CN105646448B - Pyridine compounds and their and application thereof - Google Patents

Abstract

The present invention relates to pyridine derivatives represented by general formula I and their pharmaceutically acceptable salts, hydrates or prodrugs, wherein the substituents Y, Ar, R ₁ and R ₂ have the meanings given in the specification. The present invention also relates to the compounds of general formula I which have the effect of inhibiting c-Kit, RET and FLT3 kinases, and also relates to the preparation of such compounds and their pharmaceutically acceptable salts, hydrates or prodrugs in the preparation of treatment due to c-Kit, RET and FLT3 kinases. Use in medicines for diseases caused by abnormally high expression of kinases such as RET and FLT3, especially in the preparation of medicines for treating and/or preventing cancer.

Description

Pyridine compounds and their uses

technical field

The present invention relates to novel pyridine compounds and pharmaceutically acceptable salts, hydrates or prodrugs thereof, their preparation methods and pharmaceutical compositions containing the compounds. The present invention also relates to the use of such compounds and their pharmaceutically acceptable salts, hydrates or their prodrugs in preparing medicines for treating diseases caused by abnormally high expression of tyrosine kinases such as c-Kit, RET and FLT3, Especially in the preparation of medicaments for the treatment and/or prevention of cancer.

Background technique

Malignant tumor is a disease that seriously endangers human life and health. With changes in external factors such as environmental pollution, the number of cancer cases worldwide is increasing year by year. It is particularly important to develop safer and more effective tumor treatment drugs.

Protein Kinase (PK) is an enzyme that catalyzes the phosphorylation of hydroxyl groups on tyrosine, serine and threonine residues of proteins through the transfer of terminal phosphate of ATP, and participates in the regulation of cellular physiological activities through signal transduction pathways . Tyrosine kinases are the most important class of protein kinases. When activated, they can gradually amplify biological signals through phosphorylation, and finally transmit them to the nucleus, affecting various cellular responses such as cell division. (proliferation), cell differentiation, apoptosis, and metabolism.

At present, a large number of studies have shown that abnormal activation or mutation of tyrosine kinases can lead to specific biological signal disturbances, accelerated cell proliferation, and then lead to the occurrence of proliferative diseases. A variety of tyrosine kinases, including VEGFR, EGFR, PDGFR, FGFR, RET, FLT3, c-Kit, CDK, etc., have been used as important therapeutic targets in the research and development of anti-tumor drugs, and have achieved important results. Several drugs are on the market, such as sorafenib, gefitinib, etc.

Sorafenib is a multi-target oral inhibitor developed by Bayer. It has significant inhibitory effects on various tyrosine kinases such as VEGFR, PDGFR, RET, FLT3, c-Kit, etc. Amino acid kinase Raf, and has inhibitory activity on the proliferation of various tumor cells.

The inventors have designed and synthesized a series of new pyridine derivatives on the basis of the references. The screening of pharmacological activities shows that the compounds involved in the present invention have significant effects on tyrosine kinases such as c-Kit, RET and FLT3. It has inhibitory activity and has an inhibitory effect on the proliferation of tumor cells.

SUMMARY OF THE INVENTION

The present invention relates to pyridine compounds represented by general formula I and their pharmaceutically acceptable salts, hydrates or prodrugs,

in,

Y is H, halogen;

Ar is C ₆ -C ₁₀ aryl, and Ar is optionally substituted with 1-3 identical or different R ₃ ;

R ₃ is H, hydroxy, halogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy, halogenated (C ₁ -C ₆ )alkyl or (C ₁ -C ₆ )alkyl ) alkoxy;

R ₁ and R ₂ are the same or different, and are independently selected from (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy;

Or R ₁ and R ₂ together with the nitrogen atom to which they are attached form a 5-6 membered saturated heterocyclic group, the saturated heterocyclic group optionally contains 1-4 in addition to the nitrogen atom attached to R ₁ and R ₂ a heteroatom selected from N, O and S, the saturated heterocyclic group is optionally substituted with 1-3 identical or different R ₄ ;

R ₄ is H, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy, halogen, hydroxyl;

n is an integer between 1 and 3 (1, 2 or 3), preferably 2;

represents where the substituent is attached.

The present invention preferably relates to the pyridine compounds represented by the general formula I and their pharmaceutically acceptable salts, hydrates or prodrugs, wherein,

Y is H, F;

Ar is C ₆ -C ₁₀ aryl, and Ar is optionally substituted with 1-3 identical or different R ₃ ;

R ₃ is H, hydroxy, halogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy, halogenated (C ₁ -C ₆ )alkyl or (C ₁ -C ₆ )alkyl ) alkoxy;

R ₁ and R ₂ are the same or different, and are independently selected from (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy;

Or R ₁ and R ₂ together with the nitrogen atom to which they are attached form a 5-6 membered saturated heterocyclic group, the saturated heterocyclic group optionally contains 1-4 in addition to the nitrogen atom attached to R ₁ and R ₂ a heteroatom selected from N, O and S, the saturated heterocyclic group is optionally substituted with 1-3 identical or different R ₄ ;

R ₄ is H, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy, halogen, hydroxyl;

n is an integer between 1-3 (1, 2 or 3);

The present invention preferably relates to the pyridine compounds represented by the general formula I and their pharmaceutically acceptable salts, hydrates or prodrugs, wherein,

Y is H, F;

Ar is phenyl, oxazolyl and pyrazolyl, and Ar is optionally substituted with 1-3 identical or different R ₃ ;

R ₃ is H, hydroxy, halogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy, halogenated (C ₁ -C ₆ )alkyl or (C ₁ -C ₆ )alkyl ) alkoxy;

R ₁ and R ₂ are the same or different, and are independently selected from (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy;

Or R ₁ and R ₂ together with the nitrogen atom to which they are attached form a 5-6 membered saturated heterocyclic group, the saturated heterocyclic group optionally contains 1-4 in addition to the nitrogen atom attached to R ₁ and R ₂ a heteroatom selected from N, O and S, the saturated heterocyclic group is optionally substituted with 1-3 identical or different R ₄ ;

R ₄ is H, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy, halogen, hydroxyl;

n is an integer between 1-3 (1, 2 or 3);

represents where the substituent is attached.

The present invention preferably relates to the pyridine compounds represented by the general formula I and their pharmaceutically acceptable salts, hydrates or prodrugs, wherein,

Y is H, F;

Ar is phenyl, and Ar is optionally substituted with 1-3 identical or different R ₃ ;

R ₃ is H, halogen, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy, halogenated (C ₁ -C ₆ )alkyl or (C ₁ -C ₆ )alkane Oxygen;

R ₁ and R ₂ are the same or different, and are independently selected from (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy;

Or R ₁ and R ₂ together with the nitrogen atom to which they are attached form a 5-6 membered saturated heterocyclic group, the saturated heterocyclic group optionally contains 1-4 in addition to the nitrogen atom attached to R ₁ and R ₂ a heteroatom selected from N, O and S, the saturated heterocyclic group is optionally substituted with 1-3 identical or different R ₄ ;

R ₄ is H, (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy, halogen, hydroxyl;

n is an integer between 1-3 (1, 2 or 3);

represents where the substituent is attached.

The present invention preferably relates to the pyridine compounds represented by the general formula I and their pharmaceutically acceptable salts, hydrates or prodrugs, wherein,

Y is H, F;

Ar is phenyl, and Ar is optionally substituted with 1-3 identical or different R ₃ ;

R ₃ is H, halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )alkoxy, halogenated (C ₁ -C ₄ )alkyl;

R ₁ and R ₂ are the same or different, and are independently selected from (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )alkoxy;

or R1 and R2 together with the nitrogen atom to which they are attached form ₄ -morpholinyl, ₁ -piperidinyl, 4-methyl-1-piperazinyl, 1-piperazinyl, 4-methyl- 1-Piperidinyl, 1-pyrrolidinyl, 4-thiomorpholinyl;

n is an integer between 1-3 (1, 2 or 3);

represents where the substituent is attached.

The present invention preferably relates to the pyridine compounds represented by the general formula I and their pharmaceutically acceptable salts, hydrates or prodrugs, wherein,

Y is H, F;

Ar is phenyl, and Ar is optionally substituted with 1-3 identical or different R ₃ ;

R ₃ is halogen, tert-butyl, trifluoromethyl;

R ₁ and R ₂ are the same or different, and are independently selected from (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )alkoxy;

n is an integer between 1-3 (1, 2 or 3);

represents where the substituent is attached.

The present invention preferably relates to the pyridine compounds represented by the general formula I and their pharmaceutically acceptable salts, hydrates or prodrugs, wherein,

Y is H, F;

Ar is phenyl, and Ar is optionally substituted with 1-3 identical or different R ₃ ;

R ₃ is halogen, tert-butyl, trifluoromethyl;

R ₁ and R ₂ are the same or different and are independently selected from (C ₁ -C ₄ )alkyl;

n is an integer between 1-3;

The present invention particularly preferably relates to the pyridine compounds represented by the general formula I and their pharmaceutically acceptable salts, hydrates or prodrugs, wherein,

Y is H, F;

Ar is phenyl, and Ar is optionally substituted with 1-3 identical or different R ₃ ;

R ₃ is halogen, tert-butyl, trifluoromethyl;

R ₁ and R ₂ are the same or different, and are independently selected from methyl or ethyl;

n is an integer between 1-3;

The present invention particularly preferably relates to the pyridine compounds represented by the general formula I and their pharmaceutically acceptable salts, hydrates or prodrugs, wherein,

Y is H;

Ar is phenyl, and Ar is optionally substituted with 1-3 identical or different R ₃ ;

R ₃ is halogen, tert-butyl, trifluoromethyl;

R ₁ and R ₂ are the same or different, and are independently selected from methyl or ethyl;

n is an integer between 1-3;

The present invention also particularly preferably relates to the pyridine compounds represented by the general formula I and their pharmaceutically acceptable salts, hydrates or prodrugs, wherein,

Y is F;

Ar is phenyl, and Ar is optionally substituted with 1-3 identical or different R ₃ ;

R ₃ is halogen, tert-butyl, trifluoromethyl;

R ₁ and R ₂ are the same or different, and are independently selected from methyl or ethyl;

n is an integer between 1-3;

The compounds of general formula I and their pharmaceutically acceptable salts, hydrates or prodrugs of the present invention are preferably the following compounds, but these compounds do not imply any limitation to the present invention:

1-(4-Chloro-3-trifluoromethylphenyl)-3-[4-[[2-[4-[2-(dimethylamino)ethyl]-4H-1,2,4- triazol-3-yl]pyridin-4-yl]oxy]phenyl]urea;

1-(4-Chloro-3-trifluoromethylphenyl)-3-[4-[[2-[4-[2-(diethylamino)ethyl]-4H-1,2,4- triazol-3-yl]pyridin-4-yl]oxy]phenyl]urea;

1-(4-Chloro-3-trifluoromethylphenyl)-3-[4-[[2-[4-[2-(dimethylamino)propyl]-4H-1,2,4- triazol-3-yl]pyridin-4-yl]oxy]phenyl]urea;

1-(4-Chloro-3-trifluoromethylphenyl)-3-[4-[[2-[4-[2-(dimethylamino)ethyl]-4H-1,2,4- triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea;

1-(4-Chloro-3-trifluoromethylphenyl)-3-[4-[[2-[4-[2-(diethylamino)ethyl]-4H-1,2,4- Triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

1-(4-Chloro-3-trifluoromethylphenyl)-3-[4-[[2-[4-[2-(dimethylamino)propyl]-4H-1,2,4- Triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

1-(3-Trifluoromethylphenyl)-3-[4-[[[2-[4-[2-(dimethylamino)ethyl]-4H-1,2,4-triazole-3 -yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

1-(3-Trifluoromethylphenyl)-3-[4-[[2-[4-[2-(diethylamino)ethyl]-4H-1,2,4-triazole-3 -yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

1-(3-Trifluoromethylphenyl)-3-[4-[[2-[4-[2-(dimethylamino)propyl]-4H-1,2,4-triazole-3 -yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

1-(3-Chloro-4-fluorophenyl)-3-[4-[[2-[4-[2-(dimethylamino)ethyl]-4H-1,2,4-triazole- 3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

1-(3-Chloro-4-fluorophenyl)-3-[4-[[2-[4-[2-(diethylamino)ethyl]-4H-1,2,4-triazole- 3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

1-(3-Chloro-4-fluorophenyl)-3-[4-[[2-[4-[2-(dimethylamino)propyl]-4H-1,2,4-triazole- 3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

1-(3,4-Difluorophenyl)-3-[4-[[2-[4-[2-(dimethylamino)ethyl]-4H-1,2,4-triazole-3 -yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

1-(3,4-Difluorophenyl)-3-[4-[[2-[4-[2-(diethylamino)ethyl]-4H-1,2,4-triazole-3 -yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

1-(3,4-Difluorophenyl)-3-[4-[[2-[4-[2-(dimethylamino)propyl]-4H-1,2,4-triazole-3 -yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

1-(2,4-Dichlorophenyl)-3-[4-[[2-[4-[2-(dimethylamino)ethyl]-4H-1,2,4-triazole-3 -yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

1-(2,4-Dichlorophenyl)-3-[4-[[2-[4-[2-(diethylamino)ethyl]-4H-1,2,4-triazole-3 -yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

1-(2,4-Dichlorophenyl)-3-[4-[[2-[4-[2-(dimethylamino)propyl]-4H-1,2,4-triazole-3 -yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

1-(3-tert-Butylphenyl)-3-[4-[[2-[4-[2-(dimethylamino)ethyl]-4H-1,2,4-triazole-3- yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

1-(3-tert-Butylphenyl)-3-[4-[[2-[4-[2-(diethylamino)ethyl]-4H-1,2,4-triazole-3- yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

1-(3-tert-Butylphenyl)-3-[4-[[2-[4-[2-(dimethylamino)propyl]-4H-1,2,4-triazole-3- yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

Moreover, the pyridine derivatives of the general formula I in the present invention can be formed into pharmaceutically acceptable salts with acids according to some common methods in the art to which the present invention belongs. Pharmaceutically acceptable addition salts include inorganic and organic acid addition salts, with addition salts with the following acids being particularly preferred: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid , benzenesulfonic acid, naphthalenedisulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, benzoic acid, etc.

Furthermore, the present invention also includes prodrugs of the derivatives of the present invention. Prodrugs of the derivatives of the present invention are derivatives of general formula I, which themselves may have weak or even no activity, but after administration, are destroyed under physiological conditions (eg by metabolism, solvolysis or otherwise) by into the corresponding biologically active form.

The present invention can contain the pyridine derivatives of the above formula I and their pharmaceutically acceptable salts and hydrates as active ingredients, mixed with pharmaceutically acceptable excipients to prepare a composition, and prepared into clinically acceptable In dosage form, the above-mentioned excipients refer to diluents, adjuvants or carriers that can be used in the pharmaceutical field. The above dosage forms refer to injections, tablets, capsules and the like commonly used in clinical practice.

The compounds involved in the present invention or their pharmaceutically acceptable salts, hydrates and prodrugs can be used alone as the only anti-proliferative drugs, or can be used in combination with existing anti-proliferative drugs for treatment and/or prevention Proliferative diseases such as tumors, fibrosis, etc.

Through the kinase activity test, it is found that the compounds of the present invention have significant inhibitory activities on c-Kit, RET, FLT3 and other tyrosine kinases, indicating that the compounds can be used for diseases related to overexpression or mutation of tyrosine kinases. At the same time, we found that the compound of the present invention has a proliferation inhibitory effect on various tumor cells in vitro, so it can be used as a drug for the preparation of cancer treatment and/or prevention, such as colon, prostate, liver, kidney, breast, thyroid tumors and the like. In particular, the compound of the present invention has a prominent inhibitory effect on the proliferation of colon cancer cell HT-29. Further research shows that the compound can induce HT-29 cell cycle arrest and induce cell apoptosis, and can be especially used for preparing Medications to treat/or prevent colon cancer.

The active compound of the present invention or its available salt can be used alone as the only anti-tumor drug, or can be used together with existing anti-tumor drugs such as platinum drug cisplatin, camptothecin drug irinotecan, deoxycytidine drug Combined use of gemcitabine and paclitaxel.

The examples and preparations provided hereinafter further illustrate and illustrate the compounds of the present invention and methods for their preparation. It should be understood that the scope of the following examples and preparations do not limit the scope of the invention in any way.

The following synthetic scheme (Scheme 1) summarizes and describes the preparation of the derivatives of formula I of the present invention, all starting materials were prepared in the manner described in these schemes, by methods well known to those of ordinary skill in the art of organic chemistry, or are commercially available purchase. All final derivatives of the present invention are prepared by methods described in these schemes or by methods analogous thereto, which are well known to those of ordinary skill in the art of organic chemistry. All variable factors applied in these procedures are as defined in the claims.

The compounds of formula I according to the present invention can be prepared according to the method of route 1.

Detailed ways:

In the following examples, methods for preparing some of the compounds are depicted. It will be appreciated that the following methods, as well as other methods known to those of ordinary skill in the art, may be applicable to the preparation of all compounds described herein. The examples are intended to illustrate rather than limit the scope of the invention.

Example 1: 1-(4-Chloro-3-trifluoromethylphenyl)-3-[4-[[2-[4-[2-(dimethylamino)ethyl]-4H-1, 2,4-Triazol-3-yl]pyridin-4-yl]oxy]phenyl]urea

Step 14-Chloro-pyridine-2-carboxylic acid methyl ester (C)

At room temperature, add 150g (1.22mol) pyridine-2-carboxylic acid and 20.1g (0.197mol) sodium bromide to 200ml chlorobenzene, stir for 10min, the reaction solution rises to 50°C, slowly add 355ml (4.88mol) chlorinated chloride The sulfone was stirred for 30 min, heated to reflux for 20 h, and the solvent was evaporated to obtain the intermediate 4-chloro-pyridine-2-carbonyl chloride as a yellow oil. The intermediate 1 was added to 500 mL of toluene, and 100 mL of anhydrous methanol was added dropwise at a temperature of 0-5 °C. After the drop was completed, the reaction was stirred at 15 °C for 1.5 h. Suction filtration, the filter cake was added to saturated potassium carbonate solution, extracted with dichloromethane (3×400 mL), the organic layers were combined, washed with water, dried over anhydrous sodium sulfate, evaporated to remove the solvent, and dried to obtain 150 g of yellow solid with a yield of 72.1%.

Step 24-(4-Nitrophenoxy)-pyridine-2-carboxylic acid methyl ester (D-1)

29 g (0.17 mol) of intermediate C and 24.3 g (0.175 mol) of 4-nitrophenol were dissolved in 300 mL of dry chlorobenzene, and the reaction was refluxed for 13 h. Evaporate the chlorobenzene to obtain an oily substance, add 500ml of dichloromethane to dissolve, wash with saturated potassium carbonate solution (3×300ml), evaporate the organic layer to obtain a brown solid, add 300ml of absolute ethanol to the crude product, heat up to reflux and stir for 30min, the reaction solution Naturally cooled to room temperature, allowed to stand for crystallization, suction filtered, and the filter cake was dried to obtain 24.5 g of a light yellow solid with a yield of 52.5%.

Step 34-(4-Aminophenoxy)-pyridine-2-carboxylic acid methyl ester (E-1)

At room temperature, 10 g (0.036 mol) of methyl 4-(4-nitrophenoxy)-pyridine-2-carboxylate was dissolved in 25 mL of absolute ethanol, 1 g of palladium/carbon was added, and the reaction was stirred at room temperature for 2 h. The reaction solution was filtered, and the filtrate was concentrated to obtain 6.8 g of a pale yellow solid with a yield of 76.2%.

Step 44-[4-[3-(4-Chloro-3-trifluoromethylphenyl)ureido]phenoxy]pyridine-2-carboxylic acid methyl ester (F-1)

At room temperature, 2 g (8.2 mmol) of 4-(4-aminophenoxy)-pyridine-2-carboxylic acid methyl ester was added to 5 mL of dichloromethane, and 1.8 g (8.2 mmol) of 4-chloro-3 was added in batches. -Trifluoromethyl phenyl isocyanate, after the addition was completed, the stirring reaction was continued for 3h. The reaction solution was filtered, and the filter cake was washed with 10 mL of water and 10 mL of diethyl ether each time, and the filter cake was dried to obtain 3.8 g of a white solid with a yield of 65.3%.

Step 51-(4-Chloro-3-trifluoromethylphenyl)-3-[4-[[2-(hydrazinoyl)pyridin-4-yl]oxy]phenyl]urea (G-1)

3g (6.5mmol) of 4-[4-[3-(4-chloro-3-trifluoromethylphenyl)ureido]phenoxy]pyridine-2-carboxylic acid methyl ester was added to 15mL of ethanol, and 0.8 g (13mmol) 80% hydrazine hydrate, react at 80°C for 3h. The solvent was evaporated, 10 mL of ether was added to the system, stirred for 30 min, filtered, and the filter cake was dried to obtain 1.54 g of a white solid with a yield of 51.2%.

Step 6(E)-N'-[4-[4-[3-(4-Chloro-3-trifluoromethylphenyl)ureido]phenoxy]pyridine-2-formyl]-N,N -Dimethylaminohydrazone (H-1)

1.5 g (3.2 mmol) of 1-(4-chloro-3-trifluoromethylphenyl)-3-[4-[[2-(hydrazinoyl)pyridin-4-yl]oxy]phenyl] Urea and 3 mL of DMF-DMA were added to 20 mL of dichloromethane, and the temperature was raised to reflux for 4 h. The solvent was evaporated, 20 mL of ether was added to the system, stirred for 20 min, filtered, the filter cake was washed once with 10 mL of ether, and dried to obtain 1.2 g of the product with a yield of 72.0%.

Step 71-(4-Chloro-3-trifluoromethylphenyl)-3-[4-[[2-[4-[2-(dimethylamino)ethyl]-4H-1,2,4 -Triazol-3-yl]pyridin-4-yl]oxy]phenyl]urea (I, Example 1)

0.5 g (0.96 mmol) of (E)-N'-[4-[4-[3-(4-chloro-3-trifluoromethylphenyl)ureido]phenoxy]pyridine-2-formyl ]-N,N-dimethylaminohydrazone was dissolved in 4 mL of glacial acetic acid, 1.5 mL of N,N-dimethylethylenediamine was slowly added, and the temperature was raised to 90 °C for 3 h. Most of the solvent was evaporated, the residue was poured into ice water, the pH was adjusted to 8-9 with 15% NaOH solution, and filtered to obtain an off-white solid. Using methanol: dichloromethane (1:50-1:10) as the eluent, column chromatography was used to obtain 0.17 g of a white solid with a yield of 32.6%.

ESI-MS m/z: 546.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.47 (s, 1H), 8.54 (s, 1H), 8.27 (m, 2H), 7.72 (d, J=7.2Hz, 1H), 7.57 (dd, J=7.7, 1.4Hz, 1H), 7.56 (m, 2H), 7.37 (m, 1H), 7.21 (d, J=7.3Hz, 2H) ,6.73(d,J＝7.5Hz,2H),4.35(t,J＝6.9Hz,2H),3.27(t,J＝7.4Hz,2H),2.17(s,6H).

Example 2: 1-(4-Chloro-3-trifluoromethylphenyl)-3-[4-[[2-[4-[2-(diethylamino)ethyl]-4H-1, 2,4-Triazol-3-yl]pyridin-4-yl]oxy]phenyl]urea

With intermediate H-1(E)-N'-[4-[4-[3-(4-chloro-3-trifluoromethylphenyl)ureido]phenoxy]pyridine-2-formyl] -N,N-dimethylaminohydrazone and N,N-diethylethylenediamine were used as raw materials, and were prepared according to the method of Example 1.

ESI-MS m/z: 574.1 [M+H] ⁺ ;

Example 3: 1-(4-Chloro-3-trifluoromethylphenyl)-3-[4-[[2-[4-[2-(dimethylamino)propyl]-4H-1, 2,4-Triazol-3-yl]pyridin-4-yl]oxy]phenyl]urea

With intermediate H-1(E)-N'-[4-[4-[3-(4-chloro-3-trifluoromethylphenyl)ureido]phenoxy]pyridine-2-formyl] -N,N-dimethylaminohydrazone and N,N-dimethylpropanediamine were used as raw materials, and were prepared according to the method of Example 1.

ESI-MS m/z: 560.1 [M+H] ⁺ ;

Example 4: 1-(4-Chloro-3-trifluoromethylphenyl)-3-[4-[[2-[4-[2-(dimethylamino)ethyl]-4H-1, 2,4-Triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea;

Step 14-(2-Fluoro-4-nitrophenoxy)-pyridine-2-carboxylic acid methyl ester (D-2)

29 g (0.17 mol) of intermediate C and 27.5 g (0.175 mol) of 2-fluoro-4-nitrophenol were dissolved in 200 ml of dry chlorobenzene, and the reaction was refluxed for 11 h. Evaporate the chlorobenzene to obtain an oily substance, add 500ml of dichloromethane to dissolve, wash with saturated potassium carbonate solution (2×300ml), evaporate the organic layer to obtain a brown solid, add 300ml of absolute ethanol to the crude product, heat up to reflux and stir for 30min, the reaction solution Naturally cooled to room temperature, left to stand for crystallization, suction filtered, and the filter cake was dried to obtain 22.1 g of a light yellow solid with a yield of 44.5%.

Step 24-(2-Fluoro-4-aminophenoxy)-pyridine-2-carboxylic acid methyl ester (E-2)

5 g (0.017 mol) of 4-(2-fluoro-4-nitrophenoxy)-pyridine-2-carboxylic acid methyl ester was dissolved in 20 mL of absolute ethanol, 0.5 g of palladium/carbon was added, and the reaction was carried out at room temperature for 3 h. The palladium/carbon was removed by filtration, and the filtrate was concentrated to obtain 2.9 g of a pale yellow solid with a yield of 65.1%.

Step 34-[4-[3-(4-Chloro-3-trifluoromethylphenyl)ureido]-2-fluorophenoxy]pyridine-2-carboxylic acid methyl ester (F-2)

At room temperature, 2 g (8.2 mmol) of 4-(4-aminophenoxy)-pyridine-2-carboxylic acid methyl ester was added to 5 mL of dichloromethane, and 1.8 g (8.2 mmol) of 4-chloro-3 was added in batches. -Trifluoromethyl phenyl isocyanate, after the addition was completed, the stirring reaction was continued for 3h. The reaction solution was filtered, the filter cake was rinsed once with 10 mL of ether, and the filter cake was dried to obtain 3.8 g of a white solid with a yield of 65.3%.

Step 41-(4-Chloro-3-trifluoromethylphenyl)-3-[3-fluoro-4-[[2-(hydrazinoyl)pyridin-4-yl]oxy]phenyl]urea ( G-2)

5 g (10.3 mmol) of methyl 4-[4-[3-(4-chloro-3-trifluoromethylphenyl)ureido]-2-fluorophenoxy]pyridine-2-carboxylate was added to 20 mL of acetonitrile , slowly added 1.3g (20.7mmol) of 80% hydrazine hydrate, and reacted at 80°C for 5h. Part of the solvent was concentrated, the system was cooled to room temperature, filtered, the filter cake was washed once with 10 mL of diethyl ether, and dried to obtain 3.1 g of a white solid with a yield of 62.5%.

Step 5 (E)-N'-[4-[4-[3-(4-Chloro-3-trifluoromethylphenyl)ureido]-2-fluorophenoxy]pyridine-2-formyl] -N,N-Dimethylaminohydrazone (H-2)

2 g (4.1 mmol) of 1-(4-chloro-3-trifluoromethylphenyl)-3-[3-fluoro-4-[[2-(hydrazinoyl)pyridin-4-yl]oxy] Phenyl]urea and 4 mL of DMF-DMA were added to 10 mL of dichloromethane, and the temperature was raised to reflux for 5 h. Most of the solvent was evaporated, and 30 mL of ether was added to the system, resulting in a large amount of yellow solid. After filtration, 1.5 g of product was obtained, and the yield was 68.1%.

Example 4: 1-(4-Chloro-3-trifluoromethylphenyl)-3-[4-[[2-[4-[2-(dimethylamino)ethyl]-4H-1, 2,4-Triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

0.3 g (0.56 mmol) of (E)-N'-[4-[4-[3-(4-chloro-3-trifluoromethylphenyl)ureido]-2-fluorophenoxy]pyridine- 2-Formyl]-N,N-dimethylaminohydrazone was dissolved in 3 mL of glacial acetic acid, 1 mL of N,N-dimethylethylenediamine was slowly added, and the temperature was raised to 90°C for reaction for 3 hours. Most of the solvent was evaporated, the residual liquid was poured into ice water, the pH was adjusted to 8-9 with 15% NaOH solution, and filtered to obtain an off-white solid. Using methanol: dichloromethane (1:100-1:10) as the eluent, column chromatography was used to obtain 0.13 g of a white solid with a yield of 42.1%.

ESI-MS m/z: 563.9 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.97 (s, 1H), 9.85 (s, 1H), 8.64 (s, 1H), 8.60 (d, J＝5.2Hz, 1H), 8.11(s, 1H), 7.75(d, J＝12.3Hz, 1H), 7.65(dd, J＝15.6, 8.1Hz, 2H), 7.56(s, 1H) ,7.40(t,J＝8.5Hz,1H),7.28(d,J＝8.0Hz,1H),7.13(dd,J＝5.6,2.6Hz,1H),4.61(t,J＝5.9Hz,2H) , 2.57(t, J=6.0Hz, 2H), 2.13(s, 6H).

According to the method of Example 4, with intermediate H-2(E)-N'-[4-[4-[3-(4-chloro-3-trifluoromethylphenyl)ureido]-2-fluoro Phenoxy]pyridine-2-formyl]-N,N-dimethylaminohydrazone as raw material, respectively reacted with N,N-diethylethylenediamine and N,N-dimethylpropylenediamine to prepare Example 5 and Example 6.

Example 5: 1-(4-Chloro-3-trifluoromethylphenyl)-3-[4-[[2-[4-[2-(diethylamino)ethyl]-4H-1, 2,4-Triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

ESI-MS m/z: 592.2 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.40 (s, 1H), 9.30 (s, 1H), 8.62 (s, 1H), 8.59 (d, J＝4.5Hz, 1H), 8.12(s, 1H), 7.70(m, 3H), 7.52(s, 1H), 7.36(m, 2H), 7.14(dd, J＝5.52.4Hz, 1H ),4.56(t,J＝5.9Hz,2H),2.66(t,J＝6.1Hz,2H),2.39(quartet,J＝6.3Hz,4H),0.76(t,J＝6.3Hz,6H).

Example 6: 1-(4-Chloro-3-trifluoromethylphenyl)-3-[4-[[2-[4-[2-(dimethylamino)propyl]-4H-1, 2,4-Triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

ESI-MS m/z: 578.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.63 (s, 1H), 9.53 (s, 1H), 8.66 (s, 1H), 8.60 (d, J=5.7Hz, 1H), 8.12 (d, J=2.2Hz, 1H), 7.75 (dd, J=13.1, 2.2Hz, 1H), 7.68 (dd, J=8.8, 2.1Hz, 1H) ,7.63(d,J＝8.8Hz,1H),7.56(d,J＝2.4Hz,1H),7.40(t,J＝8.9Hz,1H),7.30(d,J＝8.7Hz,1H),7.13 (dd, J=5.7, 2.5Hz, 1H), 4.52 (t, J=7.2Hz, 2H), 2.20 (t, J=6.7Hz, 2H), 2.11 (s, 6H), 1.88 (quintet, J= 7.0Hz, 2H).

According to the method of Example 4, using methyl 4-(2-fluoro-4-aminophenoxy)-pyridine-2-carboxylate as raw material, after reacting with 3-trifluoromethyl phenyl isocyanate, and then reacting with 3-trifluoromethyl phenyl isocyanate in turn The intermediate (E)-N'-[4-[4-[3-(3-trifluoromethylphenyl)ureido]-2-fluorophenoxy]pyridine- 2-Formyl]-N,N-dimethylaminohydrazone, which are respectively combined with N,N-dimethylethylenediamine, N,N-diethylethylenediamine and N,N-dimethylpropanediamine Amine cyclization afforded the compounds of Examples 7-9.

Example 7: 1-(3-Trifluoromethylphenyl)-3-[4-[[2-[4-[2-(dimethylamino)ethyl]-4H-1,2,4- Triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

ESI-MS m/z: 530.2 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.26(s, 1H), 9.24(s, 1H), 8.64(s, 1H), 8.60 (d,J＝5.7Hz,1H),8.03(s,1H),7.76(d,J＝13.2Hz,1H),7.62(d,J＝8.0Hz,1H),7.56–7.49(m,2H) ,7.40(t,J＝8.9Hz,1H),7.32(m,1H),7.15(dd,J＝5.7,2.5Hz,1H),4.63(t,J＝6.0Hz,2H),2.62(t, J=6.2Hz, 2H), 2.17(s, 6H).

Example 8: 1-(3-Trifluoromethylphenyl)-3-[4-[[2-[4-[2-(diethylamino)ethyl]-4H-1,2,4- Triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

ESI-MS m/z: 558.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.72 (s, 2H), 8.62 (s, 1H), 8.58 (d, J=5.6 Hz ,1H),8.01(s,1H),7.76(dd,J=13.0,0.9Hz,1H),7.62(d,J=8.2Hz,1H),7.52(m,2H),7.39(t,J= 8.9Hz, 1H), 7.30(dd, J＝21.4, 7.9Hz, 2H), 7.12(dd, J＝5.1, 1.9Hz, 1H), 4.55(t, J＝5.8Hz, 2H), 2.66(t, J=5.5Hz, 2H), 2.39 (dd, J=13.6, 6.6Hz, 4H), 0.76 (t, J=6.9Hz, 6H).

Example 9: 1-(3-Trifluoromethylphenyl)-3-[4-[[2-[4-[2-(dimethylamino)propyl]-4H-1,2,4- Triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

ESI-MS m/z: 544.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.92 (s, 2H), 8.65 (s, 1H), 8.59 (d, J=5.7 Hz ,1H),8.04(s,1H),7.77(d,J＝13.2Hz,1H),7.65(d,J＝8.1Hz,1H),7.55(d,J＝2.4Hz,1H),7.51(t , J=7.9Hz, 1H), 7.35 (m, 3H), 7.12 (dd, J=5.7, 2.5Hz, 1H), 4.50 (t, J=7.2Hz, 2H), 2.16 (t, J=6.7Hz) ,2H),2.08(s,6H),1.85(quintet,J=6.9Hz,2H).

According to the method of Example 4, using methyl 4-(2-fluoro-4-aminophenoxy)-pyridine-2-carboxylate as a raw material, after reacting with 3-chloro-4-fluorophenyl isocyanate, and then sequentially Reaction with hydrazine hydrate and DMF-DMA to obtain intermediate (E)-N'-[4-[4-[3-(3-chloro-4-fluorophenyl)ureido]-2-fluorophenoxy] Pyridine-2-formyl]-N,N-dimethylaminohydrazone, which are respectively combined with N,N-dimethylethylenediamine, N,N-diethylethylenediamine and N,N-dimethylethylenediamine Cyclization of propylene diamine gave the compounds of Examples 10-12.

Example 10: 1-(3-Chloro-4-fluorophenyl)-3-[4-[[2-[4-[2-(dimethylamino)ethyl]-4H-1,2,4 -Triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

ESI-MS m/z: 514.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.23 (s, 1H), 9.11 (s, 1H), 8.63 (s, 1H), 8.59 (d, J=5.6Hz, 1H), 7.81 (d, J=6.5Hz, 1H), 7.74 (d, J=12.5Hz, 1H), 7.53 (d, J=1.7Hz, 1H), 7.37 (m ,3H),7.29(d,J＝8.6Hz,1H),7.14(dd,J＝5.3,2.1Hz,1H),4.61(t,J＝5.9Hz,2H),2.57(t,J＝6.1Hz ,2H),2.12(s,6H).

Example 11: 1-(3-Chloro-4-fluorophenyl)-3-[4-[[2-[4-[2-(diethylamino)ethyl]-4H-1,2,4 -Triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

ESI-MS m/z: 542.2[M+H] ⁺ . ¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.45(s,1H), 9.34(s,1H), 8.62(s,1H), 8.58 (d, J=5.7Hz, 1H), 7.79–7.82 (dd, J=7.6, 2.0Hz, 1H), 7.74 (dd, J=13.2, 2.3Hz, 1H), 7.52 (d, J=2.4Hz, 1H), 7.39(t, J=9.0Hz, 1H), 7.35–7.36(m, 1H), 7.33(m, 1H), 7.27(dd, J=8.8, 1.3Hz, 1H), 7.13(dd, J =5.7,2.5Hz,1H),4.55(t,J=6.2Hz,2H),2.66(t,J=6.0Hz,2H),2.39(q,J=6.9Hz,4H),0.76(t,J =7.0Hz,6H).

Example 12: 1-(3-Chloro-4-fluorophenyl)-3-[4-[[2-[4-[2-(dimethylamino)propyl]-4H-1,2,4 -Triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

ESI-MS m/z: 528.3; [M+H] ⁺ ; ¹ H NMR (400MHz, DMSO) δ 9.32(s, 1H), 9.21(s, 1H), 8.66(s, 1H), 8.60(d , J＝5.7Hz, 1H), 7.81(d, J＝7.2Hz, 1H), 7.74(dd, J＝13.2, 2.0Hz, 1H), 7.55(d, J＝2.4Hz, 1H), 7.34(m ,4H),7.13(dd,J＝5.7,2.5Hz,1H),4.51(t,J＝7.3Hz,2H),2.16(t,J＝6.7Hz,2H),2.09(s,6H),1.85 (quintet, J=6.8Hz, 2H).

According to the method of Example 4, using methyl 4-(2-fluoro-4-aminophenoxy)-pyridine-2-carboxylate as raw material, after reacting with 3,4-difluorophenyl isocyanate, and then reacting with 3,4-difluorophenyl isocyanate in turn The intermediate (E)-N'-[4-[4-[3-(3,4-difluorophenyl)ureido]-2-fluorophenoxy]pyridine- 2-Formyl]-N,N-dimethylaminohydrazone, which are respectively combined with N,N-dimethylethylenediamine, N,N-diethylethylenediamine and N,N-dimethylpropanediamine Amine cyclization afforded the compounds of Examples 13-15.

Example 13: 1-(3,4-Difluorophenyl)-3-[4-[[2-[4-[2-(dimethylamino)ethyl]-4H-1,2,4- Triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

ESI-MS m/z: 498.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.13 (s, 1H), 9.05 (s, 1H), 8.59 (s, 1H), 8.56 (d, J=5.8Hz, 1H), 7.64(m, 2H), 7.50(d, J=2.6Hz, 1H), 7.35(m, 2H), 7.25(m, 1H), 7.11(m, 2H) ,4.58(t,J＝6.4Hz,2H),2.55(t,J＝6.4Hz,2H),2.10(s,6H).

Example 14: 1-(3,4-Difluorophenyl)-3-[4-[[2-[4-[2-(diethylamino)ethyl]-4H-1,2,4- Triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

ESI-MS m/z: 526.3 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.16(s, 1H), 9.08(s, 1H), 8.66(s, 1H), 8.60 (d, J＝5.8Hz, 1H), 7.73(dd, J＝13.3, 2.2Hz, 1H), 7.67(ddd, J＝13.2, 7.6, 2.5Hz, 1H), 7.54(s, 1H), 7.37( m, 2H), 7.29(d, J＝7.6Hz, 1H), 7.16(d, J＝8.0Hz, 2H), 4.64(t, J＝5.9Hz, 2H), 2.70(t, J＝6.0Hz, 2H),0.87(t,J＝6.5Hz,6H).

Example 15: 1-(3,4-Difluorophenyl)-3-[4-[[2-[4-[2-(dimethylamino)propyl]-4H-1,2,4- Triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

ESI-MS m/z: 512.1; [M+H] ⁺ ; ¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.36(s,1H), 9.28(s,1H), 8.65(s,1H), 8.60(d,J＝5.7Hz,1H),7.74(dd,J＝13.2,1.7Hz,1H),7.68(ddd,J＝13.2,7.5,2.2Hz,1H),7.55(d,J＝2.1Hz ,1H),7.38(dd,J＝18.0,9.1Hz,2H),7.30(t,J＝8.2Hz,1H),7.17(d,J＝9.0Hz,1H),7.13(dd,J＝5.6, 2.4Hz, 1H), 4.51 (t, J=7.2Hz, 2H), 2.16 (t, J=6.6Hz, 2H), 2.09 (s, 6H), 1.85 (quintet, J=7.2Hz, 2H).

According to the method of Example 4, using methyl 4-(2-fluoro-4-aminophenoxy)-pyridine-2-carboxylate as raw material, after reacting with 2,4-dichlorophenyl isocyanate, and then reacting with 2,4-dichlorophenyl isocyanate in turn The intermediate (E)-N'-[4-[4-[3-(2,4-dichlorophenyl)ureido]-2-fluorophenoxy]pyridine- 2-Formyl]-N,N-dimethylaminohydrazone, which are respectively combined with N,N-dimethylethylenediamine, N,N-diethylethylenediamine and N,N-dimethylpropanediamine Amine cyclization afforded the compounds of Examples 16-18.

Example 16: 1-(2,4-Dichlorophenyl)-3-[4-[[2-[4-[2-(dimethylamino)ethyl]-4H-1,2,4- Triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

ESI-MS m/z: 530.0 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.06 (s, 1H), 8.63 (s, 1H), 8.60 (d, J=5.4 Hz ,2H),8.18(d,J＝8.9Hz,1H),7.76(d,J＝11.5Hz,1H),7.63(s,1H),7.54(s,1H),7.40(m,2H),7.27 (d, J=8.8Hz, 1H), 7.13 (dd, J=5.6, 2.5Hz, 1H), 4.61 (t, J=6.0Hz, 2H), 2.59 (t, J=6.1Hz, 2H), 2.13 (s,6H).

Example 17: 1-(2,4-Dichlorophenyl)-3-[4-[[2-[4-[2-(diethylamino)ethyl]-4H-1,2,4- Triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

ESI-MS m/z: 558.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.87 (s, 1H), 8.62 (s, 1H), 8.59 (d, J=5.7 Hz ,1H),8.53(s,1H),8.18(d,J＝9.0Hz,1H),7.76(d,J＝13.3Hz,1H),7.64(d,J＝1.9Hz,1H),7.53(d , J＝1.4Hz, 1H), 7.40(m, 2H), 7.26(d, J＝9.1Hz, 1H), 7.14(dd, J＝5.6, 2.4Hz, 1H), 4.56(t, J＝6.0Hz) ,2H),2.68(t,J＝6.1Hz,2H),2.42(quartet,J＝5.9Hz,4H),0.78(t,J＝6.7Hz,6H).

Example 18: 1-(2,4-Dichlorophenyl)-3-[4-[[2-[4-[2-(dimethylamino)propyl]-4H-1,2,4- Triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

ESI-MS m/z: 544.1; ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.79 (s, 1H), 8.65 (s, 1H), 8.61 (d, J=5.7 Hz, 1H), 8.50 ( s, 1H), 8.18 (d, J=9.0Hz, 1H), 7.76 (dd, J=13.2, 2.4Hz, 1H), 7.64 (d, J=2.4Hz, 1H), 7.55 (d, J=2.5 Hz, 1H), 7.41(m, 2H), 7.26(d, J=8.9Hz, 1H), 7.14(dd, J=5.7, 2.6Hz, 1H), 4.52(t, J=7.2Hz, 2H), 2.28(t, J=6.7Hz, 2H), 2.17(s, 6H), 1.89(quintet, J=6.9Hz, 2H).

According to the method of Example 4, using methyl 4-(2-fluoro-4-aminophenoxy)-pyridine-2-carboxylate as raw material, after reacting with 3-tert-butylphenyl isocyanate, and then hydrating with 3-tert-butylphenyl isocyanate in turn The intermediate (E)-N'-[4-[4-[3-(tert-butylphenyl)ureido]-2-fluorophenoxy]pyridine-2-formyl was prepared by the reaction of hydrazine and DMF-DMA ]-N,N-dimethylaminohydrazone, which is cyclized with N,N-dimethylethylenediamine, N,N-diethylethylenediamine and N,N-dimethylpropylenediamine, respectively. The compounds of Examples 19-21 were obtained.

Example 19: 1-(3-tert-butylphenyl)-3-[4-[[2-[4-[2-(dimethylamino)ethyl]-4H-1,2,4-tris Azol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

ESI-MS m/z: 518.2; ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.62 (s, 1H), 8.57 (s, 1H), 8.45 (d, J=6.1 Hz, 2H), 7.77 ( d, J=7.5Hz, 1H), 7.59 (m, 1H), 7.50 (m, 2H), 7.44 (dd, J=7.5, 1.4Hz, 1H), 7.27 (dd, J=8.1, 1.4Hz, 1H) ),7.16(t,J＝8.1Hz,1H),7.04(d,J＝7.5Hz,1H),6.87(dd,J＝7.4,5.0Hz,1H),4.55(t,J＝7.3Hz,2H) ), 2.98(t, J＝6.3Hz, 2H), 2.20(s, 6H), 1.27(s, 9H).

Example 20: 1-(3-tert-Butylphenyl)-3-[4-[[2-[4-[2-(diethylamino)ethyl]-4H-1,2,4-tris Azol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

ESI-MS m/z: 546.1.

Example 21: 1-(3-tert-butylphenyl)-3-[4-[[2-[4-[2-(dimethylamino)propyl]-4H-1,2,4-tris Azol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea

ESI-MS m/z: 532.1.

Study on the biological activity of the product of the present invention

The present invention provides compounds for inhibiting tumor cell proliferation activity:

Using MTT method, the pyrimidine derivatives of the above formula I according to the present invention were screened for the activity of inhibiting lung cancer cell H460, colon cancer cell HT-29 and human breast cancer cell MDA-MB-231 in vitro. The specific operations are:

(1) After the cells were recovered and passaged 2-3 times to stabilize, they were digested from the bottom of the culture flask with trypsin solution (0.25%). After the cell digestion solution was poured into the centrifuge tube, the culture solution was added to stop the digestion. Centrifuge the centrifuge tube at 800 r/min for 10 min, discard the supernatant, add 5 mL of culture medium, mix the cells by pipetting, pipette 10 μL of the cell suspension and add it to a cell counting plate for counting, and adjust the cell concentration to 10 ⁴ cells/well. In the 96-well plate, 100 μL of cell suspension was added to the rest of the 96-well plates, except that A1 was a blank well without cells. The 96-well plate was placed in an incubator for 24 h.

(2) Dissolve the test sample with 50 μL of dimethyl sulfoxide, then add an appropriate amount of culture medium to dissolve the sample into 2 mg/mL liquid, and then dilute the sample in a 24-well plate to 20, 4, 0.8, 0.16, 0.032 μg/mL.

Each concentration was added to 3 wells, and the growth of the cells in the surrounding two rows and two columns was greatly affected by the environment, and was only used for the blank cell wells. The 96-well plate was placed in an incubator for 72 h.

(3) Discard the medicated culture medium in the 96-well plate, wash the cells twice with phosphate buffered solution (PBS), add 100 μL of MTT (0.5 mg/mL) to each well and put it in the incubator for 4 hours, then discard To the MTT solution, add 100 μL of dimethyl sulfoxide. Shake on a magnetic shaker to fully dissolve the surviving cells and the MTT reaction product formazan, and put it into a microplate reader to measure the results. The drug IC ₅₀ value can be obtained by the Bliss method.

Table 1 shows the compounds inhibiting the activity of lung cancer cells H460, colon cancer cells HT-29 and human breast cancer cells MDA-MB-231.

Table 1

It can be clearly seen from the above results that the compounds of formula I to be protected by the present invention can significantly inhibit the proliferation of H460, HT-29 and MDA-MB-231 cells, and the biological activities of some compounds are better than those of the marketed drug Sorafenib.

The present invention provides compounds for inhibiting the activity of tyrosine kinases:

The pyrimidine derivatives of the above formula I according to the present invention were screened for the activity of inhibiting c-Kit, RET and FLT3 kinases in vitro by Mobility shift assay. The specific operations are:

(1) 1X Kinase Basal Buffer with 50 mM HEPES (pH 7,5), 0.0015% Brij-35, 10 mM MgCl ₂ and 2 mM DTT; with 100 mM HEPES (pH 7, 5), 0.015% Brij-35, 0.2% Coating Reagent, 50mM EDTA make up stop buffer.

(2) The test kinase was added to the enzyme base buffer to prepare a 2.5X enzyme solution; the FAM-labeled peptide and ATP were added to the enzyme base buffer to prepare a 2.5X peptide solution.

(3) The test compound was prepared into a solution with a concentration of 0.5 mM in DMSO, and the gradient was diluted according to the test concentration; 5 μL of the test compound was added to the test plate, 10 μL of 2.5X enzyme solution was added, and shaken for 10 min, followed by 10 μL of 2.5X enzyme solution. Peptide solution, the reaction system was incubated at 28°C for 1 h; 25 μL of stop buffer was added to stop the reaction.

(4) Draw the inhibitory concentration curve and calculate the IC ₅₀ value.

The inhibitory data of example compounds on c-Kit, RET and FLT3 kinases are shown in Table 2.

Table 2

It can be clearly seen from the above test results that the compound of the general formula I to be protected by the present invention has a good in vitro antitumor activity, which is equivalent to or better than the marketed antitumor drug sorafenib.

Example 22: Tablets

Using 10 g of the compound containing the compound in claim 1 (taking the compound of Example 5 as an example), adding 20 g of auxiliary materials according to the general pharmaceutical tableting method, and mixing, then compressed into 100 tablets, each weighing 300 mg.

Example 23: Capsules

Using 10 g of the compound containing the compound in claim 1 (taking the compound of Example 7 as an example), after mixing 20 g of the auxiliary material according to the requirements of pharmaceutical capsules, it is put into hollow capsules, and each capsule weighs 300 mg.

Example 24: Injection

Using 10 g of the compound containing the compound in claim 1 (taking the compound of Example 1 as an example), according to the conventional method of pharmacy, carry out activated carbon adsorption, after filtering through a 0.65 μm microporous membrane, fill in a nitrogen tank to prepare a water injection preparation, Each bottle contains 2mL, and a total of 100 bottles are filled.

While the invention has been described in terms of specific embodiments, modifications and equivalents will be apparent to those skilled in the art and are intended to be included within the scope of the invention.

Claims (5)

1. The following compounds and their pharmaceutically acceptable salts are selected from: 1-(4-Chloro-3-trifluoromethylphenyl)-3-[4-[[2-[4-[2-(dimethylamino)ethyl]-4H-1,2,4- triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea; 1-[4-[[2-[4-[2-(dimethylamino)ethyl]-4H-1,2,4-triazol-3-yl]pyridin-4-yl]oxy]- 3-Fluorophenyl]-3-(3-trifluoromethylphenyl)urea; 1-(3-Chloro-4-fluorophenyl)-3-[4-[[2-[4-[2-(dimethylamino)ethyl]-4H-1,2,4-triazole- 3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea; 1-(4-Chloro-3-trifluoromethylphenyl)-3-[4-[[2-[4-[2-(diethylamino)ethyl]-4H-1,2,4- triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea; 1-[4-[[2-[4-[2-(diethylamino)ethyl]-4H-1,2,4-triazol-3-yl]pyridin-4-yl]oxy]- 3-Fluorophenyl]-3-(3-trifluoromethylphenyl)urea; 1-(3-Chloro-4-fluorophenyl)-3-[4-[[2-[4-[2-(diethylamino)ethyl]-4H-1,2,4-triazole- 3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea; 1-(4-Chloro-3-trifluoromethylphenyl)-3-[4-[[2-[4-[2-(dimethylamino)propyl]-4H-1,2,4- triazol-3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea; 1-[4-[[2-[4-[2-(dimethylamino)propyl]-4H-1,2,4-triazol-3-yl]pyridin-4-yl]oxy]- 3-Fluorophenyl]-3-(3-trifluoromethylphenyl)urea; 1-(3-Chloro-4-fluorophenyl)-3-[4-[[2-[4-[2-(dimethylamino)propyl]-4H-1,2,4-triazole- 3-yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea; 1-(3,4-Difluorophenyl)-3-[4-[[2-[4-[2-(dimethylamino)propyl]-4H-1,2,4-triazole-3 -yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea; 1-(2,4-Dichlorophenyl)-3-[4-[[2-[4-[2-(dimethylamino)propyl]-4H-1,2,4-triazole-3 -yl]pyridin-4-yl]oxy]-3-fluorophenyl]urea. 2. A pharmaceutical composition comprising the compound of claim 1 and a pharmaceutically acceptable salt thereof as an active ingredient and a pharmaceutically acceptable excipient. 3. Use of the compound of claim 1 and a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 2 in the preparation of a medicament for the treatment and/or prevention of proliferative diseases. 4. Use of the compound of claim 1 and a pharmaceutically acceptable salt thereof or the composition of claim 2 in the preparation of a medicament for treating and/or preventing cancer. 5. The application according to claim 4, wherein the cancer is lung cancer, colon cancer, liver cancer, leukemia or thyroid cancer.