技术领域technical field
本发明涉及新的2-吡啶甲酰胺类化合物及其药学上可接受的盐、水合物、溶剂化物或其前药的制备方法以及含有所述化合物的药物组合物，及其在制备治疗和/ 或预防癌症的药物中的用途。The present invention relates to the preparation method of new 2-pyridinecarboxamide compounds and pharmaceutically acceptable salts, hydrates, solvates or their prodrugs and pharmaceutical compositions containing said compounds, and their use in the preparation of therapeutic and/or Or use in medicines for the prevention of cancer.
背景技术Background technique
AML(急性髓性白血病)是由骨髓造血祖细胞的恶性增殖引起的疾病，其总发病率为每10万人4.1例。细胞遗传学水平的复杂发病机制和预后特征增加了治疗急性髓性白血病的风险。AML (acute myeloid leukemia) is a disease caused by malignant proliferation of bone marrow hematopoietic progenitor cells, and its overall incidence rate is 4.1 cases per 100,000 people. Complex pathogenesis and prognostic features at the cytogenetic level increase the risk of treating acute myeloid leukemia.
FLT-3(FMS样酪氨酸激酶)是干细胞酪氨酸激酶，是III型受体酪氨酸激酶家族的成员。研究人员发现FLT-3蛋白在大多数AML患者中高表达，并极大地促进了造血干细胞的存活和增殖。FLT-3蛋白的基因位于染色体13q12，属于III型受体酪氨酸激酶(RTK)家族，与KIT、FMS及血小板源生长因子受体(PDGFR)具有高度结构同源性。FLT-3在造血祖细胞的生长和分化过程中有着确切的作用。FLT-3的表达通常限定在CD34+胞中，在这种细胞中其配体几乎无处不在，像其他RTKs一样， FLT-3受体与配体的结合导致受体在细胞膜上形成二聚体，致使自身磷酸化，激活一系列下游信号传导途径如RAS/MEK/PI3K/AKT/mTOR和STAT-5通路，它们在促进细胞周期进程，抑制细胞凋亡和促进细胞分化过程中有着多种作用。突变的FLT-3 通常表现出较高的活性，并失去了与CD34+的关联性，且能在没有配体结合的情况下，自身磷酸化激活下游信号通路。FLT-3 (FMS-like tyrosine kinase) is a stem cell tyrosine kinase, a member of the type III receptor tyrosine kinase family. The researchers found that the FLT-3 protein is highly expressed in most AML patients and greatly promotes the survival and proliferation of hematopoietic stem cells. The gene of FLT-3 protein is located on chromosome 13q12, belongs to the type III receptor tyrosine kinase (RTK) family, and has high structural homology with KIT, FMS and platelet-derived growth factor receptor (PDGFR). FLT-3 has a definite role in the growth and differentiation of hematopoietic progenitor cells. FLT-3 expression is usually restricted to CD34+ cells, where its ligand is almost ubiquitous, and like other RTKs, binding of the FLT-3 receptor to the ligand results in dimerization of the receptor at the cell membrane body, resulting in autophosphorylation, activating a series of downstream signaling pathways such as RAS/MEK/PI3K/AKT/mTOR and STAT-5 pathways, which have various roles in promoting cell cycle progression, inhibiting apoptosis and promoting cell differentiation effect. Mutant FLT-3 usually exhibits higher activity, loses its association with CD34+ , and can autophosphorylate and activate downstream signaling pathways in the absence of ligand binding.
FLT-3基因的突变是AML中最常见的遗传改变，并且作为骨髓恶性肿瘤发展的重要驱动突变而众所周知。最普遍的FLT-3突变体是内部串联重复改变(FLT3-ITD 突变)，它影响着受体的近膜区域。FLT3-ITD突变有着不同的长度和位置，大约23％ AML患者中存在这一突变。其他重要突变发生在酪氨酸激酶结构域(FLT3-TKD) 中，通常发生在激活环(AL)残基D835，该位置的天冬氨酸被酪氨酸残基取代从而稳定了活化位点与ATP结合的构象，提高了其活性。这些突变存在于所有AML患者中的约30％，并导致更差的预后。鉴于FLT-3突变AML疗效普遍较差的现状，FLT-3 受体酪氨酸激酶靶向抑制剂的研究己经成为药物研究的重点。Mutations in the FLT-3 gene are the most common genetic alterations in AML and are well known as important driver mutations in the development of myeloid malignancies. The most prevalent FLT-3 mutant is an altered internal tandem duplication (FLT3-ITD mutation), which affects the juxtamembrane region of the receptor. FLT3-ITD mutations vary in length and location and are present in approximately 23% of AML patients. Other important mutations occur in the tyrosine kinase domain (FLT3-TKD), usually at activation loop (AL) residue D835, where the aspartic acid is replaced by a tyrosine residue that stabilizes the activation site A conformation that binds to ATP, enhancing its activity. These mutations are present in approximately 30% of all AML patients and lead to a worse prognosis. In view of the generally poor curative effect of FLT-3 mutant AML, research on FLT-3 receptor tyrosine kinase-targeted inhibitors has become the focus of drug research.
为了研制出新型高效的FLT-3受体酪氨酸激酶抑制剂，本发明人对吡啶类FLT-3抑制剂化合物进行了进一步研究，对多个结构位点进行修饰和改造，合成了一系列结构新颖的2-吡啶甲酰胺类衍生物。体外抗肿瘤活性筛选试验表明，该类化合物具有抗肿瘤活性。In order to develop new and efficient FLT-3 receptor tyrosine kinase inhibitors, the inventors further studied pyridine FLT-3 inhibitor compounds, modified and transformed multiple structural sites, and synthesized a series of Novel 2-pyridinecarboxamide derivatives. The antitumor activity screening test in vitro shows that the compound has antitumor activity.
发明内容Contents of the invention
本发明涉及通式Ⅰ所示的2-吡啶甲酰胺类化合物及其药学上可接受的盐、水合物、溶剂化物或前药，其中The present invention relates to 2-pyridinecarboxamide compounds represented by general formula I and pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof, wherein
R1为甲基、二甲基、乙基、正丙基、异丙基、4-甲基吗啉基、氰基；R is methyl, dimethyl, ethyl,n -propyl, isopropyl, 4-methylmorpholinyl, cyano;
R2选自1～4个相同或不同的氢、氟；R2 is selected from 1 to 4 identical or different hydrogen and fluorine;
X为O、S；X is O, S;
Y为-Ar1-Ar2；Y is -Ar1 -Ar2 ;
Ar1为6元杂芳基，此杂芳基含有2个N的杂原子和1个O杂原子；Ar1 is a 6-membered heteroaryl group, which contains 2 N heteroatoms and 1 O heteroatom;
Ar2为(C6-C10)杂芳基，Ar2除与Ar1相并外，还被有R3取代的芳基取代；Ar2 is a (C6 -C10 )heteroaryl group, and Ar2 is substituted with an aryl group substituted by R3 in addition to being combined with Ar1 ;
R3为1-2个选自氢、卤素、(C1-C4)烷基、(C1-C4)烷氧基、任选被卤代的(C1-C4) 烷基或(C1-C4)烷氧基、单或二(C1-C6烷基)取代的氨基、(C1-C6)烷基酰氨基、游离的、成盐的、酯化的和酰胺化的羧基、(C1-C6)烷基亚磺酰基、磺酰基、(C1-C6)烷基酰基、氨基甲酰基、被单或二(C1-C6烷基)取代的氨基甲酰基、(C1-C3)亚烷基二氧基的取代基。R3 is 1-2 selected from hydrogen, halogen, (C1 -C4 ) alkyl, (C1 -C4 ) alkoxy, optionally halogenated (C1 -C4 ) alkyl or (C1 -C4 )alkoxy, mono or di(C1 -C6 alkyl) substituted amino, (C1 -C6 )alkylamido, free, salified, esterified and Amidated carboxyl, (C1 -C6 )alkylsulfinyl, sulfonyl, (C1 -C6 )alkylacyl, carbamoyl, mono- or di-(C1 -C6 alkyl) substituted Substituents of carbamoyl, (C1 -C3 )alkylenedioxy.
本发明优选还涉及定义如下的通式Ⅰ化合物，或其消旋体或旋光异构体，或其药学上可接受的盐和/或水合物，其中The present invention preferably also relates to a compound of general formula I as defined below, or a racemate or an optical isomer thereof, or a pharmaceutically acceptable salt and/or hydrate thereof, wherein
R1为甲基、乙基、正丙基、4-甲基吗啉基；R is methyl, ethyl,n -propyl, 4-methylmorpholinyl;
R2为H，其取代位为苯环上与X所连碳原子的邻位；R2 is H, and its substituent position is the ortho position of the carbon atom connected to X on the benzene ring;
X为O；X is O;
Y为-Ar1-Ar2；Y is -Ar1 -Ar2 ;
Ar1为6元杂芳基，此杂芳基含有2个N的杂原子和1个O杂原子；Ar1 is a 6-membered heteroaryl group, which contains 2 N heteroatoms and 1 O heteroatom;
Ar2为(C6-C10)杂芳基，Ar2除与Ar1相并外，还被有R3取代的芳基取代；Ar2 is a (C6 -C10 )heteroaryl group, and Ar2 is substituted with an aryl group substituted by R3 in addition to being combined with Ar1 ;
R3为1-2个选自氢、卤素、(C1-C4)烷基、(C1-C4)烷氧基、任选被卤代的(C1-C4) 烷基或(C1-C4)烷氧基、单或二(C1-C6烷基)取代的氨基、(C1-C6)烷基酰氨基、游离的、成盐的、酯化的和酰胺化的羧基、(C1-C6)烷基亚磺酰基、磺酰基、(C1-C6)烷基酰基、氨基甲酰基、被单或二(C1-C6烷基)取代的氨基甲酰基、(C1-C3)亚烷基二氧基的取代基。R3 is 1-2 selected from hydrogen, halogen, (C1 -C4 ) alkyl, (C1 -C4 ) alkoxy, optionally halogenated (C1 -C4 ) alkyl or (C1 -C4 )alkoxy, mono or di(C1 -C6 alkyl) substituted amino, (C1 -C6 )alkylamido, free, salified, esterified and Amidated carboxyl, (C1 -C6 )alkylsulfinyl, sulfonyl, (C1 -C6 )alkylacyl, carbamoyl, mono- or di-(C1 -C6 alkyl) substituted Substituents of carbamoyl, (C1 -C3 )alkylenedioxy.
本发明优选还涉及定义如下的通式Ⅰ化合物，或其消旋体或旋光异构体，或其药学上可接受的盐和/或水合物，其中The present invention preferably also relates to a compound of general formula I as defined below, or a racemate or an optical isomer thereof, or a pharmaceutically acceptable salt and/or hydrate thereof, wherein
R1为甲基、乙基、正丙基、4-甲基吗啉基；R is methyl, ethyl,n -propyl, 4-methylmorpholinyl;
R2为H，其取代位为苯环上与X所连碳原子的邻位；R2 is H, and its substituent position is the ortho position of the carbon atom connected to X on the benzene ring;
X为O；X is O;
Y为Y is
R3为1-2个选自氢、卤素、(C1-C4)烷基、(C1-C4)烷氧基。R3 is 1-2 selected from hydrogen, halogen, (C1 -C4 ) alkyl, (C1 -C4 ) alkoxy.
本发明非常特别优选的下列通式Ⅰ衍生物，包括其消旋体或旋光异构体，及其药学上可接受的盐和/或水合物，但这些化合物并不意味着对本发明的任何限制：Very particularly preferred derivatives of the following general formula I of the present invention, including their racemates or optical isomers, and pharmaceutically acceptable salts and/or hydrates thereof, but these compounds do not imply any limitation to the present invention :
(1)4-甲基-N-(4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代-1-苯基-1,6- 二氢哒嗪-3-甲酰胺(1) 4-methyl-N-(4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-6-oxo-1-phenyl-1,6 - Dihydropyridazine-3-carboxamide
(2)4-甲基-N-(4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代-1-(对甲苯基)-1,6-二氢哒嗪-3-甲酰胺(2) 4-methyl-N-(4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-6-oxo-1-(p-tolyl)- 1,6-Dihydropyridazine-3-carboxamide
(3)1-(2-氯苯基)-4-甲基-N-(4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代 -1,6-二氢哒嗪-3-甲酰胺(3) 1-(2-chlorophenyl)-4-methyl-N-(4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-6-oxo Substituent-1,6-dihydropyridazine-3-carboxamide
(4)1-(3-氟苯基)-4-甲基-N-(4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代 -1,6-二氢哒嗪-3-甲酰胺(4) 1-(3-fluorophenyl)-4-methyl-N-(4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-6-oxo Substituent-1,6-dihydropyridazine-3-carboxamide
(5)4-甲基-N-(4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代-1-(3-(三氟甲基)苯基)-1,6-二氢哒嗪-3-甲酰胺(5) 4-methyl-N-(4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-6-oxo-1-(3-(trifluoro Methyl)phenyl)-1,6-dihydropyridazine-3-carboxamide
(6)4-甲基-N-(4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代-1-(3-(三氟甲氧基)苯基)-1,6-二氢哒嗪-3-甲酰胺(6) 4-methyl-N-(4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-6-oxo-1-(3-(trifluoro Methoxy)phenyl)-1,6-dihydropyridazine-3-carboxamide
(7)1-(4-溴-2-氟苯基)-4-甲基-N-(4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代-1,6-二氢哒嗪-3-甲酰胺(7) 1-(4-bromo-2-fluorophenyl)-4-methyl-N-(4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl) -6-oxo-1,6-dihydropyridazine-3-carboxamide
(8)N-(3-氟-4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-4-甲基-6-氧代-1-苯基 -1,6-二氢哒嗪-3-甲酰胺(8) N-(3-fluoro-4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-4-methyl-6-oxo-1-phenyl -1,6-dihydropyridazine-3-carboxamide
(9)N-(3-氟-4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-4-甲基-6-氧代-1-(对甲苯基)-1,6-二氢哒嗪-3-甲酰胺(9) N-(3-fluoro-4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-4-methyl-6-oxo-1-(p Tolyl)-1,6-dihydropyridazine-3-carboxamide
(10)1-(2-氯苯基)-N-(3-氟-4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-4-甲基 -6-氧代-1,6-二氢哒嗪-3-甲酰胺(10) 1-(2-chlorophenyl)-N-(3-fluoro-4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-4-methyl -6-oxo-1,6-dihydropyridazine-3-carboxamide
(11)N-(3-氟-4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-1-(3-氟苯基)-4-甲基 -6-氧代-1,6-二氢哒嗪-3-甲酰胺(11) N-(3-fluoro-4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-1-(3-fluorophenyl)-4-methyl -6-oxo-1,6-dihydropyridazine-3-carboxamide
(12)N-(3-氟-4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-4-甲基-6-氧代-1-(3-(三氟甲基)苯基)-1,6-二氢哒嗪-3-甲酰胺(12) N-(3-fluoro-4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-4-methyl-6-oxo-1-(3 -(Trifluoromethyl)phenyl)-1,6-dihydropyridazine-3-carboxamide
(13)N-(3-氟-4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-4-甲基-6-氧代-1-(3-(三氟甲氧基)苯基)-1,6-二氢哒嗪-3-甲酰胺(13) N-(3-fluoro-4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-4-methyl-6-oxo-1-(3 -(Trifluoromethoxy)phenyl)-1,6-dihydropyridazine-3-carboxamide
(14)1-(4-溴-2-氟苯基)-N-(3-氟-4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-4- 甲基-6-氧代-1,6-二氢哒嗪-3-甲酰胺(14) 1-(4-bromo-2-fluorophenyl)-N-(3-fluoro-4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)- 4-Methyl-6-oxo-1,6-dihydropyridazine-3-carboxamide
(15)N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)苯基)-4-甲基-6-氧代-1-(邻甲苯基)-1,6-二氢哒嗪-3-甲酰胺(15) N-(4-((2-(ethylcarbamoyl)pyridin-4-yl)oxy)phenyl)-4-methyl-6-oxo-1-(o-tolyl)- 1,6-Dihydropyridazine-3-carboxamide
(16)N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)苯基)-4-甲基-6-氧代-1-(对-甲苯基)-1,6-二氢哒嗪-3-甲酰胺(16) N-(4-((2-(ethylcarbamoyl)pyridin-4-yl)oxy)phenyl)-4-methyl-6-oxo-1-(p-tolyl) -1,6-dihydropyridazine-3-carboxamide
(17)N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)苯基)-1-(3-氟苯基)-4-甲基-6-氧代-1,6-二氢哒嗪-3-甲酰胺(17) N-(4-((2-(ethylcarbamoyl)pyridin-4-yl)oxy)phenyl)-1-(3-fluorophenyl)-4-methyl-6-oxo Substituent-1,6-dihydropyridazine-3-carboxamide
(18)1-(4-溴-2-氟苯基)-N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)苯基)-4-甲基 -6-氧代-1,6-二氢哒嗪-3-甲酰胺(18) 1-(4-bromo-2-fluorophenyl)-N-(4-((2-(ethylcarbamoyl)pyridin-4-yl)oxy)phenyl)-4-methyl -6-oxo-1,6-dihydropyridazine-3-carboxamide
(19)N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)苯基)-4-甲基-6-氧代-1-(3-(三氟甲氧基)苯基)-1,6-二氢哒嗪-3-甲酰胺(19) N-(4-((2-(ethylcarbamoyl)pyridin-4-yl)oxy)phenyl)-4-methyl-6-oxo-1-(3-(trifluoro Methoxy)phenyl)-1,6-dihydropyridazine-3-carboxamide
(20)N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)-3-氟苯基)-4-甲基-6-氧代-1-(邻甲苯基)-1,6-二氢哒嗪-3-甲酰胺(20) N-(4-((2-(ethylcarbamoyl)pyridin-4-yl)oxy)-3-fluorophenyl)-4-methyl-6-oxo-1-(o Tolyl)-1,6-dihydropyridazine-3-carboxamide
(21)N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)-3-氟苯基)-4-甲基-6-氧代-1-(对甲苯基)-1,6-二氢哒嗪-3-甲酰胺(21) N-(4-((2-(ethylcarbamoyl)pyridin-4-yl)oxy)-3-fluorophenyl)-4-methyl-6-oxo-1-(p Tolyl)-1,6-dihydropyridazine-3-carboxamide
(22)N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)-3-氟苯基)-1-(3-氟苯基)-4-甲基 -6-氧代-1,6-二氢哒嗪-3-甲酰胺(22) N-(4-((2-(ethylcarbamoyl)pyridin-4-yl)oxy)-3-fluorophenyl)-1-(3-fluorophenyl)-4-methyl -6-oxo-1,6-dihydropyridazine-3-carboxamide
(23)1-(4-溴-2-氟苯基)-N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)-3-氟苯基)-4- 甲基-6-氧代-1,6-二氢哒嗪-3-甲酰胺(23) 1-(4-bromo-2-fluorophenyl)-N-(4-((2-(ethylcarbamoyl)pyridin-4-yl)oxy)-3-fluorophenyl)- 4-Methyl-6-oxo-1,6-dihydropyridazine-3-carboxamide
(24)N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)-3-氟苯基)-4-甲基-6-氧代-1-(3-(三氟甲氧基)苯基)-1,6-二氢哒嗪-3-甲酰(24) N-(4-((2-(ethylcarbamoyl)pyridin-4-yl)oxy)-3-fluorophenyl)-4-methyl-6-oxo-1-(3 -(Trifluoromethoxy)phenyl)-1,6-dihydropyridazine-3-formyl
(25)4-甲基-6-氧代-N-(4-((2-(丙基氨基)吡啶-4-基)氧基)苯基)-1-(邻甲苯基)-1,6- 二氢哒嗪-3-甲酰胺(25) 4-methyl-6-oxo-N-(4-((2-(propylamino)pyridin-4-yl)oxy)phenyl)-1-(o-tolyl)-1, 6-Dihydropyridazine-3-carboxamide
(26)4-甲基-6-氧代-N-(4-((2-(丙基氨基)吡啶-4-基)氧基)苯基)-1-(对甲苯基)-1,6- 二氢哒嗪-3-甲酰胺(26) 4-methyl-6-oxo-N-(4-((2-(propylamino)pyridin-4-yl)oxy)phenyl)-1-(p-tolyl)-1, 6-Dihydropyridazine-3-carboxamide
(27)1-(3-氟苯基)-4-甲基-6-氧代-N-(4-((2-(丙基氨基)吡啶-4-基)氧基)苯基)-1,6- 二氢哒嗪-3-甲酰胺(27) 1-(3-fluorophenyl)-4-methyl-6-oxo-N-(4-((2-(propylamino)pyridin-4-yl)oxy)phenyl)- 1,6-Dihydropyridazine-3-carboxamide
(28)1-(4-溴-2-氟苯基)-4-甲基-6-氧代-N-(4-((2-(丙基氨基)吡啶-4-基)氧基)苯基)-1,6-二氢哒嗪-3-甲酰胺(28) 1-(4-bromo-2-fluorophenyl)-4-methyl-6-oxo-N-(4-((2-(propylamino)pyridin-4-yl)oxy) Phenyl)-1,6-dihydropyridazine-3-carboxamide
(29)4-甲基-6-氧代-N-(4-((2-(丙基氨基)吡啶-4-基)氧基)苯基)-1-(3-(三氟甲氧基) 苯基)-1,6-二氢哒嗪-3-甲酰胺(29) 4-methyl-6-oxo-N-(4-((2-(propylamino)pyridin-4-yl)oxy)phenyl)-1-(3-(trifluoromethoxy base) phenyl)-1,6-dihydropyridazine-3-carboxamide
(30)4-甲基-6-氧代-N-(4-((2-(丙基氨基)吡啶-4-基)氧基)苯基)-1-(3-(三氟甲基)苯基)-1,6-二氢哒嗪-3-甲酰胺(30) 4-methyl-6-oxo-N-(4-((2-(propylamino)pyridin-4-yl)oxy)phenyl)-1-(3-(trifluoromethyl )phenyl)-1,6-dihydropyridazine-3-carboxamide
(31)4-甲基-N-(4-((2-((3-吗啉代)氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代-1-(对甲苯基)-1,6-二氢哒嗪-3-甲酰胺(31) 4-methyl-N-(4-((2-((3-morpholino)carbamoyl)pyridin-4-yl)oxy)phenyl)-6-oxo-1-( p-tolyl)-1,6-dihydropyridazine-3-carboxamide
(32)1-(3-氟苯基)-4-甲基-N-(4-((2-((3-吗啉代)氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代-1,6-二氢哒嗪-3-甲酰胺(32) 1-(3-fluorophenyl)-4-methyl-N-(4-((2-((3-morpholino)carbamoyl)pyridin-4-yl)oxy)phenyl )-6-oxo-1,6-dihydropyridazine-3-carboxamide
(33)1-(4-溴-2-氟苯基)-4-甲基-N-(4-((2-((3-吗啉代)氨基甲酰基)吡啶-4-基)氧基) 苯基)-6-氧代-1,6-二氢哒嗪-3-甲酰胺(33) 1-(4-bromo-2-fluorophenyl)-4-methyl-N-(4-((2-((3-morpholino)carbamoyl)pyridin-4-yl)oxy base) phenyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide
(34)4-甲基-N-(4-((2-((3-吗啉代)氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代-1-(3-(三氟甲基)苯基)-1,6-二氢哒嗪-3-甲酰胺(34) 4-methyl-N-(4-((2-((3-morpholino)carbamoyl)pyridin-4-yl)oxy)phenyl)-6-oxo-1-( 3-(Trifluoromethyl)phenyl)-1,6-dihydropyridazine-3-carboxamide
按照本发明所属领域的一些通常方法，本发明的通式Ⅰ的2-吡啶甲酰胺类衍生物可以与酸生成它的药学上可接受的盐。酸可以包括无机酸或有机酸，与下列酸形成的盐是特别优选的：盐酸、氢溴酸、硫酸、磷酸、甲磺酸、乙磺酸、甲苯磺酸、苯磺酸、萘二磺酸、乙酸、丙酸、乳酸、三氟乙酸、马来酸、柠檬酸、富马酸、洒石酸、苯磺酸、苯甲酸或对甲苯磺酸等。此外，本发明还包括本发明化合物的前药。依据本发明，前药是通式Ⅰ化合物的衍生物，它们自身可能具有较弱的活性或甚至没有活性，但是在给药后，在生理条件下(例如通过代谢、溶剂分解或另外的方式) 被转化成相应的生物活性形式。According to some common methods in the field of the present invention, the 2-pyridinecarboxamide derivatives of the general formula I of the present invention can form its pharmaceutically acceptable salts with acids. The acids may include inorganic or organic acids, the salts with the following acids being particularly preferred: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, naphthalene disulfonic acid , acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, tartaric acid, benzenesulfonic acid, benzoic acid or p-toluenesulfonic acid, etc. Furthermore, the present invention also includes prodrugs of the compounds of the present invention. According to the invention, prodrugs are derivatives of the compounds of general formula I, which themselves may have weak activity or even no activity, but after administration, under physiological conditions (for example by metabolism, solvolysis or otherwise) is converted into the corresponding biologically active form.
除非另外指出，本发明所用的术语“卤代”是指氟代、氯代、溴代或碘代；“烷基”是指直链或支链的烷基；“环烷基”是指取代或未取代的环烷基；“烯基”是指直链或支链的烯基；“炔基”是指直链或支链的炔基；“芳基”是指除去芳烃中的一个氢原子而得的有机基团，如苯基、萘基；5-10元杂芳基包括含有一个或多个选自N、O和S的杂原子，其中每个杂芳基的环状体系可以是单环或多环的，环状体系是芳香性的，一共含有5-10个原子，可以举出例如咪唑基、吡啶基、嘧啶基、吡唑基、(1,2,3)- 和(1,2,4)-三唑基、吡嗪基、四唑基、呋喃基、噻吩基、异噁唑基、噁唑基、吡唑基、吡咯基、噻唑基、苯并噻吩基、苯并呋喃基、苯并咪唑基、苯并噻唑基、吲哚基、喹啉基等；5-10元杂环基包括含有一个或多个选自N、O和S的杂原子，其中每个杂芳基的环状体系可以是单环或多环的，但是是非芳香性的，环状体系一共含有5-10 个原子，可以任选包括1或2个碳碳双键或碳碳叁键，可以举出例如吡咯烷基、吗啉基、哌嗪基、哌啶基、噻唑啉基等。Unless otherwise indicated, the term "halo" used in the present invention refers to fluoro, chloro, bromo or iodo; "alkyl" refers to straight or branched chain alkyl; "cycloalkyl" refers to substituted or unsubstituted cycloalkyl; "alkenyl" refers to straight-chain or branched alkenyl; "alkynyl" refers to straight-chain or branched alkynyl; "aryl" refers to the removal of one hydrogen in aromatic Atom-derived organic groups, such as phenyl, naphthyl; 5-10 membered heteroaryls include one or more heteroatoms selected from N, O and S, wherein the ring system of each heteroaryl can be It is monocyclic or polycyclic, and the ring system is aromatic, containing 5-10 atoms in total, such as imidazolyl, pyridyl, pyrimidyl, pyrazolyl, (1,2,3)- and (1,2,4)-Triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, oxazolyl, pyrazolyl, pyrrolyl, thiazolyl, benzothienyl, Benzofuryl, benzimidazolyl, benzothiazolyl, indolyl, quinolinyl, etc.; 5-10 membered heterocyclic groups include one or more heteroatoms selected from N, O and S, wherein each The ring system of each heteroaryl group can be monocyclic or polycyclic, but is non-aromatic, and the ring system contains 5-10 atoms in total, and can optionally include 1 or 2 carbon-carbon double bonds or carbon-carbon triple bonds. Bonds include, for example, pyrrolidinyl, morpholinyl, piperazinyl, piperidinyl, thiazolinyl and the like.
本发明还涉及通式Ⅰ的化合物具有强的抑制FLT-3的作用，并且还涉及该类化合物及其药学上可接受的盐、水合物在制备治疗由于FLT-3激酶异常高表达所引起疾病的药物中的用途，特别是在制备治疗和/或预防癌症的药物中的用途。The present invention also relates to the compound of general formula I having a strong FLT-3 inhibitory effect, and also relates to the preparation and treatment of diseases caused by abnormally high expression of FLT-3 kinase by such compounds and their pharmaceutically acceptable salts and hydrates. The application in the medicament of the present invention, especially the application in the preparation of the medicament for treating and/or preventing cancer.
下面合成路线1-3描述了本发明的通式Ⅰ化合物的制备，所有的原料都是通过这些反应式中描述的方法、通过有机化学领域普通技术人员熟知的方法制备的或者可商购。本发明的全部最终化合物都是通过这些反应式中描述的方法或通过与其类似的方法制备的，这些方法是有机化学领域普通技术人员熟知的。这些反应式中应用的全部可变因数如下文的定义或如权利要求中的定义。The following synthetic schemes 1-3 describe the preparation of the compounds of general formula I of the present invention. All starting materials are prepared by the methods described in these schemes, by methods well known to those skilled in the art of organic chemistry or are commercially available. All final compounds of the present invention are prepared by the methods described in these schemes or by methods analogous thereto, which methods are well known to those of ordinary skill in the art of organic chemistry. All variables used in these equations are as defined below or as defined in the claims.
按照本发明的式Ⅰ化合物，Y为R3如发明内容部分所定义，均可按在路线1的方法由中间体A和中间体B通过取代反应制得。According to the compound of formula I of the present invention, Y is R3 , as defined in the Summary of the Invention, can be prepared from intermediate A and intermediate B through substitution reactions according to the method in Scheme 1.
按照本发明的式Ⅰ化合物，中间体A的制备方法如路线2，其他取代基如权利要求中所定义。According to the compound of formula I of the present invention, the preparation method of intermediate A is as shown in Scheme 2, and other substituents are as defined in the claims.
按照本发明的式Ⅰ化合物，Y为中间体B的制备方法如路线3，其他取代基如权利要求中所定义。According to the compound of formula I of the present invention, Y is The preparation method of intermediate B is as in route 3, and other substituents are as defined in the claims.
以上三条路线中所有中间体的取代基R1、R2、R3如权利要求中所定义。The substituents R1 , R2 and R3 of all intermediates in the above three routes are as defined in the claims.
具体实施方式：Detailed ways:
实施例旨在阐述而不是限制本发明的范围。化合物的核磁共振氢谱用BrukerARX-300测定，质谱用Agilent1100LC/MSD测定；所用试剂均为分析纯或化学纯。The examples are intended to illustrate, not limit, the scope of the invention. The proton nuclear magnetic resonance spectrum of the compound was determined by BrukerARX-300, and the mass spectrum was determined by Agilent1100LC/MSD; the reagents used were analytically pure or chemically pure.
按照制备通法1，分别制得实施例1-34化合物(见表一)。According to Preparation General Method 1, the compounds of Examples 1-34 were respectively prepared (see Table 1).
表一Table I
实施例1Example 1
4-甲基-N-(4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代-1-苯基-1,6-二氢哒嗪-3-甲酰胺4-Methyl-N-(4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-6-oxo-1-phenyl-1,6-dihydro Pyridazine-3-carboxamide
ESI-MSm/z:455.16.ESI-MS m/z: 455.16.
实施例2Example 2
4-甲基-N-(4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代-1-(对甲苯基)-1,6-二氢哒嗪-3-甲酰胺4-Methyl-N-(4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-6-oxo-1-(p-tolyl)-1,6 -Dihydropyridazine-3-carboxamide
ESI-MSm/z:469.18.ESI-MS m/z: 469.18.
实施例3Example 3
1-(2-氯苯基)-4-甲基-N-(4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代 -1,6-二氢哒嗪-3-甲酰胺1-(2-Chlorophenyl)-4-methyl-N-(4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-6-oxo-1 ,6-Dihydropyridazine-3-carboxamide
ESI-MSm/z:489.12.ESI-MS m/z: 489.12.
实施例4Example 4
1-(3-氟苯基)-4-甲基-N-(4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代 -1,6-二氢哒嗪-3-甲酰胺1-(3-fluorophenyl)-4-methyl-N-(4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-6-oxo-1 ,6-Dihydropyridazine-3-carboxamide
ESI-MSm/z:473.15.ESI-MS m/z: 473.15.
实施例5Example 5
4-甲基-N-(4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代-1-(3-(三氟甲基) 苯基)-1,6-二氢哒嗪-3-甲酰胺4-Methyl-N-(4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-6-oxo-1-(3-(trifluoromethyl) Phenyl)-1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:523.15.ESI-MS m/z: 523.15.
实施例6Example 6
4-甲基-N-(4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代-1-(3-(三氟甲氧基)苯基)-1,6-二氢哒嗪-3-甲酰胺4-methyl-N-(4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-6-oxo-1-(3-(trifluoromethoxy )phenyl)-1,6-dihydropyridazine-3-carboxamide
步骤一：4-氯吡啶酰氯的制备(2)Step 1: Preparation of 4-chloropyridine chloride (2)
在150mL圆底烧瓶中先后加入吡啶甲酸(10g,0.081mol),NaBr(0.1g,0.001mol)和50mL氯化亚砜进行超声溶解，85℃下回流24h。将反应液冷却至室温后浓缩，加入适量甲苯待用，得到淡黄色液体为目标产物，产率约为97.5％。Picolinic acid (10 g, 0.081 mol), NaBr (0.1 g, 0.001 mol) and 50 mL of thionyl chloride were successively added into a 150 mL round bottom flask for ultrasonic dissolution, and refluxed at 85° C. for 24 h. The reaction solution was cooled to room temperature and then concentrated, and an appropriate amount of toluene was added for later use to obtain a light yellow liquid as the target product with a yield of about 97.5%.
步骤二：4-氯吡啶甲酸乙酯的制备(3)Step 2: Preparation of ethyl 4-chloropicolinate (3)
在100mL圆底烧瓶中先后加入30mL二氯甲烷、乙醇(16.43g,0.17mol)和三乙胺(17.17g,0.17mol)，在冰浴条件下搅拌20min，逐滴加入上述关键中间体2(16.43 g,0.11mol)，冰浴条件下搅拌0.5h。将反应液加入到适量饱和食盐水中，再加入适量NaOH水溶液调节pH至9-10时，用二氯甲烷进行萃取，合并有机层，无水硫酸钠干燥，浓缩，加入少量石油醚，有大量淡黄色固体析出，抽滤得到目标产物，产率为80％。In a 100mL round bottom flask, 30mL of dichloromethane, ethanol (16.43g, 0.17mol) and triethylamine (17.17g, 0.17mol) were successively added, stirred for 20min under ice bath conditions, and the above key intermediate 2 was added dropwise ( 16.43 g, 0.11mol), stirred in ice bath for 0.5h. Add the reaction solution to an appropriate amount of saturated saline, then add an appropriate amount of NaOH aqueous solution to adjust the pH to 9-10, extract with dichloromethane, combine the organic layers, dry over anhydrous sodium sulfate, concentrate, add a small amount of petroleum ether, there is a large amount of light A yellow solid was precipitated, and the target product was obtained by suction filtration with a yield of 80%.
步骤三：4-(4-硝基苯氧基)吡啶甲酸乙酯的制备(4)Step 3: Preparation of ethyl 4-(4-nitrophenoxy)picolinate (4)
在150mL圆底烧瓶中先后加上述关键中间体3(13.22g,0.071mol)和对硝基苯酚(3.61g,0.026mol)用40mL氯苯超声溶解后升温至130℃冷凝回流4h。待反应液冷却至室温后将反应液体倒入100mL冷的石油醚溶剂中常温搅拌10min，倒去上清液留黏稠液体。将黏稠液体用适量二氯甲烷溶解后再加入适量NaOH水溶液调节 pH至9-10时，用二氯甲烷进行萃取，合并有机层，无水硫酸钠干燥，浓缩，加入少量石油醚，有大量淡黄色固体析出，过滤得到目标产物，产率为80％。The key intermediate 3 (13.22g, 0.071mol) and p-nitrophenol (3.61g, 0.026mol) were successively added to a 150mL round-bottomed flask, dissolved in 40mL of chlorobenzene by ultrasonication, and then heated to 130°C to condense and reflux for 4h. After the reaction liquid was cooled to room temperature, the reaction liquid was poured into 100 mL of cold petroleum ether solvent and stirred at room temperature for 10 min, and the supernatant was poured off to leave a viscous liquid. Dissolve the viscous liquid with an appropriate amount of dichloromethane, then add an appropriate amount of NaOH aqueous solution to adjust the pH to 9-10, extract with dichloromethane, combine the organic layers, dry over anhydrous sodium sulfate, concentrate, add a small amount of petroleum ether, there is a large amount of light A yellow solid was precipitated, and the target product was obtained by filtration with a yield of 80%.
步骤四：4-(4-硝基苯氧基)吡啶甲酸(5)Step 4: 4-(4-nitrophenoxy)picolinic acid (5)
在100mL圆底烧瓶中加入中间体4(10g,0.035mol)溶于20mL乙醇中，室温下逐滴加入10％NaOH(22.8g,0.057mol)，室温搅拌1h。将反应液旋干大部分以后加入250mL饱和食盐水，室温搅拌，逐滴加入浓盐酸调节pH值至2-3后，有淡黄色固体析出，持续搅拌3h。抽滤，留固体，烘干，得淡黄色固体7.51g，产率83.2％。Intermediate 4 (10 g, 0.035 mol) was dissolved in 20 mL of ethanol in a 100 mL round bottom flask, 10% NaOH (22.8 g, 0.057 mol) was added dropwise at room temperature, and stirred at room temperature for 1 h. After most of the reaction solution was spin-dried, 250 mL of saturated saline was added, stirred at room temperature, concentrated hydrochloric acid was added dropwise to adjust the pH value to 2-3, a light yellow solid precipitated out, and the stirring was continued for 3 h. After suction filtration, the solid was retained and dried to obtain 7.51 g of a light yellow solid with a yield of 83.2%.
步骤五：4-(4-硝基苯氧基)吡啶甲酰氯的制备(6)Step 5: Preparation of 4-(4-nitrophenoxy)picoligoyl chloride (6)
在50mL圆底烧瓶中加入上述中间体5(5g,0.019mol)用15mL氯化亚砜进行超声溶解后升温至85℃搅拌10min后滴加DMF(0.073g,0.001mol)，持续搅拌40 min。将反应液进行浓缩，加入适量甲苯待用。得到淡黄色液体为目标产物，产率约为98.0％。The above-mentioned intermediate 5 (5 g, 0.019 mol) was added into a 50 mL round-bottomed flask, dissolved in 15 mL of thionyl chloride by ultrasonic, heated to 85 °C and stirred for 10 min, then DMF (0.073 g, 0.001 mol) was added dropwise, and stirred continuously for 40 min. The reaction solution was concentrated, and an appropriate amount of toluene was added for use. The target product was obtained as a pale yellow liquid with a yield of about 98.0%.
步骤六：4-氯-N-甲基吡啶酰胺的制备(7)Step 6: Preparation of 4-chloro-N-methylpicolinamide (7)
在100mL烧杯中先后加入30mL二氯甲烷、30％甲氨水溶液(5.66g,0.0377mol) 和三乙胺(3.81g,0.0377mol)，冰浴条件下搅拌20min后逐滴加入上述关键中间体6 (5.24g,0.0188mol)，冰浴条件持续搅拌0.5h。将反应液加入到适量饱和食盐水中，再加入适量NaOH水溶液调节pH至9-10，用二氯甲烷进行萃取，合并有机层，无水硫酸钠干燥，浓缩，加入少量石油醚，有大量黄褐色固体析出，抽滤得到目标产物7，产率为85％。Add 30mL of dichloromethane, 30% methylammonia aqueous solution (5.66g, 0.0377mol) and triethylamine (3.81g, 0.0377mol) into a 100mL beaker successively, stir for 20min under ice-bath conditions, then add the key intermediate 6 dropwise (5.24g, 0.0188mol), stirring continuously in ice bath for 0.5h. Add the reaction solution to an appropriate amount of saturated saline, then add an appropriate amount of NaOH aqueous solution to adjust the pH to 9-10, extract with dichloromethane, combine the organic layers, dry over anhydrous sodium sulfate, concentrate, add a small amount of petroleum ether, a large amount of yellowish brown Solids were precipitated, and the target product 7 was obtained by suction filtration with a yield of 85%.
步骤七：4-氯-N-甲基吡啶酰胺的制备(A)Step 7: Preparation of 4-chloro-N-methylpicolinamide (A)
在50mL圆底烧瓶中先后加入上述关键中间体7(4.37g,0.016mol)、活性炭(1.92g,0.16mol)和FeCl3·6H2O(0.43g,0.0016mol)用20mL乙醇超声溶解，90℃冷凝回流0.5h后逐滴加入80％水合肼(8.0g,0.128mol)，持续搅伴3-4h。将溶液趁热抽滤，留滤液，旋干，加入适量异丙醇和石油醚有褐色固体析出，抽滤得到目标产物8，产率为70％。In a 50mL round bottom flask, the above-mentioned key intermediate 7 (4.37g, 0.016mol), activated carbon (1.92g, 0.16mol) and FeCl3 6H2 O (0.43g, 0.0016mol) were successively added and dissolved in 20mL of ethanol by ultrasonic, 90 After reflux at ℃ for 0.5 h, 80% hydrazine hydrate (8.0 g, 0.128 mol) was added dropwise, and the stirring was continued for 3-4 h. The solution was suction-filtered while it was hot, the filtrate was retained, spin-dried, an appropriate amount of isopropanol and petroleum ether were added, and a brown solid was precipitated, and the target product 8 was obtained by suction filtration with a yield of 70%.
步骤八：(E)-2-(2-(3-甲氧基苯基)亚肼基)-3-氧代丁酸乙酯(9)Step 8: (E)-2-(2-(3-methoxyphenyl)hydrazono)-3-oxobutanoic acid ethyl ester (9)
在250mL的三口瓶(A瓶)中加入10mL水、40mL乙醇和无水乙酸钠超声至溶解，加入乙酰乙酸乙酯(21.06g,0.162mol)，室温搅拌2h备用。在另一250mL(B 瓶)三口瓶中加入3-甲氧基苯胺(10g,0.081mol)和10mL水混匀，室温搅拌，逐滴加浓盐酸调节pH至2-3，滴毕后用冰盐浴降温至-5℃时滴加50％亚硝酸钠水溶液20 mL，保持溶液温度低于0℃，搅拌30min。若溶液内有固体则补加少量的水，使得重氮盐溶液保持澄清。将A瓶的混合溶液降温至0℃以下，机械搅拌10min后将B 瓶的重氮盐溶液缓慢滴入，反应液内会有大量的黄色固体产生，滴毕后补加100mL 饱和食盐水，温度保持0℃以下反应2h。抽滤，留黄色固体，用适量水洗涤，干燥，得23.4g黄色固体，产率93％。Add 10 mL of water, 40 mL of ethanol and anhydrous sodium acetate into a 250 mL three-necked flask (Bottle A) and sonicate until dissolved, add ethyl acetoacetate (21.06 g, 0.162 mol), and stir at room temperature for 2 h for later use. Add 3-methoxyaniline (10g, 0.081mol) and 10mL water into another 250mL (bottle B) three-neck flask, mix well, stir at room temperature, add concentrated hydrochloric acid drop by drop to adjust the pH to 2-3, and use ice to When the temperature of the salt bath was lowered to -5°C, 20 mL of 50% sodium nitrite aqueous solution was added dropwise, and the temperature of the solution was kept below 0°C, and stirred for 30 min. If there are solids in the solution, add a small amount of water to keep the diazonium salt solution clear. Lower the temperature of the mixed solution in bottle A to below 0°C, stir mechanically for 10 minutes, then slowly add the diazonium salt solution in bottle B, a large amount of yellow solid will be produced in the reaction solution, add 100mL saturated saline after the drop, the temperature Keep the reaction below 0°C for 2h. Suction filtration leaves a yellow solid, which is washed with an appropriate amount of water and dried to obtain 23.4 g of a yellow solid with a yield of 93%.
步骤九：1-(3-甲氧基苯基)-4-甲基-6-氧代-1,6-二氢哒嗪-3-羧酸乙酯(10)Step 9: Ethyl 1-(3-methoxyphenyl)-4-methyl-6-oxo-1,6-dihydropyridazine-3-carboxylate (10)
在100mL圆底烧瓶中先后加入中间体9(5g,0.0214mol)、乙氧羰基亚甲基三苯基磷(9.39g,0.027mol)、Et2NH(0.28mL,0.0027mol)和30mL DMSO，超声溶解后升温至85℃反应4h，中间体10反应不完。将反应液冷却至室温后倒入500mL水中，二氯甲烷萃取，合并有机层，用饱和食盐水洗涤三次，无水硫酸钠干燥，旋干，得棕色油状物，产品中会残留中间体9。产率约为70％。Intermediate 9 (5 g, 0.0214 mol), ethoxycarbonylmethylene triphenylphosphine (9.39 g, 0.027 mol), Et2 NH (0.28 mL, 0.0027 mol) and 30 mL of DMSO were successively added to a 100 mL round bottom flask, After ultrasonic dissolution, the temperature was raised to 85°C for 4 hours, but the reaction of intermediate 10 was not complete. The reaction solution was cooled to room temperature and poured into 500 mL of water, extracted with dichloromethane, combined organic layers, washed three times with saturated brine, dried over anhydrous sodium sulfate, and spin-dried to obtain a brown oil, in which intermediate 9 remained. The yield is about 70%.
步骤十：1-(3-甲氧基苯基)-4-甲基-6-氧代-1,6-二氢哒嗪-3-羧酸(11)Step 10: 1-(3-methoxyphenyl)-4-methyl-6-oxo-1,6-dihydropyridazine-3-carboxylic acid (11)
在50mL圆底烧瓶中加入中间体10(3g,0.0116mol)溶于20mL乙醇中，室温下滴加10％NaOH(7.0g,0.0174mol)，升温至60℃搅拌1h。待反应完全旋干溶剂后加入50mL饱和食盐水，室温搅拌，逐滴加入浓盐酸调节pH值至2-3，有淡黄色固体析出，抽滤，烘干，得淡黄色固体2.22g，产率83％。Intermediate 10 (3 g, 0.0116 mol) was dissolved in 20 mL of ethanol in a 50 mL round bottom flask, 10% NaOH (7.0 g, 0.0174 mol) was added dropwise at room temperature, heated to 60° C. and stirred for 1 h. After the reaction is completely spin-dried, add 50 mL of saturated saline, stir at room temperature, add concentrated hydrochloric acid dropwise to adjust the pH value to 2-3, a light yellow solid precipitates, suction filter, and dry to obtain 2.22 g of a light yellow solid, the yield 83%.
步骤十一：1-(3-甲氧基苯基)-4-甲基-6-氧代-1,6-二氢哒嗪-3-碳酰氯(B)Step 11: 1-(3-methoxyphenyl)-4-methyl-6-oxo-1,6-dihydropyridazine-3-carbonyl chloride (B)
在100mL圆底烧瓶中先后加入中间体11(1g,0.0043mol)、10mL甲苯、吡啶(0.679g,0.0086mol)和10mL氯化亚砜，超声溶解，升温至85℃反应6h。将反应液冷却至室温，减压蒸除甲苯和氯化亚砜，得到油状液体酰氯。产率约为78％Intermediate 11 (1 g, 0.0043 mol), 10 mL of toluene, pyridine (0.679 g, 0.0086 mol) and 10 mL of thionyl chloride were successively added to a 100 mL round-bottomed flask, dissolved by sonication, and heated to 85° C. for 6 h. The reaction solution was cooled to room temperature, and toluene and thionyl chloride were distilled off under reduced pressure to obtain oily liquid acid chloride. The yield is about 78%
步骤十二：4-甲基-N-(4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代-1-(3-(三氟甲氧基)苯基)-1,6-二氢哒嗪-3-甲酰胺(I)Step 12: 4-methyl-N-(4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-6-oxo-1-(3-(tri Fluoromethoxy)phenyl)-1,6-dihydropyridazine-3-carboxamide (I)
在50mL圆底烧瓶(A瓶)中先后加入将中间体A(0.05g,0.00021mol)、N,N- 二异丙基乙胺(0.5mL,0.003mol)和10mL二氯甲烷。在50mL圆底烧瓶(B瓶)中将中间体B(0.2g,0.00071mol)溶于5mL二氯甲烷中。冰浴条件下将B瓶溶液逐滴加至A瓶溶液中，滴加完毕，缓慢升至室温，反应0.5h。反应完毕后，加入5mL 5％的氢氧化钠水溶液，调节pH至9-10，用二氯甲烷进行萃取，合并有机层，无水硫酸钠干燥，浓缩，加入少量石油醚，有大量米白色固体析出，抽滤得到目标产物，产率为70％。Intermediate A (0.05 g, 0.00021 mol), N,N-diisopropylethylamine (0.5 mL, 0.003 mol) and 10 mL of dichloromethane were successively added to a 50 mL round bottom flask (Bottle A). Intermediate B (0.2 g, 0.00071 mol) was dissolved in 5 mL of dichloromethane in a 50 mL round bottom flask (B bottle). Add the solution in bottle B to the solution in bottle A dropwise under ice-bath conditions. After the dropwise addition, slowly warm up to room temperature and react for 0.5 h. After the reaction is complete, add 5mL of 5% sodium hydroxide aqueous solution, adjust the pH to 9-10, extract with dichloromethane, combine the organic layers, dry over anhydrous sodium sulfate, concentrate, add a small amount of petroleum ether, there are a lot of off-white solids The target product was obtained by suction filtration with a yield of 70%.
ESI-MSm/z:539.14；1H NMR(400MHz,DMSO-d6)δ12.35(s,1H),9.01(s,1H), 8.95(d,J＝4.1Hz,1H),8.66(d,J＝5.1Hz,1H),8.13(d,J＝10.7Hz,1H),8.03(d,J＝ 7.0Hz,1H),7.87(d,J＝7.0Hz,2H),7.82(d,J＝7.6Hz,2H),7.70(d,J＝8.6Hz,1H), 7.60(d,J＝8.7Hz,1H),7.55(s,1H),7.34(s,1H),2.92(s,3H),2.23(s,3H),1.35(s, 3H).ESI-MSm/z: 539.14;1 H NMR (400MHz, DMSO-d6 ) δ12.35(s, 1H), 9.01(s, 1H), 8.95(d, J=4.1Hz, 1H), 8.66(d ,J=5.1Hz,1H),8.13(d,J=10.7Hz,1H),8.03(d,J=7.0Hz,1H),7.87(d,J=7.0Hz,2H),7.82(d,J =7.6Hz,2H),7.70(d,J=8.6Hz,1H), 7.60(d,J=8.7Hz,1H),7.55(s,1H),7.34(s,1H),2.92(s,3H ),2.23(s,3H),1.35(s,3H).
实施例7Example 7
1-(4-溴-2-氟苯基)-4-甲基-N-(4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代-1,6-二氢哒嗪-3-甲酰胺1-(4-bromo-2-fluorophenyl)-4-methyl-N-(4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-6- Oxo-1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:551.06；1H NMR(400MHz,DMSO-d6)δ11.36(s,1H),8.93(d,J＝ 4.7Hz,1H),8.65(d,J＝5.5Hz,1H),8.10(d,J＝6.7Hz,1H),7.96(s,1H),7.94(s,1H), 7.92(s,1H),7.80(d,J＝8.6Hz,1H),7.74(d,J＝8.1Hz,1H),7.57(d,J＝9.0Hz,1H), 7.50(s,1H),7.32(d,J＝2.3Hz,1H),7.21(s,1H),2.61(s,3H),1.40(s,3H).ESI-MSm/z: 551.06;1 H NMR (400MHz, DMSO-d6 ) δ11.36(s, 1H), 8.93(d, J＝4.7Hz, 1H), 8.65(d, J＝5.5Hz, 1H ), 8.10(d, J=6.7Hz, 1H), 7.96(s, 1H), 7.94(s, 1H), 7.92(s, 1H), 7.80(d, J=8.6Hz, 1H), 7.74(d ,J＝8.1Hz,1H),7.57(d,J＝9.0Hz,1H), 7.50(s,1H),7.32(d,J＝2.3Hz,1H),7.21(s,1H),2.61(s ,3H),1.40(s,3H).
实施例8Example 8
N-(3-氟-4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-4-甲基-6-氧代-1-苯基 -1,6-二氢哒嗪-3-甲酰胺N-(3-fluoro-4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-4-methyl-6-oxo-1-phenyl-1, 6-Dihydropyridazine-3-carboxamide
ESI-MSm/z:473.15.ESI-MS m/z: 473.15.
实施例9Example 9
N-(3-氟-4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-4-甲基-6-氧代-1-(对甲苯基)-1,6-二氢哒嗪-3-甲酰胺N-(3-fluoro-4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-4-methyl-6-oxo-1-(p-tolyl) -1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:487.17.ESI-MS m/z: 487.17.
实施例10Example 10
1-(2-氯苯基)-N-(3-氟-4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-4-甲基-6-氧代-1,6-二氢哒嗪-3-甲酰胺1-(2-chlorophenyl)-N-(3-fluoro-4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-4-methyl-6- Oxo-1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:507.11.ESI-MS m/z: 507.11.
实施例11Example 11
N-(3-氟-4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-1-(3-氟苯基)-4-甲基-6-氧代-1,6-二氢哒嗪-3-甲酰胺N-(3-fluoro-4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-1-(3-fluorophenyl)-4-methyl-6- Oxo-1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:491.14.ESI-MS m/z: 491.14.
实施例12Example 12
N-(3-氟-4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-4-甲基-6-氧代-1-(3-(三氟甲基)苯基)-1,6-二氢哒嗪-3-甲酰胺N-(3-fluoro-4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-4-methyl-6-oxo-1-(3-(tri Fluoromethyl)phenyl)-1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:541.14.ESI-MS m/z: 541.14.
实施例13Example 13
N-(3-氟-4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-4-甲基-6-氧代-1-(3-(三氟甲氧基)苯基)-1,6-二氢哒嗪-3-甲酰胺N-(3-fluoro-4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-4-methyl-6-oxo-1-(3-(tri Fluoromethoxy)phenyl)-1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:557.13.ESI-MS m/z: 557.13.
实施例14Example 14
1-(4-溴-2-氟苯基)-N-(3-氟-4-((2-(甲基氨基甲酰基)吡啶-4-基)氧基)苯基)-4-甲基 -6-氧代-1,6-二氢哒嗪-3-甲酰胺1-(4-bromo-2-fluorophenyl)-N-(3-fluoro-4-((2-(methylcarbamoyl)pyridin-4-yl)oxy)phenyl)-4-methyl yl-6-oxo-1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:569.05.ESI-MS m/z: 569.05.
实施例15Example 15
N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)苯基)-4-甲基-6-氧代-1-(邻甲苯基)-1,6-二氢哒嗪-3-甲酰胺N-(4-((2-(ethylcarbamoyl)pyridin-4-yl)oxy)phenyl)-4-methyl-6-oxo-1-(o-tolyl)-1,6 -Dihydropyridazine-3-carboxamide
ESI-MSm/z:483.19.ESI-MS m/z: 483.19.
实施例16Example 16
N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)苯基)-4-甲基-6-氧代-1-(对甲苯基)-1,6-二氢哒嗪-3-甲酰胺N-(4-((2-(Ethylcarbamoyl)pyridin-4-yl)oxy)phenyl)-4-methyl-6-oxo-1-(p-tolyl)-1,6 -Dihydropyridazine-3-carboxamide
ESI-MSm/z:483.19.ESI-MS m/z: 483.19.
实施例17Example 17
N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)苯基)-1-(3-氟苯基)-4-甲基-6-氧代-1,6-二氢哒嗪-3-甲酰胺N-(4-((2-(ethylcarbamoyl)pyridin-4-yl)oxy)phenyl)-1-(3-fluorophenyl)-4-methyl-6-oxo-1 ,6-Dihydropyridazine-3-carboxamide
ESI-MSm/z:487.17.ESI-MS m/z: 487.17.
实施例18Example 18
1-(4-溴-2-氟苯基)-N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)苯基)-4-甲基-6-氧代-1,6-二氢哒嗪-3-甲酰胺1-(4-bromo-2-fluorophenyl)-N-(4-((2-(ethylcarbamoyl)pyridin-4-yl)oxy)phenyl)-4-methyl-6- Oxo-1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:565.08；1H NMR(400MHz,DMSO-d6)δ10.87(s,1H),10.14(s,1H), 8.93(s,1H),8.62(d,J＝5.5Hz,1H),7.94(d,J＝8.8Hz,2H),7.81(d,J＝8.1Hz,1H), 7.75(d,J＝8.5Hz,1H),7.48(d,J＝6.3Hz,1H),7.34(d,J＝8.8Hz,2H),7.27(d,J＝ 3.0Hz,1H),7.20(s,1H),3.40–3.37(m,2H),2.51(s,3H),1.19(t,J＝7.1Hz,3H).ESI-MSm/z: 565.08;1 H NMR (400MHz, DMSO-d6 ) δ10.87(s, 1H), 10.14(s, 1H), 8.93(s, 1H), 8.62(d, J＝5.5Hz ,1H),7.94(d,J=8.8Hz,2H),7.81(d,J=8.1Hz,1H), 7.75(d,J=8.5Hz,1H),7.48(d,J=6.3Hz,1H ),7.34(d,J=8.8Hz,2H),7.27(d,J=3.0Hz,1H),7.20(s,1H),3.40–3.37(m,2H),2.51(s,3H),1.19 (t,J=7.1Hz,3H).
实施例19Example 19
N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)苯基)-4-甲基-6-氧代-1-(3-(三氟甲氧基)苯基)-1,6-二氢哒嗪-3-甲酰胺N-(4-((2-(ethylcarbamoyl)pyridin-4-yl)oxy)phenyl)-4-methyl-6-oxo-1-(3-(trifluoromethoxy )phenyl)-1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:553.16；1H NMR(400MHz,DMSO-d6)δ10.83(s,1H),10.28(s,1H), 8.94(s,1H),8.61(d,J＝5.5Hz,1H),7.94(d,J＝8.9Hz,2H),7.76(d,J＝7.9Hz,1H), 7.70(d,J＝7.3Hz,2H),7.48(d,J＝2.2Hz,1H),7.34(d,J＝8.8Hz,2H),7.27(d,J＝ 2.4Hz,1H),7.20(s,1H),3.37(dd,J＝13.4,6.7Hz,2H),2.52(s,3H),1.19(t,J＝7.1Hz, 3H).ESI-MSm/z: 553.16;1 H NMR (400MHz, DMSO-d6 ) δ10.83(s, 1H), 10.28(s, 1H), 8.94(s, 1H), 8.61(d, J＝5.5Hz ,1H),7.94(d,J=8.9Hz,2H),7.76(d,J=7.9Hz,1H), 7.70(d,J=7.3Hz,2H),7.48(d,J=2.2Hz,1H ),7.34(d,J=8.8Hz,2H),7.27(d,J=2.4Hz,1H),7.20(s,1H),3.37(dd,J=13.4,6.7Hz,2H),2.52(s ,3H),1.19(t,J＝7.1Hz, 3H).
实施例20Example 20
N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)-3-氟苯基)-4-甲基-6-氧代-1-(邻甲苯基)-1,6-二氢哒嗪-3-甲酰胺N-(4-((2-(ethylcarbamoyl)pyridin-4-yl)oxy)-3-fluorophenyl)-4-methyl-6-oxo-1-(o-tolyl) -1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:501.18；1H NMR(400MHz,DMSO-d6)δ10.67(s,1H),8.84(t,J＝5.8 Hz,1H),8.52(d,J＝5.5Hz,1H),7.84(d,J＝8.2Hz,2H),7.44(s,1H),7.41(s,2H), 7.38(d,J＝12.4Hz,2H),7.24(d,J＝8.5Hz,2H),7.17(d,J＝3.7Hz,1H),7.07(s,1H), 3.32–3.26(m,2H),2.42(s,3H),2.14(s,3H),1.10(t,J＝7.0Hz,3H).ESI-MSm/z: 501.18;1 H NMR (400MHz, DMSO-d6 ) δ10.67(s, 1H), 8.84(t, J=5.8 Hz, 1H), 8.52(d, J=5.5Hz, 1H ),7.84(d,J＝8.2Hz,2H),7.44(s,1H),7.41(s,2H), 7.38(d,J＝12.4Hz,2H),7.24(d,J＝8.5Hz,2H ),7.17(d,J＝3.7Hz,1H),7.07(s,1H), 3.32–3.26(m,2H),2.42(s,3H),2.14(s,3H),1.10(t,J＝ 7.0Hz, 3H).
实施例21Example 21
N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)-3-氟苯基)-4-甲基-6-氧代-1-(对甲苯基)-1,6-二氢哒嗪-3-甲酰胺N-(4-((2-(ethylcarbamoyl)pyridin-4-yl)oxy)-3-fluorophenyl)-4-methyl-6-oxo-1-(p-tolyl) -1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:501.18；1H NMR(400MHz,DMSO-d6)δ10.83(s,1H),8.96(s,1H), 8.65(s,1H),7.99(s,1H),7.97(s,1H),7.72(d,J＝8.1Hz,2H),7.51(d,J＝2.0Hz,1H), 7.45(d,J＝8.0Hz,2H),7.37(d,J＝8.8Hz,2H),7.30(d,J＝2.9Hz,1H),7.17(s,1H), 3.40(dd,J＝13.2,6.7Hz,2H),2.63(s,3H),2.52(s,3H),1.22(t,J＝7.1Hz,3H).ESI-MSm/z: 501.18;1 H NMR (400MHz, DMSO-d6 ) δ10.83(s, 1H), 8.96(s, 1H), 8.65(s, 1H), 7.99(s, 1H), 7.97 (s,1H),7.72(d,J=8.1Hz,2H),7.51(d,J=2.0Hz,1H), 7.45(d,J=8.0Hz,2H),7.37(d,J=8.8Hz ,2H),7.30(d,J=2.9Hz,1H),7.17(s,1H), 3.40(dd,J=13.2,6.7Hz,2H),2.63(s,3H),2.52(s,3H) ,1.22(t,J=7.1Hz,3H).
实施例22Example 22
N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)-3-氟苯基)-1-(3-氟苯基)-4-甲基-6-氧代-1,6-二氢哒嗪-3-甲酰胺N-(4-((2-(Ethylcarbamoyl)pyridin-4-yl)oxy)-3-fluorophenyl)-1-(3-fluorophenyl)-4-methyl-6- Oxo-1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:505.16；1H NMR(400MHz,DMSO-d6)δ10.84(s,1H),8.94(s,1H), 8.63(s,1H),7.97(d,J＝8.8Hz,2H),7.81(d,J＝10.2Hz,1H),7.74(d,J＝7.9Hz,1H), 7.69(d,J＝7.7Hz,1H),7.50(d,J＝2.3Hz,1H),7.44(d,J＝7.9Hz,1H),7.36(d,J＝ 8.8Hz,2H),7.29(d,J＝2.5Hz,1H),7.19(s,1H),3.38(dd,J＝13.3,6.5Hz,2H),2.52 (s,3H),1.20(t,J＝7.1Hz,3H).ESI-MSm/z: 505.16;1 H NMR (400MHz, DMSO-d6 ) δ10.84(s, 1H), 8.94(s, 1H), 8.63(s, 1H), 7.97(d, J＝8.8Hz ,2H),7.81(d,J=10.2Hz,1H),7.74(d,J=7.9Hz,1H), 7.69(d,J=7.7Hz,1H),7.50(d,J=2.3Hz,1H ),7.44(d,J=7.9Hz,1H),7.36(d,J=8.8Hz,2H),7.29(d,J=2.5Hz,1H),7.19(s,1H),3.38(dd,J =13.3,6.5Hz,2H),2.52(s,3H),1.20(t,J=7.1Hz,3H).
实施例23Example 23
1-(4-溴-2-氟苯基)-N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)-3-氟苯基)-4-甲基 -6-氧代-1,6-二氢哒嗪-3-甲酰胺1-(4-bromo-2-fluorophenyl)-N-(4-((2-(ethylcarbamoyl)pyridin-4-yl)oxy)-3-fluorophenyl)-4-methyl yl-6-oxo-1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:583.07.ESI-MS m/z: 583.07.
实施例24Example 24
N-(4-((2-(乙基氨基甲酰基)吡啶-4-基)氧基)-3-氟苯基)-4-甲基-6-氧代-1-(3-(三氟甲氧基)苯基)-1,6-二氢哒嗪-3-甲酰胺N-(4-((2-(ethylcarbamoyl)pyridin-4-yl)oxy)-3-fluorophenyl)-4-methyl-6-oxo-1-(3-(tri Fluoromethoxy)phenyl)-1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:571.15.ESI-MS m/z: 571.15.
实施例25Example 25
4-甲基-6-氧代-N-(4-((2-(丙基氨基)吡啶-4-基)氧基)苯基)-1-(邻甲苯基)-1,6-二氢哒嗪-3-甲酰胺4-methyl-6-oxo-N-(4-((2-(propylamino)pyridin-4-yl)oxy)phenyl)-1-(o-tolyl)-1,6-di Hydropyridazine-3-carboxamide
ESI-MSm/z:497.21.ESI-MS m/z: 497.21.
实施例26Example 26
4-甲基-6-氧代-N-(4-((2-(丙基氨基)吡啶-4-基)氧基)苯基)-1-(对甲苯基)-1,6-二氢哒嗪-3-甲酰胺4-methyl-6-oxo-N-(4-((2-(propylamino)pyridin-4-yl)oxy)phenyl)-1-(p-tolyl)-1,6-di Hydropyridazine-3-carboxamide
ESI-MSm/z:497.21.ESI-MS m/z: 497.21.
实施例27Example 27
1-(3-氟苯基)-4-甲基-6-氧代-N-(4-((2-(丙基氨基)吡啶-4-基)氧基)苯基)-1,6-二氢哒嗪-3-甲酰胺1-(3-fluorophenyl)-4-methyl-6-oxo-N-(4-((2-(propylamino)pyridin-4-yl)oxy)phenyl)-1,6 -Dihydropyridazine-3-carboxamide
ESI-MSm/z:501.18.ESI-MS m/z: 501.18.
实施例28Example 28
1-(4-溴-2-氟苯基)-4-甲基-6-氧代-N-(4-((2-(丙基氨基)吡啶-4-基)氧基)苯基)-1,6- 二氢哒嗪-3-甲酰胺1-(4-bromo-2-fluorophenyl)-4-methyl-6-oxo-N-(4-((2-(propylamino)pyridin-4-yl)oxy)phenyl) -1,6-Dihydropyridazine-3-carboxamide
ESI-MSm/z:579.09；1H NMR(400MHz,DMSO-d6)δ10.99(s,1H),8.98(d,J＝ 5.9Hz,1H),8.69(d,J＝5.6Hz,1H),8.07(d,J＝12.8Hz,1H),7.99(d,J＝9.6Hz,1H), 7.82(s,1H),7.80(d,J＝2.6Hz,2H),7.75(d,J＝9.0Hz,1H),7.60(d,J＝8.9Hz,1H), 7.52(s,1H),7.37(d,J＝2.5Hz,1H),7.25(s,1H),3.41–3.33(m,2H),2.64(s,3H), 1.64(dt,J＝14.4,7.1Hz,2H),0.98(t,J＝7.4Hz,3H).ESI-MSm/z: 579.09;1 H NMR (400MHz, DMSO-d6 ) δ10.99(s, 1H), 8.98(d, J＝5.9Hz, 1H), 8.69(d, J＝5.6Hz, 1H ),8.07(d,J＝12.8Hz,1H),7.99(d,J＝9.6Hz,1H), 7.82(s,1H),7.80(d,J＝2.6Hz,2H),7.75(d,J ＝9.0Hz,1H),7.60(d,J＝8.9Hz,1H), 7.52(s,1H),7.37(d,J＝2.5Hz,1H),7.25(s,1H),3.41–3.33(m ,2H),2.64(s,3H), 1.64(dt,J=14.4,7.1Hz,2H),0.98(t,J=7.4Hz,3H).
实施例29Example 29
4-甲基-6-氧代-N-(4-((2-(丙基氨基)吡啶-4-基)氧基)苯基)-1-(3-(三氟甲氧基)苯基)-1,6-二氢哒嗪-3-甲酰胺4-methyl-6-oxo-N-(4-((2-(propylamino)pyridin-4-yl)oxy)phenyl)-1-(3-(trifluoromethoxy)benzene base)-1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:567.17.ESI-MS m/z: 567.17.
实施例30Example 30
4-甲基-6-氧代-N-(4-((2-(丙基氨基)吡啶-4-基)氧基)苯基)-1-(3-(三氟甲基)苯基)-1,6-二氢哒嗪-3-甲酰胺4-methyl-6-oxo-N-(4-((2-(propylamino)pyridin-4-yl)oxy)phenyl)-1-(3-(trifluoromethyl)phenyl )-1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:551.18.ESI-MS m/z: 551.18.
实施例31Example 31
4-甲基-N-(4-((2-((3-吗啉代)氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代-1-(对甲苯基)-1,6-二氢哒嗪-3-甲酰胺4-Methyl-N-(4-((2-((3-morpholino)carbamoyl)pyridin-4-yl)oxy)phenyl)-6-oxo-1-(p-tolyl )-1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:582.26.ESI-MS m/z: 582.26.
实施例32Example 32
1-(3-氟苯基)-4-甲基-N-(4-((2-((3-吗啉代)氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代-1,6-二氢哒嗪-3-甲酰胺1-(3-fluorophenyl)-4-methyl-N-(4-((2-((3-morpholino)carbamoyl)pyridin-4-yl)oxy)phenyl)-6 -Oxo-1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:586.62.ESI-MS m/z: 586.62.
实施例33Example 33
1-(4-溴-2-氟苯基)-4-甲基-N-(4-((2-((3-吗啉代)氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代-1,-1,6-二氢哒嗪-3-甲酰胺1-(4-bromo-2-fluorophenyl)-4-methyl-N-(4-((2-((3-morpholino)carbamoyl)pyridin-4-yl)oxy)benzene base)-6-oxo-1,-1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:664.14.ESI-MSm/z:664.14.
实施例34Example 34
4-甲基-N-(4-((2-((3-吗啉代)氨基甲酰基)吡啶-4-基)氧基)苯基)-6-氧代-1-(3-(三氟甲基)苯基)-1,6-二氢哒嗪-3-甲酰胺4-methyl-N-(4-((2-((3-morpholino)carbamoyl)pyridin-4-yl)oxy)phenyl)-6-oxo-1-(3-( Trifluoromethyl)phenyl)-1,6-dihydropyridazine-3-carboxamide
ESI-MSm/z:636.23.ESI-MSm/z:636.23.
体外抗肿瘤细胞活性In vitro anti-tumor cell activity
对按照本发明的上式Ⅰ的含吡啶类哒嗪衍生物进行了体外抑制人宫颈癌细胞Hela、结肠癌细胞HT-29、乳腺癌细胞MCF-7、肺腺癌细胞A549和白血病细胞 MV4-11活性筛选。The pyridine-containing pyridazine derivatives according to the above formula I of the present invention have inhibited human cervical cancer cell Hela, colon cancer cell HT-29, breast cancer cell MCF-7, lung adenocarcinoma cell A549 and leukemia cell MV4- 11 activity screening.
(1)细胞复苏并传代3-5次稳定后，用1mL胰蛋白酶溶液(0.25％)使其从培养瓶底部消化30s左右，加入培养液以终止消化，并转移进离心管。将离心管在1000r/min 下离心3min，弃去上清液后加入5mL培养液，吹打混匀细胞，吸取10μL细胞混悬液加入细胞计数板中计数，调整细胞浓度为104个/孔。96孔板中除A1孔为空白孔只加180μL培养基不加细胞外，其余皆加入180μL细胞混悬液。将96孔板放入培养箱中培养24h。(1) After the cells were recovered and passaged for 3-5 times, they were digested with 1 mL of trypsin solution (0.25%) for about 30 seconds from the bottom of the culture bottle, and culture medium was added to stop the digestion, and transferred into a centrifuge tube. Centrifuge the centrifuge tube at 1000r/min for 3min, discard the supernatant, add 5mL of culture medium, pipette and mix the cells, draw 10μL of the cell suspension and add it to a cell counting plate for counting, and adjust the cell concentration to 104 cells/well. In the 96-well plate, 180 μL of cell suspension was added to well A1, which was a blank well and only 180 μL of medium was added without adding cells. The 96-well plate was placed in an incubator for 24 h.
(2)用20μL二甲基亚砜溶解受试样品，然后加入适量培养液，使样品溶解成2 mg/mL药液，然后在24孔板中将样品稀释为20,4,0.8,0.16,0.032μg/mL。(2) Dissolve the test sample with 20 μL dimethyl sulfoxide, then add an appropriate amount of culture medium to dissolve the sample into a 2 mg/mL drug solution, and then dilute the sample to 20, 4, 0.8, 0.16 in a 24-well plate , 0.032 μg/mL.
每个浓度加入3孔，其中周围两行两列细胞长势受环境影响较大，只和为空白细胞孔使用。将96孔板放入培养箱中培养72h。Each concentration was added to 3 wells, and the growth of cells in the surrounding two rows and two columns was greatly affected by the environment, and only used as blank cell wells. The 96-well plate was placed in an incubator for 72 h.
(3)将96孔板中带药培养液弃去，用磷酸缓冲溶液(PBS)将细胞冲洗两遍，在每孔中加入MTT(四氮唑)(0.5mg/mL)20μL放入培养箱中3.5h后，弃去MTT溶液，加入二甲基亚砜180μL。在磁力振荡器上振荡使存活细胞与MTT反应产物甲充分溶解，放入酶标仪中测定结果。通过Bliss法可求出药物IC50值。(3) Discard the drug-carrying culture medium in the 96-well plate, wash the cells twice with phosphate buffer solution (PBS), add 20 μL of MTT (tetrazolium) (0.5 mg/mL) into each well and put it in the incubator After 3.5 h, the MTT solution was discarded, and 180 μL of dimethyl sulfoxide was added. Oscillating on a magnetic shaker makes the reaction product formazan of surviving cells and MTT Fully dissolve, put into microplate reader to determine the result. The IC50 value of the drug can be calculated by the Bliss method.
化合物的抑制宫颈癌细胞Hela、结肠癌细胞HT-29、乳腺癌细胞MCF-7、肺腺癌细胞A549和白血病细胞MV4-11活性结果(见表二)。The compounds inhibit the activity of cervical cancer cell Hela, colon cancer cell HT-29, breast cancer cell MCF-7, lung adenocarcinoma cell A549 and leukemia cell MV4-11 (see Table 2).
FLT-3酶活性试验FLT-3 Enzyme Activity Test
用于测量FLT-3激酶活性的试验基于酶联免疫吸附试验(ELISA)。具体操作是：The assay used to measure FLT-3 kinase activity is based on an enzyme-linked immunosorbent assay (ELISA). The specific operation is:
室温下，在0.25mg/mLPGT包被的板上，将实施例化合物、50pMc-Met(His- 标记的重组人FLT-3，通过杆状病毒表达和5μMATP在试验缓冲液中(25mMMOPS, PH7.4,5mMMgCl2,0.5raMMnCl2,100μM原钒酸钠，0.01％TritonX-100,1mMDTT，最后DMSO浓度1％(v/v))温育20分钟。通过冲洗除去反应混合物并用0.2μg/mL 缀合辣根过氧化物酶(HRP)的磷酸酪氨酸特异性单克隆抗体(PY20)检测磷酸化聚合物底物。加入1M磷酸终止显色后，于450nm处通过分光光度法定量显色的底物 (TMB)的颜色。实施例化合物对FLT-3激酶的抑制数据(见表二)。At room temperature, on a plate coated with 0.25 mg/mL PGT, the compound of Example, 50 pMc-Met (His-tagged recombinant human FLT-3, expressed by baculovirus and 5 μM ATP in assay buffer (25 mM MOPS, PH7. 4,5mMMgCl2 , 0.5raMMnCl2 , 100μM sodium orthovanadate, 0.01% TritonX-100, 1mMDTT, final DMSO concentration 1% (v/v)) were incubated for 20 minutes. The reaction mixture was removed by washing and admixed with 0.2μg/mL A phosphotyrosine-specific monoclonal antibody (PY20) combined with horseradish peroxidase (HRP) was used to detect the phosphorylated polymer substrate. After adding 1M phosphoric acid to stop the color development, the amount of color developed was quantified by spectrophotometry at 450nm The color of the substrate (TMB). The inhibitory data of the compounds of the examples on FLT-3 kinase (see Table 2).
表二Table II
从上述试验结果可以清楚地看出，本发明所要保护的通式Ⅰ的化合物，具有良好的体外抗肿瘤活性，相当或优于已上市的抗肿瘤药物顺铂。It can be clearly seen from the above test results that the compound of general formula I to be protected by the present invention has good antitumor activity in vitro, which is equivalent to or better than the antitumor drug cisplatin already on the market.
尽管已经通过特定实施方案描述了本发明，但修改和等价变化对于精通此领域的技术人员而言是显见的，且它们都包含在本发明范围之内。While this invention has been described in terms of specific embodiments, it is apparent that modifications and equivalents will be apparent to those skilled in the art, and these are intended to be encompassed within the scope of this invention.
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| CN110642837A (en)* | 2019-11-07 | 2020-01-03 | 江西科技师范大学 | Pyridine amide compounds containing triazole or quinolinone structure and their application |
| CN111303121A (en)* | 2020-04-20 | 2020-06-19 | 辽宁大学 | 4-phenoxypyridine compound containing quinoxalinone and application thereof |
| CN111533735A (en)* | 2020-05-08 | 2020-08-14 | 张建蒙 | Oxo-dihydropyridazine thiazole derivative and application thereof in antitumor drugs |
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| RJ01 | Rejection of invention patent application after publication | Application publication date:20191213 |