CN107793363B

CN107793363B - Substituted arylamino aromatic heterocyclic compound and application thereof as antitumor drug

Info

Publication number: CN107793363B
Application number: CN201610803077.1A
Authority: CN
Inventors: 李建其; 黄道伟; 张庆伟; 周爱南; 刘彪; 李洋
Original assignee: Shanghai Institute of Pharmaceutical Industry; China State Institute of Pharmaceutical Industry
Current assignee: Shanghai Institute of Pharmaceutical Industry; China State Institute of Pharmaceutical Industry
Priority date: 2016-09-06
Filing date: 2016-09-06
Publication date: 2021-02-26
Anticipated expiration: 2036-09-06
Also published as: CN107793363A

Abstract

The invention discloses a substituted arylamine aromatic heterocyclic compound shown in formula (I), pharmaceutically acceptable salts thereof, a preparation method thereof and application thereof in preparing antitumor drugs,

wherein: x is selected from C, N; y is selected from C, N; r₂Selected from H, C_1‑3Alkyl or halogen of (a); ar is selected from the following groups:

wherein: r₁Selected from H, C_1‑3Alkyl, substituted C_1‑3Alkyl, saturated heterocyclic group, carboxyl, alkoxyacyl, carbamoyl, N-alkylcarbamoyl; the R is₁The position of the substituent on the benzene ring is not limited; z is selected from C, N or O; n is an integer of 0 to 3. Through activity tests, the compound has multi-target inhibition activity on a tumor cell signal transduction pathway, has a good inhibition effect on target enzymes c-Met, VEGFR-2 and EGFR, shows good inhibition activity on various tumor cells, has a prospect of resisting tumor cell drug resistance, and is suitable for being used as a broad-spectrum, high-efficiency and low-toxicity anti-tumor drug.

Description

Substituted arylamino aromatic heterocyclic compound and application thereof as antitumor drug

Technical Field

The invention relates to substituted arylamine aromatic heterocyclic compounds, a preparation method thereof and application of the compounds in preventing and treating related diseases such as tumors and the like by blocking HGF/c-Met signal channels.

Background

The local invasion and metastasis of primary tumors to other tissues are one of the most important factors affecting the survival of tumor patients, and the death of most cancer patients is caused by tumor metastasis, and the metastasis process is greatly dependent on the motility and cell proliferation of tumor cells.

Hepatocyte Growth Factor (HGF), also known as Scatter Factor (SF), is a multifunctional cytokine that has the effect of promoting the growth, migration and morphogenesis of various types of cells such as hepatocytes, epithelial cells, endothelial cells, hematopoietic cells, and the like. The c-Met transmembrane protein (c-Mesenchymal-epithelia transition factor), or c-Met tyrosine kinase, is a protein product encoded by the c-Met proto-oncogene and is a high-affinity receptor for HGF. When HGF binds to the receptor c-Met, it leads to autophosphorylation of c-Met tyrosine kinase, which activates c-Met tyrosine kinase activity, further activates various downstream signaling molecules in the cell, and induces a series of biological effects in the cell, such as cell dispersion, motility, proliferation, invasion, migration, and finally metastasis and angiogenesis (Bottaro et al, Science, 1991, 251, 802-.

Numerous studies have shown that HGF/c-Met signaling plays an important role in the development, progression and secondary metastasis of malignancies, while being closely related to poor prognosis in patients (Sattler et al, Curr Oncol Rep, 2007, 9, 102-. The continuous activation of c-Met will destroy the adhesion between tumor cells, promote cell movement and the generation of tumor new blood vessels, make the tumor cells easy to enter blood circulation and obtain the ability of invasion and metastasis. In a variety of cancers, c-Met and HGF are overexpressed in relation to surrounding tissues, such as thyroid, breast, lung, stomach, colon, pancreas, prostate, kidney, liver, ovary, and brain gliomas, among others (Birchmeier et al, Nat Rev Mol Cell Biol, 2003, 4, 915-. Another study showed that MET gene amplification is closely related to 20% of epidermal growth factor inhibitors (EGFR-TKIs) acquired drug resistance (Engelman et al, Science, 2007, 316, 1039-. Therefore, small molecule kinase inhibitors targeted to c-Met have important research significance for the treatment of the above cancers.

Currently, several c-Met small molecule inhibitors are in preclinical and clinical research. Among them, crizotinib (xalkori) developed by fevery has been approved by FDA at 8 months in 2011 for treating advanced non-small cell lung cancer (NSCLC) patients expressing an abnormally-expressed Anaplastic Lymphoma Kinase (ALK) gene, and shows a good therapeutic effect. The mark indicates that the c-Met is confirmed as a novel anti-tumor target, and simultaneously indicates that the c-Met inhibitor has wide development and treatment prospects.

Clinically under-developed c-Met inhibitors differ in their binding pattern to the c-Met kinase domain and can be divided into two broad classes:

(1) u-type small molecule inhibitors: such as Crizotinib and MK-2461;

(2) linear small molecule inhibitors: such as Cabozantinib (also referred to in some literature as XL184) and Foretinib (also referred to in some literature as XL 880).

Wherein, the first U-type small molecule inhibitor acts on an ATP-binding site at the beginning of a c-Met kinase pocket, surrounds an amino acid residue Met1211 and is mainly a high-selectivity inhibitor of c-Met; the second class of linear small molecule inhibitors is a novel C-Met inhibitor developed in recent years, acting through an extended conformation on the Ile1145 hydrophobic pocket from the ATP-binding site, the kinase link chain up to the vicinity of the C-terminal helical region, mainly being multi-target inhibitors. In view of the complicated pathogenesis of tumors, the single inhibition of a signal transduction pathway is likely to cause the tumor to develop drug resistance through an escape mechanism. It has been shown that certain mutations near the c-Met active site lead to resistance in the first class of small molecule inhibitors (Berthou et al, Oncogene 2004,23, 5387. times. 5393; Buchanan et al, mol. cancer ther.2009,8, 3181. times. 3190). The second linear small molecule inhibitor not only acts on the initial end of the c-Met active site and is beneficial to preventing drug resistance from generating, but also brings better tumor inhibition effect by simultaneous inhibition of multiple targets, so that the anti-tumor drug can achieve better treatment effect. Of these, representative two drugs are Cabozantinib (2012.11 marketed) and Foretinib (phase II clinical), developed by Elixis in combination with beverest and glatiramer smith, respectively. Therefore, the design and synthesis of a novel c-Met inhibitor, especially a multi-target inhibitor are beneficial to finding out a novel anti-malignant tumor medicament which has strong anti-tumor activity and low toxic and side effects and can inhibit the growth and metastasis of tumor cells, and meanwhile, scientific guiding significance is provided for the structural diversity research of the c-Met inhibitor and the combination mode research of the c-Met inhibitor and target enzymes.

The linear small molecule C-Met inhibitor has a structural general formula shown in the specification, and is composed of an aromatic ring, an aromatic ring connecting region and a dicarbonyl side chain, and acts on an ATP-binding site, a linking region and an Ile1145 hydrophobic pocket near a C-terminal spiral region of C-Met enzyme respectively. Wherein, two carbonyl oxygen atoms in the dicarbonyl side chain can form two hydrogen bonds with amino acid residues in the hydrophobic pocket, and the compound is a basic pharmacophore with c-Met inhibition activity. The aromatic ring acting on the ATP-binding site has the functions of regulating the physicochemical property of molecules, regulating the affinity and specificity of inhibitor molecules and c-Met target enzyme and finally improving the activity of the inhibitor molecules.

In order to find compounds with equivalent or better clinical drug activity and expand the structural types of the compounds, researchers of the invention carry out a great deal of research, carry out structural modification on ATP-binding sites acting on c-Met enzyme, and substitute aromatic ring structures reported in patents by aromatic amine group aromatic heterocyclic parent nucleus; the side chain of the cyclopropyl amide is reserved, and a series of substituted arylamine aromatic heterocyclic compounds are designed and synthesized. In the structure of the compound, substituted arylamine aromatic heterocycle can theoretically form pi-pi interaction with a receptor, so that the substituted arylamine aromatic heterocycle is more suitable for being combined with c-Met enzyme; in addition, compared with the substituted quinoline structure reported in the patent, the designed compound has better physicochemical properties, and a c-Met inhibitor with stronger activity and higher selectivity can be obtained; meanwhile, on the basis of improving or retaining the antitumor activity, the influence on normal tissues or cells of a human body is reduced, so that novel antitumor active molecules with higher safety are discovered.

The search for novel anti-tumor active molecules with development value higher than that of the existing drugs entering clinical stage is an important research content in the field and is also an urgent need in clinical practice.

Disclosure of Invention

The invention aims to disclose a substituted arylamine aromatic heterocyclic compound and application thereof as an antitumor drug so as to discover a c-Met inhibitor multi-target antitumor compound with a new structure type and meet the requirement of clinical application.

The invention provides a substituted arylamine aromatic heterocyclic compound shown as a formula (I) and pharmaceutically acceptable salts thereof:

wherein:

x is selected from C, N;

y is selected from C, N;

R₂selected from H, C_1-3Alkyl orHalogen;

ar is selected from the following groups:

wherein: r₁Selected from H, C_1-3Alkyl, substituted C_1-3Alkyl, saturated heterocyclic group, carboxyl, alkoxyacyl, carbamoyl, N-alkylcarbamoyl; the R is₁The position of the substituent on the benzene ring is not limited;

z is selected from C, N or O;

n is an integer of 0 to 3.

The substituted arylamino aromatic heterocyclic compound shown in the formula (I) and pharmaceutically acceptable salts thereof are shown in the specification, wherein Ar in the structural formula is independently selected from the following groups:

the compounds of formula (I) have the following structure:

wherein: substituent groups X, Y, R involved in the structural formula₂、R₁Z, n is selected in the same manner as the corresponding substituent in formula (I).

In a preferred embodiment, the substituted arylamino aromatic heterocyclic compounds represented by the formula (I) and pharmaceutically acceptable salts thereof have the structural formula₂Selected from H, methyl, ethyl, propyl, fluoro, chloro, bromo or iodo; more preferably, R₂Selected from H, methyl, fluorine, chlorine, bromine or iodine; most preferably, R₂Selected from H or fluorine.

In a preferred embodiment, the substituted arylamino aromatic heterocycles and pharmaceutically acceptable salts thereof of formula (I) above have the structure wherein X is selected from the group consisting of N, Y is selected from the group consisting of C; or X is selected from C and Y is selected from N; or X is selected from C and Y is selected from C.

In a preferred embodiment, the substituted arylamino aromatic heterocyclic compound represented by the formula (I) and the pharmaceutically acceptable salt thereof have a structural formula in which Ar is selected from the group consisting of

Wherein R is₁Preferably selected from methyl, ethyl, propyl, substituted C_1-3Alkyl, saturated heterocyclic group, carboxyl, C_1-3Alkanoyl, carbamoyl, N-alkylcarbamoyl; the R is₁The position of the substituent on the benzene ring is not limited. More preferably, R₁Selected from methyl, ethyl, propyl, saturated heterocyclic radical, carboxyl and C_1-3Alkanoyl, carbamoyl, N-alkylcarbamoyl; the R is₁The position of the substituent on the benzene ring is not limited. Most preferably, the first and second substrates are,

r1 is selected from the group consisting of

Wherein Z is preferably selected from N or O; more preferably, Z is selected from N. In addition, n is an integer of 0 to 3, and n is preferably an integer of 1 to 3.

Wherein n is an integer of 0 to 3, and n is preferably an integer of 1 to 3.

For the convenience of understanding the present invention, the following specific compounds are preferred from the compounds of formula (I), but the substituted arylamino aromatic heterocycles of the present invention are not limited to the following compounds:

1N- [4- (tetrafluorophenyl) -N- (4- ((2- ((3-methylcarbamoyl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

1-2N- (3-fluoro-4- ((2- ((3- (methylcarbamoyl) phenyl) amino) pyridin-4-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide;

1-3N- [4- (tetrafluorophenyl) -N- (4- ((2- ((4-methylcarbamoyl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

1-4N- [4- (tetrafluorophenyl) -N- (4- ((2- ((2-methylcarbamoyl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

1-5N- (4- (tetrafluorophenyl) -N- (4- ((2- ((4- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

1-6N- (4- (tetrafluorophenyl) -N- (4- ((2- ((3- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

1-7N- (3-fluoro-4- ((2- ((3- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide;

1-8N- (4- (tetrafluorophenyl) -N- (4- ((2- ((2- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

i-94- ((4- (4- (1- ((4-fluorophenyl) carbamoyl) cyclopropanecarbonyl) phenoxy) pyrimidin-2-yl) amino) benzoic acid;

i-103- ((4- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropanecarbonyl) phenoxy) pyrimidin-2-yl) amino) benzoic acid;

i-112- ((4- (4- (1- ((4-fluorophenyl) carbamoyl) cyclopropanecarbonyl) phenoxy) pyrimidin-2-yl) amino) benzoic acid;

methyl I-124- ((4- (4- (1- ((4-fluorophenyl) carbamoyl) cyclopropanecarbonyl) phenoxy) pyrimidin-2-yl) amino) benzoate;

i-methyl 133- ((4- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropanecarbonyl) phenoxy) pyrimidin-2-yl) amino) benzoate;

methyl I-142- ((4- (4- (1- ((4-fluorophenyl) carbamoyl) cyclopropanecarbonyl) phenoxy) pyrimidin-2-yl) amino) benzoate;

1-15N- (4- ((2- ((4-cyclopropylcarbamoyl) -2-methoxyphenyl) amino) pyrimidin-4-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide;

1-16N- (4- ((2- ((3- (cyclopropylcarbamoyl) phenyl) amino) pyrimidin-4-yl) oxy) -3-fluorophenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide;

1-17N- (4- ((2- ((2- (cyclopropylcarbamoyl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide;

1-18N- (4- ((2- ((4- ((cyclopropylmethyl) carbamoyl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide;

1-19N- (4- ((2- ((3- ((cyclopropylmethyl) carbamoyl) phenyl) amino) pyrimidin-4-yl) oxy) -3-fluorophenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide;

1-20N- (4- ((2- ((2- ((cyclopropylmethyl) carbamoyl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide;

1-21N- (4-fluorophenyl) -N- (4- ((2- ((4-morpholinylphenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

1-22N- (4-fluorophenyl) -N- (4- ((2- ((3-morpholinophenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

1-23N- (4-fluorophenyl) -N- (4- ((2- ((2-morpholinophenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

I-24N- (3-fluoro-4- ((2- ((3-morpholinophenyl) amino) pyrimidin-4-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide;

1-25N- (4-fluorophenyl) -N- (4- ((2- ((3- (morpholinomethyl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

I-26N- (4-fluorophenyl) -N- (4- ((2- ((3- (2-morpholinoethyl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

1-27N- (4-fluorophenyl) -N- (4- ((6- ((3-morpholinophenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

I-28N- (4- ((5-fluoro-6- ((3-morpholinophenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

1-29N- (4-fluorophenyl) -N- (4- ((2- ((3-morpholinophenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

1-30N- (3-fluoro-4- ((2- ((3-morpholinophenyl) amino) pyrimidin-4-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide;

1-31N- (4-fluorophenyl) -N- (4- ((2- ((5-oxo-5, 6,7, 8-tetrahydronaphthalen-2-yl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

1-32N- (4-fluorophenyl) -N- (4- ((2- ((1-oxo-2, 3-dihydro-1H-inden-5-yl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

1-33N- (4-fluorophenyl) -N- (4- ((2- ((1-oxo-6, 7,8, 9-tetrahydro-5H-benzo [7] annulen-5-on-5-yl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

I-34N- (4-fluorophenyl) -N- (4- ((2- ((2-oxoindol-5-yl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

I-35N- (4-fluorophenyl) -N- (4- ((2- ((2-oxo-1, 2,3, 4-tetrahydroquinolin-6-yl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

1-36N- (4-fluorophenyl) -N- (4- ((2- ((2-oxo-2, 3,4, 5-tetrahydro-1H-benzo [ b ] azepin-7-yl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide;

the salt of any compound of the compounds I-1 to I-36 in the formula (I) is hydrochloride, hydrobromide, sulfate, phosphate, p-toluenesulfonic acid, methanesulfonate, acetate, tartrate, malate and maleate.

The structural formula of the compound is shown in the table 1.

Preferred compound numbers and corresponding structural formulas of Table 1

The invention relates to a pharmaceutically acceptable salt of a substituted arylamine aromatic heterocyclic compound shown in a formula (I), which comprises an inorganic acid salt or an organic acid; for example, hydrochloride, hydrobromide, sulphate, phosphate, p-toluenesulphonic acid, methanesulphonate, acetate, tartrate, malate, maleate.

The pharmaceutically acceptable salt of the substituted arylamine aromatic heterocyclic compound shown in the formula (I) can be formed by the salt forming reaction of the substituted arylamine aromatic heterocyclic compound shown in the formula (I) and the organic acid or the inorganic acid through acid-base.

In another aspect, the invention provides a method for synthesizing substituted arylamine aromatic heterocyclic compounds shown in formula (I), which comprises the following steps: 1) dissolving the compound 1 in a solvent, adding a chlorinated reagent compound 1, and then reacting with para-fluoroaniline to obtain a compound 3; 2) reacting the obtained compound 3 with a compound II in the presence of a condensing agent to obtain a compound III; 3) reacting the compound III with the compound IV in the presence of an alkali catalyst to obtain a compound V; 4) and reacting the compound V with a substituted aniline compound VI in the presence of an acid catalyst to obtain a target compound.

See route one:

in a preferred embodiment, the solvent in step 1) is a commonly used aprotic solvent selected from tetrahydrofuran, dichloromethane, ethyl acetate, and the like.

In a preferred embodiment, the chlorinating reagent in step 1) is selected from thionyl chloride (SOCl)₂) Phosphorus oxychloride (POCl)₃) Phosphorus pentachloride (PCl)₅) And the like.

The reaction temperature in the step 1) is as follows: 0 to 5 ℃.

In a preferred embodiment, the condensing agent in step 2) is a commonly used condensing agent selected from edc.hcl, HBTU, HATU, and the like.

The reaction temperature of the step 2) is as follows: 10 to 40 ℃.

In a preferred embodiment, the base in step 3) is a commonly used inorganic base, such as potassium carbonate, sodium carbonate, potassium tert-butoxide, sodium tert-butoxide, etc.

The reaction solvent of step 3) is selected from: DMF or acetone

The reaction temperature of the step 3) is as follows: 60 to 100 ℃.

In a preferred embodiment, the acid in step 4) is a commonly used organic acid, such as p-toluenesulfonic acid, benzenesulfonic acid, methanesulfonic acid, and the like.

The reaction solvent of the step 4) is selected from: DMF or DMSO

The reaction temperature of the step 4) is as follows: 60 to 100 ℃.

The reaction time of each step in the synthesis method of the present invention is confirmed by a chromatographic method (e.g., thin layer chromatography or high pressure liquid phase HPLC) to react the reaction substrate, i.e., the reaction can be terminated.

The compounds of the present invention can be synthesized according to the general synthetic method of the above-mentioned route one, or can be synthesized preferably by other synthetic routes.

For example: synthesis of Compound I-1 is preferably prepared by the general synthetic method described above, specifically using the following reaction scheme 1:

dissolving the intermediate 4 and the raw material 5 in DMF, adding potassium carbonate, reacting at 80 ℃ for 10 hours to obtain an intermediate 6, and finally reacting with 3-amino-N-methylbenzamide (7) to obtain a target product I-1;

for another example: synthesis of Compounds I-32 the general synthetic procedures described above are preferably used, specifically using the following scheme 2:

dissolving the intermediate 8 and the raw material 5 in DMF, adding potassium carbonate, reacting at 80 ℃ for 10 hours to obtain an intermediate 9, and finally reacting with the raw material 10 to obtain a target product I-32;

for another example, the synthesis of compound I-28 is preferably prepared by the general synthetic method described above, specifically using the following reaction scheme 3:

dissolving the intermediate 8 and the raw material 11 in DMF, adding potassium carbonate, reacting at 80 ℃ for 10 hours to obtain an intermediate 12, and finally reacting with the raw material 13 to obtain a target product I-28;

for another example, the synthesis of compounds I-29 is preferably prepared by the general synthetic method described above, specifically using the following reaction scheme 4:

compound 14 and 15 react under DMF/sodium tert-butoxide (STB) to obtain intermediate 16, which reacts with 3 to obtain intermediate 17, which reacts with compound m-morpholinylaniline (13) to obtain the target product I-29.

The compound containing the structure of the formula I obtained by the preparation route and the method can further react with inorganic acid and organic acid in a solvent, and the salt of the corresponding compound containing the structure of the formula I is separated out by cooling.

The compounds and reagents used in the above preparation methods, such as compound 1 and edc.hcl, etc., can be purchased commercially.

In vitro enzyme inhibition experiments show that the compounds of the invention all have c-Met inhibitory activity with different degrees, most of the compounds have stronger inhibitory effect on c-Met (example 48), such as compounds I-2, I-7, I-16, I-22, I-24, I-25, I-27, I-28, I-30, I-32 and I-35, and the inhibitory activity on c-Met is equivalent to or better than that of a positive control medicament Cabozantinib. Among them, Compound I-24, was 8.4 times more active than the positive control Cabozantinib.

In vitro enzyme inhibition experiments show that the compounds of the invention also show good inhibitory activity on VEGFR2 and EGFR (example 49), and the inhibitory activity of the compounds I-22, I-24, I-27 and I-28 on three target enzymes c-Met, VEGFR2 and EGFR is better than that of a positive control drug Cabozantinib.

In vitro anti-tumor cell proliferation activity tests show that the compound disclosed by the invention has stronger anti-proliferation activity on various tumor cells compared with a positive control medicament Cabozantinib (example 50). The anti-tumor cell proliferation activity of part of the compounds is better than that of a positive control medicament Cabozantinib. The compounds I-22, I-24, I-27 and I-28 have higher inhibitory activity on a human liver cancer cell line HepG2, a human prostate cancer cell line PC-3, a human pancreatic cancer cell line PANC-1, a human renal clear cell carcinoma skin metastasis cell line Caki-1 and a human colorectal cancer cell line HCT116, are superior to a positive control drug Cabozantinib, have the activity improved by 4-7 times compared with the positive control drug Cabozantinib, and have the characteristics of broad spectrum and high efficiency.

Pharmacological tests show that the compound has weak inhibitory activity on normal cells and has lower toxic and side effects compared with a control drug Cabozantinib (example 51), and the substituted arylamino aromatic heterocyclic compound has better selectivity on the inhibition and proliferation of tumor cells and normal cells and is indicated to have lower toxic and side effects when being used as an antitumor drug.

MTD test and hERG test show that the compound of the present invention is equivalent to Cabozantinib as the control drug, and has high safety.

The water-solubility test result (example 54) shows that the water solubility of the compound of the invention is superior to that of a positive control drug Cabozantinib, which indicates that the substituted arylamino aromatic heterocyclic compound of the invention has better pharmacokinetic properties and is more beneficial to the preparation and research of preparations and prescriptions.

Pharmacological experiments show that the compound has the following beneficial effects:

1) the compound has good c-Met kinase inhibition activity, has good inhibition effect on various tumor cells of a human body, and has the characteristics of broad spectrum and high efficiency.

2) The compound has multi-target inhibition activity, has good inhibition activity on c-Met, VEGFR-2 and EGFR, and has important significance for overcoming drug resistance of tumor cells.

3) Compared with a positive control medicament in a clinical period, the compound disclosed by the invention has weak inhibition effect on normal cells while effectively inhibiting tumor cells, shows better selective inhibition activity and has a good anti-tumor clinical application prospect.

4) Compared with a positive control medicament in a clinical period, the compound has better safety and physicochemical properties, and has a value of deep research.

In conclusion, when the compound is applied as an anti-tumor medicament, the compound has stronger anti-tumor activity and smaller toxic and side effects, and is easier to use as the anti-tumor medicament.

The compound of the invention can be applied to mammals (including human beings) needing tumor treatment in the form of composition by oral administration, injection and the like; especially, oral administration is preferred. The dosage is 0.0001 mg/kg-200 mg/kg body weight per day. The optimum dose depends on the individual, and usually the dose is initially smaller and then gradually increased.

The composition comprises a therapeutically effective amount of a compound of formula (I) and a pharmaceutically acceptable carrier;

the carrier refers to a carrier which is conventional in the pharmaceutical field, such as: diluents, excipients such as water, etc.; binders such as cellulose derivatives, gelatin, polyvinylpyrrolidone, etc.; fillers such as starch and the like; disintegrating agents such as calcium carbonate, sodium bicarbonate; in addition, other adjuvants such as flavoring agents and sweeteners may also be added to the composition.

The composition can be prepared into conventional solid preparations, such as tablets, capsules and the like, and is used for oral administration; it can also be made into injection.

The composition of the invention can be prepared into various dosage forms by adopting a conventional method in the field of pharmacy, wherein the content of the active ingredient compound with the structure shown in the formula I is 0.1-99.5 percent (weight ratio) of the weight of the composition.

The compound has the advantages of having multi-target inhibition activity on a tumor cell signal transduction path, having better inhibition effect on target enzymes c-Met, VEGFR-2 and EGFR, showing good inhibition activity on various tumor cells, having good physicochemical properties and being suitable for being used as broad-spectrum, high-efficiency and low-toxicity antitumor drugs.

The invention has the advantages that the compound and the medicinal preparation thereof can be used for treating diseases caused by abnormal gene expression, such as: endocrine disorders, immune system diseases, genetic diseases and neurological diseases may also have better therapeutic effects.

The invention has the advantages that the substituted arylamine aromatic heterocyclic parent nucleus is used for replacing the substituted quinoline structure, the physical and chemical properties of the obtained compound are greatly improved, the water solubility is improved, and the preparation and the research of a preparation and a prescription are facilitated.

The invention has the advantage that the compound has better safety.

In conclusion, in order to find compounds with equivalent or better clinical drug activity and expand the structural types of the compounds, researchers of the invention carry out a great deal of research, carry out structural modification on ATP-binding sites acting on c-Met enzyme, substitute aromatic ring structures reported in patents by substituted arylamine aromatic heterocyclic parent nuclei, reserve cyclopropylamide side chains, and design and synthesize a series of substituted arylamine aromatic heterocyclic compounds. In the structure of the compound, stronger pi-pi interaction is formed between the aromatic ring part and a receptor, and a plurality of hydrogen bond acting forces are formed between the amino heterocyclic ring and amide structure and amino acid residues, so that the compound is more suitable for being combined with c-Met enzyme, and a c-Met enzyme inhibitor with stronger activity and higher selectivity is obtained. The new-structure compound has stronger enzyme inhibition activity, simultaneously shows specificity and selectivity for tumor cells and normal cells, has better physicochemical property, higher safety and smaller toxic and side effect, is easier to be used as an anti-tumor medicament, is equivalent to the prior art, and has novelty, creativity and substantial scientific progress.

Detailed Description

The present invention will be described in further detail with reference to examples, but the embodiments of the invention are not limited thereto.

Example 1

Synthesis of 1- [ (4-fluorophenyl) amido ] cyclopropanecarboxylic acid (intermediate 3)

1, 3-cyclopropane dicarboxylic acid (13.01g,0.10mol) was dissolved in 120mL of THF, and triethylamine (10.12g,0.10mol) was slowly added dropwise thereto at 0 ℃ under nitrogen protection, followed by stirring for 15 min. Slowly dropwise adding SOCl into the system₂(9.95g,0.10mol), the temperature was maintained and stirring was continued for 30 min. Finally, 60mL of a THF solution containing 4-fluoroaniline (12.22g,0.11mol) was added dropwise to the reaction mixture, and the reaction was carried out in an ice bath to completion. After the reaction is finished, adjusting the pH of the reaction solution to 9.0 by using 10% sodium hydroxide solution, stirring for 10min, adjusting the pH of the system to 5.0 by using 1N HCl, precipitating a solid, stirring for 15min, and filtering. The filter cake was washed with water to neutral and dried under vacuum to give 14.71g of a white solid in 65.9% yield. The product does not need to be purified and can be directly subjected to the next reaction。

ESI-MS[M+H]⁺:m/z 224.5

¹H NMR(400MHz,DMSO-d₆)δppm:13.05(s,1H,COOH),10.58(s,1H,CONH),7.12～7.67(m,4H,Ar-H),1.47(s,4H,CH₂CH₂).

Example 2

Synthesis of N- (4-fluorophenyl) -N- (4-hydroxyphenyl) cyclopropane-1, 1-diamide (intermediate 4)

4-aminophenol (15, 0.59g,5.38mmol) and intermediate 3(1g,4.48mmol) were dissolved in 15mL of DMF, EDC.HCl (1.03g,5.38mmol) was added thereto, and the reaction was stirred at room temperature for 3 h. After the reaction is finished, adding water into the reaction solution to separate out a white solid, adding 1N HCl to adjust the pH value to 4.0-5.0, stirring for 15min, filtering, washing a filter cake to be neutral, and drying in vacuum to obtain 1.23g of the white solid with the yield of 87.9%.

ESI-MS[M+H]⁺:m/z 315.4

¹H NMR(400MHz,DMSO-d₆)δppm:10.17(s,1H,CONH),9.73(s,1H,CONH),9.23(s,1H,OH),6.68～7.83(m,8H,Ar-H),1.48(s,4H,CH₂CH₂).

Example 3

Synthesis of N- (4- ((2-chloropyrimidin-4-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-dicarboxamide (intermediate 6)

Intermediate 4(3.14g,10mmol) and starting 2, 4-dichloropyrimidine (5,1.48g,10mmol) were dissolved in DMF: to 15mL of the solution was added potassium carbonate (1.52g,11mmol), and the mixture was heated to 80 ℃ to react for 5 hours. After the reaction is finished, the reaction product is cooled to room temperature, 60mL of water is slowly added in an ice-water bath to precipitate a white solid, the white solid is stirred for 30min, filtered and dried to obtain 3.72g of an off-white solid, and the yield is 87.4%. ESI-MS [ M + H ]]⁺:m/z 427.19

Example 4

Synthesis of N- [4- (tetrafluorophenyl) -N- (4- ((2- ((3-methylcarbamoyl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide (Compound I-1)

Intermediate 6(4.69g,11mmol), 3-amino-N-methylbenzamide (1.50g,10mmol) and p-toluenesulfonic acid monohydrate (7.60g,40mmol) were dissolved in DMF 20mL and heated to 90 ℃ for reaction for 10 h. After the reaction is finished, the temperature is reduced to room temperature, the reaction solution is slowly dripped into ice water, a viscous solid is separated out, the solid is gradually solidified from the viscous state after being stirred for 20min, a light white solid is obtained by filtration, the light white solid is pulped for 2 times under the condition of absolute ethyl alcohol reflux, and the white solid is obtained by filtration and drying, wherein the yield is 63.1%.

Heating and dissolving the I-1(0.25g) in 4mol/L hydrochloric acid/ethanol (5mL), concentrating and evaporating to dryness, recrystallizing with ethanol, and cooling to separate out a solid, namely the hydrochloride compound of I-1.

ESI-MS[M+H]⁺:m/z 541.52

¹H NMR(400MHz,DMSO-d₆)δppm:10.12(d,2H,J＝28.0Hz,CONH),9.68(s,1H,CONH),8.36(d,1H,J＝5.6Hz,ArH),8.27(d,1H,J＝4.4Hz,ArH),7.96(s,1H,ArH),7.64～7.23(m,5H,ArH),7.29(d,1H,J＝7.2Hz,ArH),7.14～7.20(m,5H,ArH),6.41(d,1H,J＝5.6Hz,NH),2.76(d,3H,J＝4.4Hz,CH₃),1.48(s,4H,CH₂).

Example 5

Synthesis of N- (3-fluoro-4-hydroxyphenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-dicarboxamide (intermediate 8)

Intermediate 8 was prepared in 85.0% yield from 3 and 15' as starting materials according to the procedure for preparation of intermediate 4 in example 2.

ESI-MS[M+H]⁺:m/z 333.11

¹H NMR(400MHz,DMSO-d₆)δppm:10.14(s,1H,CONH),9.64(s,1H,CONH),9.13(s,1H,OH),6.61～7.82(m,7H,Ar-H),1.44(s,4H,CH₂CH₂).

Example 6

Synthesis of N- (4- ((2-chloropyrimidin-4-yl) oxy) -3-fluorophenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-dicarboxamide (intermediate 9)

Intermediate 9 was prepared in 81.0% yield from 8 and 5 as starting materials according to the procedure for preparation of intermediate 6 in example 3. ESI-MS [ M + H ]]⁺:m/z 444.92

Example 7

Synthesis of N- (4- ((6-chloropyrimidin-4-yl) oxy) -3-fluorophenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-dicarboxamide (intermediate 12')

Intermediate 12' was prepared according to the procedure for the preparation of intermediate 9 in example 6, starting from 8 and 11, with a yield of 84.0%. ESI-MS [ M + H ]]⁺:m/z 445.12

Example 8

Synthesis of N- (4- ((6-chloropyrimidin-4-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-dicarboxamide (intermediate 12)

Intermediate 12 was prepared in 82.0% yield by the method for the preparation of intermediate 12' in example 7 starting from 4 and 11. ESI-MS [ M + H ]]⁺:m/z 427.11

Example 9

Synthesis of N- (3-fluoro-4- ((2- ((3- (methylcarbamoyl) phenyl) amino) pyridin-4-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide (target Compound I-2)

The target compound I-2 was prepared according to example 4 using 3-amino-N-methylbenzamide and intermediate 9 as starting materials, with a yield of 62.8%.

Dissolving the I-2(0.25g) in 10mL of ethanol, adding 5mL of hydrobromic acid, heating for 30min, concentrating and evaporating the solvent, recrystallizing by ethanol, and cooling to separate out a solid, namely the hydrobromide compound of I-2.

ESI-MS[M+H]⁺:m/z 541.37

¹H NMR(400MHz,DMSO-d₆)δppm:10.15(d,2H,J＝25.2Hz,CONH),9.83(s,1H,CONH),8.37(d,1H,J＝5.6Hz,ArH),8.14(d,1H,J＝4.4Hz,ArH),7.54～7.76(m,8H,ArH),7.11～7.25(m,4H,ArH),6.42(d,1H,J＝5.6Hz,NH),2.75(d,3H,J＝4.4Hz,CH₃),1.44～1.52(m,4H,CH₂).

Example 10

Synthesis of N- [4- (tetrafluorophenyl) -N- (4- ((2- ((4-methylcarbamoyl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide (target Compound I-3)

The target compound I-3 was prepared according to example 4 using 4-amino-N-methylbenzamide and intermediate 6 as starting materials, with a yield of 61.9%.

Dissolving the I-3(0.25g) in 10mL of ethanol, adding 5mL of sulfuric acid, heating for 30min, concentrating, evaporating to dryness, recrystallizing with ethanol, and cooling to obtain solid, i.e. the sulfate compound of I-3.

ESI-MS[M+H]⁺:m/z 541.34

¹H NMR(400MHz,DMSO-d₆)δppm:10.13(d,2H,J＝25.2Hz,CONH),9.84(s,1H,CONH),8.39(d,1H,J＝5.6Hz,ArH),8.15(d,1H,J＝4.4Hz,ArH),7.58～7.72(m,8H,ArH),7.13～7.22(m,4H,ArH),6.48(d,1H,J＝5.6Hz,NH),2.73(d,3H,J＝4.4Hz,CH₃),1.47～1.53(m,4H,CH₂).

Example 11

Synthesis of N- [4- (tetrafluorophenyl) -N- (4- ((2- ((2-methylcarbamoyl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide (target Compound I-4)

The target compound I-4 was prepared according to example 4 using 2-amino-N-methylbenzamide and intermediate 6 as starting materials, with a yield of 66.2%.

Dissolving the I-4(0.25g) in 10mL of ethanol, adding 5mL of acetic acid, heating for 30min, concentrating, evaporating to dryness, recrystallizing with ethanol, and cooling to obtain solid, i.e. the acetate compound of I-4.

ESI-MS[M+H]⁺:m/z 541.32

¹H NMR(400MHz,DMSO-d₆)δppm:10.17(d,2H,J＝25.2Hz,CONH),9.85(s,1H,CONH),8.37(d,1H,J＝5.6Hz,ArH),8.13(d,1H,J＝4.4Hz,ArH),7.59～7.77(m,8H,ArH),7.16～7.29(m,4H,ArH),6.47(d,1H,J＝5.6Hz,NH),2.74(d,3H,J＝4.4Hz,CH₃),1.47～1.59(m,4H,CH₂).

Example 12

Synthesis of N- (4- (tetrafluorophenyl) -N- (4- ((2- ((4- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide (target Compound I-5)

The target compound I-5 was prepared in 67.2% yield according to example 4 using 4- (4-methylpiperazin-1-yl) aniline and intermediate 6 as starting materials.

Dissolving the I-5(0.25g) in 10mL of ethanol, adding 2mmol of lactic acid, heating for 30min, concentrating, evaporating to dryness, recrystallizing with ethanol, and cooling to separate out a solid, namely the lactate compound of I-5.

ESI-MS[M+H]⁺:m/z 582.54

¹H NMR(400MHz,DMSO-d₆)δppm:10.13(d,2H,J＝22.4Hz,CONH),9.32(s,1H,NH),8.27(d,1H,J＝5.6Hz,ArH),7.63～7.70(m,4H,ArH),7.29(s,2H,ArH),7.16(t,4H,J＝8.8Hz,ArH),6.70(d,2H,J＝8.0Hz,ArH),6.33(d,1H,J＝5.6Hz,ArH),2.99(s,4H,CH₂),2.41(s,4H,CH₂),2.19(s,4H,CH₂),1.49(m,4H,J＝6.8Hz,CH₂).

Example 13

Synthesis of N- (4- (tetrafluorophenyl) -N- (4- ((2- ((3- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide (target Compound I-6)

The target compound I-6 was prepared in 70.2% yield from 3- (4-methylpiperazin-1-yl) aniline and intermediate 6 as starting materials according to example 4.

Dissolving the I-6(0.25g) in 10mL of ethanol, adding 2mmol of tartaric acid, heating for 30min, concentrating, evaporating to dryness, recrystallizing with ethanol, and cooling to obtain solid, i.e. tartrate compound of I-6.

ESI-MS[M+H]⁺:m/z 582.54

¹H NMR(400MHz,DMSO-d₆)δppm:10.18(s,1H,CONH),10.07(s,1H,CONH),9.36(s,1H,NH),8.33(d,1H,J＝5.6Hz,ArH),7.63～7.71(m,4H,ArH),7.14～7.18(m,5H,ArH),7.03(d,1H,J＝7.6Hz,ArH),6.93(t,1H,J＝8.0Hz,ArH),6.38～6.44(m,2H,ArH),2.94(s,4H,CH₂),2.40(s,4H,CH₂),2.20(s,3H,CH₃),1.48(s,4H,CH₂).

Example 14

Synthesis of N- (3-fluoro-4- ((2- ((3- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide (target Compound I-7)

The target compound I-7 was prepared in 71.4% yield from 3- (4-methylpiperazin-1-yl) aniline and intermediate 9 as starting materials according to example 4.

Dissolving the I-7(0.25g) in 10mL of ethanol, adding 2mmol of tannic acid, heating for 30min, concentrating, evaporating to remove the solvent, recrystallizing with ethanol, and cooling to separate out a solid, namely the tannate compound of I-7.

ESI-MS[M+H]⁺:m/z 600.27

¹H NMR(400MHz,DMSO-d₆)δppm:10.14(d,2H,J＝22.7Hz,CONH),9.38(s,1H,NH),8.27(d,1H,J＝5.6Hz,ArH),7.68～7.67(m,3H,ArH),7.29(s,2H,ArH),7.32(t,4H,J＝8.8Hz,ArH),6.77(d,2H,J＝6.8Hz,ArH),6.39(d,1H,J＝5.6Hz,ArH),2.94(s,4H,CH₂),2.47(s,4H,CH₂),2.24(s,4H,CH₂),1.49(m,4H,J＝6.8Hz,CH₂).

Example 15

Synthesis of N- (4- (tetrafluorophenyl) -N- (4- ((2- ((2- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide (target Compound I-8)

The target compound I-8 was prepared in 74.4% yield from 2- (4-methylpiperazin-1-yl) aniline and intermediate 6 as starting materials according to example 4.

Dissolving the I-8(0.25g) in 10mL of ethanol, adding 2mmol of citric acid, heating for 30min, concentrating, evaporating to remove the solvent, recrystallizing with ethanol, and cooling to obtain solid, i.e. the citrate compound of I-8.

ESI-MS[M+H]⁺:m/z 582.56

¹H NMR(400MHz,DMSO-d₆)δppm:10.14(s,1H,CONH),10.11(s,1H,CONH),9.28(s,1H,NH),8.37(d,1H,J＝5.4Hz,ArH),7.69～7.78(m,4H,ArH),7.17～7.28(m,5H,ArH),7.13(d,1H,J＝7.6Hz,ArH),6.97(t,1H,J＝8.0Hz,ArH),6.48～6.47(m,2H,ArH),2.97(s,4H,CH₂),2.44(s,4H,CH₂),2.24(s,3H,CH₃),1.49(s,4H,CH₂).

Example 16

Synthesis of 4- ((4- (4- (1- ((4-fluorophenyl) carbamoyl) cyclopropylformyl) phenoxy) pyrimidin-2-yl) amino) benzoic acid (target compound I-9)

The target compound I-9 was prepared from 4-aminobenzoic acid and intermediate 6 as starting materials in 64.4% yield as in example 4.

Dissolving the I-9(0.25g) in 10mL of ethanol, adding 2mmol of trifluoroacetic acid, heating for 30min, concentrating, evaporating to dryness, recrystallizing with ethanol, and cooling to obtain a solid, namely the trifluoroacetate compound of I-9.

ESI-MS[M+H]⁺:m/z 528.18

¹H NMR(400MHz,DMSO-d₆)δppm:11.22(s,1H,COOH),10.25(s,1H,CONH),10.11(s,1H,CONH),8.33(d,1H,J＝5.6Hz,NH),7.83(s,1H,ArH),7.67～7.77(m,5H,ArH),7.47(d,1H,J＝0.8Hz,ArH),7.27(t,1H,J＝8.4Hz,ArH),7.09～7.17(m,4H,ArH),6.42(d,2H,J＝5.6Hz,ArH),1.49(s,4H,CH₂).

Example 17

Synthesis of 3- ((4- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropanecarbonyl) phenoxy) pyrimidin-2-yl) amino) benzoic acid (target compound I-10)

The target compound I-10 was prepared from 3-aminobenzoic acid and intermediate 9 as starting materials in 67.4% yield as in example 4.

Dissolving the I-10(0.25g) in 10mL of ethanol, adding 2mmol of L-malic acid, heating for 30min, concentrating, evaporating to remove the solvent, recrystallizing with ethanol, and cooling to obtain solid, i.e. the L-malate compound of I-10.

ESI-MS[M+H]⁺:m/z 546.16

¹H NMR(400MHz,DMSO-d₆)δppm:11.27(s,1H,COOH),10.24(s,1H,CONH),10.01(s,1H,CONH),8.23(d,1H,J＝5.8Hz,NH),7.87(s,1H,ArH),7.67～7.77(m,5H,ArH),7.53(d,1H,J＝4.8Hz,ArH),7.29(t,1H,J＝8.6Hz,ArH),7.09～7.14(m,4H,ArH),6.45(d,1H,J＝5.6Hz,ArH),1.69(s,4H,CH₂).

Example 18

Synthesis of 2- ((4- (4- (1- ((4-fluorophenyl) carbamoyl) cyclopropylformyl) phenoxy) pyrimidin-2-yl) amino) benzoic acid (target compound I-11)

The target compound I-11 was prepared from 2-aminobenzoic acid and intermediate 6 as starting materials in 65.2% yield as in example 4.

Dissolving the I-11(0.25g) in 10mL of ethanol, adding 2mmol of D-malic acid, heating for 30min, concentrating, evaporating to remove the solvent, recrystallizing with ethanol, and cooling to separate out a solid, namely the D-malate compound of I-11.

ESI-MS[M+H]⁺:m/z 528.19

¹H NMR(400MHz,DMSO-d₆)δppm:11.13(s,1H,COOH),10.27(s,1H,CONH),10.18(s,1H,CONH),8.39(d,1H,J＝5.6Hz,NH),7.85(s,1H,ArH),7.64～7.72(m,5H,ArH),7.58(d,1H,J＝6.8Hz,ArH),7.37(t,1H,J＝8.4Hz,ArH),7.11～7.17(m,4H,ArH),6.46(d,2H,J＝5.6Hz,ArH),1.72(s,4H,CH₂).

Example 19

Synthesis of methyl 4- ((4- (4- (1- ((4-fluorophenyl) carbamoyl) cyclopropylformyl) phenoxy) pyrimidin-2-yl) amino) benzoate (target compound I-12)

The target compound I-12 was prepared from methyl 4-aminobenzoate and intermediate 6 as starting materials in 67.4% yield as in example 4.

Dissolving the I-12(0.25g) in 10mL of ethanol, adding 2mmol of maleic acid, heating for 30min, concentrating, evaporating to dryness, recrystallizing with ethanol, and cooling to obtain solid, i.e. the maleate compound of I-12.

ESI-MS[M+H]⁺:m/z 542.19

¹H NMR(400MHz,DMSO-d₆)δppm:10.21(s,1H,CONH),10.14(s,1H,CONH),8.43(s,1H,NH),8.11(s,1H,ArH),8.02(d,1H,J＝7.6Hz,ArH),7.66～7.76(m,4H,ArH),7.38～7.46(m,2H,ArH),7.16～7.23(m,4H,ArH),6.58(s,2H,ArH),3.90(s,3H,CH₃),1.53(s,4H,CH₂).

Example 20

Synthesis of methyl 3- ((4- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropanecarbonyl) phenoxy) pyrimidin-2-yl) amino) benzoate (target Compound I-13)

The target compound I-13 was prepared according to example 4 using methyl 3-aminobenzoate and intermediate 9 as starting materials, with a yield of 71.4%.

Dissolving the I-13(0.25g) in 10mL of ethanol, adding 2mmol of succinic acid, heating for 30min, concentrating, evaporating to dryness, recrystallizing with ethanol, and cooling to obtain solid, i.e. the succinate compound of I-13.

ESI-MS[M+H]⁺:m/z 560.19

¹H NMR(400MHz,DMSO-d₆)δppm:10.27(s,1H,CONH),10.17(s,1H,CONH),8.42(s,1H,NH),8.14(s,1H,ArH),8.06(d,1H,J＝7.8Hz,ArH),7.62～7.74(m,4H,ArH),7.37～7.43(m,1H,ArH),7.18～7.25(m,4H,ArH),6.88(s,2H,ArH),3.94(s,3H,CH₃),1.57(s,4H,CH₂).

Example 21

Synthesis of methyl 2- ((4- (4- (1- ((4-fluorophenyl) carbamoyl) cyclopropylformyl) phenoxy) pyrimidin-2-yl) amino) benzoate (target Compound I-14)

The target compound I-14 was prepared according to example 4 using methyl 3-aminobenzoate and intermediate 6 as starting materials, with a yield of 68.4%.

Dissolving the I-14(0.25g) in 10mL of ethanol, adding 2mmol of p-toluenesulfonic acid, heating for 30min, concentrating, evaporating to dryness, recrystallizing with ethanol, and cooling to separate out a solid, namely the p-toluenesulfonic acid salt compound of I-14.

ESI-MS[M+H]⁺:m/z 542.20

¹H NMR(400MHz,DMSO-d₆)δppm:10.32(s,1H,CONH),10.24(s,1H,CONH),8.47(s,1H,NH),8.24(s,1H,ArH),8.12(d,1H,J＝8.6Hz,ArH),7.69～7.72(m,4H,ArH),7.34～7.44(m,2H,ArH),7.13～7.23(m,4H,ArH),6.88(s,2H,ArH),3.91(s,3H,CH₃),1.43(s,4H,CH₂).

Example 22

Synthesis of N- (4- ((2- ((4-cyclopropylcarbamoyl) -2-methoxyphenyl) amino) pyrimidin-4-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide (target compound I-15)

The target compound I-15 was prepared according to example 4 using 4-amino-N-cyclopropylbenzamide and intermediate 6 as starting materials, with a yield of 71.4%.

Dissolving the I-15(0.25g) in 10mL of ethanol, adding 2mmol of mesylate, heating for 30min, concentrating and evaporating the solvent, recrystallizing by ethanol, and cooling to separate out a solid, namely the mesylate compound of the I-15.

ESI-MS[M+H]⁺:m/z 567.22

¹H NMR(400MHz,DMSO-d₆)δppm:10.24(s,1H,CONH),10.14(s,1H,CONH),10.12(s,1H,CONH),8.45(s,1H,NH),8.14(s,2H,ArH),8.06(d,1H,J＝7.6Hz,ArH),7.69～7.77(m,4H,ArH),7.39～7.47(m,2H,ArH),7.11～7.27(m,4H,ArH),6.56(s,2H,ArH),3.17(t,2H,J＝6.0Hz,CH₂),1.48(t,4H,J＝5.6Hz,CH₂),1.35(s,2H,CH₂),1.07(m,1H,CH₂).

Example 23

Synthesis of N- (4- ((2- ((3- (cyclopropylcarbamoyl) phenyl) amino) pyrimidin-4-yl) oxy) -3-fluorophenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide (target Compound I-16)

The target compound I-16 was prepared according to example 4 using 3-amino-N-cyclopropylbenzamide and intermediate 9 as starting materials, with a yield of 74.4%.

ESI-MS[M+H]⁺:m/z 585.21

¹H NMR(400MHz,DMSO-d₆)δppm:10.26(s,1H,CONH),10.14(s,1H,CONH),10.12(s,1H,CONH),8.47(s,1H,NH),8.34(s,2H,ArH),8.12(d,1H,J＝7.8Hz,ArH),7.64～7.74(m,4H,ArH),7.37～7.49(m,1H,ArH),7.12～7.25(m,3H,ArH),6.58(s,2H,ArH),3.18(t,2H,J＝6.4Hz,CH₂),1.45(t,4H,J＝5.4Hz,CH₂),1.55(s,2H,CH₂),1.27(m,1H,CH₂).

Example 24

Synthesis of N- (4- ((2- ((2- (cyclopropylcarbamoyl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide (target Compound I-17)

The target compound I-17 was prepared according to example 4 using 2-amino-N-cyclopropylbenzamide and intermediate 6 as starting materials, with a yield of 72.8%.

ESI-MS[M+H]⁺:m/z 567.23

¹H NMR(400MHz,DMSO-d₆)δppm:10.18(s,1H,CONH),10.11(s,1H,CONH),10.07(s,1H,CONH),8.39(d,1H,J＝5.6Hz,NH),8.04(s,2H,ArH),7.97(s,2H,ArH),7.64～7.75(m,4H,ArH),7.46(d,1H,J＝1.6Hz,ArH),7.13～7.22(m,4H,ArH),6.50(d,1H,J＝5.6Hz,ArH),3.11(t,2H,J＝6.0Hz,CH₂),1.48(t,4H,J＝5.6Hz,CH₂),1.35(s,2H,CH₂),1.04(m,1H,CH₂).

Example 25

Synthesis of N- (4- ((2- ((4- ((cyclopropylmethyl) carbamoyl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide (target Compound I-18)

The target compound I-18 was prepared according to example 4 using 4-amino-N-cyclopropylmethylbenzamide and intermediate 6 as starting materials, with a yield of 68.4%.

ESI-MS[M+H]⁺:m/z 581.23

¹H NMR(400MHz,DMSO-d₆)δppm:10.16(s,1H,CONH),10.09(s,1H,CONH),10.04(s,1H,CONH),8.39(d,1H,J＝5.6Hz,NH),8.01(s,1H,ArH),7.96(s,1H,ArH),7.62～7.73(m,4H,ArH),7.46(d,1H,J＝1.6Hz,ArH),7.13～7.22(m,5H,ArH),6.50(d,2H,J＝5.6Hz,ArH),3.11(t,2H,J＝6.0Hz,CH₂),1.48(t,4H,J＝5.6Hz,CH₂),1.35(s,2H,CH₂),1.23(s,2H,CH₂),1.04(m,1H,CH₂).

Example 26

Synthesis of N- (4- ((2- ((3- ((cyclopropylmethyl) carbamoyl) phenyl) amino) pyrimidin-4-yl) oxy) -3-fluorophenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide (target Compound I-19)

The target compound I-19 was prepared according to example 4 using 3-amino-N-cyclopropylmethylbenzamide and intermediate 9 as starting materials, with a yield of 67.7%.

ESI-MS[M+H]⁺:m/z 599.22

¹H NMR(400MHz,DMSO-d₆)δppm:10.19(s,1H,CONH),10.11(s,1H,CONH),10.02(s,1H,CONH),8.42(d,1H,J＝5.6Hz,NH),8.21(s,1H,ArH),7.97(s,1H,ArH),7.65～7.76(m,4H,ArH),7.56(d,1H,J＝1.6Hz,ArH),7.16～7.23(m,4H,ArH),6.56(d,2H,J＝5.6Hz,ArH),3.17(t,2H,J＝6.8Hz,CH₂),1.54(t,4H,J＝5.6Hz,CH₂),1.37(s,2H,CH₂),1.26(s,2H,CH₂),1.14(m,1H,CH₂).

Example 27

Synthesis of N- (4- ((2- ((2- ((cyclopropylmethyl) carbamoyl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide (target compound I-20)

The target compound I-20 was prepared according to example 4 using 2-amino-N-cyclopropylmethylbenzamide and intermediate 6 as starting materials, with a yield of 77.7%.

ESI-MS[M+H]⁺:m/z 581.24

¹H NMR(400MHz,DMSO-d₆)δppm:10.22(s,1H,CONH),10.14(s,1H,CONH),10.05(s,1H,CONH),8.41(d,1H,J＝5.8Hz,NH),8.11(s,1H,ArH),7.97(s,1H,ArH),7.64～7.75(m,4H,ArH),7.49(d,1H,J＝6.6Hz,ArH),7.15～7.27(m,5H,ArH),6.58(d,2H,J＝8.6Hz,ArH),3.13(t,2H,J＝6.4Hz,CH₂),1.46(t,4H,J＝5.8Hz,CH₂),1.37(s,2H,CH₂),1.28(s,2H,CH₂),1.14(m,1H,CH₂).

Example 28

Synthesis of N- (4-fluorophenyl) -N- (4- ((2- ((4-morpholinylphenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide (target Compound I-21)

The target compound I-21 was prepared from 4-morpholinylaniline and intermediate 6 as starting materials in 78.4% yield as in example 4.

ESI-MS[M+H]⁺:m/z 569.46

¹H NMR(400MHz,DMSO-d₆)δppm:10.16(s,1H,CONH),10.09(s,1H,CONH),9.34(s,1H,NH),8.28(d,1H,J＝4.8Hz,ArH),7.65～7.70(m,4H,ArH),7.32(s,2H,ArH),7.16(t,4H,J＝8.4Hz,ArH),6.71(d,2H,J＝7.2Hz,ArH),6.34(d,1H,J＝5.2Hz,ArH),3.68(s,4H,CH₂),2.96(s,4H,CH₂),1.49(d,4H,J＝8.4Hz,CH₂).

Example 29

Synthesis of N- (4-fluorophenyl) -N- (4- ((2- ((3-morpholinophenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide (target Compound I-22)

The target compound I-22 was prepared from 3-morpholinylaniline and intermediate 6 as starting materials in 79.4% yield as in example 4.

ESI-MS[M+H]⁺:m/z 569.57

¹H NMR(400MHz,DMSO-d₆)δppm:10.17(s,1H,CONH),10.07(s,1H,CONH),9.67(s,1H,NH),8.34(d,1H,J＝6.0Hz,ArH),7.61～7.72(m,5H,ArH),7.48(d,1H,J＝8.0Hz,ArH),7.61～7.72(m,4H,ArH),7.01～7.09(m,2H,ArH),6.47(d,1H,J＝5.6Hz,ArH),3.69(t,4H,J＝4.0Hz,CH₂),2.97(s,4H,CH₂),1.47(d,4H,J＝2.4Hz,CH₂).

Example 30

Synthesis of N- (4-fluorophenyl) -N- (4- ((2- ((2-morpholinophenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide (target Compound I-23)

The target compound I-23 was prepared from 2-morpholinylaniline and intermediate 6 as starting materials in 69.4% yield as in example 4.

ESI-MS[M+H]⁺:m/z 569.47

¹H NMR(400MHz,DMSO-d₆)δppm:10.19(s,1H,CONH),10.17(s,1H,CONH),9.47(s,1H,NH),8.44(d,1H,J＝5.6Hz,ArH),7.64～7.76(m,5H,ArH),7.38(d,1H,J＝8.0Hz,ArH),7.64～7.77(m,4H,ArH),7.11～7.19(m,2H,ArH),6.57(d,1H,J＝5.6Hz,ArH),3.67(t,4H,J＝4.0Hz,CH₂),2.99(s,4H,CH₂),1.44(d,4H,J＝2.4Hz,CH₂).

Example 31

Synthesis of N- (3-fluoro-4- ((2- ((3-morpholinophenyl) amino) pyrimidin-4-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide (target Compound I-24)

The target compound I-24 was prepared from 3-morpholinylaniline and intermediate 9 in 67.4% yield as in example 4.

ESI-MS[M+H]⁺:m/z 587.23

¹H NMR(400MHz,DMSO-d₆)δppm:10.10(s,1H,CONH),10.01(s,1H,CONH),9.47(s,1H,NH),8.15(d,1H,J＝6.0Hz,ArH),7.64～7.75(m,4H,ArH),7.58(d,1H,J＝8.0Hz,ArH),7.63～7.75(m,4H,ArH),7.04～7.19(m,2H,ArH),6.57(d,1H,J＝5.6Hz,ArH),3.56(t,4H,J＝4.0Hz,CH₂),2.87(s,4H,CH₂),1.44(d,4H,J＝2.4Hz,CH₂).

Example 32

Synthesis of N- (4-fluorophenyl) -N- (4- ((2- ((3- (morpholinomethyl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide (target Compound I-25)

The target compound I-25 was prepared according to example 4 using 3- (morpholinomethyl) aniline and intermediate 6 as starting materials, with a yield of 67.4%.

ESI-MS[M+H]⁺:m/z 583.25

¹H NMR(400MHz,DMSO-d₆)δppm:10.14(s,1H,CONH),10.07(s,1H,CONH),9.67(s,1H,NH),8.64(d,1H,J＝6.0Hz,ArH),7.63～7.78(m,5H,ArH),7.68(d,1H,J＝8.0Hz,ArH),7.65～7.78(m,4H,ArH),7.04～7.09(m,2H,ArH),6.87(d,1H,J＝5.6Hz,ArH),3.79(t,4H,J＝4.0Hz,CH₂),2.97(s,4H,CH₂),1.69(s,2H,CH₂),1.47(d,4H,J＝2.4Hz,CH₂).

Example 33

Synthesis of N- (4-fluorophenyl) -N- (4- ((2- ((3- (2-morpholinoethyl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide (target Compound I-26)

The target compound I-26 was prepared from 3- (2-morpholinoethyl) aniline and intermediate 6 as starting materials in 69.4% yield as in example 4.

ESI-MS[M+H]⁺:m/z 597.39

¹H NMR(400MHz,DMSO-d₆)δppm:10.18(s,1H,CONH),10.04(s,1H,CONH),9.64(s,1H,NH),8.37(d,1H,J＝5.6Hz,ArH),7.64～7.78(m,5H,ArH),7.47(d,1H,J＝6.8Hz,ArH),7.63～7.74(m,4H,ArH),7.04～7.07(m,2H,ArH),6.44(d,1H,J＝8.0Hz,ArH),3.67(t,4H,J＝4.0Hz,CH₂),3.44(s,4H,CH₂),2.97(s,4H,CH₂),1.47(d,4H,J＝2.4Hz,CH₂).

Example 34

Synthesis of N- (4-fluorophenyl) -N- (4- ((2- ((3- (2-morpholinoethyl) phenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide (target Compound I-31)

The target compound I-31 was prepared from 5-amino-2, 3-dihydro-1H-inden-1-one and intermediate 6 as starting materials in 64.4% yield as in example 4.

ESI-MS[M+H]⁺:m/z 538.37

¹H NMR(400MHz,DMSO-d₆)δppm:10.12(s,1H,CONH),10.05(s,1H,CONH),9.66(s,1H,NH),8.34(d,1H,J＝5.8Hz,ArH),7.63～7.71(m,5H,ArH),7.44(d,1H,J＝6.6Hz,ArH),7.63～7.71(m,4H,ArH),7.06～7.09(m,2H,ArH),6.41(d,1H,J＝6.8Hz,ArH),3.69(t,4H,J＝5.6Hz,CH₂),3.42(s,4H,CH₂).

Example 35

Synthesis of N- (4-fluorophenyl) -N- (4- ((2- ((1-oxo-2, 3-dihydro-1H-inden-5-yl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide (target Compound I-32)

The target compound I-32 was prepared according to example 4 starting from 6-amino-3, 4-dihydronaphthalen-1 (2H) -one and intermediate 9 in 74.4% yield.

ESI-MS[M+H]⁺:m/z 552.20

¹H NMR(400MHz,DMSO-d₆)δppm:10.21(s,1H,NH),10.04(s,1H,CONH),10.00(s,1H,CONH),8.42(d,1H,J＝4.8Hz,ArH),7.76(d,2H,J＝8.0Hz,ArH),7.63～7.65(m,3H,ArH),7.51(s,1H,ArH),7.31(d,1H,J＝8.0Hz,ArH),7.21(d,2H,J＝8.0Hz,ArH),7.16(t,2H,J＝4.0Hz,ArH),6.57(d,1H,J＝4.0Hz,ArH),2.59(t,2H,J＝4.0Hz,CH₂),2.40(t,2H,J＝4.0Hz,CH₂),1.86～1.90(m,2H,CH₂),1.46～1.50(m,4H,CH₂).

Example 36

Synthesis of N- (4-fluorophenyl) -N- (4- ((2- ((1-oxo-6, 7,8, 9-tetrahydro-5H-benzo [7] annulen-5-on-5-yl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide (target Compound I-33)

The target compound I-33 was prepared according to example 4 starting from 6-amino-3, 4-dihydronaphthalen-1 (2H) -one and intermediate 6 in 74.4% yield.

Heating and dissolving the I-33(0.25g) in 4mol/L hydrochloric acid/ethanol (5mL), concentrating and evaporating the solvent to dryness, recrystallizing by ethanol, and cooling to separate out a solid, namely the hydrochloride compound of I-33.

ESI-MS[M+H]⁺:m/z 566.21

¹H NMR(400MHz,DMSO-d₆)δppm:10.24(s,1H,NH),10.07(s,1H,CONH),10.02(s,1H,CONH),8.44(d,1H,J＝4.0Hz,ArH),7.78(d,2H,J＝6.8Hz,ArH),7.62～7.67(m,3H,ArH),7.54(s,1H,ArH),7.36(d,1H,J＝6.0Hz,ArH),7.24(d,2H,J＝8.6Hz,ArH),7.18(t,2H,J＝4.6Hz,ArH),6.51(d,1H,J＝4.8Hz,ArH),2.57(t,2H,J＝4.6Hz,CH₂),2.44(t,2H,J＝4.4Hz,CH₂),2.07～1.98(m,2H,CH₂),1.88～1.92(m,2H,CH₂),1.48～1.54(m,4H,CH₂).

Example 37

Synthesis of N- (4-fluorophenyl) -N- (4- ((2- ((2-oxoindol-5-yl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide (target Compound I-34)

The target compound I-34 was prepared from 5-aminoindolin-2-one and intermediate 6 as starting materials in 68.0% yield as in example 4.

Dissolving the I-34(0.25g) in hot ethanol (5mL), slowly dropwise adding concentrated sulfuric acid (98%) 0.1mL, concentrating, evaporating to remove the solvent, recrystallizing with ethanol, and cooling to separate out a solid, namely the sulfate compound of I-34.

ESI-MS[M+H]⁺:m/z 539.24

¹H NMR(400MHz,DMSO-d₆)δppm:10.21(s,1H,NH),10.19(s,1H,CONH),9.67(s,1H,CONH),9.34(s,1H,CONH),8.46(d,1H,J＝4.6Hz,ArH),7.87(d,2H,J＝8.4Hz,ArH),7.66～7.78(m,2H,ArH),7.56(s,1H,ArH),7.62(s,1H,ArH),7.37(d,2H,J＝4.0Hz,ArH),7.35～7.37(m,3H,ArH),6.74(d,1H,J＝4.0Hz,ArH),2.47(s,2H,CH₂),1.49～1.52(m,4H,CH₂).

Example 38

Synthesis of N- (4-fluorophenyl) -N- (4- ((2- ((2-oxo-1, 2,3, 4-tetrahydroquinolin-6-yl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide (target Compound I-35)

The target compound I-35 was prepared from 6-amino-3, 4-dihydronaphthalen-1 (2H) -one and intermediate 6 as starting materials, according to example 4, with a yield of 78.0%.

ESI-MS[M+H]⁺:m/z 553.34

¹H NMR(400MHz,DMSO-d₆)δppm:10.27(s,1H,NH),10.09(s,1H,CONH),9.87(s,1H,CONH),9.24(s,1H,CONH),8.36(d,1H,J＝4.0Hz,ArH),7.77(d,2H,J＝8.0Hz,ArH),7.64～7.68(m,2H,ArH),7.46(s,1H,ArH),7.42(s,1H,ArH),7.27(d,2H,J＝4.0Hz,ArH),7.15～7.17(m,3H,ArH),6.54(d,1H,J＝4.0Hz,ArH),2.27(s,2H,CH₂),1.94(t,2H,J＝4.0Hz,CH₂),1.47～1.52(m,4H,CH₂).

Example 39

Synthesis of N- (3-fluoro-4- ((2- ((3-morpholinophenyl) amino) pyrimidin-4-yl) oxy) phenyl) -1, 5-dimethyl-3-oxy-2-benzene-2, 3-dihydro-1H-pyrazole-4-carboxamide (target Compound I-36)

The target compound I-36 was prepared from 7-amino-4, 5-dihydro-1H-benzo [ b ] aza-2- (3H) -one and intermediate 9 as starting materials, according to example 4, with a yield of 68.0%.

ESI-MS[M+H]⁺:m/z 567.17

¹H NMR(400MHz,DMSO-d₆)δppm:10.22(s,1H,NH),10.01(s,1H,CONH),9.80(s,1H,CONH),9.28(s,1H,CONH),8.34(d,1H,J＝4.0Hz,ArH),7.75(d,2H,J＝8.0Hz,ArH),7.63～7.65(m,2H,ArH),7.48(s,1H,ArH),7.41(s,1H,ArH),7.20(d,2H,J＝4.0Hz,ArH),7.11～7.17(m,3H,ArH),6.51(d,1H,J＝4.0Hz,ArH),2.29(s,2H,CH₂),2.04(t,2H,J＝4.0Hz,CH₂),1.95(t,2H,J＝4.0Hz,CH₂),1.46～1.52(m,4H,CH₂).

Example 40

Synthesis of N- (4-fluorophenyl) -N- (4- ((6- ((3-morpholinophenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide (target Compound I-27)

The target compound I-27 was prepared from 3-morpholinylaniline and intermediate 12 as starting materials, according to example 4, with a yield of 72.4%.

ESI-MS[M+H]⁺:m/z 569.17

¹H NMR(400MHz,DMSO-d₆)δppm:10.15(s,1H,CONH),10.06(s,1H,CONH),9.45(s,1H,NH),8.34(s,1H,ArH),7.62～7.69(m,4H,ArH),7.13～7.17(m,4H,ArH),7.02(d,1H,J＝8.0Hz,ArH),6.62(dd,1H,J₁＝1.6Hz,J₂＝8.0Hz,ArH),6.04(s,1H,ArH),2.73(t,4H,J＝4.0Hz,CH₂),3.05(t,4H,J＝4.0Hz,CH₂),1.46(s,4H,CH₂).

EXAMPLE 41

Synthesis of N- (4- ((5-fluoro-6- ((3-morpholinophenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide (target Compound I-28)

The target compound I-28 was prepared from 3-morpholinylaniline and intermediate 12' as starting materials in 77.4% yield according to example 4.

ESI-MS[M+H]⁺:m/z 587.20

¹H NMR(400MHz,DMSO-d₆)δppm:10.32(s,1H,CONH),10.02(s,1H,CONH),9.53(s,1H,NH),8.30(s,1H,ArH),6.79(dd,1H,J₁＝4Hz,J₂＝16.0Hz,ArH),7.62～7.66(m,2H,ArH),7.42(d,1H,J＝8.0Hz,ArH),7.29(t,1H,J＝8.0Hz,CH₂),7.13～7.19(m,4H,ArH),7.04(d,1H,J＝8.0Hz,ArH),6.64(dd,1H,J₁＝4Hz,J₂＝8.0Hz,ArH),6.20(s,1H,ArH),3.74(t,4H,J＝4.0Hz,CH₂),3.07(t,4H,J＝4.0Hz,CH₂),1.46(d,1H,J＝4.0Hz,CH₂).

Example 42

Synthesis of intermediate 4- ((2-chloropyridin-4-yl) oxy) aniline (16)

Dissolving raw material 2, 4-dichloropyridine (14, 1.47g, 0.01mol) in 10mL of DMF, adding p-aminophenol (15, 1.09g, 0.01mol) and potassium tert-butoxide (STB, 2.24g, 0.02mol), reacting at 100 ℃ for 8 hours, cooling to room temperature after the reaction is finished, slowly adding the reaction liquid into ice water, separating out solid, filtering to obtain a crude product, and purifying by dichloromethane/methanol (30:1) column chromatography to obtain off-white solid 16, 1.40g, 64.5%. ESI-MS [ M + H ]]⁺:m/z 221.11

Example 43

Synthesis of intermediate N- (4- ((2-chloropyridin-4-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-dicarboxamide (17)

The target compound 17 was prepared from the raw material 3 and the intermediate 16 as the starting materials in accordance with example 2, at a yield of 90.4%, ESI-MS [ M + H ]]⁺:m/z 426.09。

Example 44

Synthesis of 4- ((2-chloropyridin-4-yl) oxy) -3-fluoroaniline (18)

Starting from 14 and 15', the procedure for the preparation of intermediate 16 in example 41 was followed to give the title compound 18 in 92.0% yield. ESI-MS [ M + H ]]⁺:m/z 239.04

Example 45

Synthesis of N- (4- ((2-chloropyridin-4-yl) oxy) -3-fluorophenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-dicarboxamide (19)

Starting from 18 and 3, the procedure for the preparation of intermediate 17 in example 42 was followed to give the title compound 19 in 79% yield. ESI-MS [ M + H ]]⁺:m/z 444.12

Example 46

Synthesis of N- (4-fluorophenyl) -N- (4- ((2- ((3-morpholinophenyl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide (target Compound I-29)

The starting materials, M-morpholinoaniline (13, 0.23g, 1.29mmol), intermediate 17(0.50g, 1.18mmol), palladium acetate (0.006g, 0.029mmol) and (. + -.) -2,2 '-bis- (diphenylphosphino) -1,1' -binaphthyl (BINAP, 0.037g, 0.059mmol) were dissolved in 15mL of 1, 4-dioxane, stirred for 20 minutes, and then cesium carbonate (0.58g, 1.76mmol) was added, reflux reaction was carried out for 2 hours, after completion of the reaction, the temperature was lowered to room temperature, celite was filtered, the filtrate was concentrated, and the residue was purified by petroleum ether/ethyl acetate column chromatography (3:1) to give I-29 as a white solid, 0.33g, 50%, ESI-MS [ M + H ] +: M/z 221.11.

ESI-MS[M+H]⁺:m/z 568.14

¹H NMR(400MHz,DMSO-d₆)δppm:10.17(s,1H,CONH),10.04(s,1H,CONH),8.85(s,1H,NH),8.03(d,1H,J＝4.0Hz,ArH),7.72(d,2H,J＝8.0Hz,ArH),7.62～7.65(m,2H,ArH),7.13～7.19(m,5H,ArH),7.04(d,2H,J＝8.0Hz,ArH),6.46～6.49(m,1H,ArH),6.39(dd,1H,J₁＝1.6Hz,J₂＝4.0Hz,ArH),6.16(d,1H,J＝4.0Hz,ArH),3.72(t,4H,J＝4.0Hz,CH₂),3.02(t,4H,J＝4.0Hz,CH₂),1.46(d,1H,J＝4.0Hz,CH₂).

Example 47

Synthesis of N- (3-fluoro-4- ((2- ((3-morpholinophenyl) amino) pyrimidin-4-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-diamide (target Compound I-30)

The target compound I-30 was prepared from m-morpholinoaniline (13) and intermediate 19 by reference to the method for synthesizing the target compound I-29 in example 45 with a yield of 51.4%.

ESI-MS[M+H]⁺:m/z 586.24

¹H NMR(400MHz,DMSO-d₆)δppm:10.14(s,1H,CONH),10.07(s,1H,CONH),8.85(s,1H,NH),8.04(d,1H,J＝4.0Hz,ArH),7.74(d,2H,J＝8.0Hz,ArH),7.62～7.66(m,2H,ArH),7.11～7.15(m,5H,ArH),7.06(d,2H,J＝8.0Hz,ArH),6.47～6.52(m,1H,ArH),6.37(dd,1H,J₁＝1.6Hz,J₂＝4.0Hz,ArH),3.74(t,4H,J＝4.0Hz,CH₂),3.06(t,4H,J＝4.0Hz,CH₂),1.48(d,1H,J＝4.0Hz,CH₂).

Example 48

Compound in vitro inhibition activity assay for c-Met enzyme:

the MET enzyme (Cat:14-526) kit produced by Millipore is selected to test the inhibitory activity of the compound on c-Met, and the experimental operation is carried out according to the kit instruction. Firstly testing the inhibition percentage of the compound to the c-Met enzyme under the two concentrations of 10 mu M and 1 mu M, and selecting the compound with better enzyme inhibition activity to continue to carry out enzyme inhibition IC₅₀And (6) testing. The results are shown in Table 2.

Results of in vitro C-Met inhibitory Activity of the Compounds of Table 2

As can be seen from the above Table 2, the compounds of the present invention have strong inhibitory activity against c-Met enzyme, such as compounds I-2, I-7, I-16, I-22, I-24, I-25, I-27, I-28, I-30, I-32 and I-35, and the inhibitory activity against c-Met is equal to or better than that of the positive control drug Cabozantinib. Among them, the activity of the compound I-24 was 8.4 times that of the positive control Cabozantinib.

Example 49

Compounds tested for VEGFR2 and EGFR enzyme inhibitory activity in vitro:

partial compounds with better in vitro c-Met inhibition activity are selected and subjected to enzyme inhibition activity tests on VEGFR2 and EGFR in vitro. The VEGFR2enzyme (Cat: K2643) kit from Sigma and the EGFR (Cat: PV3872) kit from Invitrogen were used to carry out enzyme inhibition IC of the compounds on VEGFR2 and EGFR₅₀The test and experimental operation are carried out according to the kit instruction. The results are shown in Table 3.

Results of in vitro inhibitory Activity of the Compounds of Table 3 on VEGFR2 and EGFR

As can be seen from table 3 above, the tested compounds of the present invention showed good inhibitory activity against VEGFR2 and EGFR while inhibiting c-Met, and the inhibitory activity was comparable to or better than that of the positive control drug Cabozantinib. Wherein, the inhibitory activities of the compounds I-22, I-24, I-27 and I-28 on three target enzymes, namely c-Met, VEGFR2 and EGFR are all better than those of a positive control medicament, namely Cabozantinib.

Example 50

The in vitro tumor cell inhibiting activity test of the compound of the invention comprises the following steps:

the activity of the compound of the invention on human colorectal cancer cell line HCT116, human renal clear cell carcinoma skin metastatic cell line Caki-1, human pancreatic cancer cell line PANC-1, human liver cancer cell line HepG2 and human prostate cancer cell line PC-3 is determined, and IC thereof is₅₀The value was determined by the CCK-8 method (Cat # CK04-13, Dojindo) and Cabozantinib was selected as the control drug. The specific results are shown in Table 4 (unit: μ M):

TABLE 4 in vitro inhibitory Activity of Compounds on tumor cells

As can be seen from the above table 4, compared with the positive control drug Cabozantinib, the compound of the invention shows good in vitro anti-tumor cell proliferation activity on various tumor cells; the anti-tumor cell proliferation activity of part of the compounds is better than that of a positive control medicament Cabozantinib. The compounds I-22, I-24, I-27 and I-28 have higher inhibitory activity on a human liver cancer cell line HepG2, a human prostate cancer cell line PC-3, a human pancreatic cancer cell line PANC-1, a human renal clear cell carcinoma skin metastasis cell line Caki-1 and a human colorectal cancer cell line HCT116, are superior to a positive control drug Cabozantinib, have the activity improved by 4-7 times compared with the positive control drug Cabozantinib, and have the characteristics of broad spectrum and high efficiency.

Example 51

Test for in vitro inhibitory Activity of Compounds on Normal cells:

determination of the Activity of the Compounds of the invention on MRC-5 human embryonic Lung fibroblasts, IC₅₀The value was measured by the CCK-8 method (Cat # CK04-13, Dojindo). In-vitro inhibitory activity IC of normal cell strain by selecting Cabozantinib as positive control drug₅₀And (6) testing. The specific results are as follows (unit: μ M):

in vitro inhibitory Activity of Compounds of Table 5 and control drugs on Normal cells

As can be seen from table 5 above, the tested compound of the present invention has a weaker inhibitory activity on normal cells and lower toxic and side effects than the control drug Cabozantinib, which reveals that the substituted arylamino aromatic heterocyclic compound of the present invention has better selectivity in the inhibition of proliferation of tumor cells and normal cells, and indicates that the compound of the present invention has lower toxic and side effects when used as an antitumor drug compared with the existing drug Cabozantinib.

Example 52

Preliminary safety evaluation of compounds:

the test compounds I-22, I-24, I-27 and I-28 show that the Maximum Tolerated Dose (MTD) of the mice orally taken by intragastric administration once is above 720mg/kg, which is equivalent to the positive control drug Cabozantinib, the male and female properties are not obviously different, and the safety is good.

Example 53

In vitro preliminary toxic and side effect investigation:

the in vitro toxic and side effects of the compounds I-22, I-24, I-27 and I-28 are preliminarily examined through an hERG potassium ion channel inhibition test. Meanwhile, bacterial back-mutation experiments (Ames experiments) were performed.

Table 6 inhibition of hERG potassium channel by compounds and control drugs

The hERG experiment result shows that the inhibition activities of the test compounds I-22, I-24, I-27, I-28 and Cabozantinib on the hERG potassium ion channel are all larger than 30 mu M, which indicates that the potential cardiac toxicity is lower; ames experiments showed that none of the test compounds I-22, I-24, I-27, I-28 had a mutagenic effect on both the Salmonella typhimurium strain TA98 and TA 100.

Example 54

Evaluation of physicochemical Properties of the Compound:

the water solubility test of the preferred compounds of the present invention showed that the compounds of the present invention were comparable or superior in water solubility to the positive control.

Comparison of Water solubility of Compounds and control drugs in Table 7

The water solubility test result shows that: the water solubility of the test compounds I-22, I-24, I-27 and I-28 is improved compared with that of a positive control medicament Cabozantinib, wherein the water solubility of the compound I-27 is the best, is improved by nearly three times, reaches 44.6mg/mL, and has better development prospect.

Example 55

The preparation method comprises the following steps: mixing any one of the compounds I-1 to I-36 or its salt with sucrose and corn starch, moistening with water, stirring, drying, pulverizing, sieving, adding calcium stearate, mixing, and tabletting. Each tablet weighs 290mg, and the content of active ingredients is 100 mg.

Example 56

Injection preparation: 15mg of any one compound selected from I-1 to I-36 or a salt thereof

Water for injection 80mg

The preparation method comprises the following steps: dissolving any one of the compounds I-1 to I-36 or a salt thereof in water for injection, mixing well, filtering, and sub-packaging the obtained solution in ampoules under sterile conditions, wherein each ampoule contains 95mg of active ingredient and the content of active ingredient is 15mg per ampoule.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Claims

1. A substituted arylamine aromatic heterocyclic compound shown as a formula (I) and pharmaceutically acceptable salts thereof:

wherein, the specific compound is as follows,

I-36N- (4-fluorophenyl) -N- (4- ((2- ((2-oxo-2, 3,4, 5-tetrahydro-1H-benzo [ b ] azepin-7-yl) amino) pyrimidin-4-yl) oxy) phenyl) cyclopropane-1, 1-diamide.

2. The substituted arylamino aromatic heterocyclic compounds of formula (I) and pharmaceutically acceptable salts thereof as claimed in claim 1, wherein the salts include inorganic or organic acid salts of hydrochloride, hydrobromide, sulfate, phosphate, p-toluenesulfonic acid, methanesulfonate, acetate, tartrate, malate, maleate.

3. The pharmaceutically acceptable salt of the substituted arylamino aromatic heterocyclic compound of formula (I) as claimed in claim 2 wherein said salt is formed by an acid-base salt formation reaction of said substituted arylamino aromatic heterocyclic compound of formula (I) with an organic or inorganic acid.

4. A composition comprising a substituted arylamino aromatic heterocycle compound of any one of claims 1-3 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

5. The composition of claim 4, which is prepared as a tablet, capsule or injection.

6. A composition according to claim 4, wherein the substituted arylamino aromatic heterocycles or pharmaceutically acceptable salts thereof are present in an amount of from 0.1% to 99.5% by weight of the composition.

7. The use of the substituted arylamino aromatic heterocyclic compound or the pharmaceutically acceptable salt thereof in the preparation of the antitumor medicament is claimed in any one of 1-3.