CN113121429B

CN113121429B - C-Met kinase inhibitor and preparation method and application thereof

Info

Publication number: CN113121429B
Application number: CN202010044952.9A
Authority: CN
Inventors: 张贵民; 白文钦; 张君香
Original assignee: Lunan Pharmaceutical Group Corp
Current assignee: Lunan Pharmaceutical Group Corp
Priority date: 2020-01-15
Filing date: 2020-01-15
Publication date: 2024-04-26
Anticipated expiration: 2040-01-15
Also published as: CN113121429A

Abstract

The invention belongs to the technical field of biological medicines, and particularly provides a c-Met kinase inhibitor represented by a general formula I, wherein in-vitro antitumor activity screening experiments show that the compound has stronger inhibition activity on four cancer cells, namely human colon cancer cells (HT 29), human non-small cell lung cancer cells (A549), human large cell lung cancer cells (H460) and human gastric cancer cells (MKN-45), and has better clinical application prospect.

Description

C-Met kinase inhibitor and preparation method and application thereof

Technical Field

The invention belongs to the technical field of biological medicines, and particularly relates to a c-Met kinase inhibitor.

Background

Mesenchymal epithelial transforming factor (c-Met) is a tyrosine kinase receptor discovered in 1980 and is usually activated by binding to its natural ligand Hepatocyte Growth Factor (HGF), also known as the scatter factor (HGF/SF). HGF binding to c-Met induces a variety of complex signaling pathways that lead to cell proliferation, movement, migration and survival, but are likely to lead to the development of cancer when deregulated. Therefore, the c-Met signaling pathway gradually becomes an attractive target for targeted treatment of cancers, and research and development of novel antitumor drugs by inhibiting c-Met have received extensive attention from numerous researchers. In the current approach to targeting the c-Met signaling pathway, small molecule inhibitors that act directly on the ATP binding site of c-Met kinase are considered to be the most effective therapeutic agents. Over the past decade, many c-Met inhibitors have been reported, such as BMS777607 from Bai Mei Shi Guibao (Bristol-Myers Squibb), acylthioureas from Kirin Brewery (Kirin Brewery), c-Met inhibitors AM 7 from Ann corporation (Amgen), cabozantinib (Cabozantinib) and Foretinib. The structure-activity relationship shows that the inhibitor mainly consists of three parts, namely a quinoline skeleton (part A) which is easy to form hydrogen bond with c-Met kinase, an aryl fragment (part B) which can extend into a hydrophobic bag and an intermediate bridge chain, wherein the modification site of the inhibitor is mainly at the 7-position and the intermediate connecting part of the quinoline ring. However, it has been reported that certain class II c-Met inhibitors with VEGFR activity may lead to under-dosing problems of c-Met inhibition in clinical applications due to VEGFR related side effects. In addition, synthetic kinase inhibitors must be designed to have good kinase selectivity, as inhibition of a particular kinase would risk adverse reactions. Therefore, new c-Met inhibitors with good selectivity for VEGFR-2 and minimal side effects should be developed. In addition, with the use of c-Met kinase inhibitors in large quantities, secondary gene mutations or subcloning of tumor cells in the body result in the generation and expansion of drug resistant tumor cells containing the new mutated gene, ultimately resulting in failure of tumor therapy, i.e., drug resistance. Thus, there is an urgent need to study and develop c-Met kinase inhibitors having novel structures in order to find therapeutic methods for solving drug resistance.

Piperazine amide segment is an important pharmacophore, can form hydrogen bond interaction with a drug target, and is widely applied to the design of anticancer drugs. Based on the application of the fragment in the design of antitumor drugs, the research tries to take the piperazine amide fragment as the intermediate bridge chain part of the c-Met inhibitor so as to obtain the lead compound with better antitumor activity.

Disclosure of Invention

In view of the deficiencies of the prior art, one aspect of the present application is to provide a c-Met kinase inhibitor. Specifically, the present application provides a compound represented by formula I:

Wherein R is selected from phenyl; phenyl is optionally substituted with 0, 1, 2 or 3 substituents, each substituent independently selected from halogen, haloalkyl, alkyl or alkoxy.

Preferably, said R is selected from phenyl; phenyl is optionally substituted with 0 or 1 substituents; the substituent is independently selected from halogen, C1-C3 haloalkyl, C1-C3 alkane or C1-C3 alkoxy, and the halogen is selected from one of fluorine, chlorine and bromine.

Further preferred, the compound is selected from the following compounds or pharmaceutically acceptable salts thereof:

in a second aspect the present invention provides a process for the preparation of a compound of formula I, the process comprising the step of reacting a compound of formula III with a compound of formula II to obtain a compound of formula I:

wherein R is as defined above.

Preferably, the synthesis method specifically comprises the following steps: the compound of formula II reacts with the compound of formula III under the action of triphosgene to produce the compound of formula I.

Further preferably, the molar ratio of the compound of formula II to triphosgene is from 2 to 3:1; the organic solvent used in the reaction is a1, 2-dichloroethane/pyridine mixture.

The method further comprises the step of reducing the nitro group of the compound of formula IV to provide the compound of formula II:

preferably, the reducing agent used to reduce the compound of formula IV is selected from iron powder or zinc powder.

The process further comprises the step of reacting a compound of formula VI with a compound of formula V to obtain a compound of formula IV:

Preferably, the molar ratio of the compound of formula VI to the compound of formula V in the present reaction is from 1:1.0 to 1.5.

The process further comprises the step of reacting the compound of formula VII with a chlorinating agent to give the compound of formula VI:

Preferably, the chlorinating agent in the present reaction is selected from one of phosphorus oxychloride, phosphorus trichloride and phosphorus pentachloride.

The process further comprises the step of reacting a compound of formula IX with a compound of formula VIII in a basic environment to obtain a compound of formula VII:

Preferably, the base is selected from sodium methoxide or sodium ethoxide.

Further preferably, the molar ratio of the compound of formula IX to the base is from 1:1.0 to 1.2.

The following further details the preparation of the compounds of formula I of the present invention:

The compound of the formula II and triphosgene are dissolved in the mixed solution of 1, 2-dichloroethane/pyridine to react for 2 to 4 hours at the temperature of between 65 and 80 ℃, and then the compound of the formula III is added to continue the reaction.

Preferably, the molar ratio of the compound of formula II to triphosgene is from 2 to 3:1.

Preferably, the volume ratio of the 1, 2-dichloroethane to the pyridine is 20:0.5-3.

In one embodiment, the molar ratio of the compound of formula II to triphosgene is 2.5:1; the volume ratio of 1, 2-dichloroethane to pyridine was 20:1.

The preparation of the compound of formula II comprises the steps of: and (3) dissolving the compound shown in the formula IV in a proton solvent, heating, adding a saturated NH ₄ Cl solution and a reducing agent, and heating and refluxing for reaction.

Preferably, the protic solvent is selected from ethanol, water or a mixed solvent of both; the reducing agent is selected from iron powder or zinc powder.

In one embodiment, the volume ratio of ethanol to water is 9:1; the reducing agent is iron powder.

The preparation of the compound of formula IV comprises the steps of: the compound of the formula VI and the compound of the formula V are dissolved in an organic solvent and heated and refluxed for 15 to 32 hours.

Preferably, the organic solvent is selected from one of chlorobenzene, ethylene glycol dimethyl ether, paraxylene, diphenyl ether, 1, 4-dioxane, pyridine/1, 4-dioxane mixed solvent, NMP/1, 4-dioxane mixed solvent, DMF/pyridine mixed solvent.

Preferably, the molar ratio of the compound of formula VI to the compound of formula V is from 1:1.0 to 1.5; the reaction time is 28 to 32 hours.

In one embodiment, the molar ratio of the compound of formula VI to the compound of formula V is 1:1.2; the reaction time was 30 hours.

The preparation of the compound of formula VI comprises the steps of: the compound of formula VII and the chloro reagent are dissolved in a reaction solvent and heated for reflux reaction.

Preferably, the reaction solvent is POCl ₃ or acetonitrile.

Preferably, the molar ratio of the compound of formula VII to the chlorinating agent is from 1:15 to 28.

Preferably, the chlorinating agent is selected from one of phosphorus oxychloride, phosphorus trichloride and phosphorus pentachloride.

In one embodiment, the molar ratio of the compound of formula VII to the chlorinating agent is 1:21.5; the chlorinating agent is phosphorus oxychloride.

The preparation of the compound of formula VII comprises the steps of: the compound of formula IX and alkali are dissolved in organic solvent, and methylene dichloride solution of formula VIII is added with stirring to react at room temperature.

Preferably, the molar ratio of the compound of formula IX to the base is from 1:2.0 to 4.5.

Preferably, the molar ratio of the compound of formula IX to formula VIII is from 1:4 to 7.

Preferably, the organic solvent is selected from ethylene glycol dimethyl ether or methylene chloride.

In one embodiment, the base is sodium methoxide; the molar ratio of the compound of formula IX to sodium methoxide is 1:3; the molar ratio of the compound of formula IX to formula VIII is 1:5.

In a preferred embodiment, the process for the preparation of the compounds of formula I comprises the steps of: reacting a compound shown in a formula IX with a compound shown in a formula VIII under an alkaline condition to obtain a compound shown in a formula VII, reacting hydroxyl on the compound shown in the formula VII with a compound shown in a formula V after chlorination to obtain a compound shown in a formula IV, reducing nitro on the compound shown in the formula IV to obtain a compound shown in a formula II, and finally reacting the compound shown in the formula II with the compound shown in the formula III to obtain the compound shown in the formula I.

The synthetic route is as follows:

In a third aspect the present invention provides a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof.

In a fourth aspect, the present invention provides an application of a compound of formula I as an active ingredient in preparing an antitumor drug.

Preferably, the tumor is selected from: colon cancer, lung cancer, stomach cancer, liver cancer, breast cancer, prostate cancer, ovarian cancer or prostate cancer.

The in vitro anti-tumor activity screening result shows that the compound shown in the formula I has stronger inhibition activity on human colon cancer cells (HT-29), human large cell lung cancer cells (H460), human non-small cell lung cancer cells (A549) and human stomach cancer cells (MKN-45), and part of the compound has higher activity than that of a control drug Foretinib. Therefore, the compound can be used for preparing antitumor drugs. The c-Met kinase inhibitor has the advantages of novel structure, simple synthesis process, high product purity, strong inhibition effect on tumor cells and good application prospect.

Detailed Description

The advantages of the invention will now be further described by the following examples, which are given for illustrative purposes only and do not limit the scope of the invention, while variations and modifications apparent to those skilled in the art in light of the present disclosure are included within the scope of the invention.

Preparation of 6, 7-dimethoxy-4-hydroxyquinoline (VII):

Example 1:

2-amino-4, 5-dimethoxyacetophenone (IX) (5.85 g,30.0 mmol) and sodium methoxide (4.86 g,90.0 mmol) were dissolved in 50ml of dichloromethane at room temperature, and after magnetically stirring the solution, a solution of ethyl formate (VIII) (11.1 g,150.0 mmol) in dichloromethane (10 ml) was added dropwise thereto, and the mixture was reacted at room temperature for 8 hours. After completion of the TLC detection reaction, the reaction mixture was washed once with 80ml of saturated aqueous NaHCO ₃ and saturated aqueous NaCl, dried over anhydrous Na ₂SO₄, filtered, and the filtrate was transferred to a 250ml round bottom flask and concentrated under reduced pressure (35 ℃), finally a yellow solid was obtained, yield: 93.2%, HPLC purity: 98.03%. m.p.228-230 ℃; ESI-HRMS (m/z): 228.0721[ M+Na ] ⁺.

Example 2:

2-amino-4, 5-dimethoxyacetophenone (IX) (5.85 g,30.0 mmol) and sodium ethoxide (9.19 g,135.0 mmol) were dissolved in 50ml of ethylene glycol dimethyl ether at room temperature, and after magnetically stirring the solution, a solution of ethyl formate (VIII) (13.32 g,180.0 mmol) in dichloromethane (10 ml) was added dropwise thereto, and after that, the reaction was carried out at room temperature for 6 hours. After completion of the TLC detection reaction, the reaction was washed once with 80ml of saturated aqueous NaHCO ₃ and saturated aqueous NaCl, respectively, dried over anhydrous Na ₂SO₄, filtered, and the filtrate was transferred to a 250ml round bottom flask and concentrated under reduced pressure (35 ℃ C.) to give a yellow solid, 92.5% yield and 98.62% HPLC purity. m.p.228-230 ℃; ESI-HRMS (m/z): 228.0719[ M+Na ] ⁺.

Example 3:

2-amino-4, 5-dimethoxyacetophenone (IX) (5.85 g,30.0 mmol) and sodium methoxide (3.24 g,60.0 mmol) were dissolved in 50ml of dichloromethane at room temperature, and after magnetically stirring the solution, a solution of ethyl formate (VIII) (8.88 g,120.0 mmol) in dichloromethane (10 ml) was added dropwise thereto, and the mixture was reacted at room temperature for 7 hours. After completion of the TLC detection reaction, the reaction was washed once with 80ml of saturated aqueous NaHCO ₃ and saturated aqueous NaCl, respectively, dried over anhydrous Na ₂SO₄, filtered, and the filtrate was transferred to a 250ml round bottom flask and concentrated under reduced pressure (35 ℃ C.) to give a yellow solid, yield 91.7%, HPLC purity 98.83%. m.p.228-230 ℃; ESI-HRMS (m/z): 228.0720[ M+Na ] ⁺.

Example 4:

2-amino-4, 5-dimethoxyacetophenone (IX) (5.85 g,30.0 mmol) and sodium methoxide (9.72 g,180.0 mmol) were dissolved in 50ml of ethylene glycol dimethyl ether at room temperature, and after magnetically stirring the solution, a solution of ethyl formate (VIII) (15.54 g,210.0 mmol) in methylene chloride (10 ml) was added dropwise thereto and reacted at room temperature for 6 hours. After completion of the TLC detection reaction, the reaction was washed once with 80ml of saturated aqueous NaHCO ₃ and saturated aqueous NaCl, respectively, dried over anhydrous Na ₂SO₄, filtered, and the filtrate was transferred to a 250ml round bottom flask and concentrated under reduced pressure (35 ℃ C.) to give a yellow solid, yield 88.6% and HPLC purity 98.50%. m.p.228-230 ℃; ESI-HRMS (m/z): 228.0718[ M+Na ] ⁺.

Example 5:

2-amino-4, 5-dimethoxyacetophenone (IX) (5.85 g,30.0 mmol) and sodium methoxide (1.46 g,27.0 mmol) were dissolved in 50ml of dichloromethane at room temperature, and after magnetically stirring the solution, a solution of ethyl formate (VIII) (6.66 g,90.0 mmol) in dichloromethane (10 ml) was added dropwise thereto, and the mixture was reacted at room temperature for 8 hours. After completion of the TLC detection reaction, the reaction was washed once with 80ml of saturated aqueous NaHCO ₃ and saturated aqueous NaCl, respectively, dried over anhydrous Na ₂SO₄, filtered, and the filtrate was transferred to a 250ml round bottom flask and concentrated under reduced pressure (35 ℃ C.) to give a yellow solid, yield 82.3% and HPLC purity 97.78%. m.p.228-230 ℃; ESI-HRMS (m/z): 228.0722[ M+Na ] ⁺.

Example 6:

2-amino-4, 5-dimethoxyacetophenone (IX) (5.85 g,30.0 mmol) and sodium methoxide (4.86 g,90.0 mmol) were dissolved in 50ml of ethylene glycol dimethyl ether at room temperature, and after magnetically stirring the solution, a solution of ethyl formate (VIII) (22.2 g,300.0 mmol) in methylene chloride (20 ml) was added dropwise thereto, and the mixture was reacted at room temperature for 5 hours. After completion of the TLC detection reaction, the reaction was washed once with 80ml of saturated aqueous NaHCO ₃ and saturated aqueous NaCl, respectively, dried over anhydrous Na ₂SO₄, filtered, and the filtrate was transferred to a 250ml round bottom flask and concentrated under reduced pressure (35 ℃ C.) to give a yellow solid, 84.5% yield and 98.30% HPLC purity. m.p.228-230 ℃; ESI-HRMS (m/z): 228.0721[ M+Na ] ⁺.

Preparation of 6, 7-dimethoxy-4-chloroquinoline (VI):

Example 7:

VII (4.10 g,20.0 mmol) and POCl ₃ (40 ml,0.43 mol) were dissolved in dry acetonitrile (40 ml) and the flask was placed in an oil bath and heated to reflux for 4h. After completion of the TLC detection reaction, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. To the resulting residue was poured ice water (100 ml) and vigorously stirred, after which the pH was adjusted to 8 with concentrated NH ₄ OH, and the temperature was controlled to within 20 ℃. The mixture was extracted twice with dichloromethane (320 ml. Times.2). The combined organic phases were washed successively with saturated aqueous NaCl and water, dried over anhydrous Na ₂SO₄, filtered and concentrated to give a white solid with a yield of 68.3% and an HPLC purity of 98.11%. m.p.134-136 ℃; ESI-HRMS (m/z): 246.0413[ M+Na ] ⁺.

Example 8:

VII (4.10 g,20.0 mmol) and PCl ₃ (40 ml,0.460 mol) were dissolved in dry acetonitrile (40 ml) and the flask was placed in an oil bath and heated to reflux for 5h. After completion of the TLC detection reaction, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. To the resulting residue was poured ice water (100 ml) and vigorously stirred, after which the pH was adjusted to 8 with concentrated NH ₄ OH, and the temperature was controlled to within 20 ℃. The mixture was extracted twice with dichloromethane (320 ml. Times.2). The combined organic phases were washed successively with saturated aqueous NaCl and water, dried over anhydrous Na ₂SO₄, filtered and concentrated to a white solid, yield: 66.2%, HPLC purity: 98.06%. m.p.134-136 ℃; ESI-HRMS (m/z): 246.0415[ M+Na ] ⁺.

Example 9:

VII (4.10 g,20.0 mmol) and PCl ₅ (40 ml,0.300 mol) were dissolved in dry acetonitrile (40 ml) and the flask was placed in an oil bath and heated to reflux for 7h. After completion of the TLC detection reaction, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. To the resulting residue was poured ice water (100 ml) and vigorously stirred, after which the pH was adjusted to 8 with concentrated NH ₄ OH, and the temperature was controlled to within 20 ℃. The mixture was extracted twice with dichloromethane (320 ml. Times.2). The combined organic phases were washed successively with saturated aqueous NaCl and water, dried over anhydrous Na ₂SO₄, filtered and concentrated to a white solid, yield: 65.1%, HPLC purity: 98.97%. m.p.134-136 ℃; ESI-HRMS (m/z): 246.0412[ M+Na ] ⁺.

Example 10:

POCl ₃ (51 mL,560 mmol) was added to compound VII (4.10 g,20.0 mmol) and the mixture was heated to reflux for 3 hours. At the end of the reaction, cooled to room temperature, concentrated under reduced pressure, and the pale residue poured into ice water (100 mL) with vigorous stirring, the solution ph=8 was adjusted with solid NaHCO ₃, the temperature was controlled to within 20 ℃, and the mixture was extracted with CH ₂Cl₂ (3×50 mL). The organic phases were combined, washed once with saturated brine and water, dried over anhydrous Na ₂SO₄, filtered and concentrated under reduced pressure to give an off-white solid, yield: 74.3%, HPLC purity: 97.94%. m.p.134-136 ℃; ESI-HRMS (m/z): 246.0413[ M+Na ] ⁺.

Preparation of 6, 7-dimethoxy-4- (2-fluoro-4-nitrophenoxy) quinoline (IV):

example 11:

VI (4.46 g,20.0 mmol) and 2-fluoro-4-nitrophenol (V) (3.79 g,24.1 mmol) were dissolved in diphenyl ether (40 ml) and the mixture was heated to reflux for 30h. When the residual VI is less than 5%, the reaction can be stopped. And (5) cooling to room temperature, and distilling under reduced pressure. The residue was dissolved in dichloromethane (80 ml), washed twice with saturated K ₂CO₃ solution (20 ml. Times.2) and once with water (20 ml). The organic phase was dried over anhydrous Na ₂SO₄, filtered, and the solid obtained by concentration was recrystallized from absolute ethanol to give a pale yellow solid, yield: 80.4% of HPLC purity 98.90%. m.p.161-163 ℃; ESI-HRMS (m/z): 367.0809[ M+Na ] ⁺.

Example 12:

VI (4.46 g,20.0 mmol) and 2-fluoro-4-nitrophenol (V) (3.14 g,20.0 mmol) were dissolved in chlorobenzene (40 ml) and the mixture was heated to reflux for 32h. When the residual VI is less than 5%, the reaction can be stopped. And (5) cooling to room temperature, and distilling under reduced pressure. The residue was dissolved in dichloromethane (80 ml), washed twice with saturated K ₂CO₃ solution (20 ml. Times.2) and once with water (20 ml). The organic phase was dried over anhydrous Na ₂SO₄, filtered and the solid obtained by concentration was recrystallized from anhydrous ethanol to give a pale yellow solid with a yield of 78.9% and an HPLC purity of 98.09%. m.p.161-163 ℃; ESI-HRMS (m/z): 367.0811[ M+Na ] ⁺.

Example 13:

VI (4.46 g,20.0 mmol) and 2-fluoro-4-nitrophenol (V) (4.71 g,30.0 mmol) were dissolved in ethylene glycol dimethyl ether (40 ml) and the mixture was heated to reflux for 28h. When the residual VI is less than 5%, the reaction can be stopped. And (5) cooling to room temperature, and distilling under reduced pressure. The residue was dissolved in dichloromethane (80 ml), washed twice with saturated K ₂CO₃ solution (20 ml. Times.2) and once with water (20 ml). The organic phase was dried over anhydrous Na ₂SO₄, filtered and the solid obtained by concentration was recrystallized from anhydrous ethanol to give a pale yellow solid with a yield of 77.2% and an HPLC purity of 98.79%. m.p.161-163 ℃; ESI-HRMS (m/z): 367.0808[ M+Na ] ⁺.

Example 14:

VI (4.46 g,20.0 mmol) and 2-fluoro-4-nitrophenol (V) (5.34 g,34.0 mmol) were dissolved in paraxylene (40 ml) and a mixture of DMAP (4.89 g,40 mmol) was added and heated to reflux for 30h. When the residual VI is less than 5%, the reaction can be stopped. And (5) cooling to room temperature, and distilling under reduced pressure. The residue was dissolved in dichloromethane (80 ml), washed twice with saturated K ₂CO₃ solution (20 ml. Times.2) and once with water (20 ml). The organic phase was dried over anhydrous Na ₂SO₄, filtered, and the solid obtained by concentration was recrystallized from absolute ethanol to give a pale yellow solid, yield: 73.5%, HPLC purity: 98.01%. m.p.161-163 ℃; ESI-HRMS (m/z): 367.0811[ M+Na ] ⁺.

Example 15:

VI (4.46 g,20.0 mmol) and 2-fluoro-4-nitrophenol (V) (6.28 g,40.0 mmol) were dissolved in 1, 4-dioxane (40 ml) and the mixture was heated to reflux for 30h. When the residual VI is less than 5%, the reaction can be stopped. And (5) cooling to room temperature, and distilling under reduced pressure. The residue was dissolved in dichloromethane (80 ml), washed twice with saturated K ₂CO₃ solution (20 ml. Times.2) and once with water (20 ml). The organic phase was dried over anhydrous Na ₂SO₄, filtered, and the solid obtained by concentration was recrystallized from absolute ethanol to give a pale yellow solid, yield: 72.6% of HPLC purity of 98.78%. m.p.161-163 ℃; ESI-HRMS (m/z): 367.0809[ M+Na ] ⁺.

Example 16:

VI (4.46 g,20.0 mmol) and 2-fluoro-4-nitrophenol (V) (3.79 g,24.1 mmol) were dissolved in a mixture of pyridine (30 mL) and 1, 4-dioxane (15 mL), and the mixture was heated at reflux for 35h. When the residual VI is less than 5%, the reaction can be stopped. And (5) cooling to room temperature, and distilling under reduced pressure. The residue was dissolved in dichloromethane (80 ml), washed twice with saturated K ₂CO₃ solution (20 ml. Times.2) and once with water (20 ml). The organic phase was dried over anhydrous Na ₂SO₄, filtered, and the solid obtained by concentration was recrystallized from absolute ethanol to give a pale yellow solid, yield: 79.2% of HPLC purity of 98.50%. m.p.161-163 ℃; ESI-HRMS (m/z): 367.0812[ M+Na ] ⁺.

Example 17:

VI (4.46 g,20.0 mmol) and 2-fluoro-4-nitrophenol (V) (3.79 g,24.1 mmol) were dissolved in a mixture of NMP (40 mL) and 1, 4-dioxane (10 mL), and the mixture was heated at reflux for 30h. When the residual VI is less than 5%, the reaction can be stopped. And (5) cooling to room temperature, and distilling under reduced pressure. The residue was dissolved in dichloromethane (80 ml), washed twice with saturated K ₂CO₃ solution (20 ml. Times.2) and once with water (20 ml). The organic phase was dried over anhydrous Na ₂SO₄, filtered, and the solid obtained by concentration was recrystallized from absolute ethanol to give a pale yellow solid, yield: 76.8% of HPLC purity of 98.00%. m.p.161-163 ℃; ESI-HRMS (m/z): 367.0810[ M+Na ] ⁺.

Example 18:

VI (4.46 g,20.0 mmol) and 2-fluoro-4-nitrophenol (V) (3.79 g,24.1 mmol) were dissolved in a mixture of DMF (20 mL) and pyridine (20 mL) and the mixture was heated at reflux for 30h. When the residual VI is less than 5%, the reaction can be stopped. And (5) cooling to room temperature, and distilling under reduced pressure. The residue was dissolved in dichloromethane (80 ml), washed twice with saturated K ₂CO₃ solution (20 ml. Times.2) and once with water (20 ml). The organic phase was dried over anhydrous Na ₂SO₄, filtered, and the solid obtained by concentration was recrystallized from absolute ethanol to give a pale yellow solid, yield: 78.4% of HPLC purity 98.60%. m.p.161-163 ℃; ESI-HRMS (m/z): 367.0807[ M+Na ] ⁺.

Preparation of 6, 7-dimethoxy-4- (2-fluoro-4-aminophenoxy) quinoline (II)

Example 19:

IV (6.88 g,20.0 mmol) was dissolved in ethanol/water (90 ml,9:1 v/v) and saturated NH ₄ Cl solution (90 ml) and Fe powder (11.17 g,0.2 mol) were added to the mixture at reflux for 5h when the oil bath temperature was raised to 78 ℃. The reaction is immediately filtered after TLC detection is complete, filter cakes are leached by ethanol, filtrate is decompressed and concentrated to obtain white solid, and the yield is: 90.6%, HPLC purity: 99.06%. m.p.193-195 ℃; ESI-HRMS (m/z): 337.1105[ M+Na ] ⁺.

Example 20:

IV (6.88 g,20.0 mmol) was dissolved in ethanol (90 ml) and saturated NH ₄ Cl solution (90 ml) and Fe powder (11.17 g,0.2 mol) were added to the mixture at reflux for 5h when the temperature of the oil bath was raised to 78 ℃. The reaction is immediately filtered after TLC detection is complete, filter cakes are leached by ethanol, filtrate is decompressed and concentrated to obtain white solid, and the yield is: 88.2%, HPLC purity: 99.05%. m.p.193-195 ℃; ESI-HRMS (m/z): 337.1108[ M+Na ] ⁺.

Example 21:

IV (6.88 g,20.0 mmol) was dissolved in ethanol/water (90 ml,1:9 v/v) and saturated NH ₄ Cl solution (90 ml) and Zn powder (13.08 g,0.2 mol) were added to the mixture at reflux for 6h when the oil bath temperature was raised to 78 ℃. The reaction is immediately filtered after TLC detection is complete, filter cakes are leached by ethanol, filtrate is decompressed and concentrated to obtain white solid, and the yield is: 89.6%, HPLC purity: 99.16%. m.p.193-195 ℃; ESI-HRMS (m/z): 337.1109[ M+Na ] ⁺.

Preparation of Compound I:

Example 22 preparation of target compound I-a:

II (1.57 g,5.0 mmol) was dissolved with triphosgene (0.59 g,2.0 mmol) using a mixed solution of 1, 2-dichloroethane (20 ml)/pyridine (1 ml), the mixed solution was reacted at 75℃for 3 hours, then N-phenylpiperazine (5.0 mmol) was added for further reaction, after TLC detection of the reaction was completed, the mixed solution was concentrated under reduced pressure, and the product was purified by column chromatography to give a white solid to give the objective compound I-a, melting point 210-211 ℃. Yield: 88.2%, HPLC purity: 99.86%.

¹H NMR(400Hz,CDCl₃)δ:8.89(s,1H,-NH-),8.12(d,J＝8.0Hz,1H,-N＝CH-CH＝),7.58(t,J＝16.0Hz,2H,Ar-H),7.30(s,1H,Ar-H),7.21(t,J＝8.0Hz,2H,Ar-H),7.15(t,J＝4.0Hz,1H,Ar-H),6.94-6.80(m,4H,Ar-H),3.92(s,3H,-OCH₃),3.83(s,3H,-OCH₃),3.52(m,4H,-CO-N-[CH₂-CH₂]₂-N-Ph),3.29(m,4H,-CO-N-[CH2-CH₂]₂-N-Ph);¹³C NMR(100MHz,CDCl₃)δ:159.4,157.6,155.3,151.0,150.9,150.6,149.6,146.0,136.7,133.5,129.6(2C),123.1,121.9,118.2,114.5,114.3(2C),111.1,107.9,106.2,100.7,56.1(2C),52.8(2C),51.6(2C).

ESI-HRMS(m/z):525.1015[M+Na]⁺。

EXAMPLE 23 preparation of the target Compound I-b (Compound II, 2)

II (1.26 g,4.0 mmol) and triphosgene (0.59 g,2.0 mmol) were dissolved in a mixed solution of 1, 2-dichloroethane (20 ml)/pyridine (3 ml), the mixed solution was reacted at 80℃for 2 hours, then 1- (4-chlorophenyl) piperazine (5.0 mmol) was added for further reaction, after TLC detection, the mixed solution was concentrated under reduced pressure, and the product was purified by column chromatography to give a white solid, to give the objective compound I-b, melting point 159-161℃yield: 87.3% and 99.81% HPLC purity.

¹H NMR(400Hz,CDCl₃)δ:(400Hz,CDCl₃)δ:8.91(s,1H,-NH-),8.10(d,J＝8.0Hz,1H,-N＝CH-CH＝),7.58(t,J＝8.0Hz,2H,Ar-H),7.30(s,1H,Ar-H),7.21(t,J＝4.0Hz,2H,Ar-H),7.12(t,J＝8.0Hz,1H,Ar-H),6.89-6.72(t,3H,Ar-H),3.92(s,3H,-OCH₃),3.83(s,3H,-OCH₃),3.52(m,4H,-CO-N-[CH₂-CH₂]₂-N-Ph),3.29(m,4H,-CO-N-[CH₂-CH₂]₂-N-Ph);¹³C NMR(100MHz,CDCl₃)δ:159.3,157.7,151.3,155.1,150.7,150.6,147.7,146.2,136.9,133.4,129.7(2C),127.2,123.2,118.4,115.7(2C),114.6,111.5,108.0,106.0,101.1,56.2(2C),53.1(2C),51.7(2C).

ESI-HRMS(m/z):559.1507[M+Na]⁺。

EXAMPLE 24 preparation of the target Compound I-c (Compound II, 3)

II (1.88 g,6.0 mmol) was dissolved with triphosgene (0.59 g,2.0 mmol) and reacted with a mixed solution of 1, 2-dichloroethane (20 ml)/pyridine (0.5 ml) at 65℃for 4h, then 1- (4-fluorophenyl) piperazine (5.0 mmol) was added to continue the reaction, after TLC detection was completed, the mixed solution was concentrated under reduced pressure, and the product was purified by column chromatography to give the objective compound I-c, melting point 308-310℃yield 86.8%, HPLC purity 99.32%.

¹H NMR(400Hz,CDCl₃)δ:8.90(s,1H,-NH-),8.12(d,J＝8.0Hz,1H,-N＝CH-CH＝),7.59(t,1H,J＝8.0Hz,Ar-H),7.58(t,J＝4.0Hz,1H,Ar-H),7.30(s,1H,Ar-H),7.15(t,J＝8.0Hz,1H,Ar-H),7.07(t,J＝8.0Hz,3H,Ar-H),6.89-6.71(m,3H,Ar-H),3.92(s,3H,-OCH₃),3.83(s,3H,-OCH₃),3.52(m,4H,-CO-N-[CH₂-CH₂]₂-N-Ph),3.29(m,4H,-CO-N-[CH₂-CH₂]₂-N-Ph);¹³C NMR(100MHz,CDCl₃)δ:159.5,157.4,156.8,154.8,150.9,150.6,150.3,146.2,145.2,136.9,133.4,123.1,118.2,116.4(2C),115.9(2C),114.5,111.1,107.9,106.2,100.7,56.1(2C),52.8(2C),51.6(2C).

ESI-HRMS(m/z):543.0915[M+Na]⁺。

EXAMPLE 25 preparation of the target Compound I-d

II (1.57 g,5.0 mmol) was dissolved with triphosgene (0.59 g,2.0 mmol) and reacted with a mixed solution of 1, 2-dichloroethane (20 ml)/pyridine (0.3 ml) at 60℃for 5h, then 1- (4-methoxyphenyl) piperazine (5.0 mmol) was added to continue the reaction, after TLC detection the reaction was completed, the mixed solution was concentrated under reduced pressure, and the product was purified by column chromatography to give the objective compound I-d, melting point 184-186℃yield 83.6% and HPLC purity 98.31%.

¹H NMR(400Hz,CDCl₃)δ:8.90(s,1H,-NH-),8.10(d,J＝8.0Hz,1H,-N＝CH-CH＝),7.59-7.58(m 2H,Ar-H),7.30(s,1H,Ar-H),7.15(t,J＝8.0Hz,1H,Ar-H),7.07(t,J＝4.0Hz,1H,Ar-H),6.89-6.65(m,5H,Ar-H),3.92(s,3H,-OCH₃),3.83-3.81(s,6H,-OCH₃),3.52(m,4H,-CO-N-[CH₂-CH₂]₂-N-Ph),3.29(m,4H,-CO-N-[CH₂-CH₂]₂-N-Ph);¹³C NMR(100MHz,CDCl₃)δ:159.3,157.7,155.1,152.8,151.3,150.7,150.6,146.3,146.1,137.0,133.1,123.3,117.9,115.3(2C),115.1(2C),114.5,110.8,107.6,106.3,100.5,56.4(2C),55.5,53.1(2C),52.0(2C).

ESI-HRMS(m/z):555.2019[M+Na]⁺。

EXAMPLE 26 preparation of the target Compound I-e

II (1.57 g,5.0 mmol) was dissolved with triphosgene (0.44 g,1.5 mmol) and reacted with a mixed solution of 1, 2-dichloroethane (20 ml)/pyridine (5 ml) at 80℃for 1h, followed by addition of 1- (4-methylphenyl) piperazine (5.0 mmol) for further reaction, after completion of TLC detection, the mixed solution was concentrated under reduced pressure, and the product was purified by column chromatography to give the objective compound I-e in a yield of 82.2% and an HPLC purity of 97.89%.

¹H NMR(400Hz,CDCl₃)δ:8.90(s,1H,-NH-),8.10(d,J＝8.0Hz,1H,-N＝CH-CH＝),7.59-7.58(t,J＝8.0Hz,2H,Ar-H),7.30-7.29(m,3H,Ar-H),7.15(t,J＝4.0Hz,1H,Ar-H),7.07(t,J＝8.0Hz,1H,Ar-H),6.89(t,J＝8.0Hz,1H,Ar-H),6.61(t,J＝16.0Hz,2H,Ar-H),3.92(s,3H,-OCH₃),3.83(s,3H,-OCH₃),3.52(m,4H,-CO-N-[CH₂-CH₂]₂-N-Ph),3.29(m,4H,-CO-N-[CH₂-CH₂]₂-N-Ph);¹³C NMR(100MHz,CDCl₃)δ:159.5,158.0,154.8,152.9,151.5,150.7,150.3,146.2,137.3,133.6,126.2(3C),125.1,123.1,117.2,115.5,113.3(2C),111.5,107.0,106.2,101.7,56.1(2C),52.0(2C),50.6(2C).

ESI-HRMS(m/z):593.2176[M+Na]⁺。

EXAMPLE 27 preparation of the target Compounds I-f

II (1.57 g,5.0 mmol) was dissolved with triphosgene (1.18 g,4 mmol) and the mixture was reacted with 1, 2-dichloroethane (20 ml)/pyridine (3 ml) at 75℃for 3h, then 1- (4-trifluoromethylphenyl) piperazine (5.0 mmol) was added to continue the reaction, after TLC detection the reaction was completed, the mixture was concentrated under reduced pressure, and the product was purified by column chromatography to give the objective compound I-f, melting point 182-183℃yield: 83.7%, HPLC purity: 97.37%.

ESI-HRMS(m/z):593.1809[M+Na]⁺。

Example 28: synthesis of target Compound I-g

The same as in example 22, but 1- (3-methoxyphenyl) piperazine was used instead of N-phenylpiperazine. The detection data of the reaction products are as follows: white solid, melting point 213-215 ℃, yield 82.9%, HPLC purity: 97.63%.

¹H NMR(400Hz,CDCl₃)δ:8.90(s,1H,-NH-),8.10(d,J＝8.0Hz,1H,-N＝CH-CH＝),7.59-7.58(t,J＝8.0Hz,2H,Ar-H),7.30(s,1H,Ar-H),7.15-7.07(m,3H,Ar-H),6.89(t,J＝8.0Hz,1H,Ar-H),6.62(t,J＝16.0Hz,1H,Ar-H),6.38(d,1H,J＝8.0Hz,Ar-H),6.17(t,J＝4.0Hz,1H,Ar-H),3.92(s,3H,-OCH₃),3.83(s,3H,-OCH₃),3.74(s,3H,-OCH₃),3.52(m,4H,-CO-N-[CH₂-CH₂]₂-N-Ph),3.29(m,4H,-CO-N-[CH₂-CH₂]₂-N-Ph);¹³C NMR(100MHz,CDCl₃)δ:161.5,159.3,158.7,154.1,151.9,150.7,150.0,146.2,136.0,135.4,131.6,124.1,118.0,114.0,111.6,111.3,106.9,106.1,105.7,100.0,97.0,57.1(2C),55.9,51.8(2C),51.5(2C).

ESI-HRMS(m/z):555.2024[M+Na]⁺。

EXAMPLE 29 Synthesis of target Compound I-h

The same as in example 22, but 1- (3-chlorophenyl) piperazine was used instead of N-phenylpiperazine. The detection data of the reaction products are as follows: white solid, melting point 219-221 ℃, yield 83.2%, HPLC purity: 98.12%.

¹H NMR(400Hz,CDCl₃)δ:8.91(s,1H,-NH-),8.09(d,J＝8.0Hz,1H,-N＝CH-CH＝),7.59-7.58(t,J＝8.0Hz,2H,Ar-H),7.30(s,1H,Ar-H),7.15-7.06(m,3H,Ar-H),6.95-6.82(t,J＝4.0Hz,3H,Ar-H),6.66(t,J＝8.0Hz,1H,Ar-H),3.92(s,3H,-OCH₃),3.83(s,3H,-OCH₃),3.52(m,4H,-CO-N-[CH₂-CH₂]₂-N-Ph),3.29(m,4H,-CO-N-[CH₂-CH₂]₂-N-Ph);¹³C NMR(100MHz,CDCl₃)δ:163.8,159.0,157.7,155.1,151.3,151.2,150.7,150.6,145.2,136.9,133.4,130.2,123.7,118.2,114.5,110.1,110.0,108.2,107.5,104.2,101.8,100.1,56.9(2C),52.9(2C),52.6(2C).

ESI-HRMS(m/z):559.1509[M+Na]⁺

EXAMPLE 30 Synthesis of target Compound I-I

The same as in example 22, but 1- (2-methoxyphenyl) piperazine was used instead of N-phenylpiperazine. The detection data of the reaction products are as follows: white solid, melting point 211-213 ℃, yield 84.6%, HPLC purity: 98.07%.

¹H NMR(400Hz,CDCl₃)δ:8.92(s,1H,-NH-),8.11(d,J＝8.0Hz,1H,-N＝CH-CH＝),7.59-7.58(m,2H,Ar-H),7.30(s,1H,Ar-H),7.15(t,J＝8.0Hz,1H,Ar-H),7.07(t,J＝4.0Hz,1H,Ar-H),6.89-6.70(m,5H,Ar-H),3.92(s,3H,-OCH₃),3.86(s,3H,-OCH₃),3.83(s,3H,-OCH₃),3.52(m,4H,-CO-N-[CH₂-CH₂]₂-N-Ph),3.29(m,4H,-CO-N-[CH₂-CH₂]₂-N-Ph);¹³C NMR(100MHz,CDCl₃)δ:162.2,159.6,155.7,155.1,151.0,150.7,149.6,146.2,141.9,137.2,133.8,125.1,123.0,122.8,120.9,118.2,114.5,113.5,112.1,110.3,107.3,106.2,101.7,56.1(2C),56.0,52.1(2C),51.1(2C).

ESI-HRMS(m/z):555.2023[M+Na]⁺。

EXAMPLE 31 Synthesis of target Compound I-j

The same as in example 22, but 1- (2-chlorophenyl) piperazine was used instead of N-phenylpiperazine. The detection data of the reaction products are as follows: white solid, melting point 189-201 ℃, yield 81.9%, HPLC purity: 97.68%.

¹H NMR(400Hz,CDCl₃)δ:8.90(s,1H,-NH-),8.10(d,J＝8.0Hz,1H,-N＝CH-CH＝),7.59-7.58(m,2H,Ar-H),7.35(dd,J＝5.9Hz,10.1Hz,1H,Ar-H),7.30(s,1H,Ar-H),7.07-7.03(t,J＝16.0Hz,2H,Ar-H),6.89(t,J＝8.0Hz,1H,Ar-H),6.77(t,J＝8.0Hz,1H,Ar-H),6.67(t,J＝4.0Hz,1H,Ar-H),3.92(s,3H,-OCH₃),3.83(s,3H,-OCH₃),3.52(m,4H,-CO-N-[CH₂-CH₂]₂-N-Ph),3.29(m,4H,-CO-N-[CH₂-CH₂]₂-N-Ph);¹³C NMR(100MHz,CDCl₃)δ:159.8,157.0,156.1,151.3,150.8,150.7,150.4,146.5,136.6,134.4,131.8,129.5,127.3,125.2,123.5,123.0,119.2,116.5,112.1,107.1,106.3,102.7,56.1(2C),52.6(2C),51.7(2C).

ESI-HRMS(m/z):559.1576[M+Na]⁺。

Test method and results for cytotoxic Activity of Compounds I-a to I-j

In order to study the capability of the synthesized target compound to inhibit the proliferation of tumor cells in the experiment, the in vitro cytotoxicity of the compound to four tumor cells, namely human colon cancer cells (HT-29), human non-small cell lung cancer cells (A549), human large cell lung cancer cells (H460) and human gastric cancer cells (MKN-45), is measured, and Foretinib is used as a positive control. The assay used was the standard MTT assay.

The experimental method specifically comprises the following steps:

The cell cryopreservation tube was removed from liquid nitrogen, rapidly thawed at 39 ℃ and transferred to a 15mL centrifuge tube, 10mL of 10% fbs-containing medium was added, centrifuged for 5min (1000 rpm), the medium was removed, 10% fbs-containing medium and diabody-containing medium was added again, and transferred to a flask for culturing. Taking cells in logarithmic growth phase, removing culture solution in a culture flask, washing the cells once by PBS, carrying out pancreatin digestion and centrifugation collection, re-suspending the cells by using a culture medium containing 10% fetal bovine serum, counting and adjusting the cells to a proper concentration (the cell density is 5 multiplied by 104/mL, the cell activity is more than 90%), and adding the cell suspension into a 96-micro-well plate with 100 mu L per well. The target compounds were diluted with DMSO to 20. Mu.L solution, and the test target compounds were diluted with DMSO in a 3-fold gradient. 5. Mu.L of the diluted compound solution was added to 495. Mu.L of a medium containing 10% FBS, respectively, to prepare test compounds. 100. Mu.L of the solution containing the compound to be tested was added to the corresponding wells of a 96-well plate and cultured in a carbon dioxide cell incubator for 72 hours. The medium was removed, 150. Mu.L of 0.3 mg/mL ^-1 MTT working solution (0.002. Mu.g/mL ^-1 PMS) was added to each well, and the mixture was placed in a carbon dioxide incubator for 2 hours. The 96-well plate was shaken in an oscillator for 5min, and the absorbance A450 (450 nm) was read with an ELISA reader. All experiments were performed in 3 parallel groups or in triplicate. Finally, the median inhibitory concentration (IC ₅₀) of the test compound is calculated.

The results of in vitro toxicity tests of the compounds I-a to I-j on proliferation inhibition of four cancer cells, namely human colon cancer cells (HT 29), human non-small cell lung cancer cells (A549), human large cell lung cancer cells (H460) and human gastric cancer cells (MKN-45), are shown in Table 1.

TABLE 1

Note that: (1) screening method: MTT method; (2) action time: 72 hours. (3) P < 0.05.

In vitro experiments show that the compounds I-a to I-j have stronger inhibition activities on four cancer cells, namely human colon cancer cells (HT 29), human non-small cell lung cancer cells (A549), human large cell lung cancer cells (H460), and human gastric cancer cells (MKN-45), and the activity of the compounds is equivalent to that of a clinical drug Foretinib, wherein the inhibition activity of the compounds I-H on the human non-small cell lung cancer cells (A549) and the human large cell lung cancer cells (H460) is obviously better than that of a control drug, and the inhibition activity of the compounds I-j on the human colon cancer cells (HT 29), the human large cell lung cancer cells (H460) and the human gastric cancer cells (MKN-45) is obviously better than that of the control drug, so that the compounds have good application prospect.

Therefore, the compound prepared by the invention can be used for preparing antitumor drugs, and the compound has the advantages of simple synthesis method, low-cost and easily obtained raw materials and high product purity.

Claims

1. A compound represented by formula I or a pharmaceutically acceptable salt thereof:

，

wherein R is selected from phenyl; phenyl is optionally substituted with 0,1, 2 or 3 substituents, each substituent independently selected from halogen, C1-C3 haloalkyl, C1-C3 alkyl or C1-C3 alkoxy.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R is selected from phenyl; phenyl optionally substituted with 0 or 1 substituents; the substituents are independently selected from halogen, C1-C3 haloalkyl, C1-C3 alkanyl or C1-C3 alkoxy.

3. A compound according to claim 2, or a pharmaceutically acceptable salt thereof, selected from the following compounds:

。

4. A process for the preparation of a compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, which comprises the step of reacting a compound of formula III with a compound of formula II to obtain a compound of formula I:

，

wherein R is as defined in any one of claims 1 to 3; the synthesis method specifically comprises the following steps: the compound of formula II reacts with the compound of formula III under the action of triphosgene to produce the compound of formula I.

5. The method of claim 4, comprising the step of reducing the nitro group of the compound of formula IV to provide the compound of formula II:

。

6. The method of claim 5, comprising the step of reacting a compound of formula VI with a compound of formula V to provide a compound of formula IV:

。

7. The method of claim 6, comprising the step of reacting the compound of formula VII with a chlorinating agent to provide the compound of formula VI:

。

8. The process of claim 7, comprising the step of reacting a compound of formula ix with a compound of formula VIII in a basic environment to provide a compound of formula VII:

。

9. A pharmaceutical composition comprising a compound of any one of claims 1-3, or a pharmaceutically acceptable salt thereof.

10. Use of a compound of claim 1 or a pharmaceutically acceptable salt thereof in the manufacture of an antitumor medicament.