsubstituted or substituted isoxazolyl, unsubstituted or substituted benzothienyl; the substitution means that each is independently substituted by 1-5 R M ;
  • R M Two R M and the atoms connected to them form a substituted or unsubstituted 3-7 membered carbocyclic ring or heterocyclic ring, and the substitution refers to being substituted by 1-5 R 8 groups;
  • the R 8 are each independently selected from hydroxyl, cyano, amino, halogen, -NH (C1-6 alkyl), C1-6 alkyl, C1-6 heteroalkyl, C1-6 haloalkyl, C1-6 hydroxyl Alkyl, C3-6 cycloalkyl, C3-6 heterocycloalkyl.
  • the alkyl group is a saturated aliphatic linear or branched alkyl group with 1 to 8 carbon atoms; typical alkyl groups include but are not limited to: methyl, ethyl, propyl, isopropyl, and cyclopropyl , n-butyl, isobutyl, tert-butyl, etc.
  • halogens are each independently selected from F, Cl, Br, I;
  • the haloalkyl group means that at least one hydrogen atom in the alkyl group is replaced by a halogen atom. In certain embodiments, if two or more hydrogen atoms are replaced by halogen atoms, the halogen atoms are the same as or different from each other;
  • the heteroalkyl group means that at least one skeleton C atom in the alkyl group is replaced by a heteroatom (N, O, S).
  • examples of the heteroalkyl group may include alkoxy, alkylamino, alkylthio, etc.
  • the heteroatoms are the same or different from each other;
  • the cycloalkyl group is a saturated or partially unsaturated 3-10 membered monocyclic or polycyclic alicyclic ring, and can be a monovalent group or a bivalent group (ie, cycloalkylene group);
  • the heterocycloalkyl group is a saturated 3-10 membered monocyclic or polycyclic alicyclic heterocyclic ring containing one or more heteroatoms selected from N, O, and S, and can be a monovalent group or a bivalent group. (i.e. heterocycloalkylene);
  • the aryl group means that each atom constituting the ring in the aryl ring is a carbon atom, including a single ring or a condensed polycyclic ring, and it can be a monovalent group or a bivalent group (i.e., arylene group).
  • the aryl ring preferably has 5 to 10 carbon atoms, and more preferably an aryl group has 5 to 7 carbon atoms.
  • the heteroaryl group is an aromatic group containing one or more heteroatoms selected from N, O, and S on the ring.
  • a heteroaryl group can be a monovalent group or a bivalent group (i.e., a heteroarylene group).
  • heterocyclyl groups are monocyclic or polycyclic, and at least one of them is a non-aromatic ring group containing one or more heteroatoms selected from N, O, and S on the ring.
  • a heterocyclyl group may be a monovalent group or a bivalent group (i.e., a heterocyclylene group).
  • the above-mentioned compound of formula (I) is selected from the group consisting of compounds of the following formula:
  • the present invention also provides a method for preparing the above-mentioned compound of formula (I), which includes the following steps: performing a coupling reaction between a compound of formula (A) and a compound of formula (B) to obtain a compound of formula (I).
  • ring M 1 , ring M 2 , X and R 1 are each independently defined as above; TMS is -Si(CH 3 ) 3 ;
  • the coupling reaction is carried out in the presence of a palladium metal catalyst and a copper metal catalyst, and in the presence of a base.
  • the palladium metal catalyst includes Pd(PPh 3 ) 2 Cl 2 , Pd(OAc) 2 , and Pd(PPh 3 ) One or more of 4 ;
  • the copper metal catalyst includes CuI and/or CuCl;
  • the base includes CsF, Cs 2 CO 3 , KF, K 2 CO 3 , NaHCO 3 , One or more of Na 2 CO 3 , Et3N, ( i Pr) 2 EtN, and DMAP;
  • the coupling reaction is performed in the presence of a solvent, and the solvent includes one or more of acetonitrile, 1,4-dioxane, and DMF.
  • the method includes the following steps: the compound of formula (A) and the compound of formula (B) are carried out in the presence of cesium fluoride, Pd(PPh 3 ) 2 Cl 2 , CuI and triethylamine in an acetonitrile solvent.
  • the coupling reaction gives compounds of formula (I).
  • the present invention provides an instructive synthetic scheme (as shown in synthetic route I). It will be appreciated that the reagents/reaction conditions shown in the synthetic schemes can be modified or optimized using general knowledge of organic chemistry to prepare different compounds of the invention.
  • the A 1 , A 2 , ring M 2 , X, Z, R H1 and R 1 are independently as mentioned above.
  • the synthetic route I includes the following steps:
  • Step 1 Mix compounds I-1, I-2 and Et 3 N, add palladium metal catalyst and copper metal catalyst, and react under argon protection (for example, at room temperature) to obtain compound I-3;
  • Step 2 Add compounds I-3 and I-4 to a suitable solvent, add palladium metal catalyst, K 2 CO 3 , and heat the reaction under argon protection to obtain important intermediate A; in addition, suitable ligands can also be added ;
  • Step 3 Condensation reaction occurs between compounds I-5 and I-6, wherein X is as described above. After mixing compound I-5, HATU, DIPEA and DMF (for example, stirring at room temperature for 30 to 60 minutes), add compound I-6 and react (for example, at room temperature for 6 to 12 hours) to obtain intermediate B;
  • Step 4 Mix intermediates A, B, base and MeCN, add palladium metal catalyst and copper metal catalyst, and react under argon protection (for example, in the case of I substitution, react at room temperature, and in the case of Br substitution (reaction at 80°C, reaction time (for example, 2 to 6 hours)) to obtain compound I-7;
  • argon protection for example, in the case of I substitution, react at room temperature, and in the case of Br substitution (reaction at 80°C, reaction time (for example, 2 to 6 hours)
  • the palladium metal catalyst described in steps 1 and 4 is Pd(PPh 3 ) 2 Cl 2 , and the copper metal catalyst is CuI;
  • the solvent in step 2 is one or more of toluene, ethanol, ethylene glycol dimethyl ether, tetrahydrofuran, 1,4-dioxane and water;
  • the palladium metal catalyst is Pd(PPh 3 ) 4 , Pd Any one of (OAC) 2 and (dppf) PdCl 2 ;
  • the base is any one of K 2 CO 3 , Cs 2 CO 3 , NaHCO 3 and Na 2 CO 3 ;
  • the ligand is X- Any of Phos and PPh 3 ;
  • the base in step 4 is cesium fluoride and/or triethylamine
  • synthetic route I includes the following steps:
  • Step 1 Add compound I-1 and Et 3 N to the round-bottomed flask, replace the oxygen with argon, add Pd(PPh 3 ) 2 Cl 2 and CuI, repeat the operation of removing oxygen, and react at room temperature for about 15 minutes. , add I-2, and continue the reaction at room temperature for 3 hours
  • compound I-3 can be obtained after purification; among them, the equivalents of compound I-1, I-2, Pd(PPh 3 ) 2 Cl 2 and CuI can be about 1.0, 1.0 ⁇ 1.5, 0.05 ⁇ 0.1, 0.1 ⁇ respectively. 0.2.
  • Step 2 Add compounds I-3, I-4, K 2 CO 3 , THF and water to the round bottom flask, replace the oxygen with argon, add Pd(OAc) 2 and X-Phos, and repeat the operation of removing oxygen. Heating to 80°C for reaction, the reaction is completed in 4-8 hours, and compound A can be obtained after purification; the equivalents of I-3, I-4, K 2 CO 3 , Pd(OAc) 2 and X-Phos can be 1.0 respectively. 1.0-1.5, 2.0-3.0, 0.05-0.1, 0.1-0.2; the volume ratio of THF/H 2 O is 4/1.
  • Step 3 Add compound I-5, HATU, DIPEA and DMF to the round-bottomed flask, stir at room temperature for 30 minutes, then add compound I-6, and react at room temperature; after 12 hours of reaction, compound B can be obtained after purification;
  • the equivalents of compounds I-5, I-6, HATU and DIPEA may be about 1.0, 1.0 ⁇ 1.2, 1.0 ⁇ 1.5, 2.0 ⁇ 4.0 respectively.
  • Step 4 Add compounds A, B, Et 3 N, CsF and MeCN to the round-bottomed flask, replace oxygen with argon, add Pd(PPh 3 ) 2 Cl 2 , CuI, repeat the operation of removing oxygen, and heat to room temperature or to Reaction at 80°C, the reaction is completed in 3-6 hours, and compound I-7 can be obtained after purification; the equivalents of compounds A, B, cesium fluoride, Pd(PPh 3 ) 2 Cl 2 , CuI and Et 3 N can be approximately 1.0 respectively. ,1.0 ⁇ 1.5,2.5 ⁇ 3.0,0.05 ⁇ 0.1,0.1 ⁇ 0.2,2.5 ⁇ 3.0.
  • the present invention also provides a pharmaceutical composition, which includes: one selected from the group consisting of the above-mentioned compound of formula (I), its deuterated compound, pharmaceutically acceptable salt, solvate, ester, acid, metabolite and prodrug. or more, and pharmaceutically acceptable excipients.
  • the present invention also provides the above-mentioned compound of formula (I) or its deuterated compound, pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug or the above-mentioned pharmaceutical composition for preparing KIT (mutant type) inhibitor. uses in.
  • the present invention also provides the above-mentioned compound of formula (I) or its deuterated compound, pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug, or the above-mentioned pharmaceutical composition prepared for treatment, prevention or Use in a medicament to improve one or more diseases or conditions selected from the group consisting of tumors, inflammation, autoimmune and neurological diseases.
  • the tumor is a gastrointestinal stromal tumor, in particular a gastrointestinal stromal tumor involving a KIT mutation, more particularly a gastrointestinal stromal tumor caused by a KIT mutation that is resistant to Imatinib and/or Sunitinib tumor.
  • the compound of formula (I) of the present invention has a potent killing effect on tumor cells carrying primary KIT mutations and drug-resistant KIT mutations.
  • the compound containing a heteroaryl ring alkynyl structure according to the present invention shows that the tumor cell lines carrying different mutant forms of KIT, D816V, V559D, V559D-V654A, V559D/Y823D and V559D/N822K It has strong inhibitory activity on cell proliferation.
  • the compound containing a heteroaryl ring alkynyl structure according to the present invention has low cytotoxicity to normal 32D cells and the KIT wild-type cell line NCI-H526 cell line, showing the toxicity of the compound.
  • compounds containing heteroaryl ring alkynyl structures according to the present invention have low inhibitory activity against BCR-ABL, EGFR, and PDGFR-dependent cell lines, showing that such compounds have relative selectivity.
  • the compound containing a heteroaryl ring alkynyl structure according to the present invention shows obvious activity in inhibiting the growth of related tumors in a long-term animal efficacy model, which is significantly better than the positive drug Ripretinib.
  • the animals are in good condition (including no significant decrease in body weight), and there is no animal death.
  • Figure 1 is a graph showing the effects of compounds 77, 85 and Ripretinib on 32D KIT D816V cell KIT and its downstream signaling pathways.
  • Figure 2 is a graph showing the effects of compounds 27, 48 and Ripretinib on 32D KIT V559D cell KIT and its downstream signaling pathways.
  • Figure 3 is a graph showing the effects of compounds 27, 48 and Ripretinib on KIT and its downstream signaling pathways in 32D KIT V559D-V654A cells.
  • Figure 4 is a graph showing the efficacy of compounds 77, 85, and Ripretinib on subcutaneous transplanted tumors of 32D KIT D816V nude mice.
  • Figure 5 is a graph showing the effects of compounds 77, 85, and Ripretinib on the body weight of tumor-bearing (32D KIT D816V) nude mice.
  • Step 1 Add 2-amino-5-bromo-3-iodopyridine (1.0g, 3.35mmol), trimethylsilyl acetylene (427.2mg, 4.35mmol) and Et 3 N (50mL) into a round-bottomed flask.
  • Step 2 Add 2-amino-5-bromo-3-trimethylethynylpyridine (300mg, 1.11mmol) and 1-methylpyrazole-4-boronic acid pinacol ester (348mg, 1.67 mmol), K 2 CO 3 (385mg, 2.79mmol) and THF/H 2 O (8/2mL), replace oxygen with argon, add Pd(OAc) 2 (25mg, 0.11mmol) and X-Phos (106 mg, 0.22mmol), heated to 70°C for 4 hours. After the reaction, the reaction solution was extracted with ethyl acetate (15 mL ⁇ 3) and water (10 mL).
  • Step 3 Add compound 4-methyl-3-iodobenzoic acid (2.0g, 7.63mmol), HATU (3.8g, 9.92mmol), DIPEA (2.5g, 19.08mmol) and DMF (40mL) into the round-bottomed flask. After stirring at room temperature for 30 minutes, the compound benzylamine (818 mg, 7.63 mmol) was added, and the reaction was carried out at room temperature for 12 hours. After the reaction, the reaction solution was extracted with ethyl acetate (50 mL ⁇ 3) and water (40 mL), and the organic phase was washed with tap water. (30 mL g (yield: 93%).
  • Step 4 Add compound N-benzyl-3-iodo-4-methylbenzamide (100 mg, 0.28 mmol), 5-(1-methyl-1H-pyrazol-4-yl) into the round-bottomed flask. -3-((Trimethylsilyl)ethynyl)-2-aminopyridine (100 mg, 0.37 mmol), Et 3 N (86 mg, 0.85 mmol), CsF (108 mg, 0.71 mmol) and MeCN (20 mL), using Replace oxygen with argon, add Pd(PPh 3 ) 2 Cl 2 (10 mg, 0.014 mmol) and CuI (5.4 mg, 0.028 mmol), repeat the oxygen removal operation, and react at room temperature for 3 hours.
  • Pd(PPh 3 ) 2 Cl 2 10 mg, 0.014 mmol
  • CuI 5.4 mg, 0.028 mmol
  • the synthesis method is as in Example 1 except that phenylethylamine is used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 1-phenylcyclopropylamine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 5-fluoro-3-iodobenzoic acid was used instead of 4-methyl-3-iodobenzoic acid.
  • the synthesis method was as in Example 1 except that 5-fluoro-3-iodobenzoic acid was used instead of 4-methyl-3-iodobenzoic acid and 3-aminomethylthiophene was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 5-fluoro-3-iodobenzoic acid was used instead of 4-methyl-3-iodobenzoic acid and 3-aminomethylpyridine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 5-fluoro-3-iodobenzoic acid was used instead of 4-methyl-3-iodobenzoic acid and 6-fluoro-3-aminomethylpyridine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 4-chlorobenzylamine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 2-trifluoromethylbenzylamine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 2-methoxybenzylamine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 3-methoxybenzylamine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 4-methoxybenzylamine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 4-fluorobenzylamine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 2,4-difluorobenzylamine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 2-fluoro-4-chlorobenzylamine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 3-aminomethylpyridine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 4-aminomethylpyridine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 5-aminomethylpyrimidine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 3-aminomethyl-6-methoxypyridine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 3-aminomethyl-6-chloropyridine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 3-aminomethyl-6-fluoropyridine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 3-aminomethyl-6-methylaminopyridine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that (6-(1H-pyrazol-1yl)pyridin-3-yl)methanamine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 3-fluorobenzylamine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 3-aminomethylthiophene was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 2-aminomethylthiophene was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 4-aminomethylthiazole was used instead of benzylamine.
  • Step 1 Add 2-chloro-5-bromo-3-iodopyridine (1.0g, 3.14mmol) and MeNH2 (2.0M, 5mL) to the sealed tube, heat to 80°C and react overnight. After the reaction, the solvent was evaporated to dryness under reduced pressure, and 650 mg of 2-methylamino-5-bromo-3-iodopyridine was obtained by column chromatography (yield: 66%).
  • Step 2 Add 2-methylamino-5-bromo-3-iodopyridine (300 mg, 0.96 mmol), trimethylsilyl acetylene (122.4 mg, 1.25 mmol) and Et 3 N (20 mL) into a round-bottomed flask. Replace oxygen with argon, add Pd(PPh 3 ) 2 Cl 2 (34 mg, 0.048 mmol) and CuI (18 mg, 0.096 mmol), repeat the operation of removing oxygen, and react at room temperature for 4 hours; after the reaction, add 20 mL of ethyl acetate. Dilute the reaction solution and filter to obtain the product 2-methylamino-5-bromo-3-trimethyl.
  • Step 3 Add 2-amino-5-bromo-3-trimethylethynylpyridine (200mg, 0.71mmol) and 1-methylpyrazole-4-boronic acid pinacol ester (294mg, 1.41 mmol), K 2 CO 3 (244mg, 1.77mmol) and THF/H 2 O (8/2mL), replace oxygen with argon, add Pd(OAc) 2 (16mg, 0.071mmol) and X-Phos (50mg, 0.11mmol), heated to 70°C for 3 hours. After the reaction, the reaction solution was extracted with ethyl acetate (15 mL ⁇ 3) and water (10 mL).
  • Step 4 Add 4-methyl-3-iodobenzoic acid (2.0g, 7.63mmol), HATU (3.8g, 9.92mmol), DIPEA (2.5g, 19.08mmol) and DMF (40mL) into the round-bottomed flask at room temperature. After stirring for 30 minutes, add 3-aminomethylpyridine (825 mg, 7.63 mmol) and react at room temperature for 12 hours; after the reaction, extract the reaction solution with ethyl acetate (50 mL ⁇ 3) and water (40 mL), and use the organic phase with Wash with water (30 mL methyl)benzamide 2.5g (yield: 93%).
  • Step 5 Add 3-iodo-4-methyl-N-(pyridin-3-ylmethyl)benzamide (100 mg, 0.28 mmol), 5-(1-methyl-1H-pyridin) into the round-bottomed flask.
  • Azol-4-yl)-3-((trimethylsilyl)ethynyl)-2-methylaminopyridine 101 mg, 0.35 mmol
  • Et 3 N 86 mg, 0.85 mmol
  • CsF 108 mg, 0.71 mmol
  • MeCN 15 mL
  • replace oxygen with argon add Pd(PPh 3 ) 2 Cl 2 (10 mg, 0.014 mmol), CuI (5.4 mg, 0.028 mmol), repeat the operation of removing oxygen, and react at room temperature for 3 hours.
  • the synthesis method was as in Example 30 except that ethylamine was used instead of methylamine.
  • the synthesis method was as in Example 30, except that ethylamine was used instead of methylamine and 3-aminomethylthiophene was used instead of 3-aminomethylpyridine.
  • the synthesis method was as in Example 30 except that cyclopropylamine was used instead of methylamine and 3-aminomethylthiophene was used instead of 3-aminomethylpyridine.
  • the synthesis method was as in Example 1, except that 1-trifluoromethyl-1H-pyrazolyl-4-boronic acid zanacol ester was used instead of 1-methyl-1H-pyrazolyl-4-boronic acid zanacol ester.
  • the synthesis method was as in Example 1, except that 1-difluoromethyl-1H-pyrazolyl-4-boronic acid zanacol ester was used instead of 1-methyl-1H-pyrazolyl-4-boronic acid zanacol ester.
  • the synthesis method was as in Example 1 except that thiophen-3-ylmethylene-d2-amine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that 6-fluoropyridin-3-ylmethylene-d2-amine was used instead of benzylamine.
  • the synthesis method was as in Example 1 except that (5-cyclopropylthiophen-2-yl)methylamine was used instead of (5-methylthiophen-2-yl)methylamine.
  • the synthesis method was as in Example 61 except that 5-chlorothiophene-2-aldehyde was used instead of 5-cyclopropylthiophene-2-aldehyde.
  • Step 1 Add 4-methyl-3-iodobenzoic acid (2.0g, 7.63mmol), HATU (3.8g, 9.92mmol), DIPEA (2.5g, 19.08mmol) and DMF (40mL) into a round-bottomed flask at room temperature. After stirring at low temperature for 30 minutes, 3-aminomethylthiophene (864 mg, 7.63 mmol) was added, and the reaction was carried out at room temperature for 12 hours. After the reaction, the reaction solution was extracted with ethyl acetate (50 mL ⁇ 3) and water (40 mL).
  • Step 2 Add 3-iodo-4-methyl-N-(thiophen-3-ylmethyl)benzamide (100 mg, 0.28 mmol), 5-bromo-3-(trimethyl Silyl)ethynyl)-2-aminopyridine (90mg, 0.34mmol), Et 3 N (85mg, 0.84mmol), CsF (128mg, 0.84mmol) and MeCN (20mL), replace oxygen with argon, add Pd ( PPh 3 ) 2 Cl 2 (10 mg, 0.014 mmol), CuI (5.3 mg, 0.028 mmol), repeat the operation of removing oxygen, and react at room temperature for 3 hours.
  • Pd ( PPh 3 ) 2 Cl 2 10 mg, 0.014 mmol
  • CuI 5.3 mg, 0.028 mmol
  • Step 3 Add 3-((2-amino-5-bromopyridin-3-yl)ethynyl)-4-methyl-N-(thiophen-3-ylmethyl)benzamide ( 80mg, 0.19mmol), 4-cyclopropylimidazole (30mg, 0.28mmol), CuI (7mg, 0.038mmol), Cs2CO3 (122mg, 0.38mmol) and DMF (2mL), reacted at 120°C for 30h under argon protection. Cool to room temperature, add 10 mL of ethyl acetate to dilute the reaction solution, filter, and wash the filter cake with ethyl acetate.
  • the synthesis method was as in Example 1 except that 5-iodo-pyrimidin-2-amine was used instead of 5-bromo-3-iodopyridin-2-amine and 3-aminomethylthiophene was used instead of benzylamine.
  • Step 1 Removal of (2-((tert-butoxyformyl)((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)thiazole-4-boronic acid zonal ester (
  • N-methylpyrazole-4-boronic acid pinacol ester is replaced and 3-aminomethylthiophene is used instead of benzylamine
  • Intermediate 67-IM can be prepared by referring to the synthesis method of Example 1.
  • Step 2 Add intermediate 67-IM (50 mg, 0.075 mmol) and DCM (2 mL) into a round-bottomed flask, add TFA (0.5 mL), and react at room temperature overnight. Pour the reaction solution into NaHCO 3 aqueous solution, and extract with DCM. Combine the organic phases, wash with NaCl aqueous solution, dry over anhydrous Na 2 SO 4 , evaporate the solvent to dryness under reduced pressure, and separate by column chromatography to obtain Product 40 mg (light yellow solid; yield: 94%).
  • the synthesis method was as follows except that 6-chloro-3-iodoimidazole[1,2-b]pyrazine was used instead of 5-bromo-3-iodopyridin-2-amine and 3-aminomethylthiophene was used instead of benzylamine. example 1.
  • the synthesis method was as in Example 80 except that compound 83 was used instead of compound 79.
  • the synthesis method was as in Example 1, except that 3-bromoimidazole[1,2-a]pyrimidine was used instead of 5-bromo-3-iodopyridin-2-amine and 3-aminomethylthiophene was used instead of benzylamine.
  • Step 1 Add 5-iodo-7H-pyrrole[2,3-d]pyrimidin-4amine (1.0g, 3.85mmol), K 2 CO 3 (1.59g, 11.54mmol) and anhydrous to a round-bottomed flask. DMF (10 mL), add methyl iodide (0.29 mL, 4.61 mmol) dropwise, and react at room temperature for 8 hours.
  • Step 2 Add 5-iodo-7-methyl-7H-pyrrole[2,3-d]pyrimidin-4-amine (0.5g, 1.82mmol) and trimethylsilylacetylene (233mg, 2.37mmol), Et 3 N (5mL) and MeCN (20mL), replace the oxygen with argon, add Pd(PPh 3 ) 2 Cl 2 (64mg, 0.091mmol), CuI (17.4mg, 0.091mmol), and repeat the oxygen removal Operation, react overnight at room temperature; after the reaction is completed, add 50 mL of ethyl acetate to dilute the reaction solution, filter, evaporate the solvent to dryness under reduced pressure, and separate by column chromatography to obtain the product 7-methyl-5-((trimethylsilyl) Ethynyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine 420 mg (yield: 94.2%).
  • Step 3 Add 3-iodo-4-methyl-N-(thiophen-3-ylmethyl)benzamide (90 mg, 0.25 mmol), 7-methyl-5-(trimethyl (silyl)ethynyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (62 mg, 0.25 mmol), Et 3 N (76 mg, 0.76 mmol), CsF (115 mg, 0.76 mmol), and MeCN (10 mL), replace oxygen with argon, add Pd(PPh 3 ) 2 Cl 2 (9 mg, 0.013 mmol) and CuI (5 mg, 0.025 mmol), repeat the operation of removing oxygen, and react at room temperature for 3 hours.
  • Pd(PPh 3 ) 2 Cl 2 9 mg, 0.013 mmol
  • CuI 5 mg, 0.025 mmol
  • the synthesis method was as in Example 72 except that deuterated methyl iodide was used instead of methyl iodide.
  • Step 1 Add compound 90 (0.3g, 0.55mmol) and DCM (5mL) into a round-bottomed flask, add HCl.dioxane solution (0.5mL, 2mmol), and stir at room temperature overnight. After the reaction, the solvent was evaporated to dryness under reduced pressure to obtain 0.26g of yellow solid 91-IM (yield: 98.2%), which was directly used in the next step without further purification.
  • Step 2 Add 106-IM (80mg, 0.17mmol), K 2 CO 3 (115mg, 0.84mmol) and anhydrous DMF (2mL) obtained in the previous step into the round-bottomed flask, and add IEt (16 ⁇ L, 0.2mmol) dropwise. , react at room temperature overnight. After the reaction, pour the reaction solution into water, extract with ethyl acetate, combine the organic phases, wash with H 2 O and NaCl aqueous solution respectively, dry over anhydrous Na 2 SO 4 , evaporate the solvent to dryness under reduced pressure, and perform column chromatography.
  • Step 1 Add 7H-pyrrolo[2,3-d]pyrimidin-4-amine (1.0g, 7.45mmol), DMF-DMA (1.07g, 8.95mmol) and DMF (20mL) into a round-bottomed flask at room temperature. Reaction was allowed to take place overnight. The solvent was evaporated to dryness under reduced pressure to obtain a crude product, which was redissolved in DCM, washed with H 2 O and NaCl aqueous solutions, dried over anhydrous Na 2 SO 4 , and separated by column chromatography to obtain 950 mg of compound 115-2 (yield: 67.4%). .
  • Step 2 Add compound 115-2 (250 mg, 1.32 mmol), 3,6-dihydro-2H-pyran-4-boronic acid pinacol ester (333 mg, 1.59 mmol), and copper acetate (291 mg) into the round-bottomed flask. ,1.59mmol), 2,2'-bipyridine (248mg, 1.59 mmol), anhydrous Na 2 CO 3 (420 mg, 3.96 mmol) and DMAC (10 mL). Stir at 90°C for 4 hours, add DCM and wash with water three times. The organic phase was dried over anhydrous Na 2 SO 4 and evaporated to dryness under reduced pressure to obtain intermediate 115-3 without further purification.
  • Step 3 Add EtOH (10 mL) and ethylenediamine (0.2 mL) to intermediate 115-3 in the previous step, and react under reflux for 16 h. After the reaction, the solvent was evaporated to dryness under reduced pressure, redissolved in DCM, washed with NaCl aqueous solution, and dried over anhydrous Na 2 SO 4. The solvent was evaporated to dryness under reduced pressure and separated by column chromatography to obtain 160 mg of compound 115-4 (two-step total product). rate: 56%).
  • Step 4 Add compound 115-4 (160 mg, 0.74 mmol), NIS (183 mg, 0.81 mmol) and DMF (3 mL) into the round-bottomed flask, and react at room temperature for 4 hours. After the reaction, the solvent was evaporated to dryness under reduced pressure, and 7-(3,6-dihydro-2H-pyran-4-yl)-5-iodo-7H-pyrrolo[2,3-d] was separated by column chromatography. Pyrimidine-4-amine 180 mg (yield: 71%).
  • Step 1 Add 115-2 (0.5g, 2.64mmol) and anhydrous THF (30mL) to the round-bottomed flask, add NaH (60%, 211mg) under ice bath, stir for 30 minutes, add ethyl chloroformate (430mg, 3.96mmol), react at room temperature for 2 hours. After the reaction, the reaction was quenched by adding dropwise a saturated aqueous ammonium chloride solution in an ice bath, extracted with ethyl acetate, the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and separated by column chromatography to obtain 142-1 (400 mg, Yellow oil, yield 57.9%).
  • Step 2 and step 3 can be obtained by referring to step 3 and step 4 of Example 115 respectively to obtain 126-3 as a brown solid.
  • MTT assay was used to detect the effects of compounds on cell proliferation. Collect the cells in good condition, inoculate them into a 96-well plate, add different concentrations of compounds, and culture them in a 37°C, 5% CO 2 , saturated humidity incubator for 72 hours (h). After the drug effect ends, add MTT to each well and continue culturing for 4 hours in a 37°C, 5% CO 2 , saturated humidity incubator. Add triple solution (10% SDS, 5% isobutanol, 0.01mol/L HCl) and place it in a 37°C incubator for 12 hours to ensure that the blue-violet formazan is completely dissolved; measure the OD value at 570nm and 690nm wavelengths on a microplate reader. . Calculate the inhibitory rate of the compound on cells according to the following formula:
  • Inhibition rate (%) (OD value of control hole - OD value of administration hole)/OD value of control hole ⁇ 100%
  • Compound IC 50 was calculated using Graphpad Prism 8.0 software, and a concentration (log)-inhibition rate plot was generated.
  • the heteroaromatic alkynyl compounds in the embodiments of the present invention all have significant inhibitory activity on the proliferation of 32D KIT D816V cells, and 33 of them are better than Ripretinib.
  • the IC 50 value of Examples 76, 77, 99 and 120 against the proliferation of 32D KIT D816V cells is less than 1 nM, which is significantly better than the positive drug Ripretinib, showing that this type of compound has a strong advantage in inhibiting the proliferation of cells carrying KIT D816V drug-resistant mutations. .
  • KIT mutant cells 32D KIT V559D, 32D KIT V559D-V654A, 32D KIT V559D-Y823D and 32D KIT V559D-N822K
  • KIT wild-type cells NCI-H526, Mo7e , HMC-1
  • 32D cell proliferation inhibitory activity the results are shown in Table 3.
  • the representative compounds have strong inhibitory activity on the proliferation of cells carrying different mutant KITs of V559D, V559D-V654A, V559D-Y823D and V559D-N822K, reflecting the advantages of this type of compound in inhibiting mutant KIT in a broad spectrum; at the same time, Representative compounds have weak activity against wild-type KIT-dependent cell lines and have a high therapeutic index compared with mutant KIT. For example, the selection index for KIT D861V is higher than 490 times and is better than the positive drug ripretinib. In addition, the compounds are not cytotoxic to 32D normal cells, indicating that these compounds have good selectivity and can avoid potential off-target toxic side effects.
  • results show that the representative compounds are not cytotoxic to wild-type PDGFR and EGFR-dependent cell lines, and initially show good target selectivity; while the positive drug ripretinib has relatively obvious inhibitory activity against PDGFR-dependent U118MG cells, and treatment The index is lower.
  • Cells were seeded in six-well plates, added with different concentrations of drugs, and treated in a 37°C, 5% CO 2 , saturated humidity incubator for 4 hours. Cells were lysed on ice with 1 ⁇ SDS gel loading buffer (50mM Tris-HCl (pH 6.8), 100mM DTT, 2% SDS, 10% glycerol, 0.1% bromophenol blue). The cell lysate was denatured by heating in a boiling water bath for 10 minutes. Perform SDS-PAGE electrophoresis. After electrophoresis, transfer the protein to a PVDF membrane and place it in blocking solution (5% skimmed milk powder diluted in TBST) at room temperature for 1 hour. After washing the membrane, add the corresponding I and II antibodies and use ECL to The reagent develops color, and is finally observed and photographed in the ECL chemiluminescence imaging system.
  • 1 ⁇ SDS gel loading buffer 50mM Tris-HCl (pH 6.8), 100mM DTT, 2% S
  • mice BALB/c-nu nude mice, 5-6 weeks, ⁇ . Each nude mouse was subcutaneously inoculated with 32D KIT D816V cells. After the average tumor volume reached ⁇ 100 mm 3 , the animals were divided into groups (D0) according to tumor volume. Mice were administered intragastrically (ig) (twice a day) with a volume of 10 mL/kg; the solvent group was given the same volume of "solvent"; the tumor volume was measured twice a week, the mice were weighed, and the data were recorded.
  • Compound 85 (30, 100 mg/kg, i.g., BID ⁇ 12) dose-dependently inhibited the growth of subcutaneous transplanted tumors in 32D KIT D816V nude mice, with tumor inhibition rates of 20% and 50% respectively; compound 77 (20, 40 mg/kg, i.g., BID ⁇ 12) also dose-dependently inhibited the growth of subcutaneous transplanted tumors in 32D KIT D816V nude mice, with tumor inhibition rates of 45% and 71% respectively; Ripretinib (30, 100 mg/kg, i.g., BID ⁇ 12) has an inhibitory effect on 32D KIT The tumor inhibition rates of subcutaneously transplanted tumors in D816V nude mice were 13% and 21% respectively; the tumor-bearing mice could tolerate the above drugs well, and no obvious symptoms such as weight loss occurred. In comparison, the efficacy of compounds 77 and 85 on subcutaneous transplanted tumors in 32D KIT D816V nude mice were significantly stronger than Ripret
  • the compounds containing heteroaryl ring alkynyl groups in the embodiments of the present invention have strong inhibitory activity against 32D KIT D816V cells; at the same time, representative compounds 27, 48, 77 and 85 have strong inhibitory activity against V559D, V559D-V654A, V559D/Y823D and V559D /N822K Different mutant forms of KIT all have potent inhibitory activity, indicating that this type of compound has the advantage of potent and broad-spectrum inhibition of different types of mutant KIT.
  • Representative compounds 27, 48, 77 and 85 have weak inhibitory effects on 32D normal cells and cells carrying wild-type KIT, PDGRF ⁇ and EGFR, indicating that these compounds have high selectivity and can avoid related off-target toxic side effects; the important thing is , the representative compounds 77 and 85 have obvious medicinal effects on the 32D KIT D816V nude mouse subcutaneous transplant tumor model; their medicinal efficacy is significantly stronger than the reference compound Ripretinib (P ⁇ 0.05), indicating that the compounds containing heteroaryl ring alkynyl groups It has the advantage of stronger medicinal effect in the body.

Abstract

本发明涉及一类含杂芳环炔基化合物及其制备方法和用途，所述含杂芳环炔基化合物为式(I)的化合物，或其氘代化合物、药学上可接受的盐、溶剂化物、酯、酸、代谢物或前药。本发明所述的式(I)化合物具有对原发性KIT突变和耐药型KIT突变携带的肿瘤细胞具有强效的杀伤效果。

Description

一类含杂芳环炔基化合物及其制备方法和用途 技术领域
本发明涉及KIT突变抑制剂，具体涉及一类含杂芳环炔基化合物及其制备方法和用途。更具体地，本发明涉及一种能抑制KIT突变细胞株增殖的化合物、该类化合物制备方法以及用于突变型KIT相关疾病的制药用途。
背景技术
KIT(又称CD117或干细胞因子受体)是一个145KDa的跨膜受体酪氨酸激酶(Receptor Tyrosine Kinase,RTK)。KIT属于第三型RTK，由含有5个免疫球蛋白样结构域(D1～D5)、1个跨膜结构域及1个含有近膜结构域(JMD)和酪氨酸激酶(TK)结构域的胞质区域构成。同家族的激酶还包括PDGFRα(血小板源性生长因子受体α)，PDGFRβ(血小板源性生长因子受体β)，FLT3(FMS样酪氨酸激酶3)和CSF-lR(集落刺激因子1受体)。正常情况下，配体干细胞因子(SCF)与胞外结构域结合使受体二聚化，导致胞质区TK结构域内酪氨酸自身磷酸化，进一步引起下游信号通路(如PI3K、JAK-STAT、Ras-Erk、Src家族激酶及PLC等信号通路)的转导，并引发多种生理过程，如细胞增殖、***以及组织的生长、存活。
KIT功能获得性突变与肿瘤、炎症、自身免疫性疾病等多种疾病相关。大量研究表明KIT突变与胃肠道间质瘤(GIST)、***性肥大细胞增多症、肥大细胞白血病、少量的急性髓性白血病、神经胶质瘤及肺癌发生、发展密切相关，其中，突变KIT在GIST中作用研究得最为广泛、深入，约85％GIST由KIT突变所致。GIST是胃肠道最常见的间叶源性肿瘤，发病率约为1/10万～2/10万，占全部消化道肿瘤的1～3％，其中大约80％的GIST患者具有位于近膜结构域(exon 11)及胞外结构域(exon 9)的原发性KIT突变。在这些原发性突变中，发生在JM区的突变(例如V560D)最为常见，并可导致配体非依赖性的KIT组成性激活，进而诱导GIST发生(Chen,L.L.et al.Clin.Cancer Res.2005,11,pg.3668-3677；Mol,C.D.,et al.J.Biol.Chem.2004,279,pg.31655-31663)。Imatinib作为治疗GIST的一线药物，对大多数原发性KIT突变的GIST均有效，但90％患者最终均会由于KIT发生次级突变而产生耐药，导致肿瘤复发。这些耐药突变主要发生于KIT的ATP结合口袋(exon 13,如K642E,V654A；exon 14,如T670I)和活化环(A-loop；exon 17,如D816H,D816V,D820A,N822K,N822H,N822V)。Sunitinib和Regorafenib作为GIST的二线和三线用药，只能克服少数Imatinib耐药性突变(如发生于ATP结合口袋的V654A和T670I突变)，且临床响应率较低。据报道，在Imatinib耐药性病例中，50％患者具有A-loop的继发性突变(Demetri GD,et al.Lancet.2006,368:1329-1338；Nishida T,et al.Int.J.Clin.Oncol.2009,14:143-149)。因此，开发能逆转A-loop(exon 17)突变所致耐药性的新型KIT抑制剂具有重要的临床意义。
目前，开发对耐药性GIST具有疗效的KIT抑制剂进展缓慢，特别对发生于A-loop的耐药性突变具有强效抑制活性的小分子KIT抑制剂更为困难。Ripretinib于2020年被 FDA批准用于GIST的治疗，能克服包括exon 17在内的多种KIT耐药性突变，为耐药性的GIST患者提供了新的治疗手段。但Ripretinib疗效还有待提高，其用于GIST二线治疗试验未能优效sunitinib而失败；另外，ripretinib还存在高血压、腹痛等严重不良反应,与其激酶选择性差相关，特别是对VEGFR2具有较强抑制活性。因此，研制出活性更强、对KIT选择性更好、对耐药突变KIT更有效的药物，有效治疗GIST具有重要意义且十分迫切。
CN104211639A、CN108456163A和CN111662227A均涉及杂芳环炔基类化合物，其中涉及连接链L为-C(O)NH-、-NHC(O)-、醚链或氨基链的化合物。WO2015089210A1公开的杂芳环炔基类化合物，虽然连接链包括-C(O)NHCH2-的情况，但其吡啶环间位必须有砜亚胺作为优势结构，并且中间苯环必须为无取代基的情况；此外，其关注点在VEGFR和PDGFR上，而不涉及对KIT突变细胞株的抑制活性。
发明内容
本申请发明人在CN104211639A、CN108456163A和CN111662227A的基础上，在对连接链L为-C(O)NH-、-NHC(O)-、醚链或氨基链的研究中发现，对L改构成-C(O)NHC(Ra)(Rb)-后，可显著提高化合物对KIT突变细胞株的抑制活性(如表格1所示)。本申请基于实施例1发现该类新颖母核结构对KIT突变细胞株均体现优异的抑制剂活性，具有选择性。
表1结构L对活性的影响
本发明提供一类结构新颖的杂芳环炔基类小分子激酶抑制剂，该类化合物对KIT突变细胞株(包括KIT D816V在内的多种突变形式)具有非常强的增殖抑制活性。同时，该类化合物具有高选择性，对非肿瘤细胞(32D细胞)及KIT野生型细胞(NCI-H526、Moe7)均没有明显细胞毒性，对EGFR、PDGFR依赖的细胞抑制活性弱，体现出该类化合物具有较高选择性。此外，代表性化合物体内抑瘤效果显著，并且毒性低。
本发明提供一种式(I)的化合物，或其氘代化合物、药学上可接受的盐、溶剂化物、酯、酸、代谢物或前药：
其中：
X选自-(C(Ra)(Rb))q-；
其中Ra、Rb各自独立地选自氢、氘、卤素、C1-4烷基；或者连接在同一个碳原子上的Ra、Rb与该碳原子一起形成3-5元碳环；
q选自1、2；
R1选自氢、卤素、C1-6烷基；
环M1选自以下结构：
其中：
A1、A2各自独立CRH4或N原子；
A3为CRH2或N原子；
A4为CRH3、NRH3或N原子；
Z选自未取代或取代的5-10元杂芳基、未取代或取代的5-10元杂环基；所述取代是指上述基团各自独立地被1-5个R2基团取代；或者Z和与其相连的碳原子以及相邻的A1或A2一起形成取代或未取代的环W1，所述取代是指环W1被1-5个R2基团取代；在杂芳基、杂环基，或者环W1中，一个或多个环C原子任选地被相应数目的C(＝O)基团替代，一个或多个环S或N原子任选地被氧化形成S-氧化物或N-氧化物；
W为未取代或取代的C6-10芳基、未取代或取代的5-7元杂芳基、未取代或取代的5-7元杂环基；所述取代是指被1-5个R2基团取代；在芳基、杂芳基或杂环基中，一个或多个环C原子任选地被相应数目的C(＝O)基团替代，一个或多个环S或N原子任选地被氧化形成S-氧化物或N-氧化物；
所述R2各自独立选自卤素、氰基、C1-6烷基、C1-6杂烷基、C1-6烷氧羰基、C2-6烯基、C2-6炔基、C3-6环烷基、3-6元杂环烷基、C4-8环烯基、4-8元杂环烯基、C6-10芳基、5-7元杂芳基、5-7元杂环基、-C(＝O)(C1-6烷基)、-C(＝O)(C3-6环烷基)、-OR3、-N(R3)2、-(C1-6烷基)OR3、-C(＝O)N(R3)2、-(C1-6烷基)-C(＝O)N(R3)2、-SR3、-S(＝O)R3、-S(＝O)2R3、-N(R3)C(＝O)R3、-(C1-6烷基)-N(R3)C(＝O)R3、-N(R3)S(＝O)2R3、-P(＝O)(R3)(R3)；其中所述C1-6烷基、C1-6杂烷基、C1-6烷氧羰基、C2-6烯基、C2-6炔基、C3-6环烷基、3-6元杂环烷基、4-8元环烯基、4-8元杂环烯基、C6-10芳基、5-7元杂芳基、5-7元杂环基任选被1-5个R3取代；或者
两个R2和与其相连的原子形成取代或未取代的3-7元碳环或杂环，所述取代是指被1-5个R3取代；
所述R3各自独立选自羟基、氰基、氨基、卤素、C1-6烷基、C1-6烷氧羰基、C3-C6环烷基、C1-6杂烷基、C1-6卤代烷基、羟基C1-6烷基、-NH(C1-6烷基)、-NH(C3-6环烷基)、-NHC(＝O)(C1-6烷基)、被氰基取代的包含选自O、N、S中的一个或多个的5-7元杂环基甲基；
RH1、RH2和RH4各自独立选自氢、卤素、氨基、氰基、羟基、-N(R4)(R5)；
所述R4、R5各自独立选自氢、C1-6烷基、C1-6杂烷基、C3-6环烷基、C3-6杂环烷基、C1-6烷基酰基、C3-6环烷基酰基；
RH3选自氢、未取代或取代的C1-6烷基、未取代或取代的C1-6杂烷基、-C(＝O)OC1-6烷基、-C(＝O)OC3-6环烷基、-C(＝O)C1-6烷基、-C(＝O)C3-6环烷基、未取代或取代的C3-6环烷基、未取代或取代的C2-6烯基、未取代或取代的C4-6环烯基、未取代或取代的C6-10芳基、未取代或取代的5-7元杂芳基、未取代或取代的4-7元杂环基；所述取代是指各自独立地被1-5个R6所取代；在环烷基、环烯基、芳基、杂芳基、杂环基中，一个或多个环C原子任选地被相应数目的C(＝O)基团替代，一个或多个环S或N原子任选地被氧化形成S-氧化物或N-氧化物；
所述R6各自独立选自卤素、氰基、C1-6烷基、C1-6杂烷基、C2-6烯基、C2-6炔基、C3-6环烷基、C4-8环烯基、C6-10芳基、5-7元杂芳基、5-7元杂环基、-C(＝O)R7、OR7、-N(R7)2、-(C1-6烷基)OR7、-C(＝O)N(R7)2、-(C1-6烷基)-C(＝O)N(R7)2、-SR7、-S(＝O)R7、-S(＝O)2R7、-N(R7)C(＝O)R7、-(C1-6烷基)-N(R7)C(＝O)R7、-N(R7)S(＝O)2(R7)、-P(＝O)(R7)(R7)；其中所述C1-6烷基、C1-6杂烷基、C2-6烯基、C2-6炔基、C3-6环烷基、C4-8环烯基、C6-10芳基、5-7元杂芳基、5-7元杂环基任选被1-5个R7取代；或者
两个R6与其相连的原子一起形成取代或未取代的5-7元碳环或5-7元杂环，所述取代是指被1-5个R7基团取代；
所述R7各自独立选自卤素、羟基、氰基、氨基、C1-6烷基、C2-6烯基、C2-6炔基、C3-6环烷基、C1-6杂烷基、C4-6杂环烷基、C1-6卤代烷基、羟基C1-6烷基、NH(C1-6环烷基)、-NHC(＝O)(C1-6烷基)、C(＝O)(C1-6烷基)、C(＝O)(C3-6环烷基)
环M2选自未取代或取代的C6-10芳基、未取代或取代的5-7元杂芳基、未取代或取代的5-7杂环基；所述取代是指各自独立地被1-5个RM所取代；
所述RM各自独立地选自卤素、氰基、C1-6杂烷基、C2-6烯基、C2-6炔基、C3-6环烷基、3-6元杂环烷基、C4-6环烯基、C6-10芳基、5-7元杂芳基、-C(＝O)(C1-6烷基)、-C(＝O)(C3-6环烷基)、-(C1-6烷基)-R8、-OR8、-N(R8)2、-NH(C1-6烷基)-R8、-O(C1-6烷基)-R8、-C(＝O)N(R8)2、-SR8、-S(＝O)R8、-S(＝O)2R8；其中所述C1-6烷基、C3-6环烷基、3-6元杂环烷基、C4-6环烯基、4-6元杂环烯基、C6-10芳基、5-7元杂芳基任选各自独立地被1-5个R8取代；或者
两个RM和与其相连的原子一起形成取代或未取代的3-7元碳环或杂环，所述取代是指被1-5个R8基团取代；
所述R8各自独立选自羟基、氰基、氨基、卤素、-NH(C1-6烷基)、C1-6烷基、C1-6杂烷基、C1-6卤代烷基、C1-6羟基烷基、C3-6环烷基、C3-6杂环烷基。
在优选实施方式中，化合物具有下述式(II)的结构：
其中，
R1选自卤素、C1-3烷基；优选选自甲基、F、Cl；
Ra、Rb各自独立选自氢、氘、卤素、甲基；或者Ra和Rb和与它们连接的碳原子一起形成三元碳环；
环M1选自以下结构：
其中A1、A2、A3、A4、环W、Z和RH1的定义各自如上述所述；以及
环M2的定义如上所述。
在优选实施方式中，环M1选自下述的结构：
其中，环原子上的“C,N”表示该处为CH或者N，
R2、RH1、RH2和RH3的定义各自如上述所述。
在优选实施方式中，
环M2选自未取代或取代的苯基、未取代或取代的噻吩基、未取代或取代的吡唑基、未取代或取代的噻唑基、未取代或取代的吡啶基、未取代或取代的噁唑基、未取代或取代的异噁唑基、未取代或取代的苯并噻吩基；所述取代是指各自独立地被1-5个RM所取代；
所述RM各自独立地选自卤素、羟基、氨基、氰基、C1-6烷基、C1-6杂烷基、C2-6烯基、C2-6炔基、C3-6环烷基、3-6元杂环烷基、C4-6环烯基、4-6元杂环烯基、C6-10芳基、5-7元杂芳基、-C(＝O)(C1-6烷基)、-C(＝O)(C3-6环烷基)、-(C1-6烷基)-R8、-OR8、-N(R8)2、-NH(C1-6烷基)-R8、-O(C1-6烷基)-R8、-C(＝O)N(R8)2、-SR8、-S(＝O)R8、-S(＝O)2R8； 其中所述C1-6烷基、C3-6环烷基、3-6元杂环烷基、C4-6环烯基、4-6元杂环烯基、C6-10芳基、5-7元杂芳基任选各自独立地被1-5个R8取代；或者
两个RM和与其相连的原子形成取代或未取代的3-7元碳环或杂环，所述取代是指被1-5个R8基团取代；
所述R8各自独立选自羟基、氰基、氨基、卤素、-NH(C1-6烷基)、C1-6烷基、C1-6杂烷基、C1-6卤代烷基、C1-6羟基烷基、C3-6环烷基、C3-6杂环烷基。
在本发明中，
所述烷基为具有1-8个碳原子的饱和脂肪族直链或支链的烷基；典型的烷基包括但不限于：甲基、乙基、丙基、异丙基、环丙基、正丁基、异丁基、叔丁基等。
所述卤素各自独立地选自F、Cl、Br、I；
所述卤代烷基为烷基中至少一个氢原子被卤原子置换。在某些实施方式中，如果两个或更多氢原子被卤原子置换，所述卤原子彼此相同或不同；
所述杂烷基为烷基中至少一个骨架C原子被杂原子(N、O、S)置换，例如，杂烷基的例子可以包括烷氧基、烷基氨基、烷基硫基等。在某些实施方式中，如果两个或更多C原子被杂原子置换，所述杂原子彼此相同或不同；
所述环烷基为饱和或部分不饱和的3-10元单环或多环脂环，同时可以是单价基团或双价基团(即亚环烷基)；
所述杂环烷基为环上含有选自N、O、S中一个或多个杂原子的饱和3-10元单环或多环脂杂环，同时可以是单价基团或双价基团(即亚杂环烷基)；
所述芳基是指芳香基环中每一个构成环的原子都是碳原子，包括单环或稠环多环，同时可以是单价基团或双价基团(即亚芳基)。在本发明中，芳基环优选5-10个碳原子，更优选具有5-7个碳原子的芳基。
所述杂芳基为环上含有选自N、O、S中一个或多个杂原子的芳香基。根据结构，杂芳基可以是单价基团或双价基团(即亚杂芳基)。
所述杂环基为单环或多环，并且至少一个为环上含有选自N、O、S中一个或多个杂原子的非芳香性环基团。根据结构，杂环基可以是单价基团或双价基团(即亚杂环基)。
优选地，上述的式(I)化合物选自下式化合物：








本发明还提供一种制备上述的式(I)化合物的方法，包括如下步骤：式(A)化合物与式(B)化合物进行偶联反应得到式(I)化合物。
其中，环M1、环M2、X及R1的定义各自独立地如上所述；TMS为-Si(CH3)3；
优选地，所述偶联反应在钯金属催化剂和铜金属催化剂存在下，在碱存在下进行，所述钯金属催化剂包括Pd(PPh3)2Cl2、Pd(OAc)2、和Pd(PPh3)4中的一种或多种；优选地，所述铜金属催化剂包括CuI和/或CuCl；优选地，所述碱包括CsF、Cs2CO3、KF、K2CO3、NaHCO3、Na2CO3、Et3N、(iPr)2EtN、DMAP中的一种或者两种以上；
优选地，所述偶联反应在溶剂存在下进行，所述的溶剂包括乙腈、1,4-二氧六环、DMF中的一种或两种以上。
进一步优选地，所述方法包括如下步骤：式(A)化合物与式(B)化合物在氟化铯、Pd(PPh3)2Cl2、CuI和三乙胺的存在下，在乙腈溶剂中进行偶联反应得到式(I)化合物。
进一步优选地，本发明提供一个指导性的合成方案(如合成路线I所示)。应当理解的是，可以利用有机化学的一般知识对合成方案中所示的试剂/反应条件进行修改或优化，以制备本发明中不同的化合物。
合成路线I
所述A1、A2、环M2、X、Z、RH1及R1分别独立如前所述，合成路线I包括以下步骤：
步骤1：将化合物I-1、I-2和Et3N混合，加入钯金属催化剂、铜金属催化剂，氩气保护下反应(例如在室温条件下)得到化合物I-3；
步骤2：将化合物I-3、I-4加入合适的溶剂中，加入钯金属催化剂、K2CO3，氩气保护下，加热反应得到重要中间体A；此外，也可加入合适的配体；
步骤3：化合物I-5及I-6发生缩合反应，其中所述X如前所述。将化合物I-5、HATU、DIPEA和DMF混合(例如，室温下搅拌30～60分钟)后，加入化合物I-6，反应(例如室温下反应6～12小时)得到中间体B；
步骤4：将中间体A、B、碱和MeCN混合，加入钯金属催化剂、铜金属催化剂，氩气保护下反应(例如在I取代的情况下，在室温条件下反应，而在Br取代的情况下，在80℃反应，反应时间例如2～6小时)得到化合物I-7；
优选地，
步骤1、4所述的钯金属催化剂为Pd(PPh3)2Cl2，铜金属催化剂为CuI；
步骤2所述溶剂为甲苯、乙醇、乙二醇二甲醚、四氢呋喃、1,4-二氧六环和水中的一种或多种；所述钯金属催化剂为Pd(PPh3)4,Pd(OAC)2和(dppf)PdCl2中的任意一种；所述碱为K2CO3,Cs2CO3,NaHCO3和Na2CO3中的任意一种；所述配体为X-Phos和PPh3中的任意一种；
步骤4所述碱为氟化铯和/或三乙胺；
进一步优选地，合成路线I包括如下步骤：
步骤1：向圆底烧瓶中加入化合物I-1、和Et3N，用氩气置换氧气，加入Pd(PPh3)2Cl2、CuI，重复除氧气的操作，室温条件下反应约15分钟，加入I-2，室温继续反应3小时 结束，纯化即可得到化合物I-3；其中，化合物I-1、I-2、Pd(PPh3)2Cl2和CuI的当量可以分别约为1.0,1.0～1.5,0.05～0.1,0.1～0.2。
步骤2：向圆底烧瓶中加入化合物I-3、I-4、K2CO3、THF和水，用氩气置换氧气，加入Pd(OAc)2和X-Phos,重复除氧气的操作，加热至80℃反应，反应4-8小时结束，纯化即可得到化合物A；其中I-3、I-4、K2CO3、Pd(OAc)2和X-Phos的当量可以分别为1.0，1.0-1.5,2.0-3.0，0.05-0.1,0.1-0.2；THF/H2O的体积比为4/1。
步骤3：向圆底烧瓶中加入化合物I-5、HATU、DIPEA和DMF，室温下搅拌30分钟后，加入化合物I-6，室温下进行反应；反应12小时结束，纯化即可得到化合物B；化合物I-5、I-6、HATU和DIPEA的当量可以分别约为1.0,1.0～1.2,1.0～1.5,2.0～4.0。
步骤4：向圆底烧瓶中加入化合物A、B、Et3N、CsF和MeCN，用氩气置换氧气，加入Pd(PPh3)2Cl2、CuI，重复除氧气的操作，室温或加热至80℃反应，反应3-6小时结束，纯化即可得到化合物I-7；化合物A、B、氟化铯、Pd(PPh3)2Cl2、CuI和Et3N的当量可以分别约为1.0,1.0～1.5,2.5～3.0,0.05～0.1,0.1～0.2,2.5～3.0。
本发明还提供一种药物组合物，其包括：选自上述式(I)化合物、其氘代化合物、药学上可接受的盐、溶剂化物、酯、酸、代谢物和前药中的一种或多种，以及药学上可接受的辅料。
本发明还提供上述的式(I)化合物或其氘代化合物、医药上可接受的盐、溶剂化物、酯、酸、代谢物或前药或者上述药物组合物在制备KIT(突变型)抑制剂中的用途。
本发明还提供上述的式(I)化合物或其氘代化合物、药学上可接受的盐、溶剂化物、酯、酸、代谢物或前药，或上述药物组合物在制备用于治疗、预防或改善一种或多种疾病或病症的药物中的用途，所述疾病或病症选自肿瘤、炎症、自身免疫性和神经***性疾病。
优选地，所述肿瘤是胃肠道间质瘤，特别是涉及KIT突变的胃肠道间质瘤，更特别地是对Imatinib和/或Sunitinib耐药的由KIT突变引起的胃肠道间质瘤。
本发明所述的式(I)化合物具有对原发性KIT突变和耐药型KIT突变携带的肿瘤细胞具有强效的杀伤效果。
根据本发明另一具体实施方式，根据本发明的含杂芳环炔基结构的化合物对携带D816V、V559D、V559D-V654A、V559D/Y823D和V559D/N822K不同突变形式KIT的肿瘤细胞株均显示出了很强的抑制细胞增殖活性。
根据本发明另一具体实施方式，根据本发明的含杂芳环炔基结构的化合物对正常的32D细胞及KIT野生型细胞株NCI-H526细胞株细胞毒性低，显示出所述化合物具有的毒性选择性高的优点。
根据本发明另一具体实施方式，根据本发明的含杂芳环炔基结构的化合物对BCR-ABL、EGFR、PDGFR依赖的细胞株具有较低的抑制活性，显示出该类化合物具有相对选择性的优点。
根据本发明另一具体实施方式，根据本发明的含杂芳环炔基结构的化合物在长期的动物药效模型上表现出明显的抑制相关肿瘤生长的活性，显著优于阳性药Ripretinib。
根据本发明另一具体实施方式，在药效剂量下，动物状态好(包括体重无明显下降)，无动物死亡的现象。
附图说明
图1是表现出化合物77、85和Ripretinib对32D KIT D816V细胞KIT及其下游信号通路的影响的图。
图2是表现出化合物27、48和Ripretinib对32D KIT V559D细胞KIT及其下游信号通路的影响的图。
图3是表现出化合物27、48和Ripretinib对32D KIT V559D-V654A细胞KIT及其下游信号通路的影响的图。
图4是表现出化合物77、85、Ripretinib对32D KIT D816V裸小鼠皮下移植瘤的疗效的图。
图5是表现出化合物77、85、Ripretinib对荷瘤(32D KIT D816V)裸小鼠体重的影响的图。
具体实施方式
以下对本发明的具体实施方式进行详细说明。应当理解的是，此处所描述的具体实施方式仅用于示例性地对本发明进行说明，并不用于限制本发明。
(一)化合物制备实施例
实施例1 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-苄基苯甲酰胺的制备
步骤1：向圆底烧瓶中加入2-氨基-5-溴-3-碘吡啶(1.0g,3.35mmol)、三甲基硅乙炔(427.2mg,4.35mmol)和Et3N(50mL)，用氩气置换氧气，加入Pd(PPh3)2Cl2(117.4mg,0.17mmol)、CuI(63.7mg,0.33mmol)，重复除氧气的操作，室温条件下反应6小时；反应结束后加入50mL乙酸乙酯稀释反应液，过滤即可得到产物2-氨基-5-溴-3-三甲基乙炔基吡啶900mg(产率：99.9％)。
步骤2：向圆底烧瓶中加入2-氨基-5-溴-3-三甲基乙炔基吡啶(300mg,1.11mmol)、1-甲基吡唑-4-硼酸频哪醇酯(348mg,1.67mmol)、K2CO3(385mg,2.79mmol)和THF/H2O(8/2mL)，用氩气置换氧气，加入Pd(OAc)2(25mg,0.11mmol)和X-Phos(106 mg,0.22mmol)，加热至70℃反应4小时。反应结束后，用乙酸乙酯(15mL×3)和水(10mL)萃取反应液，有机相用饱和NaCl水溶液洗(10mL×3)，无水Na2SO4干燥，减压蒸干溶剂，柱层析分离得5-(1-甲基-1H-吡唑-4-基)-3-((三甲基硅基)乙炔基)-2-氨基吡啶245.0mg(产率：81.31％)。
步骤3：向圆底烧瓶中加入化合物4-甲基-3碘苯甲酸(2.0g,7.63mmol)、HATU(3.8g,9.92mmol)、DIPEA(2.5g,19.08mmol)和DMF(40mL)，室温下搅拌30分钟后，加入化合物苄胺(818mg,7.63mmol)，室温反应12小时；反应结束后，乙酸乙酯(50mL×3)和水(40mL)萃取反应液，有机相分别用自来水洗(30mL×3)，饱和NaCl溶液洗(30mL×3)，无水硫酸钠干燥，减压蒸干溶剂，柱层析分离得N-苄基-3-碘-4-甲基苯甲酰胺2.5g(产率：93％)。
步骤4：向圆底烧瓶中加入化合物N-苄基-3-碘-4-甲基苯甲酰胺(100mg,0.28mmol)、5-(1-甲基-1H-吡唑-4-基)-3-((三甲基硅基)乙炔基)-2-氨基吡啶(100mg,0.37mmol)、Et3N(86mg,0.85mmol)、CsF(108mg,0.71mmol)和MeCN(20mL)，用氩气置换氧气，加入Pd(PPh3)2Cl2(10mg,0.014mmol)、CuI(5.4mg,0.028mmol)，重复除氧气的操作，室温条件下反应3小时。反应结束后，乙酸乙酯(30mL×3)和水(20mL)萃取反应液，有机相用饱和NaCl溶液洗(10mL×3)，经无水硫酸钠干燥后，减压蒸干溶剂，柱层析分离得产物3-(2-氨基吡啶基-5-溴-3-乙炔基)-4-甲基-N-苄基苯甲酰胺78mg(产率：65％)。1H NMR(400MHz,DMSO-d6)δ9.08(t,J＝6.0Hz,1H),8.26(d,J＝2.3Hz,1H),8.17(d,J＝1.9Hz,1H),8.08(s,1H),7.84(d,J＝2.4Hz,1H),7.83–7.79(m,2H),7.43(d,J＝8.0Hz,1H),7.35–7.30(m,4H),7.27–7.21(m,1H),6.28(s,2H),4.48(d,J＝5.9Hz,2H),3.84(s,3H),2.53(s,3H).LR-MS 422.1(M+1).
实施例2 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-苯基乙基苯甲酰胺的制备
除用苯乙胺代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ8.60(t,J＝5.8Hz,1H),8.26(d,J＝2.4Hz,1H),8.09(d,J＝4.0Hz,2H),7.85(d,J＝2.3Hz,1H),7.82(s,1H),7.74(dd,J＝1.8,8.0Hz,1H),7.41(d,J＝8.1Hz,1H),7.33–7.23(m,4H),7.20(t,J＝7.2Hz,1H),6.28(s,2H),3.84(s,3H),3.49(q,J＝6.8Hz,2H),2.85(t,J＝7.5Hz,2H),2.53(s,3H).LR-MS 436.1(M+1).
实施例3 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(1-苯基环丙基)苯甲酰胺的制备
除用1-苯基环丙基胺代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.21(s,1H),8.26(d,J＝2.4Hz,1H),8.16(d,J＝1.8Hz,1H),8.08(s,1H),7.84(d,J＝2.3Hz,1H),7.82(s,1H),7.79(d,J＝7.5Hz,1H),7.42(d,J＝8.0Hz,1H),7.27(t,J＝7.6Hz,2H),7.22–7.12(m,3H),6.28(s,2H),3.84(s,3H),2.54(s,3H),1.06(br s,4H).LR-MS 448.2(M+1).
实施例4 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-苄基-5-氟苯甲酰胺的制备
除用5-氟-3-碘苯甲酸代替4-甲基-3-碘苯甲酸外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.23(t,J＝6.0Hz,1H),8.27(d,J＝2.3Hz,1H),8.06(s,1H),8.01(s,1H),7.82(d,J＝2.3Hz,1H),7.82–7.75(m,3H),7.71(d,J＝9.1Hz,1H),7.34(d,J＝4.5Hz,4H),7.30–7.22(m,1H),6.46(s,2H),4.50(d,J＝5.9Hz,2H),3.84(s, 3H).LR-MS 426.2(M+1).
实施例5 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-5-氟-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用5-氟-3-碘苯甲酸代替4-甲基-3-碘苯甲酸和3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.25(t,J＝5.9Hz,1H),8.18(d,J＝1.3Hz,1H),8.13–8.06(m,1H),8.01(d,J＝1.1Hz,1H),7.96(t,J＝2.0Hz,1H),7.75(d,J＝1.5Hz,1H),7.62–7.53(m,1H),7.36(d,J＝7.5Hz,1H),7.05(dd,J＝1.6,7.5Hz,1H),7.00(d,J＝1.5Hz,1H),6.97(d,J＝1.6Hz,1H),6.29(s,2H),4.55(d,J＝5.7Hz,2H),3.87(s,3H).LR-MS432.1(M+1).
实施例6 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-5-氟-N-(吡啶-3-基甲基)苯甲酰胺的制备
除用5-氟-3-碘苯甲酸代替4-甲基-3-碘苯甲酸和3-氨甲基吡啶代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.23(t,J＝6.0Hz,1H),8.64(d,J＝1.3Hz,1H),8.47(dd,J＝1.3,5.0Hz,1H),8.13(d,J＝1.3Hz,1H),8.12–8.07(m,2H),7.90–7.83(m,3H),7.61–7.54(m,1H),7.50(dd,J＝5.0,8.0Hz,1H),7.05(d,J＝1.5Hz,1H),6.28(s,2H),4.50(d,J＝5.8Hz,2H),3.85(s,3H).LR-MS 427.2(M+1).
实施例7 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-5-氟-N-((6-氟吡啶-3-基)甲基)苯甲酰胺的制备
除用5-氟-3-碘苯甲酸代替4-甲基-3-碘苯甲酸和6-氟-3-氨甲基吡啶代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.26(t,J＝5.9Hz,1H),8.38(d,J＝1.4Hz,1H),8.14(d,J＝1.3Hz,1H),8.08(dd,J＝1.6,8.7Hz,2H),8.04–7.99(m,1H),7.95(t,J＝2.0Hz,1H),7.85(d,J＝1.6Hz,1H),7.53(dt,J＝2.0,8.8Hz,1H),7.16(t,J＝8.0Hz,1H),7.06(d,J＝1.6Hz,1H),6.27(s,2H),4.50(d,J＝5.6Hz,2H),3.83(s,3H).LR-MS 445.4(M+1).
实施例8 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-(4-氯苄基)-4-甲基苯甲酰胺的制备
除用4-氯苄胺代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.10(t,J＝6.3Hz,1H),8.26(d,J＝2.2Hz,1H),8.17(s,1H),8.09(s,1H),7.88(d,J＝2.3Hz,1H),7.81(d,J＝9.9Hz,2H),7.44(d,J＝8.0Hz,1H),7.41–7.32(m,4H),6.38(s,2H),4.46(d,J＝6.0Hz,2H),3.84(s,3H),2.54(s,3H).LR-MS 456.7(M+1).
实施例9 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-(2-三氟甲基苄基)-4-甲基苯甲酰胺的制备
除用2-三氟甲基苄胺代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.15(t,J＝5.7Hz,1H),8.26(d,J＝2.2Hz,1H),8.20(s,1H),8.08(s,1H),7.88–7.79(m,3H),7.74(d,J＝7.8Hz,1H),7.66(t,J＝7.6Hz,1H),7.54(d,J＝7.8Hz,1H),7.52–7.43(m,2H),6.27(s,2H),4.67(d,J＝5.6Hz,2H),3.84(s,3H),2.55(s,3H).LR-MS 490.5(M+1).
实施例10 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-(2-甲氧基苄基)-4-甲基苯甲酰胺的制备
除用2-甲氧基苄胺代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ8.88(t,J＝5.9Hz,1H),8.26(d,J＝2.3Hz,1H),8.18(s,1H),8.08(s,1H),7.86–7.79(m,3H),7.43(d,J＝8.0Hz,1H),7.24(t,J＝7.8Hz,1H),7.19(d,J＝7.4Hz,1H),7.00(d,J＝8.2Hz,1H),6.91(t,J＝7.4Hz,1H),6.26(s,2H),4.45(d,J＝5.7Hz,2H),3.84(s,3H),3.83(s,3H),2.54(s,3H).LR-MS 452.3(M+1).
实施例11 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-(3-甲氧基苄基)-4-甲基苯甲酰胺的制备
除用3-甲氧基苄胺代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.04(d,J＝5.9Hz,1H),8.26(d,J＝2.2Hz,1H),8.16(s,1H),8.08(s,1H),7.86–7.77(m,3H),7.43(d,J＝8.0Hz,1H),7.24(t,J＝8.0Hz,1H),6.93–6.86(m,2H),6.82(d,J＝8.3Hz,1H),6.26(s,2H),4.45(d,J＝5.9Hz,2H),3.84(s,3H),3.73(s,3H),2.53(s,3H).LR-MS 452.3(M+1).
实施例12 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-(3-甲氧基苄基)-4-甲基苯甲酰胺的制备
除用4-甲氧基苄胺代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.01(t,J＝6.1Hz,1H),8.27(d,J＝2.3Hz,1H),8.17(s,1H),8.10(s,1H),7.90(s,1H),7.84(s,1H),7.81(d,J＝8.3Hz,1H),7.43(d,J＝8.0Hz,1H),7.26(d,J＝8.3Hz,2H),6.90(d,J＝8.3Hz,2H),6.42(s,2H),4.41(d,J＝5.8Hz,2H),3.85(s,3H),3.73(s,3H),2.54(s,3H).LR-MS 452.3(M+1).
实施例13 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-(4-氟苄基)-4-甲基苯甲酰胺的制备
除用4-氟苄胺代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.08(t,J＝6.0Hz,1H),8.26(s,1H),8.16(s,1H),8.09(s,1H),7.87(s,1H),7.84–7.78(m,2H),7.43(d,J＝8.0Hz,1H),7.40–7.32(m,2H),7.16(t,J＝8.7Hz,2H),6.36(s,2H),4.46(d,J＝5.9Hz,2H),3.84(s,4H),2.53(s,3H).LR-MS 440.2(M+1).
实施例14 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-(2,4-二氟苄基)-4-甲基苯甲酰胺的制备
除用2,4-二氟苄胺代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.05(t,J＝5.8Hz,1H),8.26(d,J＝2.3Hz,1H),8.15(s,1H),8.07(s,1H),7.83(d,J＝2.2Hz,1H),7.80(d,J＝9.5Hz,2H),7.43(q,J＝8.1Hz,2H),7.23(t,J＝10.0Hz,1H),7.07(td,J＝2.4,8.6Hz,1H),6.26(s,2H),4.48(d,J＝5.6Hz,2H),3.84(s,3H),2.53(s,3H).LR-MS 458.1(M+1).
实施例15 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-(2-氟-4-氯苄基)-4-甲基苯甲酰胺的制备
除用2-氟-4-氯苄胺代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.07(t,J＝5.0Hz,1H),8.26(s,1H),8.15(s,1H),8.07(s,1H),7.84–7.77(m,3H),7.46–7.37(m,3H),7.28(d,J＝8.5Hz,1H),6.26(s,2H),4.48(d,J＝5.6Hz,2H),3.84(d,J＝1.4Hz,3H),2.53(s,3H).LR-MS 474.7(M+1).
实施例16 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(吡啶-3-基甲基)苯甲酰胺的制备
除用3-氨甲基吡啶代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.13(t,J＝5.9Hz,1H),8.57(s,1H),8.47(d,J＝3.9Hz,1H),8.26(d,J＝1.8Hz,1H),8.15(s,1H),8.07(s,1H),7.84(d,J＝2.3Hz,1H),7.82–7.77(m,2H),7.74(d,J＝8.0Hz,1H),7.43(d,J＝8.0Hz,1H),7.37(dd,J＝4.8,7.8Hz,1H),6.27(s,2H),4.50(d,J＝5.8Hz,2H),3.84(s,3H),2.53(s,3H).LR-MS 474.7(M+1).LR-MS 423.5(M+1).
实施例17 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(吡啶-4-基甲基)苯甲酰胺的制备
除用4-氨甲基吡啶代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.17(t,J＝6.1Hz,1H),8.51(d,J＝4.8Hz,2H),8.26(d,J＝2.3Hz,1H),8.18(s,1H),8.08(s,1H),7.86–7.79(m,3H),7.45(d,J＝8.1Hz,1H),7.32(d,J＝5.3Hz,2H),6.28(s,2H),4.50(d,J＝5.9Hz,2H),3.84(s,3H),2.54(s,3H).LR-MS 423.5(M+1).
实施例18 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(嘧啶-5-基甲基)苯甲酰胺的制备
除用5-氨甲基嘧啶代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.17(t,J＝5.8Hz,1H),9.10(s,1H),8.79(s,2H),8.26(d,J＝2.3Hz,1H),8.15(s,1H),8.07(s,1H),7.95–7.81(m,2H),7.79(d,J＝8.3Hz,1H),7.43(d,J＝8.0Hz,1H),6.28(s,2H),4.50(d,J＝5.7Hz,2H),3.84(s,3H),2.53(s,3H).LR-MS 424.5(M+1).
实施例19 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-((6-甲氧基吡啶-3-基)甲基)-4-甲基苯甲酰胺的制备
除用3-氨甲基-6-甲氧基吡啶代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.04(t,J＝5.7Hz,1H),8.26(d,J＝2.4Hz,1H),8.14(s,2H),8.08(s,1H),7.87–7.80(m,2H),7.78(dd,J＝1.9,8.0Hz,1H),7.68(dd,J＝2.5,8.5Hz,1H),7.42(d,J＝8.0Hz,1H),6.80(d,J＝8.5Hz,1H),6.27(s,2H),4.40(d,J＝5.8Hz,2H),3.84(s,3H),3.82(s,3H),2.53(s,3H).LR-MS 453.2(M+1).
实施例20 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-((6-氯吡啶-3-基)甲基)-4-甲基苯甲酰胺的制备
除用3-氨甲基-6-氯吡啶代替苄胺外，合成方法如实施例1。
1H NMR(600MHz,DMSO-d6)δ9.13(t,J＝5.9Hz,1H),8.40(d,J＝2.5Hz,1H),8.26(d,J＝2.4Hz,1H),8.14(d,J＝1.9Hz,1H),8.07(s,1H),7.83(d,J＝2.4Hz,1H),7.82–7.80(m,2H),7.79(dd,J＝2.0,8.0Hz,1H),7.50(d,J＝8.2Hz,1H),7.43(d,J＝8.0Hz,1H),6.25(s,2H),4.49(d,J＝5.8Hz,2H),3.84(s,3H),2.54(s,3H).LR-MS 457.1(M+1).
实施例21 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-((6-氟吡啶-3-基)甲基)-4-甲基苯甲酰胺的制备
除用3-氨甲基-6-氟吡啶代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.13(t,J＝5.8Hz,1H),8.26(d,J＝2.4Hz,1H),8.21(d,J＝2.5Hz,1H),8.15(d,J＝1.9Hz,1H),8.08(s,1H),7.94(td,J＝2.6,8.2Hz,1H),7.83(d,J＝2.4Hz,1H),7.82(s,1H),7.79(dd,J＝2.0,8.0Hz,1H),7.43(d,J＝8.1Hz,1H),7.16(dd,J＝2.8,8.5Hz,1H),6.27(s,2H),4.49(d,J＝5.8Hz,2H),3.84(s,3H),2.53(s,3H).LR-MS 441.5(M+1).
实施例22 3-((2-氨基-5-(1-环丙基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-((6-氟吡啶 -3-基)甲基)-4-甲基苯甲酰胺的制备
除用3-氨甲基-6-氟吡啶代替苄胺和用1-环丙基-1H-吡唑-4-硼酸频哪醇酯代替1-甲基-1H-吡唑-4-硼酸频哪醇酯外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.15(t,J＝5.7Hz,1H),8.49(d,J＝1.4Hz,1H),8.31–8.22(m,1H),8.12(d,J＝2.2Hz,1H),7.98(d,J＝1.3Hz,2H),7.82(d,J＝1.3Hz,1H),7.80(dd,J＝2.0,7.5Hz,1H),7.30(dd,J＝1.2,7.4Hz,1H),7.08(d,J＝1.5Hz,1H),6.92(t,J＝8.1Hz,1H),6.39(s,2H),4.50(t,J＝5.7Hz,2H),2.52(s,3H),2.44–2.37(m,1H),0.85–0.74(m,2H),0.61–0.49(m,2H).LR-MS 467.2(M+1).
实施例23 3-((2-氨基-5-(1-异丙基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-((6-氟吡啶-3-基)甲基)-4-甲基苯甲酰胺的制备
除用3-氨甲基-6-氟吡啶代替苄胺和用1-异丙基-1H-吡唑-4-硼酸频哪醇酯代替1-甲基-1H-吡唑-4-硼酸频哪醇酯外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.13(t,J＝5.7Hz,1H),8.45(d,J＝1.3Hz,1H),8.23(d,J＝1.3Hz,1H),8.12–8.04(m,3H),7.86(d,J＝1.6Hz,1H),7.75(dd,J＝2.0,7.5Hz,1H),7.37(dd,J＝1.2,7.5Hz,1H),7.14(t,J＝8.0Hz,1H),7.06(d,J＝1.5Hz,1H),5.05(s,2H),4.62–4.52(m,1H),4.52(t,J＝5.6Hz,2H),2.54(s,3H),1.30(d,J＝6.8Hz,6H).LR-MS 469.5(M+1).
实施例24 3-((2-氨基-5-(1-异丙基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-((6-氟吡啶-3-基)甲基)-4-甲基苯甲酰胺的制备
除用3-氨甲基-6-甲氨基吡啶代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ8.90(t,J＝5.8Hz,1H),8.26(d,J＝2.4Hz,1H),8.12(d,J＝1.9Hz,1H),8.08(s,1H),7.95(d,J＝2.4Hz,1H),7.83(d,J＝2.4Hz,1H),7.82(s,1H),7.77(dd,J＝1.9,7.9Hz,1H),7.41(d,J＝8.1Hz,1H),7.37(dd,J＝2.4,8.5Hz,1H),6.43–6.37(m,2H),6.26(s,2H),4.28(d,J＝5.7Hz,2H),3.84(s,3H),2.73(d,J＝4.8Hz,3H),2.52(s,3H).LR-MS 452.2(M+1).
实施例25 N-((6-(1H-吡唑-1-基)吡唑-3-基)甲基)-3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基苯甲酰胺的制备
除用(6-(1H-吡唑-1基)吡啶-3-基)甲胺代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.16(t,J＝5.9Hz,1H),8.60(d,J＝2.5Hz,1H),8.45(d,J＝1.4Hz,1H),8.26(d,J＝2.4Hz,1H),8.16(d,J＝1.9Hz,1H),8.07(s,1H),7.95(dd,J＝2.2,8.5Hz,1H),7.91(dd,J＝0.8,8.5Hz,1H),7.83(d,J＝2.4Hz,1H),7.82–7.79(m,3H),7.44(d,J＝8.1Hz,1H),6.57(dd,J＝1.7,2.6Hz,1H),6.27(s,2H),4.53(d,J＝5.8Hz,2H),3.84(s,3H),2.54(s,3H).LR-MS 489.6(M+1).
实施例26 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-(3-氟苄基)-4-甲基苯甲酰胺的制备
除用3-氟苄胺代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.10(t,J＝6.0Hz,1H),8.26(d,J＝2.4Hz,1H),8.17(d,J＝1.9Hz,1H),8.07(s,1H),7.83(d,J＝2.4Hz,1H),7.83–7.79(m,2H),7.44(d,J＝8.1Hz,1H),7.38(td,J＝6.1,7.9Hz,1H),7.17(d,J＝7.8Hz,1H),7.16–7.11(m,1H),7.08(td,J＝2.7,8.9Hz,1H),6.25(s,2H),4.49(d,J＝5.9Hz,2H),3.84(s,3H),2.54(s,3H).LR-MS 440.5(M+1).
实施例27 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩 -3-基甲基)苯甲酰胺的制备
除用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.01(t,J＝5.9Hz,1H),8.26(d,J＝2.4Hz,1H),8.15(d,J＝1.9Hz,1H),8.08(s,1H),7.84(d,J＝2.4Hz,1H),7.82(s,1H),7.80(dd,J＝2.0,8.0Hz,1H),7.49(dd,J＝3.0,4.9Hz,1H),7.42(d,J＝8.1Hz,1H),7.34(dd,J＝1.2,3.0Hz,1H),7.09(dd,J＝1.2,4.9Hz,1H),6.27(s,2H),4.47(d,J＝5.8Hz,2H),3.84(s,3H),2.53(s,3H).LR-MS 427.1(M+1).
实施例28 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-2-基甲基)苯甲酰胺的制备
除用2-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ8.97(t,J＝5.8Hz,1H),8.30(d,J＝1.3Hz,1H),8.17(d,J＝2.0Hz,1H),8.05(d,J＝1.3Hz,1H),7.75(d,J＝1.6Hz,1H),7.73(dd,J＝2.1,7.6Hz,1H),7.44(dd,J＝1.6,7.5Hz,1H),7.29(dd,J＝1.1,7.7Hz,1H),7.12(dd,J＝1.7,7.6Hz,1H),7.06–6.97(m,2H),6.29(s,2H),4.42(d,J＝5.8Hz,2H),3.92(s,3H),2.53(s,3H).LR-MS 428.3(M+1).
实施例29 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻唑-4-基甲基)苯甲酰胺的制备
除用4-氨甲基噻唑代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.10(t,J＝5.9Hz,1H),9.07(d,J＝2.0Hz,1H),8.26(d,J＝2.4Hz,1H),8.18(d,J＝1.9Hz,1H),8.08(s,1H),7.84(d,J＝2.5Hz,1H),7.83–7.80(m,2H),7.50–7.46(m,1H),7.43(d,J＝8.1Hz,1H),6.27(s,2H),4.62(d,J＝5.7Hz,2H),3.84(s,3H),2.54(s,3H).LR-MS 429.5(M+1).
实施例30 4-甲基-3-((5-(1-甲基-1H-吡唑-4-基)-2-(甲基氨基)吡啶-3-基)乙炔基)-N-(吡啶-3-基甲基)苯甲酰胺的制备
步骤1：向封管加入2-氯-5-溴-3-碘吡啶(1.0g,3.14mmol)和MeNH2(2.0M,5mL)，加热至80℃反应过夜。反应结束后，减压蒸干溶剂，柱层析分离得2-甲氨基-5-溴-3-碘吡啶650mg(产率：66％)。1H NMR(400MHz,DMSO-d6)δ8.10(d,J＝2.2Hz,1H),8.07(d,J＝2.2Hz,1H),6.32–6.24(m,1H),2.78(d,J＝4.6Hz,3H).LR-MS 313.1(M+1).
步骤2：向圆底烧瓶中加入2-甲氨基-5-溴-3-碘吡啶(300mg,0.96mmol)、三甲基硅乙炔(122.4mg,1.25mmol)和Et3N(20mL)，用氩气置换氧气，加入Pd(PPh3)2Cl2(34mg,0.048mmol)、CuI(18mg,0.096mmol)，重复除氧气的操作，室温条件下反应4小时；反应结束后加入20mL乙酸乙酯稀释反应液，过滤即可得到产物2-甲氨基-5-溴-3-三甲 基乙炔基吡啶260mg(产率：96％)。1H NMR(400MHz,DMSO-d6)δ8.11(d,J＝2.5Hz,1H),7.67(d,J＝2.4Hz,1H),6.34(q,J＝4.3Hz,1H),2.85(d,J＝4.7Hz,3H),0.25(s,9H).LR-MS 284.2(M+1).
步骤3：向圆底烧瓶中加入2-氨基-5-溴-3-三甲基乙炔基吡啶(200mg,0.71mmol)、1-甲基吡唑-4-硼酸频哪醇酯(294mg,1.41mmol)、K2CO3(244mg,1.77mmol)和THF/H2O(8/2mL)，用氩气置换氧气，加入Pd(OAc)2(16mg,0.071mmol)和X-Phos(50mg,0.11mmol)，加热至70℃反应3小时。反应结束后，用乙酸乙酯(15mL×3)和水(10mL)萃取反应液，有机相用饱和NaCl水溶液洗(10mL×3)，无水Na2SO4干燥，减压蒸干溶剂，柱层析分离得5-(1-甲基-1H-吡唑-4-基)-3-((三甲基硅基)乙炔基)-2-甲氨基吡啶150mg(产率：74.7％)。LR-MS 285.4(M+1).
步骤4：向圆底烧瓶中加入4-甲基-3碘苯甲酸(2.0g,7.63mmol)、HATU(3.8g,9.92mmol)、DIPEA(2.5g,19.08mmol)和DMF(40mL)，室温下搅拌30分钟后，加入3-氨甲基吡啶(825mg,7.63mmol)，室温反应12小时；反应结束后，乙酸乙酯(50mL×3)和水(40mL)萃取反应液，有机相分别用水洗(30mL×3)，饱和NaCl溶液洗(30mL×3)，无水硫酸钠干燥，减压蒸干溶剂，柱层析分离得3-碘-4-甲基-N-(吡啶-3-基甲基)苯甲酰胺2.5g(产率：93％)。LR-MS 353.1(M+1).
步骤5：向圆底烧瓶中加入3-碘-4-甲基-N-(吡啶-3-基甲基)苯甲酰胺(100mg,0.28mmol)、5-(1-甲基-1H-吡唑-4-基)-3-((三甲基硅基)乙炔基)-2-甲氨基吡啶(101mg,0.35mmol)、Et3N(86mg,0.85mmol)、CsF(108mg,0.71mmol)和MeCN(15mL)，用氩气置换氧气，加入Pd(PPh3)2Cl2(10mg,0.014mmol)、CuI(5.4mg,0.028mmol)，重复除氧气的操作，室温条件下反应3小时。反应结束后，乙酸乙酯(20mL×3)和水(20mL)萃取反应液，有机相用饱和NaCl溶液洗(10mL×3)，经无水硫酸钠干燥后，减压蒸干溶剂，柱层析分离得产物4-甲基-3-((5-(1-甲基-1H-吡唑-4-基)-2-(甲基氨基)吡啶-3-基)乙炔基)-N-(吡啶-3-基甲基)苯甲酰胺90mg(产率：72.6％)。1H NMR(400MHz,DMSO-d6)δ9.15(t,J＝5.9Hz,1H),8.56(d,J＝2.2Hz,1H),8.47(dd,J＝1.6,4.8Hz,1H),8.33(d,J＝2.4Hz,1H),8.17(d,J＝1.9Hz,1H),8.06(s,1H),7.83(d,J＝2.3Hz,1H),7.82–7.78(m,2H),7.74(dt,J＝2.0,8.0Hz,1H),7.44(d,J＝8.1Hz,1H),7.37(dd,J＝4.7,7.8Hz,1H),6.45(q,J＝4.3Hz,1H),4.50(d,J＝5.8Hz,2H),3.84(s,3H),2.91(d,J＝4.6Hz,3H),2.53(s,3H).LR-MS 437.5(M+1).
实施例31 4-甲基-3-((5-(1-甲基-1H-吡唑-4-基)-2-(乙基氨基)吡啶-3-基)乙炔基)-N-(吡啶-3-基甲基)苯甲酰胺的制备
除用乙胺代替甲胺外，合成方法如实施例30。
1H NMR(400MHz,DMSO-d6)δ9.16(t,J＝6.0Hz,1H),8.57(s,1H),8.47(s,1H),8.32(d,J＝2.4Hz,1H),8.15(d,J＝1.9Hz,1H),8.06(s,1H),7.83(d,J＝2.4Hz,1H),7.83–7.78(m,2H),7.74(d,J＝8.0Hz,1H),7.44(d,J＝8.0Hz,1H),7.37(dd,J＝4.7,7.9Hz,1H),6.34(t,J＝5.8Hz,1H),4.50(d,J＝5.8Hz,2H),3.84(s,3H),3.50–3.39(m,3H),2.54(s,3H),1.17(t,J＝7.0Hz,3H).LR-MS 451.2(M+1).
实施例32 3-((2-乙基氨基)-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-((6-氟吡啶-3-基)甲基)-4-甲基苯甲酰胺的制备
除用乙胺代替甲胺和用3-氨甲基-6-氟吡啶代替3-氨甲基吡啶外，合成方法如实施 例37。
1H NMR(400MHz,DMSO-d6)δ9.20(t,J＝6.0Hz,1H),8.39(d,J＝1.3Hz,1H),8.17(dd,J＝1.6,3.2Hz,2H),8.08(d,J＝1.3Hz,1H),8.04–7.98(m,1H),7.93(s,1H),7.84(d,J＝1.6Hz,1H),7.66(dd,J＝2.0,7.5Hz,1H),7.36(dd,J＝1.2,7.6Hz,1H),7.15(t,J＝7.9Hz,1H),7.06(d,J＝1.6Hz,1H),4.50(d,J＝5.8Hz,2H),3.85(s,3H),3.47(q,J＝8.0Hz,2H),2.54(s,3H),1.09(t,J＝8.0Hz,3H).LR-MS 469.3(M+1).
实施例33 3-((2-乙基氨基)-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用乙胺代替甲胺和用3-氨甲基噻吩代替3-氨甲基吡啶外，合成方法如实施例30。
1H NMR(400MHz,DMSO-d6)δ9.14(t,J＝6.0Hz,1H),8.22(d,J＝1.0Hz,1H),8.16(d,J＝2.1Hz,1H),8.00(d,J＝1.3Hz,1H),7.93(s,1H),7.75(d,J＝1.5Hz,1H),7.70(dd,J＝2.0,7.5Hz,1H),7.36(d,J＝8.1Hz,1H),7.28(dd,J＝1.2,7.6Hz,1H),7.06–6.99(m,2H),6.96(d,J＝1.6Hz,1H),4.55(t,J＝6.0Hz,2H),3.90(s,3H),3.47(q,J＝8.0Hz,2H),2.51(s,3H),1.07(t,J＝8.0Hz,3H).LR-MS 456.7(M+1).
实施例34 3-((2-环丙基氨基)-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用环丙胺代替甲胺和用3-氨甲基噻吩代替3-氨甲基吡啶外，合成方法如实施例30。
1H NMR(400MHz,DMSO-d6)δ9.07(t,J＝5.8Hz,1H),8.25(d,J＝1.3Hz,1H),8.17(d,J＝2.0Hz,1H),8.16(d,J＝1.3Hz,1H),7.88(s,1H),7.85(d,J＝1.5Hz,1H),7.68(dd,J＝2.0,7.5Hz,1H),7.43–7.32(m,2H),7.09(d,J＝1.6Hz,1H),7.03(dd,J＝1.6,7.5Hz,1H),7.00(d,J＝1.5Hz,1H),6.28(s,2H),3.93(s,3H),2.57(s,3H),2.29(m,1H),0.75–0.47(m,4H).LR-MS 468.4(M+1).
实施例35 3-((2-(N-乙酰氨基)-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺
向圆底烧瓶中加入化合物27(80mg,0.187mmol)、Et3N(57mg,0.561mmol)和THF(3mL)，冰浴下滴加乙酰氯(26μL,0.374mmol)，室温搅拌3h。加入甲醇淬灭反应，随后减压蒸干溶剂，柱层析分离纯化得到产物3-((2-(N-乙酰氨基)-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺70mg(浅黄色固体；产率：73％)。1H NMR(400MHz,DMSO-d6)δ9.24(t,J＝5.9Hz,1H),8.87(d,J＝2.3Hz,1H),8.48–8.38(m,2H),8.14(s,1H),8.01(d,J＝1.9Hz,1H),7.85(dd,J＝8.0,2.0Hz,1H),7.46(d,J＝8.1Hz,1H),7.40(dd,J＝5.1,1.3Hz,1H),7.03(d,J＝3.5Hz,1H),6.97(dd,J＝5.1,3.4Hz,1H),4.63(d,J＝5.8Hz,2H),3.91(s,3H),2.44(s,3H),2.25(s,6H).LR-MS 512.2(M+1).
实施例36 3-((2-氨基-5-(3,5-二甲基异噁唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用3,5-二甲基异噁唑-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑-4-硼酸嚬那醇酯和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.01(t,J＝5.9Hz,1H),8.15(d,J＝1.9Hz,1H),7.99(d,J＝2.3Hz,1H),7.80(dd,J＝8.0,1.9Hz,1H),7.66(d,J＝2.3Hz,1H),7.49(dd,J＝4.9,3.0Hz,1H),7.42(d,J＝8.1Hz,1H),7.35–7.31(m,1H),7.09(dd,J＝4.9,1.3Hz,1H),6.50(s,2H),4.46(d,J＝5.8Hz,2H),2.51(s,3H),2.37(s,3H),2.20(s,3H).LR-MS 443.1(M+1).
实施例37 3-((2-氨基-5-(1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用1H-吡唑-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑-4-硼酸嚬那醇酯和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ11.42(s,1H),9.07(t,J＝5.8Hz,1H),8.20(d,J＝1.3Hz,1H),8.17(d,J＝2.0Hz,1H),8.02(d,J＝1.3Hz,1H),7.70(dd,J＝2.0,7.5Hz,1H),7.37(d,J＝7.3Hz,1H),7.28(dd,J＝1.1,7.4Hz,1H),7.23(s,2H),7.02(dd,J＝1.6,7.3Hz,1H),7.01(s,1H),6.26(s,2H),4.55(d,J＝5.8Hz,2H),2.52(s,3H).LR-MS 414.5(M+1).
实施例38 3-((2-氨基-5-(3,5-二甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用3,5-二甲基-1H-吡唑-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑-4-硼酸嚬那醇酯和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ11.95(s,1H),8.96(t,J＝5.8Hz,1H),8.18(d,J＝1.3Hz,1H),8.16(d,J＝2.0Hz,1H),7.95(d,J＝1.1Hz,1H),7.70(dd,J＝2.1,7.6Hz,1H),7.37(d,J＝7.4Hz,1H),7.28(dd,J＝1.2,7.2Hz,1H),7.07–6.97(m,2H),6.29(s,2H),4.42(d,J＝5.7Hz,2H),2.53(s,3H),2.39(s,3H),2.36(s,3H).LR-MS 442.3(M+1).
实施例39 3-((2-氨基-5-(1-甲基-1H-吡唑-5-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用1-甲基-1H-吡唑-5-硼酸嚬那醇酯代替1-甲基-1H-吡唑-4-硼酸嚬那醇酯和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.14(t,J＝5.7Hz,1H),8.79(d,J＝1.3Hz,1H),8.08(d,J＝2.0Hz,1H),7.98(d,J＝1.1Hz,1H),7.78(dd,J＝2.0,7.5Hz,1H),7.64(d,J＝7.5Hz,1H),7.37(d,J＝7.4Hz,1H),7.28(dd,J＝1.1,7.4Hz,1H),7.07–6.97(m,2H),6.35(d,J＝7.5Hz,1H),6.29(s,2H),4.43(d,J＝5.7Hz,2H),3.88(s,3H),2.54(s,3H).LR-MS428.1(M+1).
实施例40 3-((2-氨基-5-(1-甲基-1H-咪唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用1-甲基-1H-咪唑-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑-4-硼酸嚬那醇酯和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ8.76(t,J＝5.8Hz,1H),8.88(d,J＝1.3Hz,1H),8.17(d,J＝2.0Hz,1H),8.05(d,J＝1.3Hz,1H),7.83(s,1H),7.70(dd,J＝2.0,7.5Hz,1H),7.36(d,J＝7.6Hz,1H),7.33(s,1H),7.28(dd,J＝1.1,7.4Hz,1H),7.05(dd,J＝1.3,7.5Hz,1H),7.00(d,J＝1.5Hz,1H),6.31(s,2H),4.41(d,J＝5.7Hz,2H),3.69(s,3H),2.53(s,3H).LR-MS 428.1(M+1).
实施例41 3-((2-氨基-5-(1,3,5-三甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用1,3,5-三甲基-1H-吡唑-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑-4-硼酸嚬那醇酯和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.12(t,J＝5.7Hz,1H),8.18(d,J＝1.3Hz,1H),8.09(d,J＝2.0Hz,1H),7.95(d,J＝1.1Hz,1H),7.77(dd,J＝2.1,7.6Hz,1H),7.37(d,J＝7.4Hz,1H),7.28(dd,J＝1.1,7.6Hz,1H),7.06–6.99(m,2H),6.28(s,2H),4.42(d,J＝5.8Hz,2H),3.84(s,3H),2.54(s,3H),2.52(s,3H),2.38(s,3H).LR-MS 456.6(M+1).
实施例42 3-((2-氨基-5-(1-环丙基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用1-环丙基-1H-吡唑-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑-4-硼酸嚬那醇酯和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.04(t,J＝6.0Hz,1H),8.12(d,J＝2.0Hz,1H),7.98(s,1H),7.97(s,1H),7.82(d,J＝1.3Hz,1H),7.80(dd,J＝2.0,7.5Hz,1H),7.30(dd,J＝1.2,7.5Hz,1H),7.19(d,J＝8.5Hz,1H),7.08(d,J＝1.5Hz,1H),6.88(d,J＝11.9Hz,4H),4.46(d,J＝5.8Hz,2H),2.45(s,3H),2.41(q,J＝7.0Hz,1H),0.79(ddd,J＝4.2,6.0,7.2Hz,2H),0.59–0.49(m,2H).LR-MS 454.2(M+1).
实施例43 3-((2-氨基-5-(1-异丙基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用1-异丙基-1H-吡唑-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑-4-硼酸嚬那醇酯和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.01(t,J＝5.8Hz,1H),8.29(d,J＝2.4Hz,1H),8.19(s,1H),8.15(d,J＝1.9Hz,1H),7.86(d,J＝2.4Hz,1H),7.82(d,J＝0.8Hz,1H),7.80(dd,J＝8.0,1.9Hz,1H),7.49(dd,J＝4.9,2.9Hz,1H),7.42(d,J＝8.0Hz,1H),7.34(dd,J＝3.0,1.3Hz,1H),7.09(dd,J＝5.0,1.3Hz,1H),6.24(s,2H),4.51–4.43(m,3H),2.54(s,3H),1.43(d,J＝6.6Hz,6H).LR-MS 456.2(M+1).
实施例44 3-((2-氨基-5-(1-(氧杂环丁基-3-基)-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用1-(氧杂环丁基-3-基)-1H-吡唑-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑-4-硼酸嚬那醇酯和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.07(t,J＝5.8Hz,1H),8.12(d,J＝2.0Hz,1H),7.98(s,1H),7.96(s,1H),7.82(d,J＝1.3Hz,1H),7.80(dd,J＝2.0,7.5Hz,1H),7.30(dd,J＝1.1,7.4Hz,1H),7.22–7.16(m,1H),7.08(d,J＝1.5Hz,1H),6.88–6.82(m,2H),6.39(s,2H),5.33–5.26(m,2H),5.08–5.01(m,2H),4.97–4.87(m,1H),4.55(d,J＝5.7Hz,2H),2.54(s,3H).LR-MS 470.2(M+1).
实施例45 3-((2-氨基-5-(1-(哌啶-4-基)-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用1-(哌啶-4-基)-1H-吡唑-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑-4-硼酸嚬那醇酯和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.10(t,J＝5.7Hz,1H),8.24(d,J＝1.3Hz,1H),8.17(d,J＝2.1Hz,1H),8.13(d,J＝1.3Hz,1H),7.86(d,J＝1.5Hz,1H),7.70(dd,J＝2.0,7.5Hz,1H),7.42–7.32(m,2H),7.13(d,J＝1.3Hz,1H),7.06–6.99(m,2H),6.28(s,2H),4.46(d,J＝5.8Hz,2H),4.30–4.18(m,1H),3.17–3.02(m,2H),2.78–2.62(m,2H),2.53(s,2H),2.44(s,1H),1.87–1.72(m,2H),1.70–1.51(m,2H).LR-MS 497.3(M+1).
实施例46 3-((2-氨基-5-(1-(1-甲基哌啶-4-基)-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用1-(1-甲基哌啶-4-基)-1H-吡唑-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑-4-硼酸嚬那醇酯和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.06(t,J＝5.7Hz,1H),8.24(d,J＝1.3Hz,1H),8.17(d,J＝2.0Hz,1H),8.13(d,J＝1.3Hz,1H),7.86(d,J＝1.6Hz,1H),7.70(dd,J＝2.1,7.6 Hz,1H),7.41–7.34(m,2H),7.13(d,J＝1.3Hz,1H),7.03(dd,J＝1.6,7.5Hz,1H),7.00(d,J＝1.5Hz,1H),6.27(s,2H),4.45(d,J＝5.8Hz,2H),4.33–4.15(m,1H),3.02–2.83(m,2H),2.53(s,3H),2.25(s,3H),2.22–1.87(m,4H),1.76–1.54(m,2H).LR-MS 511.3(M+1).
实施例47 3-((2-氨基-5-(1-(四氢-2H-吡喃-4-基)-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用1-(四氢-2H-吡喃-4-基)-1H-吡唑-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑-4-硼酸嚬那醇酯和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝6.0Hz,1H),8.24(d,J＝1.3Hz,1H),8.18(d,J＝2.0Hz,1H),8.13(d,J＝1.1Hz,1H),7.87(d,J＝1.5Hz,1H),7.70(dd,J＝2.0,7.5Hz,1H),7.43–7.33(m,2H),7.13(d,J＝1.6Hz,1H),7.03(dd,J＝1.5,7.5Hz,1H),7.00(d,J＝1.5Hz,1H),6.29(s,2H),4.45(d,J＝5.8Hz,2H),4.12–3.95(m,1H),3.95–3.79(m,2H),3.63–3.37(m,2H),2.53(s,3H),2.19–1.91(m,2H),1.89–1.66(m,2H).LR-MS498.6(M+1).
实施例48 3-((2-氨基-5-(1-(2-羟基乙基)-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用1-(2-羟基乙基)-1H-吡唑-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑-4-硼酸嚬那醇酯和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.00(t,J＝5.8Hz,1H),8.27(d,J＝2.4Hz,1H),8.15(d,J＝1.9Hz,1H),8.10(s,1H),7.85(d,J＝2.4Hz,1H),7.84(s,1H),7.80(dd,J＝1.9,8.1Hz,1H),7.49(dd,J＝3.0,5.0Hz,1H),7.42(d,J＝8.0Hz,1H),7.34(s,1H),7.09(d,J＝4.4Hz,1H),6.24(s,2H),4.92(t,J＝5.4Hz,1H),4.47(d,J＝5.8Hz,2H),4.13(t,J＝5.7Hz,2H),3.75(q,J＝5.5Hz,2H),2.53(s,3H).LR-MS 458.1(M+1).
实施例49 3-((2-氨基-5-(1-(2-羟基丙基)-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用1-(2-羟基丙基)-1H-吡唑-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑-4-硼酸嚬那醇酯和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.04(t,J＝5.9Hz,1H),8.26(d,J＝1.3Hz,1H),8.17(d,J＝2.0Hz,1H),8.03(d,J＝1.3Hz,1H),7.76(d,J＝1.5Hz,1H),7.70(dd,J＝2.0,7.5Hz,1H),7.37(d,J＝7.4Hz,1H),7.28(d,J＝8.0Hz,1H),7.06–6.98(m,3H),6.28(s,2H),4.50(d,J＝4.8Hz,1H),4.42(d,J＝5.8Hz,1H),4.21(dd,J＝6.9,12.4Hz,1H),4.07(dd,J＝6.9,12.3Hz,1H),4.01–3.90(m,1H),2.53(s,3H),1.16(d,J＝6.8Hz,3H).LR-MS 472.6(M+1).
实施例50 3-((2-氨基-5-(1-(2-(二甲氨基)乙基)-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用1-(2-(二甲氨基)乙基)-1H-吡唑-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑-4-硼酸嚬那醇酯和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.04(t,J＝5.8Hz,1H),8.26(d,J＝1.3Hz,1H),8.17(d,J＝2.1Hz,1H),8.04(d,J＝1.3Hz,1H),7.77(d,J＝1.6Hz,1H),7.70(dd,J＝2.0,7.5Hz,1H),7.37(d,J＝7.6Hz,1H),7.28(dd,J＝1.2,7.3Hz,1H),7.09–6.96(m,3H),4.66(t,J＝7.5Hz,2H),6.26(s,2H),4.44(d,J＝5.8Hz,2H),3.00(t,J＝7.5Hz,2H),2.52(s,3H),2.37(s,6H).LR-MS 485.5(M+1).
实施例51 3-((2-氨基-5-(1-(2-氟乙基)-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用1-(2-氟乙基)-1H-吡唑-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑-4-硼酸嚬那醇酯和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.06(t,J＝5.7Hz,1H),8.25(d,J＝1.3Hz,1H),8.17(d,J＝2.0Hz,1H),8.05(d,J＝1.1Hz,1H),7.78(d,J＝1.5Hz,1H),7.70(dd,J＝2.0,7.5Hz,1H),7.37(d,J＝7.5Hz,1H),7.28(dd,J＝1.2,7.4Hz,1H),7.08(d,J＝1.6Hz,1H),7.05–6.97(m,2H),4.63(m,2H),4.55(s,2H),4.42(d,J＝5.6Hz,2H),3.96(t,J＝3.0Hz,2H),2.54(s,3H).LR-MS 460.5(M+1).
实施例52 2-(4-(6-氨基-5-((2-甲基-5-((噻吩-3-基甲基)氨基甲酰基)苯基)乙炔基)吡啶-3-基)-1H-吡唑-1-基)乙酸乙酯的制备
除用2-(4-硼酸嚬那醇酯-1H-吡唑-1-基)乙酸乙酯代替1-甲基-1H-吡唑-4-硼酸嚬那醇酯和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.01(t,J＝5.8Hz,1H),8.28(d,J＝2.4Hz,1H),8.14(d,J＝1.9Hz,1H),8.12(s,1H),7.90(s,1H),7.86(d,J＝2.4Hz,1H),7.79(dd,J＝2.0,7.9Hz,1H),7.48(dd,J＝3.0,4.9Hz,1H),7.42(d,J＝8.1Hz,1H),7.36–7.31(m,1H),7.09(dd,J＝1.3,4.9Hz,1H),6.27(s,2H),5.05(s,2H),4.46(d,J＝5.8Hz,2H),4.16(q,J＝7.1Hz,2H),2.53(s,3H),1.21(t,J＝7.1Hz,3H).LR-MS 500.2(M+1).
实施例53 3-((2-氨基-5-(1-(2-二甲基氨基)-2-氧代乙基)-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
向圆底烧瓶中加入化合物52(100mg,0.20mmol)和THF/MeOH/H2O(2/0.5/0.5mL)，加入LiOH.H2O(21mg,0.50mmol)，室温反应2小时后，减压旋干直接投下一步。将上述固体重新溶于DMF(2mL)中,加入HATU(99mg,0.26mmol)和DIPEA(77mg,0.60mmol)，室温搅拌20分钟后，加入二甲胺的四氢呋喃溶液(1.0M,0.4mL)，继续反应3小时后，用乙酸乙酯(10mL×3)和水(10mL)萃取，合并有机相，分别用水(10mL×3)和饱和NaCl溶液(10mL×2)洗，经无水硫酸钠干燥后，减压蒸干溶剂，柱层析分离得3-((2-氨基-5-(1-(2-二甲基氨基)-2-氧代乙基)-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺55mg(产率：55.3％)。1H NMR(600MHz,氯仿-d)δ8.17(d, J＝2.3Hz,1H),7.91(d,J＝1.9Hz,1H),7.71–7.68(m,3H),7.66(d,J＝2.3Hz,1H),7.32–7.28(m,2H),7.23–7.21(m,1H),7.10(dd,J＝1.3,5.0Hz,1H),6.67(t,J＝5.6Hz,1H),5.09(s,2H),4.99(s,2H),4.64(d,J＝5.6Hz,2H),3.09(s,3H),2.99(s,3H),2.53(s,3H).LR-MS 499.5(M+1).
实施例54 3-((2-氨基-5-(1-三氟甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用1-三氟甲基-1H-吡唑基-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑基-4-硼酸嚬那醇酯外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.06(t,J＝5.7Hz,1H),8.26(d,J＝1.3Hz,1H),8.16(d,J＝2.0Hz,1H),8.05(d,J＝1.3Hz,1H),7.82(d,J＝1.5Hz,1H),7.71(dd,J＝2.0,7.5Hz,1H),7.37(d,J＝7.4Hz,1H),7.28(dd,J＝1.2,7.4Hz,1H),7.22(d,J＝1.5Hz,1H),7.03(dd,J＝1.5,7.5Hz,1H),7.00(d,J＝1.5Hz,1H),6.25(s,2H),4.42(d,J＝5.8Hz,2H),2.52(s,3H).LR-MS 482.1(M+1).
实施例55 3-((2-氨基-5-(1-三氟甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-((6-氟吡啶-3-基)甲基)-4-甲基苯甲酰胺的制备
除用1-三氟甲基-1H-吡唑基-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑基-4-硼酸嚬那醇酯和用3-氨甲基-6-氟吡啶代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝6.1Hz,1H),8.38(d,J＝1.3Hz,1H),8.16(d,J＝2.0Hz,1H),8.13(d,J＝1.3Hz,1H),8.10(d,J＝1.4Hz,1H),8.07–8.00(m,1H),7.92(d,J＝1.5Hz,1H),7.64(dd,J＝2.0,7.5Hz,1H),7.37(dd,J＝1.2,7.6Hz,1H),7.28(d,J＝1.5Hz,1H),7.15(t,J＝8.0Hz,1H),6.27(s,2H),4.50(d,J＝5.9Hz,2H),2.55(s,3H).LR-MS 495.3(M+1).
实施例56 3-((2-氨基-5-(1-二氟甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用1-二氟甲基-1H-吡唑基-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑基-4-硼酸嚬那醇酯外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝6.0Hz,1H),8.20(d,J＝1.3Hz,1H),8.16(d,J＝2.0Hz,1H),8.03(d,J＝1.1Hz,1H),7.81(t,J＝51.4Hz,1H),7.79(d,J＝1.5Hz,1H),7.73–7.67(m,1H),7.37(d,J＝7.4Hz,1H),7.28(dd,J＝1.2,7.5Hz,1H),7.13(d,J＝1.6Hz,1H),7.06–6.97(m,2H),6.29(s,2H),4.55(d,J＝5.9Hz,2H),2.54(s,3H).LR-MS464.4(M+1).
实施例57 3-((2-氨基-5-(1-(甲基-d3)-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用1-(甲基-d3)-1H-吡唑基-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑基-4-硼酸嚬那醇酯外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ8.98(t,J＝5.9Hz,1H),8.12(d,J＝2.0Hz,1H),7.98(d,J＝1.3Hz,1H),7.97(d,J＝1.6Hz,1H),7.82(d,J＝1.3Hz,1H),7.80(dd,J＝2.0,7.5Hz,1H),7.30(dd,J＝1.1,7.5Hz,1H),7.23–7.15(m,1H),7.05(d,J＝1.3Hz,1H),6.91–6.84(m,4H),4.55(d,J＝5.7Hz,2H),2.45(s,3H).LR-MS 431.5(M+1).
其中1-(甲基-d3)-1H-吡唑基-4-硼酸嚬那醇酯的合成如下所述：
向圆底烧瓶中加入1H-吡唑基-4-硼酸嚬那醇酯(0.5g,2.58mmol)、碳酸铯(2.5g,7.73mmol)和DMF(10mL),冰浴，滴加氘代碘甲烷(411mg,2.83mmol)，室温反应过夜，倒入水(20mL)，乙酸乙酯(25mL×3)萃取，合并有机相，分别用水(20mL×3)和饱和NaCl溶液(20mL×2)洗，经无水硫酸钠干燥后，减压蒸干溶剂，柱层析分离得1-(甲基-d3)-1H-吡唑基-4-硼酸嚬那醇酯467mg(产率：85.9％)。LR-MS(ESI)m/z 212.3(M+1).
实施例58 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基亚甲基-d2)苯甲酰胺的制备
除用噻吩-3-基亚甲基-d2-胺代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.03(s,1H),,8.12(d,J＝2.0Hz,1H),7.97(dd,J＝1.4,4.1Hz,2H),7.82(d,J＝1.3Hz,1H),7.80(dd,J＝2.0,7.5Hz,1H),7.30(d,J＝7.5Hz,1H),7.19(d,J＝8.5Hz,1H),7.05(d,J＝1.3Hz,1H),6.88(m,2H),6.28(s,2H),3.94(s,3H),2.48(s,3H).LR-MS(ESI)m/z 430.3(M+1).
其中噻吩-3-基亚甲基-d2-胺的制备方法如下所述：
向圆底烧瓶中加入LiAlD4(962mg,22.91mmol)，氩气充分置换气体，加入无水***(60mL)，冰浴，氩气保护下滴加3-氰基噻吩(1.0g,9.16mmol)的无水***溶液(10mL)，完毕后升温至室温，加热回流反应过夜。冷却至室温后，加入***(60mL)稀释，冰浴下滴加10％NaOH水溶液(2mL)和水(5mL)，硅藻土过滤，滤饼用***洗涤(25mL×3)，减压蒸干溶剂，柱层析分离得噻吩-3-基亚甲基-d2-胺750mg(产率：71.06％)。LR-MS(ESI)m/z 116.2(M+1).
实施例59 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-((6-氟吡啶-3-基)亚甲基-d2)-4-甲基-苯甲酰胺的制备
除用6-氟吡啶-3-基亚甲基-d2-胺代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ8.49(d,J＝1.4Hz,1H),8.30–8.23(m,1H),8.20(s,1H),8.12(d,J＝2.0Hz,1H),7.97(dd,J＝1.4,4.1Hz,2H),7.82(d,J＝1.3Hz,1H),7.80(dd,J＝2.0,7.5Hz,1H),7.30(dd,J＝1.1,7.5Hz,1H),7.05(d,J＝1.3Hz,1H),6.94–6.87(m,3H),3.94(s,3H),2.45(s,3H).LR-MS(ESI)m/z 443.2(M+1).
实施例60 甲基-4-(6-氨基-5-((2-甲基-5-((噻吩-3-基甲基)甲酰基)苯基)乙炔基)吡啶-3-基)-1H-吡唑-1-羧酸酯的制备
除用1-(甲氧基甲酰基)-1H-吡唑-4-硼酸嚬那醇酯代替1-甲基-1H-吡唑-4-硼酸嚬那醇酯和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.02(t,J＝5.9Hz,1H),8.81(s,1H),8.46(d,J＝2.4Hz,1H),8.37(s,1H),8.17(d,J＝1.9Hz,1H),8.09(d,J＝2.4Hz,1H),7.80(dd,J＝8.0,2.0Hz,1H),7.49(dd,J＝5.0,2.9Hz,1H),7.43(d,J＝8.1Hz,1H),7.34(dd,J＝3.0,1.2Hz,1H),7.09(dd,J＝4.9,1.3Hz,1H),6.48(s,2H),4.47(d,J＝5.8Hz,2H),4.00(s,3H),2.54(s,3H).LR-MS(ESI)m/z 472.2(M+1).
其中1-(甲氧基甲酰基)-1H-吡唑-4-硼酸嚬那醇酯的制备方法如下所述：
向圆底烧瓶中加入1H-吡唑-4-硼酸嚬那醇酯(2.0g,10.31mmol)和DMF(30mL),冰浴下分批加入NaH(60％wt,618mg,15.46mmol)，搅拌15分钟后加入氯甲酸甲酯(1.95g,20.61mmol)，室温搅拌反应5h。反应结束后，冰浴缓慢滴加饱和NH4Cl水溶液淬灭反应，倒入水(20mL)中，乙酸乙酯(30mL×3)萃取，合并有机相，分别用水(20mL×3)和饱和NaCl溶液(20mL×2)洗，经无水硫酸钠干燥后，减压蒸干溶剂，柱层析分离得1-(甲氧基甲酰基)-1H-吡唑-4-硼酸嚬那醇酯640mg(产率：24.6％)。1H NMR(400MHz,氯仿-d)δ8.44(s,1H),7.93(s,1H),4.06(s,3H),1.31(s,12H).LR-MS(ESI)m/z 253.1(M+1).
实施例61 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-((5-环丙基噻吩-2基)甲基)-4-甲基-苯甲酰胺的制备
除用(5-环丙基噻吩-2-基)甲基胺代替(5-甲基噻吩-2-基)甲基胺外，合成方法如实施例1。
1H NMR(600MHz,DMSO-d6)δ9.08(t,J＝5.9Hz,1H),8.26(d,J＝2.4Hz,1H),8.13(d,J＝1.9Hz,1H),8.07(s,1H),7.83(d,J＝2.4Hz,1H),7.81(d,J＝0.8Hz,1H),7.78(dd,J＝7.9,1.9Hz,1H),7.42(d,J＝8.1Hz,1H),6.78(d,J＝3.5Hz,1H),6.63(dd,J＝3.5,0.8Hz,1H),6.25(s,2H),4.52(d,J＝5.8Hz,2H),3.84(s,3H),2.53(s,3H),2.09–2.00(m,1H),0.96–0.90(m,2H),0.63–0.56(m,2H).LR-MS(ESI)m/z 468.3(M+1).
其中，(5-环丙基噻吩-2-基)甲基胺的制备方法如下所述：
向圆底烧瓶中加入5-环丙基噻吩-2-甲醛(1.0g,6.57mmol),NH4OAc(2.53g,32.85mmol)和MeOH(20mL),加入NaBH3CN(1.24g,19.71mmol)，50℃搅拌反应12h。反应结束后，倒入饱和NaHCO3水溶液中，减压蒸干大部分甲醇，乙酸乙酯(30mL×3)萃取，合并有机相，饱和NaCl溶液(20mL×2)洗，经无水硫酸钠干燥后，减压蒸干溶剂，柱层析分离得(5-环丙基噻吩-2-基)甲基胺300mg(产率：29.8％)。1H NMR(400MHz,DMSO-d6)δ7.05(d,J＝6.2Hz,1H),6.82(d,J＝6.1Hz,1H),5.03(br s,2H),4.30(s,2H),2.29–2.15(m,1H),1.03–0.85(m,4H).LR-MS(ESI)m/z 128.1(M+1).
实施例62 3-((2-氨基-5-(1-甲基-1H-吡唑-4-基)吡啶-3-基)乙炔基)-N-((5-氯噻吩-2基)甲基)-4-甲基-苯甲酰胺的制备
除用5-氯噻吩-2-醛代替5-环丙基噻吩-2-醛外，合成方法如实施例61。
1H NMR(400MHz,DMSO-d6)δ8.83(t,J＝8.0Hz,1H),8.18(s,1H),8.12(d,J＝2.1Hz,1H),7.98(d,J＝2.0Hz,1H),7.82(d,J＝2.0Hz,1H),7.77(dd,J＝7.6,1.9Hz,1H),7.33(dq,J＝7.6,1.0Hz,1H),7.30(s,1H),7.09(s,2H),6.92(d,J＝6.4Hz,1H),6.60(d,J＝6.4Hz,1H),4.75(d,J＝8.0Hz,2H),3.94(s,3H),2.45(s,3H).LR-MS(ESI)m/z 463.0(M+1).
实施例63 3-((2-氨基-5-(4-环丙基-1H-咪唑-1-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
步骤1：向圆底烧瓶中加入4-甲基-3碘苯甲酸(2.0g,7.63mmol)、HATU(3.8g,9.92mmol)、DIPEA(2.5g,19.08mmol)和DMF(40mL)，室温下搅拌30分钟后，加入3-氨甲基噻吩(864mg,7.63mmol)，室温反应12小时。反应结束后，乙酸乙酯(50mL×3)和水(40mL)萃取反应液，有机相分别用水洗(30mL×3)，饱和NaCl水溶液洗(30mL×3)，无水硫酸钠干燥，减压蒸干溶剂，柱层析分离得3-碘-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺2.4g(浅黄色固体；产率：88％)。
步骤2：向圆底烧瓶中加入3-碘-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺(100mg,0.28mmol)、5-溴-3-((三甲基硅基)乙炔基)-2-氨基吡啶(90mg,0.34mmol)、Et3N(85mg,0.84mmol)、CsF(128mg,0.84mmol)和MeCN(20mL)，用氩气置换氧气，加入Pd(PPh3)2Cl2(10mg,0.014mmol)、CuI(5.3mg,0.028mmol)，重复除氧气的操作，室温条件下反应3小时。反应结束后，乙酸乙酯(30mL×3)和水(20mL)萃取反应液，有机相用饱和NaCl溶液洗(10mL×3)，经无水硫酸钠干燥后，减压蒸干溶剂，柱层析分离得产物3-((2-氨基-5-溴吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺85mg(黄色固体；产率：71％)。
步骤3：向圆底烧瓶中加入3-((2-氨基-5-溴吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺(80mg,0.19mmol)、4-环丙基咪唑(30mg,0.28mmol)、CuI(7mg,0.038mmol)、Cs2CO3(122mg,0.38mmol)和DMF(2mL)，氩气保护下，120℃反应30h。冷却至室温，加入10mL乙酸乙酯稀释反应液，过滤，并用乙酸乙酯洗涤滤饼，减压蒸干溶剂，柱层析分离得产物3-((2-氨基-5-(4-环丙基-1H-咪唑-1-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺50mg(黄色固体；产率：59％)。1H NMR(400MHz,DMSO-d6)δ8.82(t,J＝6.0Hz,1H),8.52(d,J＝2.0Hz,1H),8.12(d,J＝2.0Hz,1H),7.80(d,J＝1.5Hz,1H),7.77(dd,J＝7.6,1.9Hz,1H),7.60(d,J＝2.0Hz,1H),7.33(dq,J＝7.6,1.1Hz,1H),7.22(dd,J＝4.8,1.8Hz,1H),7.16(d,J＝2.2Hz,1H),7.07–7.02(m,2H),6.25(s,2H),4.55(d,J＝5.9Hz,2H),2.45(d,J＝0.9Hz,3H),2.14(pd,J＝7.9,0.7Hz,1H),1.33–1.11(m,4H).
实施例64 3-((3-氨基异喹啉-4-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用4-碘-异喹啉-3-胺代替5-溴-3-碘吡啶-2-胺和用3-氨甲基噻吩代替苄胺外，合 成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝6.1Hz,1H),8.91(s,1H),8.26(d,J＝1.9Hz,1H),7.93(t,J＝8.3Hz,2H),7.80(dd,J＝8.0,2.0Hz,1H),7.71–7.66(m,1H),7.50(dd,J＝5.0,2.9Hz,1H),7.45(d,J＝8.1Hz,1H),7.35(d,J＝1.4Hz,1H),7.30(dd,J＝8.3,7.1Hz,1H),7.11(dd,J＝5.0,1.3Hz,1H),6.54(s,2H),4.49(d,J＝5.9Hz,2H),2.61(s,3H).LR-MS(ESI)m/z 398.1(M+1).
实施例65 3-((2-氨基嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用5-碘-嘧啶-2-胺代替5-溴-3-碘吡啶-2-胺和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.9Hz,1H),8.45(s,2H),7.99(d,J＝1.9Hz,1H),7.78(dd,J＝8.0,1.9Hz,1H),7.48(dd,J＝4.9,2.9Hz,1H),7.41(d,J＝8.1Hz,1H),7.36–7.31(m,1H),7.19(s,2H),7.08(dd,J＝4.9,1.3Hz,1H),4.45(d,J＝5.8Hz,2H),2.48(s,3H).LR-MS(ESI)m/z 349.1(M+1).
实施例66 3-((2-氨-5-(5,5-二甲基-5,6-二氢-4H-吡咯[1,2-b]吡唑-3-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用5,5-二甲基-5,6-二氢-4H-吡咯[1,2-b]吡唑-3-硼酸频哪醇酯(合成方法参考J.Med.Chem.2020,63,24,15564–15590)代替N-甲基吡唑-4-硼酸频哪醇酯和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,氯仿-d)δ8.11(d,J＝2.3Hz,1H),7.94(d,J＝2.0Hz,1H),7.71–7.66(m,2H),7.61(d,J＝2.3Hz,1H),7.32–7.27(m,2H),7.22–7.19(m,1H),7.08(dd,J＝5.0,1.3Hz,1H),6.80(t,J＝5.7Hz,1H),5.07(s,2H),4.63(d,J＝5.6Hz,2H),3.87(s,2H),2.83(s,2H),2.53(s,3H),1.32(s,6H).LR-MS(ESI)m/z 482.2(M+1).
实施例67 3-((2-氨基-5-(2-(((4-氰基四氢-2H-吡喃-4-基)甲基)氨基)噻唑-4-基)吡啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
步骤1：除用(2-((叔丁氧基甲酰基)((4-氰基四氢-2H-吡喃-4-基)甲基)氨基)噻唑-4-硼酸嚬那醇酯(合成方法参考Eur.J.Med.Chem.,158(2018):896-916.)代替N-甲基吡唑-4-硼酸频哪醇酯和用3-氨甲基噻吩代替苄胺外，参考实施例1的合成方法可制备中间体67-IM。1H NMR(400MHz,氯仿-d)δ8.54(s,1H),8.02–7.99(m,1H),7.93(d,J＝2.0Hz,1H),7.70(dd,J＝8.0,1.9Hz,1H),7.34–7.29(m,2H),7.22(s,1H),7.12–7.07(m,1H),7.01(s,1H),6.54(s,1H),5.27(s,2H),4.65(d,J＝5.5Hz,2H),4.51(s,2H),3.98–3.89(m,2H),3.70(td,J＝11.8,3.3Hz,2H),2.55(s,3H),1.93–1.83(m,4H),1.64(s,9H).LR-MS(ESI)m/z 669.2(M+1).
步骤2：向圆底烧瓶中加入中间体67-IM(50mg,0.075mmol)和DCM(2mL)，加入TFA(0.5mL)，室温反应过夜。将反应液倒入NaHCO3水溶液中，并用DCM萃取，合并有机相，用NaCl水溶液洗涤，经无水Na2SO4干燥，减压蒸干溶剂，柱层析分离得到 产物40mg(浅黄色固体；产率：94％)。1H NMR(400MHz,DMSO-d6)δ9.01(t,J＝5.9Hz,1H),8.52(d,J＝2.3Hz,1H),8.16(d,J＝2.0Hz,1H),8.09–8.01(m,2H),7.80(dd,J＝8.0,2.0Hz,1H),7.49(dd,J＝5.0,3.0Hz,1H),7.42(d,J＝8.0Hz,1H),7.34(dd,J＝2.9,1.2Hz,1H),7.09(dd,J＝5.0,1.3Hz,1H),6.99(s,1H),6.44(s,2H),4.47(d,J＝5.8Hz,2H),3.92(d,J＝11.2Hz,2H),3.66(d,J＝6.3Hz,2H),3.55–3.41(m,2H),2.53(s,3H),1.89(d,J＝13.5Hz,2H),1.71(td,J＝13.1,12.7,4.4Hz,2H).LR-MS(ESI)m/z 569.2(M+1).
实施例68 4-甲基-3-((6-(1-甲基-1H-吡唑-4-基)咪唑[1,2-b]并吡嗪-3-基)乙炔基)-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用6-氯-3-碘咪唑[1,2-b]并吡嗪代替5-溴-3-碘吡啶-2-胺和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,DMSO-d6)δ9.11(t,J＝6.6Hz,1H),8.47(s,1H),8.24(d,J＝9.5Hz,1H),8.18–8.05(m,3H),7.86(d,J＝8.0Hz,1H),7.72(d,J＝9.4Hz,1H),7.50(d,J＝7.0Hz,2H),7.35(s,1H),7.11(d,J＝5.0Hz,1H),4.48(d,J＝5.7Hz,2H),3.94(s,3H),2.67(s,3H).LR-MS(ESI)m/z 453.1(M+1).
实施例69叔丁基-5-((2-甲基-5-((噻吩-3-基甲基)氨基甲酰)苯基)乙炔基)-1H-吡唑[3,4-b]并吡啶-1-碳酸酯的制备
除用叔丁基-5-溴-1H-吡唑[3,4-b]并吡啶-1-碳酸酯代替5-溴-3-碘吡啶-2-胺和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,氯仿-d)δ8.88(d,J＝2.0Hz,1H),8.23(d,J＝2.0Hz,1H),8.19(s,1H),7.94(d,J＝2.0Hz,1H),7.71(dd,J＝8.0,2.0Hz,1H),7.37–7.31(m,3H),7.27–7.21(m,15H),7.11(dd,J＝4.9,1.3Hz,1H),6.34(s,1H),4.67(d,J＝5.5Hz,2H),2.58(s,3H),1.74(s,9H).LR-MS(ESI)m/z 473.2(M+1).
实施例70 3-((1H-吡唑[3,4-b]并吡啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用化合物83代替化合物79外，合成方法如实施例80。
1H NMR(400MHz,DMSO-d6)δ13.94(s,1H),9.07(t,J＝5.9Hz,1H),8.72(d,J＝2.0Hz,1H),8.51(d,J＝2.0Hz,1H),8.21(d,J＝1.4Hz,1H),8.08(d,J＝1.9Hz,1H),7.83(dd,J＝8.0,1.9Hz,1H),7.49(dd,J＝5.0,3.0Hz,1H),7.45(d,J＝8.1Hz,1H),7.34(dd,J＝2.9,1.3Hz,1H),7.09(dd,J＝4.9,1.3Hz,1H),4.47(d,J＝5.8Hz,2H),2.55(s,3H).LR-MS(ESI)m/z 373.1(M+1).
实施例71 3-(咪唑[1,2-a]并嘧啶-3-基乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用3-溴咪唑[1,2-a]并嘧啶代替5-溴-3-碘吡啶-2-胺和用3-氨甲基噻吩代替苄胺外，合成方法如实施例1。
1H NMR(400MHz,氯仿-d)δ8.81(dd,J＝4.6,1.6Hz,1H),8.21(dd,J＝8.0,1.7Hz,1H),8.01(d,J＝2.0Hz,1H),7.76(dd,J＝8.1,2.0Hz,1H),7.40(dd,J＝8.0,4.6Hz,1H),7.38–7.33(m,2H),7.25(s,2H),7.12(dd,J＝4.9,1.3Hz,1H),6.36(t,J＝6.5Hz,1H),4.67(d,J＝5.5Hz,2H),2.63(s,3H).LR-MS(ESI)m/z 373.2(M+1).
实施例72 3-((4-氨基-7-甲基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
步骤1：向圆底烧瓶中加入5-碘-7H-吡咯[2,3-d]并嘧啶-4胺(1.0g,3.85mmol)、K2CO3(1.59g,11.54mmol)和无水DMF(10mL)，滴加碘甲烷(0.29mL,4.61mmol)，室温反应8小时。反应结束后，加入2mL硫代硫酸钠水溶液，室温搅拌15min，随后倒入水中，DCM萃取，合并有机相并分别用水、NaCl水溶液洗涤，经无水Na2SO4干燥后，减压蒸干溶剂，柱层析分离得5-碘-7-甲基-7H-吡咯[2,3-d]嘧啶-4-胺600mg(产率：60％)。通过更换不同的卤代物及调整反应条件(如碱、温度等)，可获得不同N-取代物。
步骤2：向圆底烧瓶中加入5-碘-7-甲基-7H-吡咯[2,3-d]并嘧啶-4-胺(0.5g,1.82mmol)、三甲基硅乙炔(233mg,2.37mmol)、Et3N(5mL)和MeCN(20mL)，用氩气置换氧气，加入Pd(PPh3)2Cl2(64mg,0.091mmol)、CuI(17.4mg,0.091mmol)，重复除氧气的操作，室温条件下反应过夜；反应结束后加入50mL乙酸乙酯稀释反应液，过滤，减压蒸干溶剂，柱层析分离得产物7-甲基-5-((三甲基硅基)乙炔基)-7H-吡咯[2,3-d]并嘧啶-4-胺420mg(产率：94.2％)。
步骤3：向圆底烧瓶中加入3-碘-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺(90mg,0.25mmol)、7-甲基-5-((三甲基硅基)乙炔基)-7H-吡咯[2,3-d]并嘧啶-4-胺(62mg,0.25mmol)、Et3N(76mg,0.76mmol)、CsF(115mg,0.76mmol)和MeCN(10mL)，用氩气置换氧气，加入Pd(PPh3)2Cl2(9mg,0.013mmol)、CuI(5mg,0.025mmol)，重复除氧气的操作，室温条件下反应3小时。反应结束后，将反应液倒入水中，并用乙酸乙酯萃取，有机相用饱和NaCl溶液洗(10mL×3)，经无水硫酸钠干燥后，减压蒸干溶剂，柱层析分离得产物3-((4-氨基-7-甲基-7H-吡咯[2,3-d]嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺60mg(浅黄色固体；产率：59％)。1H NMR(400MHz,DMSO-d6)δ9.04(t,J＝5.7Hz,1H),8.17(s,1H),8.06(s,1H),7.79(d,J＝8.0Hz,1H),7.70(s,1H),7.49(s,1H),7.42(d,J＝8.0Hz,1H),7.34(s,1H),7.09(d,J＝5.0Hz,1H),6.71(s,2H),4.47(d,J＝5.8Hz,2H),3.73(s,3H),2.52(s,3H).LR-MS(ESI)m/z 402.1(M+1).
实施例73 3-((4-氨基-7-(甲基-d3)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用氘代碘甲烷代替碘甲烷外，合成方法如实施例72。
1H NMR(400MHz,DMSO-d6)δ9.02(t,J＝6.0Hz,1H),8.20(s,1H),8.08(s,1H),7.79(d,J＝7.8Hz,1H),7.70(s,1H),7.50(s,1H),7.42(d,J＝8.0Hz,1H),7.26-7.19(m,2H),6.63(s,2H),4.45(d,J＝5.9Hz,2H),2.53(s,3H).LR-MS(ESI)m/z 405.1(M+1).
实施例74叔丁基4-氨基-5-((2-甲基-5-(噻吩-3-基甲基)氨基甲酰基)苯基)乙炔 基)-7H-吡咯[2,3-d]并嘧啶-7-碳酸酯的制备
除用叔丁基4-氨基-5-碘-7H-吡咯[2,3-d]并嘧啶-7-碳酸酯代替5-碘-7-甲基-7H-吡咯[2,3-d]并嘧啶-4-胺外，合成方法如实施例72。
1H NMR(400MHz,DMSO-d6)δ8.96(t,J＝5.9Hz,1H),8.13(s,1H),8.12(d,J＝2.0Hz,1H),7.77(dd,J＝7.6,1.9Hz,1H),7.33(d,J＝7.6Hz,1H),7.22(dd,J＝4.8,1.8Hz,1H),7.13(s,1H),7.07–7.02(m,2H),6.63(s,2H),4.55(d,J＝5.7Hz,2H),2.52(s,3H),1.52(s,9H).LR-MS(ESI)m/z 488.2(M+1).
实施例75 3-((4-氨基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用化合物74代替67-IM外，合成方法如实施例67。
1H NMR(400MHz,DMSO-d6)δ13.15(s,1H),9.06(t,J＝6.0Hz,1H),8.41(s,1H),8.14(d,J＝1.9Hz,1H),7.94(d,J＝2.4Hz,1H),7.82(dd,J＝8.0,2.0Hz,1H),7.49(dd,J＝5.0,3.0Hz,1H),7.43(d,J＝8.1Hz,1H),7.33(d,J＝2.9Hz,1H),7.11–7.06(m,1H),6.70(s,2H),4.46(d,J＝5.8Hz,2H),2.51(s,3H).LR-MS(ESI)m/z 388.1(M+1).
实施例76 3-((4-氨基-7-乙基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中用碘乙烷代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.8Hz,1H),8.19(s,1H),8.05(s,1H),7.84–7.71(m,2H),7.49(dd,J＝5.0,2.9Hz,1H),7.42(d,J＝8.0Hz,1H),7.33(s,1H),7.09(d,J＝5.4Hz,1H),6.68(s,2H),4.46(d,J＝5.8Hz,2H),4.18(q,J＝7.2Hz,2H),2.52(s,3H),1.36(t,J＝7.2Hz,3H).LR-MS(ESI)m/z 416.1(M+1).
实施例77 3-((4-氨基-7-异丙基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中用2-碘丙烷代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.9Hz,1H),8.15(s,1H),8.05(s,1H),7.86(s,1H),7.78(dd,J＝8.0,1.9Hz,1H),7.48(dd,J＝5.0,3.0Hz,1H),7.41(d,J＝8.0Hz,1H),7.33(d,J＝2.8Hz,1H),7.09(d,J＝4.9Hz,1H),6.66(s,2H),4.92(hept,J＝7.7Hz,1H),4.46(d,J＝5.8Hz,2H),2.52(s,3H),1.45(d,J＝6.7Hz,6H).LR-MS(ESI)m/z 430.2(M+1).
实施例78 3-((4-氨基-7-异丙基-6-甲基-7H-吡咯[2,3-d]并嘧啶-5基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺
除用5-碘-7-异丙基-6-甲基-7H-吡咯[2,3-d]并嘧啶-4-胺(合成方法参考WO2014184069)代替5-碘-7-甲基-7H-吡咯[2,3-d]并嘧啶-4-胺外，合成方法如实施例72。
1H NMR(400MHz,DMSO-d6)δ8.96(t,J＝5.7Hz,1H),8.12(d,J＝2.0Hz,1H),8.10(s,1H),7.77(dd,J＝7.6,1.9Hz,1H),7.36–7.30(m,1H),7.22(dd,J＝4.8,1.8Hz,1H),7.09–7.01(m,2H),6.62(s,2H),4.85(hept,J＝4.5Hz,1H),4.55(d,J＝5.6Hz,2H),2.51(s,3H),2.27(s,3H),1.32(d,J＝4.4Hz,6H).LR-MS(ESI)m/z 444.2(M+1).
实施例79 3-((4-乙酰氨基-7-甲基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
向圆底烧瓶中加入化合物72(80mg,0.2mmol)和AcOH/Ac2O(1mL/1mL)，80℃反应8h，反应结束后，减压蒸干溶剂，柱层析分离得3-((4-乙酰氨基-7-甲基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺40mg(产率：45％)。
1H NMR(400MHz,DMSO-d6)δ10.22(s,1H),9.02(t,J＝6.0Hz,1H),8.65(s,1H), 7.99(s,2H),7.78(d,J＝9.0Hz,1H),7.49(dd,J＝5.4,2.9Hz,1H),7.41(d,J＝8.1Hz,1H),7.33(s,1H),7.09(d,J＝5.4Hz,1H),4.46(d,J＝5.7Hz,2H),3.84(s,3H),2.51(s,3H),2.16(s,3H).LR-MS(ESI)m/z 444.1(M+1).
实施例80 4-甲基-3-((7-甲基-4-(甲氨基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用5-碘-N,7-二甲基-7H-吡咯[2,3-d]并嘧啶-4胺代替5-碘-7-甲基-7H-吡咯[2,3-d]并嘧啶-4-胺外，合成方法如实施例72。
1H NMR(400MHz,DMSO-d6)δ9.01(t,J＝5.9Hz,1H),8.15(s,1H),8.05(s,1H),7.86(s,1H),7.78(dd,J＝8.0,1.9Hz,1H),7.48(dd,J＝5.0,3.0Hz,1H),7.41(d,J＝8.0Hz,1H),7.33(d,J＝2.8Hz,1H),7.09(d,J＝4.9Hz,1H),5.66(br s,1H),4.46(d,J＝5.7Hz,2H),3.95(s,3H),2.73(d,J＝2.6Hz,3H),2.52(s,3H).LR-MS(ESI)m/z 416.1(M+1).
实施例81 4-甲基-3-((7-甲基-4-(异丙氨基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用5-碘-N-异丙基-7-甲基-7H-吡咯[2,3-d]并嘧啶-4胺代替5-碘-7-甲基-7H-吡咯[2,3-d]并嘧啶-4-胺外，合成方法如实施例72。
1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝6.0Hz,1H),8.16(s,1H),8.05(s,1H),7.86(s,1H),7.78(dd,J＝8.0,1.9Hz,1H),7.48(dd,J＝5.0,3.0Hz,1H),7.41(d,J＝8.0Hz,1H),7.33(d,J＝2.8Hz,1H),7.09(d,J＝4.9Hz,1H),5.46(br s,1H),4.45(d,J＝5.9Hz,2H),3.15–2.98(m,1H),2.73(d,J＝2.6Hz,3H),2.52(s,3H),1.17(d,J＝5.9Hz,6H).LR-MS(ESI)m/z 444.2(M+1).
实施例82 3-((4-氨基-1-甲基-1H-吡唑[3,4-d]并嘧啶-3-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用3-碘-1-甲基-1H-吡唑[3,4-d]并嘧啶-4-胺代替5-碘-7-甲基-7H-吡咯[2,3-d]并嘧啶-4-胺外，合成方法如实施例72。
1H NMR(400MHz,DMSO-d6)δ9.06(t,J＝5.7Hz,1H),8.85(s,2H),8.27(s,1H),8.20(s,1H),7.86(d,J＝8.0Hz,1H),7.52–7.41(m,2H),7.34(s,1H),7.09(d,J＝5.0Hz,1H),4.47(d,J＝5.8Hz,2H),3.94(s,3H),2.53(s,3H).LR-MS(ESI)m/z 403.1(M+1).
实施例83 3-((4-氨基-7-(2,2,2-三氟乙基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物(BrCH2CF3)代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.8Hz,1H),8.23(s,1H),8.09(d,J＝1.9Hz,1H),7.81(dd,J＝7.9,2.0Hz,1H),7.76(s,1H),7.49(dd,J＝5.0,3.0Hz,1H),7.43(d,J＝8.1Hz,1H),7.33(dd,J＝2.9,1.2Hz,1H),7.09(dd,J＝4.9,1.3Hz,1H),6.82(s,2H),5.10(q,J＝9.1Hz,2H),4.47(d,J＝5.8Hz,2H),2.51(s,3H).LR-MS(ESI)m/z 470.1(M+1).
实施例84 3-((4-氨基-7-(1,1,1-三氟丙烷-2-基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物(CH3CHBrCF3)代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.02(t,J＝5.9Hz,1H),8.23(s,1H),8.11(d,J＝1.9Hz,1H),7.82(dd,J＝7.9,2.0Hz,1H),7.76(s,1H),7.49(dd,J＝5.0,3.0Hz,1H),7.43(d,J＝8.1Hz,1H),7.33(dd,J＝2.8,1.3Hz,1H),7.09(dd,J＝4.9,1.3Hz,1H),6.66(s,2H),5.02–4.89(m,1H),4.47(d,J＝5.8Hz,2H),2.52(s,3H),1.39(d,J＝6.4Hz,3H).LR-MS (ESI)m/z 484.1(M+1).
实施例85 3-((4-氨基-7-(2-甲氧乙基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物(CH3OCH2CH2Br)代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.8Hz,1H),8.24(s,1H),8.06(d,J＝1.9Hz,1H),7.79(dd,J＝8.0,1.9Hz,1H),7.70(s,1H),7.49(dd,J＝4.9,2.9Hz,1H),7.42(d,J＝8.1Hz,1H),7.33(dd,J＝3.0,1.2Hz,1H),7.09(dd,J＝5.0,1.3Hz,1H),6.69(s,2H),4.46(d,J＝5.8Hz,2H),4.31(t,J＝5.3Hz,2H),3.69(t,J＝5.3Hz,2H),3.24(s,3H),2.51(s,3H).LR-MS(ESI)m/z 446.2(M+1).
实施例86 3-((4-氨基-7-(1-甲氧丙烷-2-基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物(CH3OCH2CH(CH3)Br)代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ8.96(t,J＝5.9Hz,1H),8.12(d,J＝2.0Hz,1H),8.10(s,1H),7.77(dd,J＝7.6,1.9Hz,1H),7.54(s,1H),7.33(dd,J＝7.5,1.0Hz,1H),7.22(dd,J＝4.8,1.8Hz,1H),7.08–7.02(m,2H),6.66(s,2H),4.55(d,J＝5.7Hz,2H),4.27–4.12(m,1H),4.10–3.95(m,1H),3.27(s,3H),3.19–3.04(m,1H),2.51(s,3H),1.34(d,J＝6.2Hz,3H).LR-MS(ESI)m/z 460.2(M+1).
实施例87 3-((4-氨基-7-(2-(二甲氨基)乙基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物((CH3)2NCH2CH2Br)代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.04(t,J＝5.9Hz,1H),8.21(s,1H),8.06(d,J＝2.1Hz,1H),7.85–7.72(m,2H),7.49(dd,J＝4.9,2.9Hz,1H),7.43(d,J＝8.0Hz,1H),7.33(s,1H),7.09(d,J＝5.0Hz,1H),6.78(s,2H),4.56–4.48(m,2H),4.47(d,J＝5.8Hz,2H),3.61–3.42(m,2H),2.77(s,6H),2.51(s,3H).LR-MS(ESI)m/z 459.2(M+1).
实施例88 3-((4-氨基-7-(1-(二甲氨基)丙烷-2-基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物((CH3)2NCH2CH(CH3)Br)代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.9Hz,1H),8.15(s,1H),8.06(d,J＝1.9Hz,1H),7.79(dd,J＝8.0,1.9Hz,1H),7.70(s,1H),7.48(dd,J＝5.0,3.0Hz,1H),7.42(d,J＝8.0Hz,1H),7.36–7.30(m,1H),7.09(dd,J＝5.0,1.3Hz,1H),6.66(s,2H),4.46(d,J＝5.8Hz,2H),4.27–4.12(m,1H),4.10–3.95(m,1H),3.19–3.04(m,1H),2.18(s,6H),0.87(s,3H).LR-MS(ESI)m/z 473.2(M+1).
实施例89 3-((4-氨基-7-环丁基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.9Hz,1H),8.15(s,1H),8.06(d,J＝1.9Hz,1H),8.00(s,1H),7.79(dd,J＝8.0,1.9Hz,1H),7.49(dd,J＝5.0,3.0Hz,1H),7.42(d,J＝8.1Hz,1H),7.33(dd,J＝3.0,1.3Hz,1H),7.09(dd,J＝4.9,1.3Hz,1H),6.68(s,2H),5.15(p,J＝8.6Hz,1H),4.47(d,J＝5.8Hz,2H),2.61–2.52(m,2H),2.51(s,3H),2.44–2.35(m,2H),1.88–1.78(m,2H).LR-MS(ESI)m/z 442.2(M+1).
实施例90叔丁基-3-(4-氨基-5-((2-甲基-5-((噻吩-3-基甲基)氨基甲酰基)苯基)乙炔基)-7H-吡咯[2,3-d]并嘧啶-7-基)氮杂环丁烷-1-碳酸酯的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,氯仿-d)δ8.29(s,1H),7.90(d,J＝2.0Hz,1H),7.66(dd,J＝8.0,2.0Hz,1H),7.55(s,1H),7.36–7.29(m,2H),7.24(dd,J＝3.0,1.2Hz,4H),7.10(dd,J＝4.9,1.3Hz,1H),6.39(t,J＝5.5Hz,1H),5.70(s,2H),5.60–5.49(m,1H),4.66(d,J＝5.5Hz,2H),4.49(dd,J＝9.6,8.1Hz,2H),4.20(dd,J＝9.6,5.0Hz,2H),2.55(s,3H),1.48(s,9H).LR-MS(ESI)m/z543.2(M+1).
实施例91 3-((4-氨基-7-(1-乙基氮杂环丁烷-3-基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
步骤1：向圆底烧瓶中加入化合物90(0.3g,0.55mmol)和DCM(5mL)，加入HCl.dioxane溶液(0.5mL,2mmol)，室温搅拌过夜。反应结束后，减压蒸干溶剂得黄色固体91-IM 0.26g(产率：98.2％)，直接用于下一步，无需进一步纯化。
步骤2：向圆底烧瓶中加入上一步得到的106-IM(80mg,0.17mmol)、K2CO3(115mg,0.84mmol)和无水DMF(2mL)，滴加IEt(16μL,0.2mmol)，室温反应过夜。反应结束后，将反应液倒入水中，用乙酸乙酯萃取，合并有机相，并分别用H2O、NaCl水溶液洗涤，经无水Na2SO4干燥，减压蒸干溶剂，柱层析分离得到3-((4-氨基-7-(1-乙基氮杂 环丁烷-3-基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺25mg(白色固体；产率：32％)。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.9Hz,1H),8.16(s,1H),8.07(d,J＝1.9Hz,1H),8.03(s,1H),7.80(dd,J＝8.0,2.0Hz,1H),7.49(dd,J＝4.9,2.9Hz,1H),7.42(d,J＝8.0Hz,1H),7.33(dd,J＝2.9,1.2Hz,1H),7.09(dd,J＝5.0,1.3Hz,1H),6.73(s,2H),5.29–5.18(m,1H),4.47(d,J＝5.8Hz,2H),3.83–3.67(m,2H),3.56–3.37(m,2H),2.66–2.54(m,2H),2.52(s,3H),0.95(t,J＝7.1Hz,3H).LR-MS(ESI)m/z 471.2(M+1).
实施例92 3-((4-氨基-7-(1-异丙基氮杂环丁烷-3-基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
合成方法如实施例91。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝6.0Hz,1H),8.17(s,1H),8.05(d,J＝2.0Hz,1H),8.03(s,1H),7.81(d,J＝8.0Hz,1H),7.51(dd,J＝4.9,2.9Hz,1H),7.42(d,J＝8.0Hz,1H),7.33(dd,J＝2.9,1.2Hz,1H),7.09(dd,J＝5.0,1.3Hz,1H),6.67(s,2H),5.27–5.18(m,1H),4.45(d,J＝5.9Hz,2H),3.83–3.67(m,2H),3.52–3.34(m,2H),2.88–2.65(m,1H),2.52(s,3H),1.05(d,J＝5.1Hz,6H).LR-MS(ESI)m/z485.2(M+1).
实施例93 3-((4-氨基-7-(1-环丙基氮杂环丁烷-3-基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
合成方法如实施例91。1H NMR(400MHz,DMSO-d6)δ9.01(t,J＝6.0Hz,1H),8.14(s,1H),8.09(s,1H),8.03(s,1H),7.80(dd,J＝8.0,2.0Hz,1H),7.51(dd,J＝5.1,2.5Hz,1H),7.42(d,J＝8.0Hz,1H),7.33(d,J＝2.9Hz,1H),7.11(d,J＝5.0Hz,1H),6.68(s,2H),5.27–5.16(m,1H),4.45(d,J＝5.9Hz,2H),3.83–3.67(m,2H),3.56–3.37(m,2H),2.52(s,3H),2.46–2.34(m,1H),0.90–0.76(m,2H),0.64–0.52(m,2H).LR-MS(ESI)m/z483.2(M+1).
实施例94 3-((4-氨基-7-(1-乙酰基氮杂环丁烷-3-基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
向圆底烧瓶中加入化合物91-IM(80mg,0.17mmol)、Et3N(70μL,0.5mmol)和无水DCM(3mL)，冰浴滴加乙酰氯(13μL,0.18mmol)，室温反应1h。加入0.2mL甲醇淬灭反应后，减压蒸干溶剂，柱层析分离得3-((4-氨基-7-(1-乙酰基氮杂环丁烷-3-基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺15mg(浅黄色固体；产率：18.5％)。1H NMR(400MHz,DMSO-d6)δ9.04(t,J＝5.7Hz,1H),8.17(s,1H),8.06(s,1H),7.79(d,J＝8.0Hz,1H),7.70(s,1H),7.49(s,1H),7.42(d,J＝8.0Hz,1H),7.34(s,1H),7.09(d,J＝5.0Hz,1H),6.71(s,2H),4.96–4.80(m,1H),4.47(d,J＝5.8Hz,2H),4.12–3.96(m,2H),3.93–3.76(m,2H),2.52(s,3H),2.01(s,3H).LR-MS(ESI)m/z484.2(M+1).
实施例95 3-((4-氨基-7-(1-甲磺酰基氮杂环丁烷-3-基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用甲磺酰氯代替乙酰氯外，合成方法如实施例94。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.9Hz,1H),8.15(s,1H),8.05(s,1H),7.86(s,1H),7.78(dd,J＝8.0,1.9Hz,1H),7.48(dd,J＝5.0,3.0Hz,1H),7.41(d,J＝8.0Hz,1H),7.33(d,J＝2.8Hz,1H),7.09(d,J＝4.9Hz,1H),6.66(s,2H),4.92–4.79(m,1H),4.46(d,J＝5.8Hz,2H),4.15–3.99(m,2H),3.94–3.78(m,2H),2.89(s,3H),2.52(s,3H).LR-MS(ESI)m/z 521.1(M+1).
实施例96 3-((4-氨基-7-(3-氧代环丁基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基 -N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,氯仿-d)δ8.31(s,1H),7.91(d,J＝2.0Hz,1H),7.65(dd,J＝8.0,2.0Hz,1H),7.37–7.30(m,3H),7.24(dd,J＝2.9,1.2Hz,1H),7.10(dd,J＝4.9,1.3Hz,1H),6.35(t,J＝5.7Hz,1H),5.66(s,2H),5.39–5.32(m,1H),4.66(d,J＝5.5Hz,2H),3.87–3.63(m,4H),2.54(s,3H).LR-MS(ESI)m/z 456.1(M+1).
实施例97 3-((4-氨基-7-(3-羟基环丁基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
向圆底烧瓶中加入化合物96(50mg,0.11mmol)和无水THF(2mL)，加入NaBH4(12mg,0.33mmol)，室温反应2h。加入NH4Cl水溶液，减压蒸干溶剂，柱层析分离得3-((4-氨基-7-(3-羟基环丁基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺20mg(白色固体；产率：40％)。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.9Hz,1H),8.14(s,1H),8.06(d,J＝1.9Hz,1H),7.93(s,1H),7.79(d,J＝8.1Hz,1H),7.49(dd,J＝4.9,2.9Hz,1H),7.42(d,J＝8.1Hz,1H),7.33(s,1H),7.09(d,J＝4.9Hz,1H),6.69(s,2H),5.28(d,J＝7.0Hz,1H),4.73–4.63(m,1H),4.47(d,J＝5.8Hz,2H),4.08–3.92(m,1H),2.84–2.73(m,2H),2.52(s,3H),2.38–2.27(m,2H).LR-MS(ESI)m/z 458.2(M+1).
实施例98 3-((4-氨基-7-(氧杂环丁烷-3-基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ8.96(t,J＝6.0Hz,1H),8.12(d,J＝2.0Hz,1H),8.10(s,1H),7.77(dd,J＝7.6,1.9Hz,1H),7.54(s,1H),7.33(dd,J＝7.5,1.0Hz,1H),7.22(dd,J＝4.8,1.8Hz,1H),7.06(t,J＝1.6Hz,1H),7.04(dd,J＝4.9,1.6Hz,1H),6.69(s,2H),5.13–5.04(m,2H),4.96–4.88(m,1H),4.88–4.81(m,2H),4.55(d,J＝5.9Hz,2H),2.52(s,3H).LR-MS(ESI)m/z444.1(M+1).
实施例99 3-((4-氨基-7-环戊基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.04(t,J＝5.9Hz,1H),8.17(s,1H),8.05(d,J＝1.9Hz,1H),7.83(s,1H),7.79(dd,J＝8.0,1.9Hz,1H),7.49(dd,J＝4.9,2.9Hz,1H),7.42(d,J＝8.0Hz,1H),7.36–7.31(m,1H),7.09(dd,J＝5.0,1.3Hz,1H),6.68(s,2H),5.04(m,1H),4.47(d,J＝5.8Hz,2H),2.52(s,3H),2.11(m,2H),1.95–1.82(m,4H),1.69(m,2H).LR-MS(ESI)m/z456.2(M+1).
实施例100 3-((4-氨基-7-(四氢呋喃-3-基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ8.96(t,J＝6.0Hz,1H),8.12(d,J＝2.0Hz,1H),8.10(s,1H),7.77(dd,J＝7.6,1.9Hz,1H),7.54(s,1H),7.33(dd,J＝7.5,1.0Hz,1H),7.22(dd,J＝4.8,1.8Hz,1H),7.07–7.01(m,2H),6.68(s,2H),4.55(d,J＝5.9Hz,2H),4.18–4.00(m,2H),3.98–3.76(m,3H),2.52(s,3H),2.26–2.14(m,1H),2.02–1.88(m,1H).LR-MS(ESI)m/z 458.2(M+1).
实施例101叔丁基-4-(4-氨基-5-((2-甲基-5((噻吩-3-基)氨基甲酰基)苯基)乙炔基)-7H-吡咯[2,3-d]并嘧啶-7-基)哌啶-1-碳酸酯的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,氯仿-d)δ8.29(s,1H),7.90(d,J＝2.0Hz,1H),7.65(dd,J＝8.0,2.0Hz,1H),7.32–7.27(m,2H),7.27(s,1H),7.22–7.20(m,1H),7.08(dd,J＝5.0,1.3Hz,1H),6.61(t,J＝5.6Hz,1H),5.79(s,2H),4.79(tt,J＝12.1,4.0Hz,1H),4.64(d,J＝5.6Hz,2H),4.41–4.19(m,2H),2.98–2.81(m,2H),2.52(s,3H),2.08–2.01(m,2H),1.89–1.80(m,2H),1.48(s,9H).LR-MS(ESI)m/z 571.2(M+1).
实施例102 3-((4-氨基-7-(1-甲基哌啶4-基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.9Hz,1H),8.15(s,1H),8.05(d,J＝1.9Hz,1H),7.85(s,1H),7.79(dd,J＝8.0,1.9Hz,1H),7.48(dd,J＝5.0,3.0Hz,1H),7.42(d,J＝8.0Hz,1H),7.33(d,J＝2.9Hz,1H),7.09(d,J＝4.9Hz,1H),6.67(s,2H),4.56–4.48(m,1H),4.46(d,J＝5.8Hz,2H),2.90(d,J＝7.1Hz,2H),2.51(s,3H),2.22(s,3H),2.12–1.96(m,4H),1.91–1.80(m,2H).LR-MS(ESI)m/z 485.2(M+1).
实施例103 3-((4-氨基-7-(四氢-2H-吡喃-4-基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ8.96(t,J＝6.0Hz,1H),8.10(s,1H),8.02(d,J＝2.0Hz,1H),7.77(dd,J＝7.6,1.9Hz,1H),7.54(s,1H),7.33(dd,J＝7.5,1.0Hz,1H),7.26–7.18(m,1H),7.09–7.01(m,2H),6.64(s,2H),4.48(d,J＝5.9Hz,2H),3.90–3.73(m,3H),3.52–3.34(m,2H),2.53(s,3H),2.04–1.88(m,2H),1.79–1.64(m,2H).LR-MS(ESI)m/z 472.2(M+1).
实施例104 3-((4-氨基-7-环丙基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.9Hz,1H),8.15(s,1H),8.05(s,1H),7.86(s,1H),7.78(dd,J＝8.0,1.9Hz,1H),7.48(dd,J＝5.0,3.0Hz,1H),7.41(d,J＝8.0Hz,1H),7.33(d,J＝2.8Hz,1H),7.09(d,J＝4.9Hz,1H),6.62(s,2H),5.01-4.92(m,1H),4.46(d,J＝5.8Hz,2H),2.52(s,3H),0.64–0.51(m,2H),0.41–0.26(m,2H).LR-MS(ESI)m/z 428.1(M+1).
实施例105 3-((4-氨基-7-(戊烷-3-基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.9Hz,1H),8.15(s,1H),8.05(d,J＝1.9Hz,1H),7.85(s,1H),7.79(dd,J＝8.0,1.9Hz,1H),7.48(dd,J＝5.0,3.0Hz,1H),7.42(d,J＝8.0Hz,1H),7.33(d,J＝2.9Hz,1H),7.09(d,J＝4.9Hz,1H),6.67(s,2H),4.56–4.47(m,1H),2.52(s,3H),1.47–1.36(m,4H),0.83(t,J＝4.5Hz,6H).LR-MS(ESI)m/z 458.2(M+1).
实施例106 3-((4-氨基-7-异丁基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.02(t,J＝5.8Hz,1H),8.19(s,1H),8.05(s,1H),7.84–7.71(m,2H),7.49(dd,J＝5.0,2.9Hz,1H),7.42(d,J＝8.0Hz,1H),7.33(s,1H),7.09(d,J＝5.4Hz,1H),6.68(s,2H),4.44(d,J＝5.8Hz,2H),4.02(d,J＝6.9Hz,2H),2.51(s,3H),2.05–1.89(m,1H),0.91(d,J＝4.6Hz,6H).LR-MS(ESI)m/z 444.2(M+1).
实施例107 3-((4-氨基-7-((3-甲基氧杂环丁烷-3-基)甲基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.9Hz,1H),8.12(s,1H),8.05(s,1H),7.86(s,1H),7.77(dd,J＝8.0,1.9Hz,1H),7.48(dd,J＝5.0,3.0Hz,1H),7.41(d,J＝8.0Hz,1H),7.33(d,J＝2.8Hz,1H),7.09(d,J＝4.9Hz,1H),6.64(s,2H),4.46(d,J＝5.8Hz,2H),4.29(d,J＝9.3Hz,2H),4.14(d,J＝9.4Hz,2H),3.91(s,2H),2.52(s,3H),0.97(s,3H).LR-MS(ESI)m/z472.2(M+1).
实施例108 3-((7-(2-乙酰氨基乙基)-4-氨基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.04(t,J＝5.9Hz,1H),8.21(s,1H),8.06(d,J＝2.1Hz,1H),7.97(t,J＝5.6Hz,1H),7.85–7.72(m,2H),7.49(dd,J＝4.9,2.9Hz,1H),7.43(d,J＝8.0Hz,1H),7.33(s,1H),7.09(d,J＝5.0Hz,1H),6.78(s,2H),4.69(t,J＝6.5Hz,2H),4.45(d,J＝5.8Hz,2H),3.35–3.23(m,2H),2.51(s,3H),1.89(s,3H).LR-MS(ESI)m/z 473.2(M+1).
实施例109 3-((4-氨基-7-(3-二甲氨基)-3-氧代丙基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.01(t,J＝6.0Hz,1H),8.17(s,1H),8.06(d,J＝2.1Hz,1H),7.85–7.72(m,2H),7.49(dd,J＝4.9,2.9Hz,1H),7.43(d,J＝8.0Hz,1H),7.31(s,1H),7.09(d,J＝5.0Hz,1H),6.68(s,2H),4.83(t,J＝10.2Hz,2H),4.47(d,J＝6.0Hz,2H),2.76(s,6H),2.52(s,6H),2.41(t,J＝10.0Hz,2H).LR-MS(ESI)m/z 487.2(M+1).
实施例110 3-((4-氨基-7-(2-氟乙基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.8Hz,1H),8.24(s,1H),8.06(d,J＝1.9Hz,1H),7.79(dd,J＝8.0,1.9Hz,1H),7.70(s,1H),7.49(dd,J＝4.9,2.9Hz,1H),7.42(d,J＝8.1Hz,1H),7.33(dd,J＝3.0,1.2Hz,1H),7.09(dd,J＝5.0,1.3Hz,1H),6.69(s,2H),4.46–4.26(m,2H),4.31(t,J＝5.5Hz,2H),4.12–3.98(m,2H),2.51(s,3H).LR-MS(ESI)m/z 434.1(M+1).
实施例111 3-((4-氨基-7-羟乙基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.9Hz,1H),8.21(s,1H),8.06(d,J＝1.9Hz,1H),7.77(dd,J＝8.0,2.0Hz,1H),7.70(s,1H),7.49(dd,J＝4.9,2.9Hz,1H),7.42(d,J＝8.1Hz,1H),7.33(dd,J＝3.0,1.2Hz,1H),7.09(dd,J＝5.0,1.3Hz,1H),6.69(s,2H),4.96(t,J＝6.5Hz,1H),4.46(d,J＝5.8Hz,2H),4.32(t,J＝7.1Hz,2H),3.52–3.43(m,2H),2.53(s,3H).LR-MS(ESI)m/z 432.1(M+1).
实施例112 3-((4-氨基-7-(2-羟基-2-甲基丙基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.9Hz,1H),8.15(s,1H),8.06(d,J＝1.9Hz,1H),8.00(s,1H),7.79(dd,J＝8.0,1.9Hz,1H),7.49(dd,J＝5.0,3.0Hz,1H),7.42(d,J＝8.1Hz,1H), 7.33(dd,J＝3.0,1.3Hz,1H),7.09(dd,J＝4.9,1.3Hz,1H),6.68(s,2H),4.67(s,1H),4.47(d,J＝5.8Hz,2H),3.92(s,2H),2.51(s,3H),1.23(s,6H).LR-MS(ESI)m/z 460.2(M+1).
实施例113 3-((4-氨基-7-((2R,5S)-5-(羟甲基)四氢呋喃-2-基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ8.96(t,J＝6.0Hz,1H),8.12(d,J＝2.0Hz,1H),8.10(s,1H),7.77(dd,J＝7.6,1.9Hz,1H),7.54(s,1H),7.33(dd,J＝7.5,1.0Hz,1H),7.22(dd,J＝4.8,1.8Hz,1H),7.07–7.01(m,2H),6.62(s,2H),4.92–4.83(m,1H),4.55(d,J＝5.9Hz,2H),4.50(t,J＝7.4Hz,1H),4.42–4.31(m,1H),3.79(ddd,J＝11.8,7.4,6.3Hz,1H),3.54(ddd,J＝11.9,7.3,6.2Hz,1H),2.52(s,3H),2.22–2.06(m,2H),1.98–1.85(m,2H).LR-MS(ESI)m/z488.2(M+1).
实施例114 3-((4-氨基-7-(3,3-二氟环丁基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.9Hz,1H),8.14(s,1H),8.06(d,J＝1.9Hz,1H),7.93(s,1H),7.79(d,J＝8.1Hz,1H),7.49(dd,J＝4.9,2.9Hz,1H),7.42(d,J＝8.1Hz,1H),7.33(s,1H),7.09(d,J＝4.9Hz,1H),6.69(s,2H),4.73–4.63(m,1H),4.47(d,J＝5.8Hz,2H),2.73–2.54(m,2H),2.51(s,3H),2.47–2.25(m,2H).LR-MS(ESI)m/z 478.1(M+1).
实施例115 3-((4-氨基-7-(3,6-二氢-2H-吡喃-4-基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除用7-(3,6-二氢-2H-吡喃-4-基)-5-碘-7H-吡咯[2,3-d]并嘧啶-4-胺代替5-碘-7-甲基-7H-吡咯[2,3-d]并嘧啶-4-胺外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.9Hz,1H),8.14(s,1H),8.06(d,J＝1.9Hz,1H),7.93(s,1H),7.79(d,J＝8.1Hz,1H),7.49(dd,J＝4.9,2.9Hz,1H),7.42(d,J＝8.1Hz,1H),7.33(s,1H),7.09(d,J＝4.9Hz,1H),6.69(s,2H),5.67–5.52(m,1H),4.47(d,J＝5.8Hz,2H),4.17(d,J＝4.6Hz,2H),3.92–3.71(m,2H),2.51(s,3H),2.01–1.84(m,2H).LR-MS(ESI)m/z 470.2(M+1).
其中，中间体7-(3,6-二氢-2H-吡喃-4-基)-5-碘-7H-吡咯[2,3-d]并嘧啶-4-胺的合成路线如下所示。
步骤1：向圆底烧瓶中加入7H-吡咯[2,3-d]并嘧啶-4-胺(1.0g,7.45mmol)、DMF-DMA(1.07g,8.95mmol)和DMF(20mL)，室温反应过夜。减压蒸干溶剂得粗品，加入DCM重新溶解，分别用H2O和NaCl水溶液洗涤，经无水Na2SO4干燥后，柱层析分离得化合物115-2 950mg(产率：67.4％)。1H NMR(400MHz,DMSO-d6)δ12.06(s,1H),8.73(s,1H),8.20(s,1H),7.19(d,J＝2.7Hz,1H),6.62(dd,J＝3.4,1.9Hz,1H),2.87(s,3H),2.83(s,3H).LR-MS(ESI)m/z 190.1(M+1).
步骤2：向圆底烧瓶中加入化合物115-2(250mg,1.32mmol)、3,6-二氢-2H-吡喃-4-硼酸频哪醇酯(333mg,1.59mmol)、醋酸铜(291mg,1.59mmol)、2,2’-联吡啶(248mg, 1.59mmol)、无水Na2CO3(420mg,3.96mmol)和DMAC(10mL)。90℃搅拌4h,加入DCM并用水洗三次。有机相经无水Na2SO4干燥后，减压蒸干得中间体115-3，无需进一步纯化。
步骤3：向上一步中间体115-3加入EtOH(10mL)和乙二胺(0.2mL)，回流反应16h。反应结束后，减压蒸干溶剂，并用DCM重新溶解，NaCl水溶液洗涤，经无水Na2SO4干燥后，减压蒸干溶剂，柱层析分离得化合物115-4 160mg(两步总产率：56％)。1H NMR(400MHz,DMSO-d6)δ8.71(s,1H),7.70(d,J＝5.7Hz,1H),7.52(d,J＝6.0Hz,1H),6.72(s,2H),5.20–4.97(m,1H),4.20–4.10(m,2H),3.88–3.82(m,2H),2.17–2.05(m,2H).LR-MS(ESI)m/z 217.1(M+1).
步骤4：向圆底烧瓶中加入化合物115-4(160mg,0.74mmol)、NIS(183mg,0.81mmol)和DMF(3mL)，室温反应4h。反应结束后，减压蒸干溶剂，柱层析分离得7-(3,6-二氢-2H-吡喃-4-基)-5-碘-7H-吡咯[2,3-d]并嘧啶-4-胺180mg(产率：71％)。1H NMR(400MHz,DMSO-d6)δ8.73(s,1H),7.68(s,1H),6.68(s,2H),5.13–4.96(m,1H),4.21–3.88(m,4H),2.13–2.01(m,2H).LR-MS(ESI)m/z 343.1(M+1).
实施例116 3-((4-氨基-7-((1-甲基-1H-吡唑-4-基)甲基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的卤化物代替碘甲烷外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.02(t,J＝5.9Hz,1H),8.15(s,1H),8.05(s,1H),7.86(s,1H),7.78(dd,J＝8.0,1.9Hz,1H),7.48(dd,J＝5.0,3.0Hz,1H),7.41(d,J＝8.0Hz,1H),7.36(s,2H),7.33(d,J＝2.8Hz,1H),7.09(d,J＝4.9Hz,1H),6.64(s,2H),5.57(s,2H),4.46(d,J＝6.0Hz,2H),3.94(s,3H),2.52(s,3H).LR-MS(ESI)m/z 482.2(M+1).
实施例117 3-((4-氨基-7-(1-甲基-1H-吡唑-4-基)-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除了在步骤2中使用相应的二唑衍生物代替3,6-二氢-2H-吡喃-4-硼酸频哪醇酯以外，合成方法如实施例115。1H NMR(400MHz,DMSO-d6)δ9.02(t,J＝6.0Hz,1H),8.14(s,1H),8.05(d,J＝1.9Hz,1H),7.93(s,1H),7.79(d,J＝8.1Hz,1H),7.52(d,J＝1.8Hz,1H),7.49(dd,J＝4.9,2.9Hz,1H),7.42(d,J＝8.1Hz,1H),7.33(s,1H),7.30(d,J＝1.6Hz,1H),7.09(d,J＝4.9Hz,1H),6.69(s,2H),4.47(d,J＝5.8Hz,2H),2.51(s,3H),3.89(s,3H).LR-MS(ESI)m/z 468.2(M+1).
实施例118 3-((4-氨基-7-异丙基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(1-(噻吩-3-基)环丙基)苯甲酰胺的制备
除在步骤1中使用相应的2-碘丙烷代替碘甲烷，并参考实施例1步骤3的方法得到相应的中间体外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ8.86(s,1H),8.12(d,J＝2.0Hz,1H),8.10(s,1H),7.77(dd,J＝7.6,1.9Hz,1H),7.54(s,1H),7.33(d,J＝7.6,1H),7.25(dd,J＝5.2,1.9Hz,1H),7.10–7.05(m,2H),6.64(s,2H),4.94–4.75(m,1H),2.45(d,J＝0.9Hz,3H),1.32(d,J＝6.5Hz,6H),0.95–0.79(m,2H),0.71–0.54(m,2H).LR-MS(ESI)m/z 456.2(M+1).
实施例119 3-((4-氨基-7-异丙基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基-d2)苯甲酰胺的制备
除在步骤1中使用相应的2-碘丙烷代替碘甲烷，并参考实施例1步骤3的方法得到相应的中间体外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.03(s,1H), 8.17(s,1H),8.05(s,1H),7.86(s,1H),7.78(dd,J＝8.0,1.9Hz,1H),7.48(dd,J＝5.0,3.0Hz,1H),7.41(d,J＝8.0Hz,1H),7.33(d,J＝2.8Hz,1H),7.09(d,J＝4.9Hz,1H),6.65(s,2H),4.92(hept,J＝7.7Hz,1H),2.52(s,3H),1.45(d,J＝6.7Hz,6H).LR-MS(ESI)m/z432.2(M+1).
实施例120 3-((4-氨基-7-异丙基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-2-基甲基)苯甲酰胺的制备
除在步骤1中使用相应的2-碘丙烷代替碘甲烷，并参考实施例1步骤3的方法得到相应的中间体外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ8.98(t,J＝6.0Hz,1H),8.12(d,J＝2.0Hz,1H),8.10(s,1H),7.77(dd,J＝7.6,1.9Hz,1H),7.54(s,1H),7.44(dd,J＝5.1,1.8Hz,1H),7.33(dd,J＝7.5,1.0Hz,1H),7.12(dd,J＝6.4,1.8Hz,1H),7.02(dd,J＝6.4,5.1Hz,1H),6.64(s,2H),4.89–4.81(m,1H),4.45(d,J＝5.9Hz,2H),2.53(s,3H),1.34(d,J＝4.6Hz,6H).LR-MS(ESI)m/z 430.2(M+1).
实施例121 3-((4-氨基-7-异丙基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-N-((5-氯噻吩-2-基)甲基)-4-甲基苯甲酰胺的制备
除在步骤1中使用相应的2-碘丙烷代替碘甲烷，并参考实施例1步骤3的方法得到相应的中间体外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.04(t,J＝8.0Hz,1H),8.36–8.29(m,2H),7.98(dd,J＝7.6,1.9Hz,1H),7.75(s,1H),7.54(d,J＝7.5Hz,1H),7.33(d,J＝6.4Hz,1H),7.27(d,J＝6.4Hz,1H),6.67(s,2H),5.12–5.03(m,1H),4.56(d,J＝8.0Hz,2H),2.54(s,3H),1.43(d,J＝4.6Hz,6H).LR-MS(ESI)m/z 464.1(M+1).
实施例122 3-((4-氨基-7-异丙基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-N-((5-环丙基噻吩-2-基)甲基)-4-甲基苯甲酰胺的制备
除在步骤1中使用相应的2-碘丙烷代替碘甲烷，并参考实施例1步骤3的方法得到相应的中间体外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.02(t,J＝5.8Hz,1H),8.31(d,J＝2.0Hz,1H),8.29(s,1H),7.96(dd,J＝7.6,1.9Hz,1H),7.73(s,1H),7.52(dd,J＝7.5,1.0Hz,1H),7.15–7.07(m,2H),6.62(s,2H),5.10–4.98(m,1H),4.54(d,J＝6.0Hz,2H),2.64(s,3H),2.05–1.93(m,1H),1.51(d,J＝4.6Hz,6H),0.91–0.70(m,4H).LR-MS(ESI)m/z 470.2(M+1).
实施例123 3-((4-氨基-7-异丙基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-N-((6-氟吡啶-3-基)甲基)-4-甲基苯甲酰胺的制备
除在步骤1中使用相应的2-碘丙烷代替碘甲烷，并参考实施例1步骤3的方法得到相应的中间体外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.03(t,J＝5.8Hz,1H),8.42–8.37(m,2H),8.19(d,J＝2.0Hz,1H),8.17(s,1H),7.84(dd,J＝7.6,1.9Hz,1H),7.61(s,1H),7.40(dd,J＝7.5,1.0Hz,1H),7.23–7.08(m,1H),6.72(s,2H),4.97–4.85(m,1H),4.47(d,J＝5.7Hz,2H),2.52(s,3H),1.39(d,J＝4.7Hz,6H).LR-MS(ESI)m/z 443.2(M+1).
实施例124 3-((4-氨基-7-异丙基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-((6-甲基吡啶-3-基)甲基)苯甲酰胺的制备
除在步骤1中使用相应的2-碘丙烷代替碘甲烷，并参考实施例1步骤3的方法得到相应的中间体外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ8.96(t,J＝6.0Hz,1H),8.55(d,J＝2.3Hz,1H),8.12(d,J＝2.0Hz,1H),8.10(s,1H),7.92(dd,J＝7.5,2.2Hz,1H),7.77(dd,J＝7.6,1.9Hz,1H),7.54(s,1H),7.33(dd,J＝7.5,1.1Hz,1H),7.29(d,J＝7.5Hz,1H),6.64(s,2H),4.89–4.82(m,1H),4.50(d,J＝5.7Hz,2H),2.58(s,3H),2.52 (s,3H),1.34(d,J＝4.9Hz,6H).LR-MS(ESI)m/z 439.2(M+1).
实施例125 3-((4-氨基-7-异丙基-7H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-N-((6-环丙基吡啶-3-基)甲基)-4-甲基苯甲酰胺的制备
除在步骤1中使用相应的2-碘丙烷代替碘甲烷，并参考实施例1步骤3的方法得到相应的中间体外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ9.02(t,J＝6.0Hz,1H),8.52(d,J＝2.1Hz,1H),8.09(d,J＝2.0Hz,1H),8.04(s,1H),7.92(dd,J＝7.5,1.9Hz,1H),7.77(dd,J＝7.6,1.9Hz,1H),7.54(s,1H),7.33(d,J＝7.5Hz,1H),7.29(d,J＝7.5Hz,1H),6.64(s,2H),4.89–4.82(m,1H),4.45(d,J＝5.7Hz,2H),3.31–2.89(m,1H),2.53(s,3H),1.34(d,J＝4.9Hz,6H),0.98–0.73(m,4H).LR-MS(ESI)m/z 465.2(M+1).
实施例126乙基-4-氨基-5-((2-甲基-5-((噻吩-3-基甲基)氨基甲酰基)苯基)乙炔基)-7H-吡咯[2,3-d]并嘧啶-7-羧酸酯的制备
除用乙基-4-氨基-5-碘-7H-吡咯[2,3-d]并嘧啶-7-碳酸酯(126-3)代替5-碘-7-甲基-7H-吡咯[2,3-d]并嘧啶-4-胺外，合成方法如实施例72。1H NMR(400MHz,DMSO-d6)δ8.96(t,J＝5.8Hz,1H),8.13(s,1H),8.12(d,J＝2.0Hz,1H),7.77(dd,J＝7.6,1.9Hz,1H),7.72(s,1H),7.33(dd,J＝7.5,1.0Hz,1H),7.22(dd,J＝4.8,1.8Hz,1H),7.07–7.02(m,2H),6.70(s,2H),4.55(d,J＝5.9Hz,2H),4.36(q,J＝6.2Hz,2H),2.45(d,J＝0.9Hz,3H),1.26(t,J＝5.9Hz,3H).LR-MS(ESI)m/z 460.1(M+1).
其中，中间体126-3的合成方法如下所示：
步骤1：向圆底烧瓶中加入115-2(0.5g,2.64mmol)及无水THF(30mL)，冰浴下加入NaH(60％,211mg)，搅拌30分钟后，加入氯甲酰乙酯(430mg,3.96mmol)，室温反应2小时。反应结束后，冰浴滴加饱和氯化铵水溶液淬灭反应，乙酸乙酯萃取，合并有机相并用饱和食盐水洗涤，经无水硫酸钠干燥后，柱层析分离得142-1(400mg,黄色油状物，产率57.9％)。1H NMR(400MHz,DMSO-d6)δ9.03(s,1H),8.20(s,1H),7.23(d,J＝2.7Hz,1H),6.62(dd,J＝3.4,1.9Hz,1H),4.36(q,J＝4.2Hz,2H),2.83(s,6H),1.26(t,J＝4.3Hz,3H).LR-MS(ESI)m/z 388.1(M+1).
步骤2及步骤3分别通过参考实施例115的步骤3和步骤4可获得126-3为棕色固体。1H NMR(400MHz,DMSO-d6)δ8.63(s,1H),7.16(s,1H),6.65(s,2H),4.36(q,J＝4.2Hz,2H),1.26(t,J＝4.3Hz,3H).LR-MS(ESI)m/z 333.1(M+1).
实施例127 3-((4-氨基-7-(环丙甲酰基)-7-H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺的制备
除在步骤1中用氯甲酰环丙烷代替氯甲酰乙酯外，合成方法如实施例126。1H NMR(400MHz,DMSO-d6)δ8.96(t,J＝6.0Hz,1H),8.13(s,1H),8.12(d,J＝2.0Hz,1H),7.77(dd,J＝7.6,1.9Hz,1H),7.37–7.29(m,1H),7.30(s,1H),7.22(dd,J＝4.8,1.8Hz,1H),7.06(t,J＝1.6Hz,1H),7.04(dd,J＝4.9,1.6Hz,1H),6.62(s,2H),4.55(d,J＝5.9Hz,2H), 2.53(s,3H),1.92–1.80(m,1H),1.20–0.99(m,4H).LR-MS(ESI)m/z 456.1(M+1).
实施例128 3-((4-氨基-7-异丙基-7-H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-((5-甲基噻吩-3-基)甲基)苯甲酰胺的制备
除了在步骤1中以2-碘丙烷代替碘甲烷，在步骤3中以3-碘-4-甲基-N-(5-甲基噻吩-3-基甲基)苯甲酰胺代替3-碘-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺以外，合成方法参考实施例72。1H NMR(500MHz,氯仿-d)δ8.84(t,J＝5.8Hz,1H),8.18(s,1H),7.87(d,J＝1.9Hz,1H),7.77(dd,J＝8.1,2.0Hz,1H),7.55(s,1H),7.32(dd,J＝8.1,1.0Hz,1H),6.90(d,J＝1.6Hz,1H),6.68(dd,J＝1.6,0.7Hz,1H),5.96(s,2H),4.74–4.69(m,1H),4.51(d,J＝5.7Hz,2H),2.52(s,3H),2.36(s,3H),1.53(d,J＝4.9Hz,6H).LR-MS(ESI)m/z 444.2(M+1).
实施例129 3-((4-氨基-7-异丙基-7-H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-N-((5-氯噻吩-3-基)甲基)-4-甲基苯甲酰胺的制备
除了在步骤1中以2-碘丙烷代替碘甲烷，在步骤3中以3-碘-4-甲基-N-(5-氯噻吩-3-基甲基)苯甲酰胺代替3-碘-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺以外，合成方法参考实施例72。1H NMR(500MHz,DMSO-d6)δ8.92(t,J＝5.9Hz,1H),8.19(s,1H),7.87(d,J＝1.9Hz,1H),7.80(dd,J＝8.1,1.9Hz,1H),7.55(s,1H),7.33(dd,J＝8.1,1.0Hz,1H),7.03(d,J＝1.8Hz,1H),6.87(d,J＝1.6Hz,1H),6.58(s,2H),4.77–4.68(m,1H),4.54(d,J＝6.0Hz,2H),253(s,3H),1.50(d,J＝4.9Hz,6H).LR-MS(ESI)m/z 464.1(M+1).
实施例130 3-((4-氨基-7-异丙基-7-H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-N-((5-甲氧基噻吩-2-基)甲基)-4-甲基苯甲酰胺的制备
除了在步骤1中以2-碘丙烷代替碘甲烷，在步骤3中以3-碘-4-甲基-N-(5-甲氧基噻吩-2-基甲基)苯甲酰胺代替3-碘-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺以外，合成方法参考实施例72。1H NMR(500MHz,DMSO-d6)δ9.02(t,J＝6.0Hz,1H),8.25(s,1H),7.94(d,J＝1.9Hz,1H),7.87(dd,J＝8.2,1.9Hz,1H),7.62(s,1H),7.41–7.37(m,1H),6.97(d,J＝7.0Hz,1H),6.87(d,J＝7.0Hz,1H),6.61(s,2H),4.85–4.77(m,1H),4.58(d,J＝5.6Hz,2H),3.93(s,3H),2.52(s,3H),1.56(d,J＝4.9Hz,6H).LR-MS(ESI)m/z 460.2(M+1).
实施例131 3-((4-氨基-7-异丙基-7-H-吡咯[2,3-d]并嘧啶-5-基)乙炔基)-4-甲基-N-((5-甲基噻吩-2-基)甲基)苯甲酰胺的制备
除了在步骤1中以2-碘丙烷代替碘甲烷，在步骤3中以3-碘-4-甲基-N-(5-甲基噻吩-2-基甲基)苯甲酰胺代替3-碘-4-甲基-N-(噻吩-3-基甲基)苯甲酰胺以外，合成方法参考实施例72。1H NMR(400MHz,DMSO-d6)δ8.90(t,J＝5.8Hz,1H),8.19(s,1H),7.87(d,J＝1.9Hz,1H),7.80(dd,J＝8.1,1.9Hz,1H),7.55(d,J＝0.7Hz,1H),7.33(dd,J＝8.1,1.0Hz,1H),6.99(d,J＝6.3Hz,1H),6.89(d,J＝6.3Hz,1H),6.61(s,2H),4.79–4.69(m,1H),4.54(d,J＝5.7Hz,2H),2.52(s,3H),2.35(s,3H),1.50(d,J＝4.9Hz,6H).LR-MS(ESI)m/z 444.2(M+1).
实施例132乙基(7-异丙基-5-((2-甲基-5-((噻吩-3-基甲基)氨基甲酰基)苯基)乙炔基)-7H-吡咯[2,3-d]并嘧啶-4-基)氨基甲酸酯的制备
向圆底烧瓶中加入化合物77(80mg,0.186mmol)，DIPEA(72mg,0.559mmol)和无水DMF(2mL)，冰浴下滴加氯甲酸乙酯(30mg,0.279mmol)，搅拌1小时后滴加甲醇淬灭反应。减压蒸干溶剂，柱层析分离得化合物132(30mg，黄色固体，产率：32％)。1H NMR(400MHz,氯仿-d)δ8.71(s,1H),8.42(s,1H),8.02(s,1H),7.74(d,J＝8.1Hz,1H),7.47(s,1H),7.37–7.29(m,2H),7.26–7.22(m,1H),7.11(d,J＝5.0Hz,1H),6.63(br s,1H),5.12(p,J＝6.6Hz,1H),4.67(d,J＝5.5Hz,2H),4.29(q,J＝7.0Hz,2H),2.58(s,3H),1.55(d,J＝6.8Hz,6H),1.30(t,J＝7.3Hz,3H).LR-MS(ESI)m/z 502.2(M+1).
实施例133异丙基(7-异丙基-5-((2-甲基-5-((噻吩-3-基甲基)氨基甲酰基)苯基)乙炔基)-7H-吡咯[2,3-d]并嘧啶-4-基)氨基甲酸酯的制备
除用氯甲酸异丙酯代替氯甲酸乙酯外，合成方法如实施例132。1H NMR(400MHz,DMSO-d6)δ9.92(s,1H),8.90(t,J＝6.0Hz,1H),8.29(s,1H),7.87(d,J＝1.9Hz,1H),7.80(dd,J＝8.1,1.9Hz,1H),7.57(s,1H),7.36–7.29(m,2H),7.24(t,J＝1.7Hz,1H),6.98(dd,J＝5.0,1.7Hz,1H),5.03–4.92(m,1H),4.78–4.69(m,1H),4.55(d,J＝5.9Hz,2H),2.52(s,3H),1.50(d,J＝4.9Hz,6H),1.29(d,J＝5.7Hz,6H).LR-MS(ESI)m/z 516.2(M+1).
(二)、生物活性检测实施例
实验例一：化合物对携带不同耐药突变KIT细胞的增殖抑制活性
实验方法：
采用MTT法检测化合物对细胞增殖的影响。将状态良好的细胞收集，接种到96孔板，加入不同浓度的化合物；于37℃、含5％CO2、饱和湿度培养箱中培养72小时(h)。药物作用结束后，每孔加入MTT，继续于37℃、5％CO2、饱和湿度培养箱中培养4小时。加入三联液(10％SDS，5％异丁醇，0.01mol/L HCl)，置于37℃温箱中12h，保证蓝紫色甲臜完全溶解；酶标仪上570nm和690nm波长下测定OD值。按以下公式计算化合物对细胞的抑制率：
抑制率(％)＝(对照孔OD值－给药孔OD值)/对照孔OD值×100％
以Graphpad Prism 8.0软件计算化合物IC50，并生成浓度(对数)-抑制率图。
实验结果：
本发明实施例化合物、阳性药Ripretinib对32D KIT D816V细胞增殖抑制活性见表2：
表2化合物对32D KIT D816V细胞增殖的抑制作用


实验结论：由表2可知，本发明实施例的杂芳环炔基类化合物对32D KIT D816V细胞增殖均具有显著抑制活性，其中有33个化合物优于Ripretinib。实施例76、77、99和120对32D KIT D816V细胞增殖抑制活性IC50值小于1nM，显著优于阳性药Ripretinib，显示出该类化合物具有很强的抑制携带KIT D816V耐药突变细胞增殖的优势。
挑选代表性化合物和阳性药Ripretinib进一步评价其对其他KIT突变细胞(32D KIT V559D、32D KIT V559D-V654A、32D KIT V559D-Y823D和32D KIT V559D-N822K)、KIT野生型细胞(NCI-H526、Mo7e、HMC-1)和32D细胞增殖抑制活性，结果如表3所示。
表3代表性化合物对携带不同突变KIT、KIT野生型及正常细胞增殖抑制活性比较

NT:not test.
结果表明，代表性化合物对携带V559D、V559D-V654A、V559D-Y823D和V559D-N822K不同突变KIT的细胞增殖均有强效抑制活性，体现出该类化合物具有广谱抑制突变KIT的优点；同时，代表性化合物对野生型KIT依赖的细胞株活性较弱，与突变型KIT相比具有高的治疗指数，如对KITD861V的选择指数高于490倍，并优于阳性药ripretinib。此外，化合物对32D正常细胞不具有细胞毒性，显示该类化合物具有很好的选择性，可避免潜在的脱靶毒副作用。
进一步测试代表性化合物对其他野生型靶点依赖的细胞株的增殖抑制活性，包括PDGFRα(U118MG)、EGFR(A431)，结果如表4所示。
结果表明代表性化合物对野生型PDGFR及EGFR依赖的细胞株均不具有细胞毒性，初步显示出了良好的靶点选择性；而阳性药ripretinib对PDGFR依赖的U118MG细胞具有较为明显的抑制活性，治疗指数较低。
表4代表性化合物对PDGFRα及EGFR依赖的细胞株的增殖抑制活性
实验例二：代表性化合物27、48、77和85对KIT及其介导的下游信号影响
蛋白免疫印迹(Western Blot)检测化合物对32D KIT D816V，32D KIT V559D和32D KIT V559D-V654A细胞中KIT及其下游信号通路影响
实验方法：
细胞接种于六孔板，加入不同浓度药物，于37℃、5％CO2、饱和湿度培养箱中处理4小时。用1×SDS凝胶上样缓冲液(50mM Tris-HCl(pH 6.8)，100mM DTT，2％SDS，10％甘油，0.1％溴酚蓝)于冰上裂解细胞。细胞裂解物在沸水浴中加热10min变性， 进行SDS-PAGE电泳，电泳结束后，将蛋白转移至PVDF膜，置于封闭液(5％脱脂奶粉稀释于TBST)中室温封闭1小时，洗膜后，加入相应I抗、II抗，用ECL试剂显色，最后在ECL化学发光成像***观察、拍照。
实验结论：
代表性化合物和阳性药Ripretinib抑制携带不同突变型KIT的肿瘤细胞中KIT及其下游信号通路结果见图1-3。由图1-3可知，本发明实施例的含杂芳环炔基的化合物在细胞水平上可显著抑制不同突变形式KIT及其下游信号通路的活化。
实验例三：小鼠体内药效评价试验
实验方法：
BALB/c-nu裸小鼠，5-6周，♀。每只裸小鼠皮下接种32D KIT D816V细胞，待平均肿瘤体积达到～100mm3后，根据肿瘤体积将动物分组(D0)。小鼠灌胃(i.g.)给药(每天2次)，给药体积10mL/kg；溶剂组给予相同体积的“溶剂”；每周测2次肿瘤体积，称小鼠体重，记录数据。
化合物77、85、Repritinib均用“溶剂”(5％DMSO/5％Etoh/40％PEG 400/50％H2O配制。
实验结果：
化合物85(30、100mg/kg,i.g.,BID×12)剂量依赖性抑制32D KIT D816V裸小鼠皮下移植瘤生长，抑瘤率分别为20％和50％；化合物77(20、40mg/kg,i.g.,BID×12)同样剂量依赖性抑制32D KIT D816V裸小鼠皮下移植瘤生长，抑瘤率分别为45％和71％；Ripretinib(30、100mg/kg,i.g.,BID×12)对32D KIT D816V裸小鼠皮下移植瘤的抑瘤率分别为13％和21％；荷瘤小鼠对以上药物均能较好耐受，没有明显体重下降等症状发生。相比较，化合物77、85对32D KIT D816V裸小鼠皮下移植瘤的疗效均显著强于Ripretinib(P<0.05，高剂量组比较)。
综上可知，本发明实施例的含杂芳环炔基的化合物对32D KIT D816V细胞抑制活性强；同时，代表性化合物27、48、77和85对V559D、V559D-V654A、V559D/Y823D和V559D/N822K不同突变形式的KIT均具有强效抑制活性，表明该类化合物具有强效、广谱抑制不同类型突变KIT的优点。代表性化合物27、48、77和85对32D正常细胞、及携带野生型KIT、PDGRFα及EGFR细胞抑制作用弱，表明该类化合物具有高的选择性，可避免相关脱靶的毒副作用；重要的是，代表性化合物77、85对32D KIT D816V裸小鼠皮下移植瘤模型具有明显药效；其药效显著强于参比化合物Ripretinib(P<0.05)，表明所述含杂芳环炔基的化合物具有体内药效更强的优点。
以上结合附图详细描述了本发明的优选实施方式，但是，本发明并不限于上述实施方式中的具体细节，在本发明的技术构思范围内，可以对本发明的技术方案进行多种简单变型，这种简单变型均属于本发明的保护范围。

Claims (10)

  1. 一种式(I)的化合物，或其氘代化合物、药学上可接受的盐、溶剂化物、酯、酸、代谢物或前药：
    其中：
    X选自-(C(Ra)(Rb))q-；
    其中Ra、Rb各自独立地选自氢、氘、卤素、C1-4烷基；或者连接在同一个碳原子上的Ra、Rb与该碳原子一起形成3-5元碳环；
    q选自1、2；
    R1选自氢、卤素、C1-6烷基；
    环M1选自以下结构：
    其中：
    A1、A2各自独立为CRH4或N原子；
    A3为CRH2或N原子；
    A4为CRH3、NRH3或N原子；
    Z选自未取代或取代的5-10元杂芳基、未取代或取代的5-10元杂环基；所述取代是指上述基团各自独立地被1-5个R2基团取代；或者Z和与其相连的碳原子以及相邻的A1或A2一起形成取代或未取代的环W1，所述取代是指环W1被1-5个R2基团取代；在杂芳基、杂环基、或者环W1中，一个或多个环C原子任选地被相应数目的C(＝O)基团替代，一个或多个环S或N原子任选地被氧化形成S-氧化物或N-氧化物；
    W为未取代或取代的C6-10芳基、未取代或取代的5-7元杂芳基或未取代或取代的5-7元杂环基；所述取代是指被1-5个R2基团取代；在芳基、杂芳基或杂环基中，一个或多个环C原子任选地被相应数目的C(＝O)基团替代，一个或多个环S或N原子任选地被氧化形成S-氧化物或N-氧化物；
    所述R2各自独立选自卤素、氰基、C1-6烷基、C1-6杂烷基、C1-6烷氧羰基、C2-6烯基、C2-6炔基、C3-6环烷基、3-6元杂环烷基、C4-8环烯基、4-8元杂环烯基、C6-10芳基、5-7元杂芳基、5-7元杂环基、-C(＝O)(C1-6烷基)、-C(＝O)(C3-6环烷基)、-OR3、-N(R3)2、-(C1-6烷基)OR3、-C(＝O)N(R3)2、-(C1-6烷基)-C(＝O)N(R3)2、-SR3、-S(＝O)R3、-S(＝O)2R3、-N(R3)C(＝O)R3、-(C1-6烷基)-N(R3)C(＝O)R3、-N(R3)S(＝O)2R3、-P(＝O)(R3)(R3)；其中所述C1-6烷基、C1-6杂烷基、C1-6烷氧羰基、C2-6烯基、C2-6炔基、C3-6环烷基、3-6元杂环烷基、4-8元环烯基、4-8元杂环烯基、C6-10芳基、5-7元杂芳基、5-7 元杂环基任选被1-5个R3取代；或者
    两个R2和与其相连的原子一起形成取代或未取代的3-7元碳环或杂环，所述取代是指被1-5个R3取代；
    所述R3各自独立选自羟基、氰基、氨基、卤素、C1-6烷基、C1-6烷氧羰基、C3-C6环烷基、C1-6卤代烷基、羟基C1-6烷基、-NH(C3-6环烷基)、-NHC(＝O)(C1-6烷基)、被氰基取代的包含选自O、N、S中的一个或多个的5-7元杂环基甲基；
    RH1、RH2和RH4各自独立选自氢、卤素、氨基、氰基、羟基、-N(R4)(R5)；
    所述R4、R5各自独立选自氢、C1-6烷基、C1-6杂烷基、C3-6环烷基、C3-6杂环烷基、C1-6烷基酰基、C3-6环烷基酰基；
    RH3选自氢、未取代或取代的C1-6烷基、未取代或取代的C1-6杂烷基、-C(＝O)OC1-6烷基、-C(＝O)OC3-6环烷基、-C(＝O)C1-6烷基、-C(＝O)C3-6环烷基、未取代或取代的C3-6环烷基、未取代或取代的C2-6烯基、未取代或取代的C4-6环烯基、未取代或取代的C6-10芳基、未取代或取代的5-7元杂芳基、未取代或取代的4-7元杂环基；所述取代是指各自独立地被1-5个R6所取代；在环烷基、环烯基、芳基、杂芳基、杂环基中，一个或多个环C原子任选地被相应数目的C(＝O)基团替代，一个或多个环S或N原子任选地被氧化形成S-氧化物或N-氧化物；
    所述R6各自独立选自卤素、氰基、C1-6烷基、C1-6杂烷基、C2-6烯基、C2-6炔基、C3-6环烷基、C4-8环烯基、C6-10芳基、5-7元杂芳基、5-7元杂环基、-C(＝O)R7、OR7、-N(R7)2、-(C1-6烷基)OR7、-C(＝O)N(R7)2、-(C1-6烷基)-C(＝O)N(R7)2、-SR7、-S(＝O)R7、-S(＝O)2R7、-N(R7)C(＝O)R7、-(C1-6烷基)-N(R7)C(＝O)R7、-N(R7)S(＝O)2(R7)、-P(＝O)(R7)(R7)；其中所述C1-6烷基、C1-6杂烷基、C2-6烯基、C2-6炔基、C3-6环烷基、C4-8环烯基、C6-10芳基、5-7元杂芳基、5-7元杂环基任选被1-5个R7取代；或者
    两个R6和与其相连的原子一起形成取代或未取代的5-7元碳环或5-7元杂环，所述取代是指被1-5个R7基团取代；
    所述R7各自独立选自卤素、羟基、氰基、氨基、C1-6烷基、C2-6烯基、C2-6炔基、C3-6环烷基、C1-6杂烷基、C4-6杂环烷基、C1-6卤代烷基、羟基C1-6烷基、NH(C1-6环烷基)、-NHC(＝O)(C1-6烷基)、C(＝O)(C1-6烷基)、C(＝O)(C3-6环烷基)；
    环M2选自未取代或取代的C6-10芳基、未取代或取代的5-7元杂芳基、未取代或取代的5-7杂环基；所述取代是指各自独立地被1-5个RM所取代；
    所述RM各自独立地选自卤素、氰基、C1-6杂烷基、C2-6烯基、C2-6炔基、C3-6环烷基、3-6元杂环烷基、C4-6环烯基、C6-10芳基、5-7元杂芳基、-C(＝O)(C1-6烷基)、-C(＝O)(C3-6环烷基)、-(C1-6烷基)-R8、-OR8、-N(R8)2、-NH(C1-6烷基)-R8、-O(C1-6烷基)-R8、-C(＝O)N(R8)2、-SR8、-S(＝O)R8、-S(＝O)2R8；其中所述C1-6烷基、C3-6环烷基、3-6元杂环烷基、C4-6环烯基、4-6元杂环烯基、C6-10芳基、5-7元杂芳基任选各自独立地被1-5个R8取代；或者
    两个RM和与其相连的原子一起形成取代或未取代的3-7元碳环或杂环，所述取代是指被1-5个R8基团取代；
    所述R8各自独立选自羟基、氰基、氨基、卤素、-NH(C1-6烷基)、C1-6烷基、C1-6 杂烷基、C1-6卤代烷基、C1-6羟基烷基、C3-6环烷基、C3-6杂环烷基。
  2. 根据权利要求1所述的化合物，或其氘代化合物、药学上可接受的盐、溶剂化物、酯、酸、代谢物或前药：其中，所述化合物具有下述式(II)的结构：
    其中，
    R1选自卤素、C1-3烷基；优选选自甲基、F、Cl；
    Ra、Rb各自独立地选自氢、氘、卤素、甲基；或者Ra和Rb和与它们连接的碳原子一起形成三元碳环；
    环M1选自以下结构：
    其中A1、A2、A3、A4、环W、Z和RH1的定义如权利要求1中所述；以及
    环M2的定义如权利要求1中所述。
  3. 根据权利要求1所述的化合物，或其氘代化合物、药学上可接受的盐、溶剂化物、酯、酸、代谢物或前药，
    其中，环M1选自下述的结构：
    其中环原子上的“C,N”表示该处为CH或者N，
    R2、RH1、RH2和RH3的定义如权利要求1中所述。
  4. 根据权利要求1-3中任一项所述的化合物，或其氘代化合物、药学上可接受的盐、溶剂化物、酯、酸、代谢物或前药，其中
    环M2选自未取代或取代的苯基、未取代或取代的噻吩基、未取代或取代的吡唑基、未取代或取代的噻唑基、未取代或取代的吡啶基、未取代或取代的噁唑基、未取代或取 代的异噁唑基、未取代或取代的苯并噻吩基；所述取代是指各自独立地被1-5个RM所取代；
    所述RM各自独立地选自卤素、氰基、C1-6烷基、C1-6杂烷基、C2-6烯基、C2-6炔基、C3-6环烷基、3-6元杂环烷基、C4-6环烯基、4-6元杂环烯基、C6-10芳基、5-7元杂芳基、-C(＝O)(C1-6烷基)、-C(＝O)(C3-6环烷基)、-(C1-6烷基)-R8、-C(＝O)N(R8)2、-SR8、-S(＝O)R8、-S(＝O)2R8；其中所述C1-6烷基、C3-6环烷基、3-6元杂环烷基、C4-6环烯基、4-6元杂环烯基、C6-10芳基、5-7元杂芳基任选各自独立地被1-5个R8取代；或者
    两个RM和与其相连的原子形成取代或未取代的3-7元碳环或杂环，所述取代是指被1-5个R8基团取代；
    所述R8各自独立选自羟基、氰基、氨基、卤素、-NH(C1-6烷基)、C1-6烷基、C1-6杂烷基、C1-6卤代烷基、C1-6羟基烷基、C3-6环烷基、C3-6杂环烷基。
  5. 根据权利要求1-3中任一项所述的化合物，或其氘代化合物、药学上可接受的盐、溶剂化物、酯、酸、代谢物或前药，其中，所述化合物选自以下的化合物：








  6. 一种制备权利要求1-5任一项所述的化合物的方法，包括如下步骤：
    式(A)化合物与式(B)化合物进行偶联反应得到式(I)化合物：
    其中，环M1、环M2、X及R1的定义如相应权利要求所述；TMS为-Si(CH3)3。
  7. 根据权利要求6所述的方法，其中
    所述偶联反应在钯金属催化剂和铜金属催化剂存在下在碱存在下在溶剂中进行；
    所述钯金属催化剂包括Pd(PPh3)2Cl2、Pd(OAc)2、和Pd(PPh3)4中的一种或多种；
    所述铜金属催化剂包括CuI和/或CuCl；
    所述碱包括CsF、Cs2CO3、KF、K2CO3、NaHCO3、Na2CO3、Et3N、(iPr)2EtN、DMAP中的一种或者两种以上；
    所述的溶剂包括乙腈、1,4-二氧六环、DMF中的一种或两种以上。
  8. 一种药物组合物，其包括选自根据权利要求1-5任一项所述的化合物、其氘代化合物、药学上可接受的盐、溶剂化物、酯、酸、代谢物和前药中的一种或多种，以及药学上可接受的辅料。
  9. 根据权利要求1-5任一项所述的化合物，或其氘代化合物、药学上可接受的盐、溶剂化物、酯、酸、代谢物或前药或者根据权利要求8所述的药物组合物在制备KIT抑制剂中的用途。
  10. 根据权利要求1-5任一项所述的化合物，或其氘代化合物、其药学上可接受的盐、溶剂化物、酯、酸、代谢物或前药或者根据权利要求8所述的药物组合物在制备用于治疗、预防或改善一种或多种疾病或病症的药物中的用途，所述疾病或病症选自肿瘤、炎症、自身免疫性和神经***性疾病。
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