WO2023193795A1

WO2023193795A1 - Compound targeting focal adhesion kinase and preparation method for and application of compound

Info

Publication number: WO2023193795A1
Application number: PCT/CN2023/086924
Authority: WO
Inventors: 张华北; 李晔
Original assignee: 北京师范大学
Priority date: 2022-04-08
Filing date: 2023-04-07
Publication date: 2023-10-12
Also published as: CN114716385B; CN114716385A

Abstract

The present invention relates to a compound targeting focal adhesion kinase and a preparation method for and an application of the compound. The compound targeting focal adhesion kinase of the present invention has the following general formula (I). The compound targeting focal adhesion kinase of the present invention is simple in structure and can be used as a labeling precursor, preferably as a radioactive labeling precursor. The compound targeting focal adhesion kinase of the present invention further has an application in targeting focal adhesion kinase, or a use in preparation of a tumor treatment drug, or a use in preparation of a tumor diagnosis imaging agent, and has a good application effect.

Description

Compounds targeting focal adhesion kinase and preparation methods and applications

Technical field

The present invention relates to the field of pharmaceutical technology, and in particular to a compound targeting focal adhesion kinase and its preparation method and application.

technical background

Focal adhesion kinase (Fak) plays a key role in tumor invasion and metastasis. It is a cytoplasmic tyrosine kinase that participates in multiple functions of cells. It plays a key role in tumor cell invasion, metastasis, angiogenesis and signaling. It plays an important role in transduction and other aspects, and is overexpressed in various tumor cells and is related to the malignancy of tumor cells, so it can become a molecular marker for tumor diagnosis. FAK is located in integrin and receptor tyrosine kinase signaling. The transduction junction can transmit extracellular matrix signals into cells, and inhibiting the function of FAK can effectively block some tumor-related signaling pathways. The overexpression of FAK may be related to its gene amplification, and it has also been shown to play an important role in cell survival, migration and invasion. Therefore, FAK has become an important target for tumor diagnosis and treatment.

Contents of the invention

In view of the above technical limitations, the present invention proposes a compound targeting focal adhesion kinase and its preparation method and application. The compound of the present invention that targets focal adhesion kinase has a simple structure and better effect.

In order to achieve the above objects, the present invention adopts the following technical solutions:

One of the objects of the present invention is to provide a compound that targets focal adhesion kinase, and the compound that targets focal adhesion kinase has the following structural formula of Formula I:

R ₁ is selected from

R ₂ is selected from hydrogen or

The R ₃ is selected from alkoxy, alkoxy substituted by alkyl, or hydroxyl;

R ₇ in R ₁ , R ₂ or R ₃ is the same or different, and each is independently selected from C1-C5 alkyl;

The R ₄ is selected from hydrogen, alkoxy, nitro,

The R ₈ is selected from a C1-C5 linear alkyl group or a cycloalkyl group; the R ₉ or R ₁₀ are the same or different, and each is independently selected from a methyl group or a heteroaryl group containing a halogen substituent; the X Selected from halogen;

The R ₅ is selected from halogen or an alkyl group completely substituted by halogen.

Preferably, in the compound targeting focal adhesion kinase,

R ₇ in R ₁ , R ₂ or R ₃ is the same or different, and each is independently selected from C1-C2 alkyl; and/or,

The alkyl group in the alkoxy group in R ₃ or R ₄ is selected from C1-C2 alkyl groups;

In the R ₃ , the alkoxy group substituted by the alkyl group has the following general formula: The R ₁₁ is selected from halogen, hydroxyl, and OTs group; the n is selected from 1-5;

The R ₈ is selected from C1-C3 alkyl or C3-C5 cycloalkyl;

The R _{9 and} R ₁₀ are independently selected from F-substituted pyrazolyl or pyridinyl.

Preferably,

R ₇ in R ₁ , R ₂ or R ₃ are all selected from methyl;

The alkyl group in the alkoxy group in R ₃ or R ₄ is selected from methyl;

The R ₅ is selected from bromine or CF ₃ ;

The R ₁₁ is selected from F or ¹⁸ F; the n is selected from 1-3;

The R ₈ is selected from ethyl or cyclopropyl;

The R _{9 and} R ₁₀ are independently selected from

Preferably, the compound targeting focal adhesion kinase has the following structural formula:

The n is selected from 1-3.

Preferably,

The R ₄ is selected from hydrogen, and preferably the R ₂ is also selected from hydrogen.

Preferably, the compound targeting focal adhesion kinase includes at least one of the following compounds:

Another object of the present invention is to provide a method for preparing a compound targeting focal adhesion kinase according to one of the objects of the present invention, which includes the following steps:

The hydroxyl group in is substituted by R ₃ group; said R ₃ is selected from alkoxy group, alkoxy group in which alkyl group is substituted, optionally hydroxyl;

(2) Optionally, The nitro group in is further substituted by R ₄ group; the R ₄ is selected The process of substitution of nitro group by R ₄ group includes the process of reduction of nitro group to amino group and then substitution of amino hydrogen;

The second method: React with the compound represented by the general formula R'-NH ₂ , and synthesize the compound of the general formula (IV) through p-toluenesulfonic acid catalysis where R' is selected from _R'1 is selected from _R3 group or nitro group.

The preparation method of the compound targeting focal adhesion kinase of the present invention can be prepared according to the existing conventional preparation method, and can also be prepared by the method given by the present invention, specifically including: preparing the structure such as formula (Ⅰ-Ⅰ) For the compound shown, the synthesis route is roughly as follows (there are two methods, denoted as method A and method B):

Method A:

The specific preparation method of method A includes the following steps:

A1) Dissolve 5-bromo-2,4-dichloropyrimidine, 2-amino-N-methylbenzamide, and potassium carbonate in DMF, and place the system at 70°C for 15 hours to react to obtain Compound 1.

A2) Dissolve compound 1 (1eq), p-methoxyaniline (1.2eq), and p-toluenesulfonic acid monohydrate (0.4eq) in 1.4-dioxane, and stir the reaction system at 70°C overnight. , compound 2 can be obtained.

A3) Compound 2 is demethoxylated under the conditions of boron tribromide and dichloromethane to obtain compound 3.

A4) Compounds 4a-4c can be obtained by reacting compound 3 with compounds 9a-9c in potassium carbonate and DMF at 70°C.

A5) Compounds 4a-4c can be obtained by reacting compounds 4a-4c with p-toluenesulfonyl chloride, triethylamine, and dichloromethane under stirring at room temperature.

A6) Compounds 5a-5c can be obtained by heating compounds 5a-5c to 50°C under the conditions of TBAF and tetrahydrofuran.

The specific preparation method of method B includes the following steps:

A1) Compound 7a-7c can be obtained by reacting compound 4-aminophenol and compound 9a-9c in potassium carbonate and DMF at 70°C.

A2) Compounds 7a-7c can be obtained by stirring compounds 7a-7c in hydrogen and palladium carbon at room temperature.

The target compound prepared by A3) is the same as step A4) of method 1, but is different from step A4. The specific method is as follows: heat compounds 8a-8c and compound 1 in p-toluenesulfonic acid monohydrate and 1.4-dioxane to 70 Compounds 4a-4c were obtained at ℃.

A4) Compound 1 and 4-aminophenylene ether are heated to 70°C with p-toluenesulfonic acid monohydrate and 1.4-dioxane to obtain compound LY-4.

When preparing the compound represented by the structural formula (I-II), the synthetic route of the present invention is as follows, which is recorded as method C:

The specific preparation method of method C includes the following steps:

B1) Dissolve 5-bromo-2,4-dichloropyrimidine in THF, slowly add ammonia water dropwise, and stir at room temperature for two hours to obtain the compound 10.

B2) Dissolve compound 10 and potassium carbonate in DMF, then slowly add dinitrofluorobenzene and react at 70°C for 5-7 hours to obtain compound 11.

B3) Compound 11 is reduced to an amino group by reducing the nitro group to an amino group under iron powder ammonium chloride conditions to obtain compound 12.

B4) Mix compound 12 and acetyl chloride in triethylamine as the base and anhydrous dichloromethane as the solvent. Stir in an ice bath for half an hour to obtain compound 13.

B5) Compound 14 can be obtained by adding compound 13, 4-methoxy-3-nitroaniline to p-toluenesulfonic acid monohydrate and 1.4-dioxane at 70°C.

B6) The synthesis method of compound 15 is the same as step B3.

B7) Compound 16 can be obtained by stirring compound 15, N-acetyl-β-alanine, HATU, DIPEA, and DMF at room temperature.

B8) The synthesis method of compound 17 is the same as step A3.

B9) The synthesis method of compounds 18a-18c and 20 is the same as step A4 of method 1.

B10) The synthesis method of compounds 19a-19c and 21 is the same as step A6 of method 1.

B11) The synthesis method of compounds 22a, 22b, 24a and 24b is the same as step B7.

B12) The synthesis method of compounds 23a and 23b is the same as step A4 of method 2.

B13) The synthesis method of compound 26 is the same as step A1 of method 1.

B14) The synthesis method of compound 27 is the same as step B3.

B15) The synthesis method of compound 28 is the same as step B7.

B16) The synthesis method of compound 29 is the same as step A1 of method 2.

B17) Compound 30a-30c is obtained by using compound diol in TBSCl, imidazole and dichloromethane as solvents under room temperature stirring conditions.

B18) The synthesis method of compounds 31a-31c is the same as step A5 of method 1.

B19) The synthesis method of compound 32 is the same as step B7.

B20) The synthesis method of compounds 33a-33c is the same as the steps

B21) Dissolve compounds 33a-33c in tetrahydrofuran, add potassium tert-butoxide, add methyl iodide after 30 minutes, and stir for 1 hour to obtain compounds 34a-34c.

B22) The synthesis method of compounds 35a-35c is the same as step A2 of method 1.

B23) The synthesis method of compounds 36a-36c is the same as step A4 of method 2.

B24) Dissolve compounds 36a-36c in ethyl acetate, add hydrochloric acid in ethyl acetate, and stir at room temperature overnight to obtain compounds 37a-37c.

B25) The synthesis method of compounds 38a-38c is the same as step A5 of method 1.

B26) The synthesis method of compounds 39a-39c is the same as step A6 of method 1.

B27) The synthesis method of compound 40 is the same as step A4 of method 1.

B28) The synthesis method of compound 41 is the same as step A1 or B3 of method 2.

B29) Compound 42 can be obtained by combining compound 41 with 2,4-dichloro-5-trifluoromethylpyrimidine under zinc chloride conditions.

B30) The synthesis method of compound 43 is the same as step A1 of method 1.

B31) The synthesis method of compound 44 is the same as steps B3 and B4, with a total of two steps.

Synthetic route diagram of compounds LY-47 to LY-52:

Raw materials and reaction conditions: (a) K ₂ CO ₃ , DMF, 70°C; (b) H ₂ , 10% Pd/C, MeOH, 50°C; (c) K ₂ CO ₃ , THF, 60°C; (d) )1)SOCl ₂ ,DCM, reflux; 2) THF, 0℃; (e) Fe powder:NH ₄ Cl＝1:1(n:n), MeOH:THF:H ₂ O＝5:5:2( v:v:v).

Raw materials and reaction conditions: (a) isopropanol, p-Toluenesulfonic acid monohydrate, 90℃; (b) TsCl, Et ₃ N, anhydrous DCM, RT; (c) K ₂₂₂ , KF, K ₂ CO ₃ , DMF.

In the present invention, there are no special requirements for the amount of each substance added in the preparation method of the compound targeting focal adhesion kinase. It can be adjusted according to the conventional dosage for similar reactions in this field or by conventional means. The synthesis method is the synthesis method disclosed above. You can also choose from synthetic methods with similar structures or similar functional group transformations that have been disclosed in the art.

The second object of the present invention is to provide the use of the compound described in one of the objects of the present invention as a labeling precursor, preferably as a radioactive labeling precursor.

The third object of the present invention is to provide the application of the compound described in one of the objects of the present invention in targeting focal adhesion kinase or in the preparation of tumor therapeutic drugs or in the combined use of drugs to treat tumors or in the preparation of tumor diagnostic displays. For use in imaging agents, it is preferred that the tumor is selected from the group consisting of lung cancer, liver cancer, ovarian cancer, pancreatic cancer, colon cancer, prostate cancer, and brain glioma.

The present invention also provides a pharmaceutical composition, which contains a therapeutically effective amount of the compound described in one of the objects of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, diluent and excipient. Pharmaceutical compositions can be formulated for specific routes of administration, such as oral, parenteral, rectal, and the like. Orally administered, such as tablets, capsules (including sustained release or timed release formulations), pills, powders, granules, relaxants, tinctures, suspensions (including nanosuspensions, micron suspensions, spray-dried dispersions) , syrups and emulsions; sublingual; buccal; parenteral, e.g. by subcutaneous, intravenous, intramuscular or intrasternal injection, or infusion techniques (e.g. as sterile injectable aqueous or non-aqueous solutions or mixed suspension); nasally, including administration to the nasal mucosa, for example by inhalation spray; topically, for example in the form of a cream, cream or ointment; or rectally, for example in the form of a suppository. They may be administered alone, but will generally be administered with a pharmaceutical carrier selected based on the chosen route of administration and standard pharmaceutical practice.

Compared with the prior art, the present invention at least has the following advantages:

1) The present invention designs and synthesizes a series of compounds targeting FAK, which can be used as small molecule compounds to inhibit tumor growth, and can also be used as small molecule compounds as inhibitors of some rare diseases;

2) The compounds described in the present invention can be used to label the radionuclide F-18 for the diagnosis of tumors and can be used as tumor imaging agents;

3) The present invention proposes that FAK-targeting small molecule compounds can be labeled in one step when preparing related radioactive drugs. The labeling rate is high. In the existing technology, most of the compounds in the patents applied by this research team are labeled by a two-step method, and the labeling rate is very low.

4) The present invention has made a large number of FAK inhibitors. Through the kinase activity test, it was found that the kinase inhibitory activity is generally high when R ₄ and R ₂ are hydrogen, and from the theoretical model analysis, it can be seen that such compounds are more likely to bind to the protein cavity. good.

5) The FAK inhibitor of the present invention has good activity. Taking compound LY-3 as an example, its kinase inhibitory activity is higher than the current clinical phase II FAK drug VS-6063, and the anti-tumor effect of compound LY-3 was found through tumor inhibition experiments. It is better than VS-6063 and the lung cancer drug erlotinib currently on the market, which shows that its single drug effect is very good.

6) The FAK inhibitors of the present invention can be used in combination, and have better anti-tumor effects than single drugs. Taking compound LY-21 as an example, its combination with erlotinib showed better results in lung cancer mouse models. Single use of the drug has better anti-tumor effect, and the mice are also very active and do not lose weight. Indicates low toxicity. It can achieve the effect of 1+1 greater than 2. It can be seen that the combination of drugs is of great significance.

7) The radioactive labeling of the FAK inhibitor of the present invention can achieve one-step labeling, and the labeling rate is high. Moreover, its tumor uptake is high and its imaging is clear, which is of great significance in tumor diagnosis.

Description of the drawings

Figure 1 shows the tumor inhibition experiments of LY-21 and existing compounds;

Figure 2 is a mass diagram of tumors taken out and weighed after 28 days of administration of LY-21 and existing compounds;

Figure 3 is a graph showing the tumor inhibition experiments of LY-3 and existing compounds;

Figure 4 is a mass diagram of tumors taken out and weighed after 28 days of administration of LY-3 and existing compounds;

Figure 5 is a diagram showing the radioactive diagnostic effect of LY-10.

Reference numbers: 1-Tumor site

Detailed ways

In order to make the purpose, technical solutions and advantages of the present invention clearer, the present invention will be described in further detail below. However, it should be understood that the description here is only used to explain the present invention and is not used to limit the scope of the present invention.

Unless otherwise defined, all technical and scientific terms used herein have the same meanings as commonly understood by those skilled in the technical field of the present invention. The terms used herein in the description of the present invention are only for describing specific implementations. The examples are not intended to limit the invention. The reagents and instruments used in this article are all commercially available. For the characterization methods involved, please refer to the relevant descriptions in the prior art, and will not be described again here.

In order to further understand the present invention, the present invention will be further described in detail below in conjunction with the best embodiments.

Example 1

Synthetic procedure as shown in Method C, synthesis of compound 10

Dissolve 5-bromo-2,4-dichloropyrimidine (30g, 131.65mmol) in THF (150mL), slowly add ammonia water (150ml) dropwise, stir at room temperature for two hours, and detect that the reaction is complete by TLC, add 150mL Water, extracted with EA (100ml*3), the organic phase was dried with anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain 24.7g of white solid product, yield: 90%. 1H NMR (600MHz, DMSO-D6) δ8.18 (d, J＝43.5Hz, 2H), 7.31 (s, 1H).

Synthesis of Compound 11

Add compound 10 (20g, 95.95mmol) and potassium carbonate (15.91g, 115.14mmol) into the reaction flask, add DMF (150mL), stir at room temperature for 15 minutes, and then slowly add dinitrofluorobenzene (21.4g, 115.14 mmol), heat the reaction system to 70°C. After five hours, the reaction is complete by TLC. Bring the reaction system to room temperature, add 20 mL of water, a yellow solid will precipitate, filter, wash the filter cake with water, collect the filter cake, and use it with ethyl acetate. The ester petroleum ether was recrystallized to obtain 31.19g of yellow solid product, yield: 86.8%. 1H NMR (400MHz, DMSO-D6) δ10.58(s,1H),8.83(s,1H),8.75(s,1H),8.63(d,J＝10.2Hz,1H),8.41(d,J＝ 9.7Hz,1H).13C NMR(151MHz,DMSO-D6)δ160.66,157.77,143.00,140.10,138.46,129.49,126.17,122.03,106.04.MS(ESI+):m/z calcd for C ₁₀ H ₅ BrClN ₅ O _4,372.9213 ; found,373.9293(M+H+)

Synthesis of Compound 13

Compound 11 (20g, 53.4mmol) was dissolved in tetrahydrofuran (150mL), methanol (150mL), and water (50mL), and iron powder (7.45g, 133.5mmol) and ammonium chloride (7.14g, 133.5mmol) were added. Stir for 6 hours at 70°C. The reaction is complete by TLC. Filter and concentrate the filtrate. Then dissolve it in tetrahydrofuran (150 mL) and place it in an ice water bath. Add triethylamine (13.5 g, 133.5 mmol) and place in an ice water bath. Stir for 40 minutes, then slowly add acetyl chloride (9.22g, 117.48mmol) dropwise, After 1 hour, the reaction was complete by TLC. Water (150 mL) was slowly added to quench the reaction, extracted with ethyl acetate, the organic phase was collected, dried over anhydrous magnesium sulfate, concentrated, and subjected to column chromatography (PE:EA=5 :1 to 1:1) to obtain 7.66g of white solid, two-step yield: 36%.

Synthesis of compound 25

Add compound 13 (3g, 7.56mmol), p-toluenesulfonic acid (520mg, 3.02mmol), and 4-amino-2-nitrophenol (1.75g, 11.34mmol) into a 250mL round-bottomed flask, and add 1,4- Dissolve the raw materials with dioxane (100 mL), replace the argon gas, and then place the reaction system at 70°C and stir for 24 hours. The reaction is complete by TLC. When the reaction system reaches room temperature, add saturated sodium bicarbonate solution to quench the reaction. , extracted with ethyl acetate, concentrated and recrystallized to obtain 3.11g of yellow solid product, yield: 79.6%.

Example 2

Synthesis of compound 26

Add compound 2-bromo-N,N-dimethylacetamide (2g, 12.05mmol), potassium carbonate (2g, 14.46mmol), and potassium iodide (200.9mg, 1.21mmol) into a 100mL round-bottom flask and add DMF (50mL ) was dissolved. Compound 25 (6.22g, 12.05mmol) was dissolved in DMF (20mL) and slowly added dropwise to the above reaction system. After the dropwise addition was completed, the compound was placed at 50°C to react for 4 hours. The reaction was completed by TLC. Bring the reaction to room temperature, add 100 mL of water, and a yellow solid will precipitate. Filter, wash the filter cake with water, and dry to obtain 5.8 g of product. Yield: 80%.

Example 3

Synthesis of compound LY-23

Add compound 26 (100 mg, 0.17 mmol), potassium carbonate (35.93 mg, 0.26 mmol), and sodium iodide (2.5 mg, 0.017 mmol) into a 10 mL round-bottom flask, add 1 mL DMF, and slowly add p-toluenesulfonic acid 2- Fluoroethyl ester (74.2 mg, 0.34 mmol) in DMF solution (1 mL). After the dropwise addition, the reaction system was placed at 70°C and stirred overnight. The reaction was brought to room temperature, 3 mL of water was added, extracted with ethyl acetate, and the organic phase was collected. , drying, concentration, and column chromatography (dichloromethane: methanol = 50:1 to 20:1) to obtain 74.1 mg of white solid product, yield: 70.59%. 1H NMR (400MHz, DMSO-d6) δ10.27(d,J＝28.2Hz,2H),8.86(s,1H),8.11(d,J＝8.3Hz,2H),7.66(m,1H),7.60 (d,J＝8.8Hz,1H),7.44(d,J＝8.4Hz,1H),6.84(d,J＝8.7Hz,1H),6.74(s,1H),6.60(d,J＝8.6Hz ,1H),5.07(d,J＝6.2Hz,1H),4.68(s,2H),4.57(t,J＝5.1Hz,1H),4.45(t,J＝5.1Hz,1H),3.13(m ,2H),2.96(s,3H),2.83(s,3H),2.07(d,J＝9.7Hz,6H).13C NMR(101MHz,DMSO)δ169.98,168.77,168.23,158.77,157.36,156.86,141.37 ,138.64,136.92,135.41,131.58,127.58,127.30,116.92,115.40,113.33,107.53,102.59,83.54,81.90,68.10,43.44,36.03,35.44,24.38,2 3.38.

Example 4

Synthesis of compound 28

Under an ice bath, dissolve the compounds pyrazole-3-carboxylic acid (300 mg, 0.45 mmol) and HATU (190.12 mg, 0.5 mmol) in 2 mL DMF, add DIPEA (116.3 mg, 0.9 mmol), and react under an ice bath for 40 min. , then dissolve compound 27 (285.7mg, 0.5mmol) in 2mL DMF and slowly drip it into the above reaction system. After the dropwise addition is completed, react at room temperature overnight. The reaction is complete by TLC. Add 5mL water and extract with ethyl acetate. , collect the organic phase, dry, concentrate, and perform column chromatography (dichloromethane: methanol = 30:1 to 15:1) to obtain 101.8 mg of gray-black solid product, yield: 34%. MS(ESI+):m/z calcd for C ₂₈ H ₂₉ BrN ₁₀ O ₅ , 664.1506; found,665.1585(M+H+).

Example 5

Synthesis of compound 29(LY-33)

The synthesis method was the same as that of compound 26, except that compound 28 was used as the starting material. 35 mg of white solid was obtained through column chromatography (dichloromethane: methanol = 40:1 to 20:1), with a yield of 66.69%. ¹ H NMR (400MHz, DMSO-d6) δ9.99 (d, J = 14.8Hz, 2H), 9.72 (s, 1H), 9.12 (s, 1H), 8.23 (s, 1H), 8.10 (s, 2H) ),7.90(s,1H),7.64(s,1H),7.58(s,1H),7.36(s,2H),6.83(s,1H),6.76(s,1H),4.88(s,3H) ,4.78(s,1H),4.54(s,1H),4.47(s,1H),2.96(s,3H),2.82(s,3H),2.04(d,J＝15.9Hz,6H).13C NMR (101MHz, DMSO-D6) δ170.06,168.84,168.13,159.38,158.89,157.44,157.07,147.03,146.23,143.49,138.15,136.90,134.77,133.74,131.79,128. 51,128.46,127.56,126.01,106.86,106.81,93.37 ,83.18,81.52,68.45,52.98,35.57,24.50,23.53,21.30.MS(ESI+):m/z calcd for C ₃₀ H ₃₂ BrFN ₁₀ O ₅ ,710.1725; found,711.1806(M+H+)

Example 6

Synthesis of compound 14

The synthesis method was the same as that of compound 25, except that compound 13 was used as raw material. After the reaction was completed, it was obtained by recrystallization (5 g, yellow solid, yield 78.8%). 1H NMR(600MHz,DMSO-D6)δ10.21(s,1H),10.15(s,1H),9.46(s,1H),8.24(m,1H),8.01(s,1H),7.81(s, 1H),7.69(s,1H),7.54(s,1H),7.45(s,1H),7.14(s,1H),3.86(s,3H),2.08(s,6H).13C NMR(151MHz, DMSO-D6)δ170.03,168.88,158.40,157.43,146.87,139.21,137.49,134.17,132.33,127.97,127.11,125.40,116.90,115.25,115.07,114.85,94. 14,57.24,24.50,23.54.MS(ESI+):m /z calcd for C21H20BrN7O5,529.0709; found,530.0776(M+H+).

Example 7

Synthesis of compound 17

Add compound 16 (2g, 3.26mmol) to the high-pressure reaction kettle, add anhydrous dichloromethane (15mL) and boron tribromide (1.63g, 6.52mmol), heat the reaction system to 60°C and react overnight. The reaction system was brought to room temperature, slowly decompressed, and saturated sodium bicarbonate solution was slowly added dropwise until the reaction system became slightly alkaline. Extract with dichloromethane, collect the organic phase, dry, concentrate, and perform column chromatography (dichloromethane:methanol). =50:1 to 25:1) to obtain 1.17g of white solid product, yield: 59.8%. 1H NMR (400MHz, DMSO-d6) δ9.99 (d, J＝9.6Hz, 2H), 9.27 (s, 1H), 9.18 (s, 1H), 8.96 (s, 1H), 8.06 (d, J＝ 5.6Hz,2H),7.88(s,1H),7.63(d,J＝10.9Hz,3H),7.30(d,J＝9.3Hz,1H),7.21(d,J＝8.7Hz,1H),6.59 (d,J＝9.6Hz,1H),3.25(s,2H),2.49(s,2H),2.04(d,J＝8.4Hz,6H),1.75(s,3H).

Example 8

Synthesis of compound 20

Add the above compound (500mg, 0.66mmol), (2-bromoethoxy)-tert-butyldimethylsilane (189mg, 0.79mmol), and potassium carbonate (109.2mg, 0.79mmol) into a 25mL round-bottomed flask, Add 5mLDMF to dissolve the raw materials and place it at 70℃ The reaction was carried out overnight. The reaction was completed by TLC. 10 mL of water was added and extracted with ethyl acetate. The organic phase was collected, dried, concentrated, and subjected to column chromatography (dichloromethane: methanol = 50:1 to 20:1) to obtain 400.1 mg of white color. Solid product, yield: 80.3%. 1H NMR(400MHz,DMSO-d6)δ10.02(s,2H),9.08(s,1H),8.73(s,1H),8.11(s,1H),7.90(s,2H),7.62(s, 2H),7.42(d,J＝7.2Hz,1H),7.34(d,J＝7.4Hz,1H),6.79(m,1H),4.00(s,2H),3.92(s,2H),3.28( s,2H),2.49(s,2H),2.07(d,J＝7.3Hz,6H),1.78(s,3H),0.87(s,9H),0.06(s,6H).13C NMR(151MHz, DMSO-D6)δ170.00,169.93,169.69,169.61,169.52,168.75,168.66,158.87,158.78,157.46,157.06,144.60,136.88,133.87,130.18,127.60,116 .78,115.09,113.10,100.00,70.96,62.11,35.63, 29.51,26.35,24.47,23.11,18.55,-4.73.MS(ESI+):m/z calcd for C33H45BrN8O6Si,756.2415; found,757.2484(M+H ⁺ )

Add the product from the previous step (400 mg, 0.53 mmol) into a 10 mL round-bottomed flask, add anhydrous dichloromethane (2 mL) and trifluoroacetic acid (2 mL), react at room temperature for 5 hours, and detect that the reaction is complete by TLC. Concentrate The solvent was removed and column chromatography (dichloromethane: methanol = 50:1 to 20:1) was performed to obtain 272.6 mg of white solid product, yield: 75%. ¹ H NMR (400MHz, DMSO-d6) δ10.12(dd,J＝16.5,51.9Hz,3H),9.27(d,J＝48.2Hz,1H),9.03(d,J＝11.8Hz,1H), 8.29(s,1H),7.99(s,1H),7.94(s,1H),7.71(s,1H),7.44(d,J＝14.6Hz,2H),7.09(s,1H),6.79(s ,1H),3.93(s,2H),3.69(s,2H),3.26(s,2H),2.55(s,2H),2.05(s,6H),1.74(s,3H).

Add the product from the previous step (200 mg, 0.3 mmol) into a 10 mL round-bottomed flask, add anhydrous dichloromethane (3 mL) and triethylamine (91.1 mg, 0.9 mmol), and sonicate the system under ultrasound for 20 minutes. Dissolve some of the raw materials as much as possible, then add p-toluenesulfonyl chloride (114.4 mg, 0.6 mmol) and stir at room temperature overnight. Concentrate the reaction solution and perform column chromatography (dichloromethane: methanol = 50:1 to 25:1) to obtain 30 mg of white color. Solid product, yield: 12.54%. 1H NMR(600MHz,Methanol-d4)δ8.02(s,2H),7.71(s,2H),7.64(s,1H),7.45(s,1H),7.29(s,2H),7.19(s, 1H),6.68(s,1H),4.39(s,2H),4.19(s,2H),3.47(s,2H),2.62(s,2H),2.34(s,3H),2.12(s,6H ),1.90(s,3H).MS(ESI+):m/z calcd for C34H37BrN8O8S,796.1638; found,797.1704(M+H+).

Example 9

Synthesis of Compound 21 (LY-22)

Compound 17 (60 mg, 0.1 mmol), 1-bromo-3-fluoropropane (16.9 mg, 0.12 mmol), and potassium carbonate (16.6 mg, 0.12 mmol) were dissolved in DMF (2 mL), and the reaction system was heated to 70°C. The reaction was carried out overnight. The reaction was completed by TLC. Water was added and extracted with ethyl acetate. The organic phase was collected, dried, concentrated, and subjected to column chromatography (dichloromethane: methanol = 50:1 to 20:1) to obtain 40 mg of white solid, yield 60.65%. ¹ H NMR(600MHz,DMSO-d6)δ10.05(m,2H),9.05(s,1H),8.84(s,1H),8.08(s,2H),7.88(s,1H),7.82(s ,1H),7.59(s,2H),7.41(s,1H),7.30(s,1H),6.75(s,1H),4.65(s,1H),4.57(s,1H),3.99(s, 2H),3.24(s,2H),2.09(s,2H),2.04(s,6H),1.74(s,3H). ¹³ C NMR(151MHz,DMSO-D6)δ170.00,169.85,169.74,168.84,158.84 ,157.40,157.16,144.92,137.04,133.84,131.91,127.82,127.53,116.91,115.76,115.29,112.69,100.00,82.14,81.07,65.18,36.69,35.74 ,30.37,24.47,23.53,23.10.MS(ESI+): m/z calcd for C28H32BrFN8O5,658.1663; found,659.1724(M+H+)

Example 10

Synthesis of compound 18b

Compound 17 (200 mg, 0.33 mmol) was dissolved in 2 mL of DMF, diethylene glycol bis-p-toluenesulfonate (207.2 mg, 0.5 mmol) and potassium carbonate (69.1 mg, 0.5 mmol) were added, and the reaction system was placed at 70°C React for 10 hours. The reaction is complete by TLC. When the reaction system reaches room temperature, add 4 mL of water, extract with ethyl acetate, collect the organic phase, dry with anhydrous sodium sulfate, and concentrate. Condensation and column chromatography (dichloromethane: methanol = 60:1 to 35:1) obtained 143.1 mg of white solid product, yield: 51.52%. 1H NMR (600MHz, DMSO-d6) δ10.07(s,2H),9.15(s,1H),8.79(s,1H),8.17(d,J＝16.2Hz,1H),7.91(s,2H) ,7.77(s,2H),7.65(s,2H),7.43(s,3H),7.35(s,1H),6.78(s,1H),4.16(s,2H),3.98(s,2H), 3.69(s,4H),3.27(s,2H),2.36(s,3H),2.07(s,6H),1.77(s,3H).MS(ESI+):m/z calcd for C36H41BrN8O9S,840.1901; found ,841.1981(M+H+).

Example 11

Dissolve the compound diethylene glycol bis-p-toluenesulfonate (2g, 4.83mmol) in 15mL tetrahydrofuran, add 5.8mL TBAF (1M tetrahydrofuran solution), place the reaction system at 50°C for 9 hours, and detect by TLC The reaction was complete, the solvent was removed by rotary evaporation, and 708.2 mg of colorless oily liquid was obtained by column chromatography (petroleum ether: ethyl acetate = 10:1 to 5:1), yield: 55.9%. 1H NMR(400MHz,Chloroform-d)δ7.80(t,J＝6.4Hz,2H),7.34(d,J＝6.1Hz,2H),4.54(s,1H),4.42(s,1H),4.17 (s,2H),3.71(s,3H),3.63(s,1H),2.44(s,3H).13C NMR(151MHz,Chloroform-d)δ144.92,133.08,129.89,128.06,82.49,70.62,70.49, 69.23,68.97.MS(ESI+):m/z calcd for C11H15FO4S,262.0675; found,285.0583(M+Na+).

Example 12

Synthesis of compound 19b

The synthesis method was the same as that of compound 20. Using compound 17 as raw material, 36 mg of white solid was obtained through column chromatography (dichloromethane: methanol = 50:1 to 20:1), with a yield of 56%. 1H NMR (400MHz, DMSO-d6) δ10.09(d,J＝18.0Hz,2H),9.07(s,1H),8.78(s,1H),8.08(s,2H),7.89(d,J＝ 11.2Hz,2H),7.59(d,J＝12.0Hz,2H),7.41(s,1H),7.31(s,1H),6.77(d,J＝9.2Hz,1H),4.58(s,1H) ,4.46(s,1H),4.04(s,2H),3.75(s,3H),3.67(s,1H),3.24(s,2H),2.48(s,2H),2.04(s,6H), 1.74(s,3H).13C NMR(151MHz,DMSO-D6)δ170.02,169.74,168.85,158.87,157.47,157.10,144.69,137.02,134.28,131.82,130.18,128.09,127.57, 116.92,116.65,115.31,113.93, 84.16,83.06,70.43,70.30,69.48,36.73,35.71,24.48,23.51,23.10.MS(ESI+):m/z calcd forC29H34BrFN8O6,688.1769; found,689.1849(M+H+).

Example 13

Synthesis of compound 18c

The synthesis method was the same as that of compound 20. Using compound 17 as raw material, 50 mg of white solid was obtained through column chromatography (dichloromethane: methanol = 50:1 to 20:1), with a yield of 49%. 1H NMR(400MHz,DMSO-d6)δ9.99(s,2H),9.06(s,1H),8.75(s,1H),8.08(s,2H),7.86(d,J＝16.5Hz,2H) ,7.73(t,J＝6.0Hz,2H),7.60(s,2H),7.42(d,J＝10.3Hz,2H),7.36(d,J＝9.8Hz,1H),7.31(d,J＝ 9.6Hz,1H),6.77(d,J=8.9Hz,1H),4.06(s,2H),4.00(s,2H),3.67(s,2H),3.54(s,2H),3.51(s, 2H),3.45(s,2H),3.23(d,J＝11.9Hz,2H),2.43(s,2H),2.35(s,3H),2.03(d,J＝10.3Hz,6H),1.74( s,3H).MS(ESI+):m/z calcd for C38H45BrN8O10S,884.2163; found,885.2238(M+H+).

Example 14

Synthesis of compound 19c

The synthesis method was the same as that of compound 20. Using compound 17 as raw material, 26 mg of white solid was obtained through column chromatography (dichloromethane: methanol = 50:1 to 20:1), with a yield of 65%. 1H NMR(400MHz,DMSO-d6)δ10.26(s,1H),10.16(s,1H),9.06(s,1H),8.81(s,1H),8.08(s,2H),7.90(d, J＝26.0Hz,2H),7.60(d,J＝14.3Hz,2H),7.44(s,1H),7.30(s,1H),6.78(s,1H),4.52(s,1H),4.40( s,1H),4.02(s,2H),3.71(s,2H),3.64(s,1H),3.57(d,J＝14.0Hz,6H),3.25(s,2H),3.12(s,1H ),2.04(s,6H),1.74(s,3H).MS(ESI+):m/z calcd for C ₃₁ H ₃₈ BrFN ₈ O ₇ ,732.2031; found,755.1894(M+Na ⁺ ).

Example 15

Synthesis of compound LY-14

Add compound 13 (3g, 7.53mmol), p-methoxyaniline (1.11g, 9.04mmol), and p-toluenesulfonic acid monohydrate (572.56mg, 3.01mmol) into a 100mL round-bottomed flask, and add 40mL isopropyl alcohol As the reaction solvent, place the reaction system at 90°C for overnight reaction. The reaction is complete by TLC. Bring the reaction system to room temperature and slowly pour it into 200 mL of ethyl acetate. A solid will precipitate. Filter and wash with ethyl acetate. Filter cake, dry the filter cake to obtain 2.6g of white solid product, yield: 71.18%. 1H NMR(400MHz,DMSO-d6)δ10.17(s,1H),10.09(s,1H),9.71(s,1H),8.78(s,1H),8.24(s,1H),7.74(s, 1H),7.54(s,1H),7.48(s,1H),7.34(d,J＝9.5Hz,2H),6.75(d,J＝6.5Hz,2H),3.70(s,3H),2.08( s,6H).MS(ESI+):m/z calcd for C ₂₁ H ₂₁ BrN ₆ O ₃ ,484.0859; found,485.0935(M+H ⁺ ).

Example 16

Synthesis of compound LY-15

Add the raw material compound LY-14 (2g, 4.12mmol) into the high-pressure reaction kettle, add anhydrous dichloromethane (15mL) and boron tribromide (1.55g, 6.18mmol), and heat the reaction system to 60°C for reaction. Wait overnight, wait until the reaction system reaches room temperature, slowly reduce the pressure, slowly add the reaction system dropwise to the saturated sodium bicarbonate solution to make the final solution slightly alkaline, extract with dichloromethane, collect the organic phase, dry, concentrate, and column Chromatography (dichloromethane: methanol = 50:1 to 35:1) obtained 1.23 g of white solid product, yield: 63.11%. 1H NMR(400MHz,DMSO-d6)δ10.10(s,1H),8.94(s,1H),8.09(s,1H),7.73(s,1H),7.64(d,J＝8.9Hz,1H) ,7.39(m,1H),7.30(d,J＝9.9Hz,2H),6.54(q,J＝9.1,11.7Hz,2H),2.08(s,6H).

Example 17

Synthesis of compound LY-16

The synthesis method is the same as that of compound 20, using compound LY-15 (1g, 2.12mmol) and (2-bromoethoxy)-tert-butyldimethylsilane as raw materials, column chromatography (petroleum ether: ethyl acetate = 5:1 to 1:1) to obtain 1.04g of white solid product, yield: 78%.

Add the product from the previous step (1.04g, 1.65mmol) into a 25mL round-bottomed flask, add 1,4-dioxane solution (10mL) and 4 molar hydrogen chloride-dioxane solution (10mL) respectively, and then Stir at room temperature for 6 hours, check that the reaction is complete by TLC, filter, wash the filter cake with 1,4-dioxane, and dry to obtain the product (747.3 mg, white solid, yield: 88%). 1H NMR(400MHz,DMSO-d6)δ10.11(s,1H),10.06(s,1H),9.11(s,1H),8.13(s,2H),7.72(s,1H),7.60(d, J＝9.2Hz,1H),7.42(d,J＝9.0Hz,3H),6.72(m,2H),4.03(m,1H),3.90(s,2H),3.69(s,2H),2.09( s,6H).MS(ESI+):m/z calcd for C ₂₂ H ₂₃ BrN ₆ O ₄ ,514.0964; found,515.1030(M+H ⁺ ).

Example 18

Synthesis of compound LY-17

Add compound LY-16 (700mg, 1.36mmol) into a 10mL round-bottomed flask, add anhydrous dichloromethane (2mL) and anhydrous triethylamine (688.1mg, 6.8mmol), p-toluenesulfonyl chloride (518.6mg , 2.72mmol) was dissolved in anhydrous dichloromethane (1mL) and slowly added dropwise to the reaction system. After the dropwise addition was completed, the mixture was stirred at room temperature for 12 hours. The reaction was completed by TLC, and 5mL of water was added to quench the reaction. Extract with dichloromethane, collect the organic phase, dry, concentrate and recrystallize (methanol) to obtain 602.6 mg of white solid product, yield: 66.18%. ¹ H NMR(600MHz,DMSO-d6)δ10.03(m,1H),9.98(s,1H),9.08(s,1H),8.11(s,1H),7.75(s,2H),7.67(s ,1H),7.55(s,1H),7.41(s,3H),7.35(s,2H),6.59(s,2H),4.26(s,2H),4.04(s,2H),2.35(s, 3H), 2.03 (s, 6H). ¹³ C NMR (101MHz, DMSO-D6) δ170.01,168.87,158.65,157.52,157.31,152.77,145.53,137.25,134.58,132.70,132.26,130.68,128 .16,127.34,120.74, 116.72,115.07,114.71,100.00,69.69,65.96,24.52,23.57,21.60.MS(ESI+):m/z calcd for C29H29BrN6O6S,668.1053; found,669.1119(M+H+).

Example 19

Synthesis of compound LY-18

To a solution of compound LY-17 (200 mg, 0.3 mmol) in DMF (2 mL), potassium carbonate (4.1 mg, 0.03 mmol), K _2.2.2 (112.9 mg, 0.3 mmol) and KF (34.9 mg, 0.6 mmol) were added , and then the reaction system was placed at 100°C for 1 h. The reaction solution was concentrated and subjected to column chromatography (dichloromethane: methanol = 60:1 to 30:1) to obtain the product (128.2 mg, white solid, yield 82.6%); 1H NMR (600MHz, DMSO-d6) δ 10.09 (s,1H),10.04(s,1H),9.08(s,1H),8.10(s,2H),7.67(s,1H),7.56(s,1H),7.41(s,3H),6.71( s,2H),4.71(s,1H),4.63(s,1H),4.14(s,1H),4.09(s,1H),2.04(s,6H).13C NMR(151MHz,DMSO-D6)δ170 .00,168.88,158.72,157.56,157.32,153.29,137.26,134.51,132.21,127.42,120.90,116.78,115.19,114.73,83.32,82.22,67.78,67.66,24 .52,23.54.MS(ESI+):m/z calcd for C ₂₂ H ₂₂ BrFN ₆ O ₃ ,516.0921; found, 517.0988(M+H+).

Example 19

Add 5-bromo-2,4-dichloropyrimidine (15g, 65.83mmol), 2-amino-N-methylbenzamide (11.86g, 79mmol), and potassium carbonate (10.92g, 79mmol) into a 500mL round bottom In the flask, add DMF (200mL) and place the system at 70°C for 15 hours to react. The reaction is complete by TLC. When the reaction system reaches room temperature, add 500mL of water. A yellow solid will precipitate. Filter and wash the filter cake with water. After drying, a yellow solid product (19.94g, yield: 88.68%) was obtained.

Add the product from the previous step (6g, 17.56mmol), p-methoxyaniline (2.59g, 21.07mmol), and p-toluenesulfonic acid monohydrate (1.34g, 7.02mmol) into a 250mL round-bottomed flask, and add 100mL of isopropyl Use propanol as the reaction solvent. Place the reaction system at 90°C for overnight reaction. The reaction is complete after TLC test. Bring the reaction system to room temperature and slowly pour it into 200 mL of ethyl acetate. If a solid will precipitate, filter it and use acetic acid. Wash the filter cake with ethyl ester and dry it to obtain 6.02g of light yellow solid without purification. Add the obtained filter cake to the high-pressure reaction kettle, add anhydrous dichloromethane (40mL) and boron tribromide (5.28g, 21.09mmol) , place the reaction system at 60°C for 4 hours, wait until the reaction kettle reaches room temperature, slowly reduce the pressure, slowly add the reaction system dropwise into the saturated sodium bicarbonate solution to make the final solution alkaline, and extract with dichloromethane , collect the organic phase, dry, concentrate, and perform column chromatography (dichloromethane: methanol = 60:1 to 35:1) to obtain 2.62g of white solid product, two-step yield: 36.02%. ,1H NMR(600MHz,DMSO-d6)δ11.28(s,1H),9.08(s,1H),9.03(s,1H),8.67(s,1H),8.61(s,1H),8.17(s ,1H),7.68(s,1H),7.37(s,1H), 7.34(s,2H),7.08(s,1H),6.66(s,2H),2.77(s,3H).MS(ESI+):m/z calcd for C18H16BrN5O2,413.0487; found,414.0557(M+H+) .

The synthesis method is the same as that of compound 20. The product of the previous step (2g, 4.83mmol) and (2-bromoethoxy)-tert-butyldimethylsilane are used as raw materials, and a two-step reaction is carried out. The first step is through column chromatography. The white solid product obtained was separated and purified. After the second step of reaction, the compound (930.4 mg, white solid, two-step yield: 42.01%) was obtained by recrystallization. 1H NMR(600MHz,DMSO-d6)δ11.32(s,1H),9.22(s,1H),8.68(s,1H),8.61(s,1H),8.20(s,1H),7.69(s, 1H),7.47(s,2H),7.41(s,1H),7.08(s,1H),6.82(s,2H),3.95(s,2H),3.87(s,2H),2.77(s,3H ),0.84(s,9H),0.04(s,6H).MS(ESI+):m/z calcd for C26H34BrN5O3Si,571.1614; found,572.1692(M+H+).

Using the product from the above step (800 mg, 1.75 mmol) as raw material, 673.8 mg of white solid product was obtained through column chromatography (petroleum ether: ethyl acetate = 3:1 to 1:2), with a yield of 63%. 1H NMR(400MHz,DMSO-d6)δ11.34(s,1H),9.29(s,1H),8.73(s,1H),8.63(s,1H),8.24(s,1H),7.80(m, 2H),7.72(d,J＝6.7Hz,1H),7.50(d,J＝10.6Hz,5H),7.13(d,J＝8.3Hz,1H),6.77(t,J＝7.2Hz,2H) ,4.33(s,2H),4.13(s,2H),2.80(s,3H),2.41(s,3H).13C NMR(151MHz,DMSO-D6)δ169.34,158.86,158.06,156.30,153.48,145.53, 139.78,134.27,132.83,131.81,130.70,128.44,128.20,122.48,122.19,122.00,121.64,114.98,114.81,69.76,66.16,26.84,21.63.MS(ESI+) :m/z calcd for C ₂₇ H ₂₆ BrN ₅ O ₅ S,611.0838; found,612.0916(M+H+).

Synthesis of compound LY-3:

The synthesis method is the same as that of compound LY-18. The nuclear magnetic data of LY-3 is 1H NMR (400MHz, DMSO-d6) δ11.34(s,1H),9.29(s,1H),8.73(s,1H),8.65(s,1H),8.25(s, 1H),7.72(m,1H),7.54(d,J＝6.6Hz,2H),7.48(m,1H),7.13(s,1H),6.91(d,J＝6.2Hz,2H),4.80( s,1H),4.68(s,1H),4.24(s,1H),4.16(s,1H),2.81(s,3H).13C NMR(101MHz,DMSO-D6)δ169.34,158.88,158.09,156.30, 153.93,139.80,134.13,131.85,128.45,122.49,122.18,122.12,121.61,114.91,83.64,81.98,67.79,26.85.MS(ESI+):m/z calcd for C ₂₀ H ₁₉ BrFN ₅ O ₂ ,459.0706; found ,460.0783(M+H+).

Add compound 1 (1g, 2.93mmol), 4-aminophenylene ether (441.7mg, 3.22mmol), and p-toluenesulfonic acid monohydrate (222.6mg, 1.17mmol) into the reaction bottle, and add isopropanol (20mL) , heated to 80°C and stirred overnight. After the reaction is completed, bring the reaction solution to room temperature, slowly pour into water (100mL), a white solid will precipitate, filter and dry to obtain the product (white solid, yield: 76.65%). ¹ H NMR(600MHz,DMSO-d6)δ11.30(s,1H),9.22(s,1H),8.69(s,1H),8.61(s,1H),8.20(s,1H),7.68(s ,1H),7.44(d,J＝25.6Hz,3H),7.09(s,1H),6.80(s,2H),3.95(s,2H),2.77(s,3H),1.28(s,3H) .

Example 20

Dissolve 4-nitrophenol (5g, 45.82mmol), bromoethanol (6.87g, 54.98mmol), and potassium carbonate (7.6g, 54.98mmol) in DMF (100mL), and pass it through column chromatography (petroleum ether: ethyl acetate). Ester = 5:1 to 2:1) was separated and purified to obtain 6.23g of yellow solid product, yield: 72%. MS (ESI ⁺ ): m/z calcd for C ₈ H ₉ NO ₄ , 183.0532; found, 184.1054 (M+H ⁺ ).

The synthesis method is the same as above, using 4-nitrophenol (5g, 45.82mmol) as raw material, and obtaining 6.87g of yellow solid product through column chromatography (petroleum ether: ethyl acetate = 5:1 to 2:1), yield: 66% . 1H NMR (400MHz, Chloroform-d) δ8.18 (q, J＝10.2, 13.6Hz, 2H), 6.98 (d, J＝8.3Hz, 2H), 4.22 (s, 2H), 3.89 (s, 2H) ,3.76(s,2H),3.67(s,2H).MS(ESI+):m/z calcd for C ₁₀ H ₁₃ NO ₅ ,227.0794; found,228.1395(M+H+).

The synthesis method is the same as that of compound 25. Compound 1 (1g, 2.93mmol) is used as raw material, and compound 4b (white solid, yield: 59.8) is obtained through column chromatography (dichloromethane: methanol = 50:1 to 20:1). %). 1H NMR(400MHz,DMSO-d6)δ11.33(s,1H),9.27(s,1H),8.72(s,1H),8.64(s,1H),8.24(s,1H),7.73(s, 1H),7.51(s,2H),7.48(d,J＝8.9Hz,1H),7.13(s,1H),6.87(s,2H),4.60(s,1H),4.06(s,2H), 3.73(s,2H),3.51(s,4H),2.81(s,3H).

Compound 4b (502.4 mg, 1 mmol), p-toluenesulfonyl chloride (190.7 mg, 1 mmol), and triethylamine (121.3 mg, 1.2 mmol) were added to the reaction flask, dissolved in anhydrous dichloromethane (6 mL), and the reaction was The system was stirred at room temperature for 16 hours, 15 mL of water was added to quench the reaction, extracted with dichloromethane, the organic phase was collected, dried and concentrated, and the product was obtained through column chromatography (dichloromethane: methanol = 50:1 to 30:1). 5b (white solid, yield: 56%). 1H NMR (400MHz, Methanol-d4) δ8.57(s,1H),8.08(s,1H),7.75(s,2H),7.60(s,1H),7.36(d,J＝24.1Hz,5H) ,7.08(s,1H),6.83(s,2H),4.16(s,2H),4.00(s,2H),3.71(s,4H),2.89(s,3H),2.35(s,3H). 13C NMR (101MHz, CHLOROFORM-D) δ169.46,156.78,155.19,144.93,139.17,133.04,132.18,131.73,129.91,128.05,126.76,122.94,122.91,122.86,122 .14,114.94,100.00,70.03,69.31,69.01,67.85 ,27.02,21.71.MS(ESI+):m/z calcd for C ₂₉ H ₃₀ BrN ₅ O ₆ S,655.1100; found,656.1162(M+H+).

The synthesis method is the same as that of LY-18. Using compound 5b as raw material, compound 6b (white solid, yield: 70.6%) can be obtained through column chromatography (dichloromethane: methanol = 100:1 to 35:1). ¹ H NMR (400MHz, DMSO-d6) δ11.29(s,1H),9.22(s,1H),8.67(s,1H),8.61(d,J＝8.4Hz,1H),8.20(s,1H ),7.68(d,J＝8.5Hz,1H),7.48(d,J＝8.6Hz,2H),7.42(d,J＝8.2Hz,1H),7.08(t,J＝5.8Hz,1H), 6.84(d,J＝8.7Hz,2H),4.57(s,1H),4.45(s,1H),4.04(s,2H),3.73(s,3H),3.65(s,1H),2.77(s ,3H). ¹³ C NMR (101MHz, DMSO-D6) δ169.34,158.90,158.07,156.29,154.24,139.80,133.87,131.83,128.44,122.46,122.15,121.60,114.83,84.41, 82.77,70.33,69.61,67.82, 26.84.

Example 21

Add nitro compound (2g, 7.37mmol), iron powder (617.7mg, 11.06mmol), ammonium chloride (591.63mg, 11.06mmol) into the reaction flask, add methanol (10mL), water (5mL), tetrahydrofuran ( 10 mL), stir the reaction system at 73°C for 6 hours, detect the reaction is complete by TLC, filter while hot, concentrate the filtrate, and perform column chromatography (petroleum ether: ethyl acetate = 3:1 to 1:1) to obtain Product (black solid, yield: 70.5%). The obtained product (black solid) was filtered and concentrated through diatomaceous earth, yield: 80%. MS (ESI+): m/z calcd for C ₁₂ H ₁₉ NO ₄ , 241.1314; found, 242.1922 (M+H ⁺ ).

The synthesis method was the same as that of compound 4b. Using compound 1 as the starting material, compound 4c (white solid, yield: 59.8%) was obtained through column chromatography (dichloromethane: methanol = 50:1 to 20:1). MS(ESI+): m/z calcd for C ₂₄ H ₂₈ BrN ₅ O _5, 545.1274; found, 546.1349(M+H ⁺ ).

The synthesis method is the same as that of compound 5b. Compound 4c is used as raw material, and the product (white solid, yield: 60%) is obtained through column chromatography (dichloromethane: methanol = 50:1 to 30:1). ¹ H NMR(600MHz,Chloroform-d)δ10.93(s,1H),8.53(s,1H),8.11(s,1H),7.76(s,2H),7.44(m,1H),7.37(s ,2H),7.33(s,1H),7.28(s,3H),7.01(s,1H),6.83(s,2H),6.39(s,1H),4.13(s,2H),4.08(s, 2H),3.79(s,2H),3.65(s,4H),3.59(s,2H),2.97(s,3H),2.39(s,3H). ¹³ C NMR(151MHz,CHLOROFORM-D)δ169. 61,158.51,156.95,156.57,154.92,144.97,139.52,133.00,132.77,131.58,129.93,128.03,128.01,126.90,122.59,122.38,121.89,114.92 ,95.03,70.86,70.81,69.94,69.38,68.80,67.87,26.95, 21.70.MS(ESI+):m/z calcd for C ₃₁ H ₃₄ BrN ₅ O ₇ S,699.1362; found,700.1400(M+H ⁺ ).

The synthesis method of compound LY-10 is the same as that of compound 6b. Compound 5c is used as raw material and a white solid product is obtained through column chromatography (dichloromethane: methanol = 50:1 to 25:1). The yield is 65%. ¹ H NMR (400MHz, DMSO-d6) δ11.29(s,1H),9.22(s,1H),8.68(s,1H),8.61(s,1H),8.20(s,1H),7.69(s ,1H),7.47(s,2H),7.43(s,1H),7.08(d,J＝7.7Hz,1H),6.83(s,2H),4.54(s,1H),4.42(s,1H) ,4.02(s,2H),3.64(m,8H),2.76(s,3H). ¹³ C NMR(101MHz,DMSO-D6)δ169.34,158.89,156.29,154.26,139.80,133.83,131.82,128.43,122.45, 122.14,121.60,114.81,84.39,82.75,70.45,70.39,70.14,69.60,67.82,26.84.MS(ESI+):m/z calcd for C ₂₄ H ₂₇ BrFN ₅ O ₄ ,547.1230; found, 548.1299 (M+H ⁺ ).

Example 21

Under ice bath, add triethylene glycol (10g, 66.59mmol) and imidazole (2.72g, 39.95mmol) into a 500mL round-bottom flask, dissolve the raw materials with anhydrous dichloromethane (150mL), and slowly add tert. Butyldimethylsilyl chloride solution in dichloromethane (5.02g TBS-Cl dissolved in 50mL anhydrous dichloromethane). After the dropwise addition, the reaction system was raised to room temperature and stirred for 12 hours. The reaction was completed by TLC. Pour the reaction system into 200 mL of water, extract with dichloromethane, collect the organic phase, wash the organic phase with saturated ammonium chloride and saturated brine respectively, dry the organic phase, concentrate, and column chromatography (petroleum ether: ethyl acetate = 3 :1 to 1:1) to obtain 11.62g of colorless oil, yield: 66%. MS (ESI+): m/z calcd for C ₁₂ H ₂₈ O ₄ Si, 264.1757; found, 265.2788 (M+H ⁺ ).

Compound 30c (1g, 3.78mmol), p-toluenesulfonyl chloride (865.6mg, 4.54mmol), and triethylamine (459.4mg, 4.54mmol) were dissolved in dichloromethane (15mL), and the reaction system was stirred at room temperature for 15 hours. , add 30 mL of water to quench the reaction, extract with dichloromethane, collect the organic phase, dry, concentrate, and column chromatography (petroleum ether: ethyl acetate = 10:1 to 5:1) to obtain the product (colorless oil, product rate: 66.8%). MS (ESI+): m/z calcd for C ₂₃ H ₃₉ N ₃ O ₈ Si, 513.2506; found, 514.2583 (M+H ⁺ ).

Compound 33c (1g, 1.95mmol) was dissolved in DMF (10mL), potassium tert-butoxide (547.6mg, 4.88mmol) was slowly added, and the mixture was stirred at 0°C for 40 minutes. Methyl iodide (692.7mg, 4.88mmol) was added mmol) was dissolved in DMF (5 mL) and slowly added dropwise to the above reaction system. After the dropwise addition was completed, the reaction system was placed at 30°C for 3 hours to react. The reaction was completed by TLC. Pour the reaction system into ice water and use it with acetic acid. Extract with ethyl ester, collect the organic phase, concentrate, and perform column chromatography (dichloromethane: methanol = 50:1 to 30:1) to obtain compound 34c (yellow solid, yield: 76%).

Add compound 34c (2g, 3.69mmol), iron powder (309.4mg, 5.54mmol), and ammonium chloride (296.33mg, 5.54mmol) into the reaction flask, then add methanol (10mL), water (5mL), and tetrahydrofuran (10mL). ), stir the reaction system at 73°C for 6 hours, detect the reaction is complete by TLC, filter while hot, concentrate the filtrate, and perform column chromatography (petroleum ether: ethyl acetate = 3:1 to 1:1) to obtain the compound 35c (black solid, yield: 70.5%).

Add compound 13 (1g, 2.51mmol), 35c (1.28g, 2.51mmol), and p-toluenesulfonic acid monohydrate (190.22mg, 1mmol) into the reaction bottle, add isopropyl alcohol (20mL), and set the reaction system Stir overnight at 80°C. When the reaction of the raw materials is complete, lower the reaction system to room temperature and pour into water (100 mL). A light yellow solid will precipitate. Filter and dry to obtain compound 36c. Dissolve the dried compound in To anhydrous dichloromethane (10 mL), trifluoroacetic acid (10 mL) was slowly added, and the reaction system was stirred at room temperature for 6 hours. After the reaction was complete, the solvent was removed by rotary evaporation and subjected to column chromatography (dichloromethane:methanol= 50:1 to 20:1) gave compound 37c (white solid, two-step yield: 66%). 1H NMR(600MHz,DMSO-d6)δ10.05(s,2H),9.16(s,1H),8.13(s,2H),7.66(s,1H),7.49(s,2H),7.36(d, J＝35.0Hz,2H),6.89(s,1H),4.52(s,1H),4.03(s,2H),3.65(s,2H),3.51(s,2H),3.44(s,2H), 3.37(s,6H),2.91(s,3H),2.64(d,J＝131.7Hz,3H),2.05(s,6H),1.81(s,3H).MS(ESI+):m/z calcd for C ₃₃ H ₄₃ BrN ₈ O ₈ ,758.2387; found, 759.2467(M+H+).

Compound 37c (100 mg, 0.13 mmol), p-toluenesulfonyl chloride (30.5 mg, 0.16 mmol), and triethylamine (26.29 mg, 0.26 mmol) were dissolved in anhydrous dichloromethane (2 mL), and reacted at room temperature for 16 hours. , the reaction of the raw materials was complete, the solvent was concentrated to remove, and compound 38c (white solid, yield: 56%) was obtained through column chromatography (dichloromethane: methanol = 50:1 to 20:1). ¹ H NMR (400MHz, DMSO-d ₆ ) δ10.02 (s, 1H), 9.95 (s, 1H), 9.22 (s, 1H), 8.18 (d, J = 5.8Hz, 2H), 7.77 (s, 2H),7.70(s,1H),7.54(d,J＝10.9Hz,2H),7.46(s,2H),7.38(s,2H),6.91(m,1H),4.09(s,2H), 4.04(s,2H),3.65(s,2H),3.55(s,2H),3.48(s,2H),3.44(s,2H),3.36(s,2H),2.93(s,3H),2.55 -2.77(s,3H),2.40(s,3H),2.07(s,8H),1.83(s,3H).MS(ESI+):m/z calcd for C ₄₀ H ₄₉ BrN ₈ O ₁₀ S,912.2476 ;found,913.2554(M+H ⁺ ).

Compound 38c (40 mg, 43.77 μmol) was dissolved in tetrahydrofuran (0.5 mL), 0.5 mL of 1M TBAF solution was added, the reaction system was stirred at 50°C overnight, the reaction system was cooled to room temperature, 2 mL of water was added, and the mixture was stirred with ethyl acetate. The ester was extracted, the organic phase was collected, concentrated, and subjected to column chromatography (dichloromethane: methanol = 30:1 to 15:1) to obtain compound 39c (white solid, yield: 62%). ¹ H NMR (400MHz, DMSO-d6) δ9.98 (s, 1H), 9.91 (s, 1H), 9.18 (s, 1H), 8.14 (d, J = 4.6Hz, 2H), 7.66 (s, 1H ),7.47(d,J＝18.3Hz,2H),7.37(d,J＝13.0Hz,2H),6.89(s,1H),4.52(s,1H),4.40(s,1H),4.03(s ,2H),3.64(d,J＝11.7Hz,3H),3.53(d,J＝12.1Hz,5H),3.33(s,2H),2.90(s,3H),2.64(d,J＝88.9Hz ,3H),2.04(s,8H),1.80(s,3H).MS(ESI ⁺ ):m/z calcd for C ₃₃ H ₄₂ BrFN ₈ O ₇ ,760.2344; found,761.2425(M+H ⁺ ).

Example 22

Compound 32 (2g, 7.48mmol), fluoroethyl p-toluenesulfonate (1.96g, 8.98mmol), and potassium carbonate (1.24g, 8.98mmol) were dissolved in DMF (20mL), and the reaction system was placed at 70°C. React for 6 hours. The reaction is complete by TLC. Add 50 mL of water, extract with ethyl acetate, collect the organic phase, dry, concentrate, and perform column chromatography (petroleum ether: ethyl acetate = 5:1 to 2:1) to obtain compound 40. (yellow solid, yield: 76.5%).

Compound 2,4-dichloro-5-trifluoromethylpyrimidine (164.9mg, 0.76mmol) was dissolved in 1,2-dichloroethane (1.5mL) and tert-butanol (1.5mL) at 0°C. , add 1M zinc chloride ether solution (1.67mL) and stir at this temperature for 1 hour. Compound 41 (180mg, 0.76mmol) is added to the reaction system, and triethylamine (85mg, 0.84mmol) is dissolved in 1 , 2-dichloroethane (0.75mL) and tert-butyl alcohol (0.75mL) were slowly added to the reaction system. After the dropwise addition was completed, it was raised to room temperature and stirred overnight. After testing that the raw material reaction was complete, add water The reaction was quenched, extracted with dichloromethane, the organic phase was dried, concentrated, and subjected to column chromatography (petroleum ether: ethyl acetate = 5:1 to 1:1) to obtain the product (white solid, yield: 36.5%). ¹ H NMR (400MHz, DMSO-d6) δ10.46(s,1H),9.00(s,1H),8.69(s,1H),8.15(s,1H),7.87(s,1H),7.33(s ,1H),7.04(s,1H),4.80(s,1H),4.68(s,1H),4.27(s,1H),4.20(s,1H),3.21(s,2H),1.75(s, 3H).MS(ESI ⁺ ):m/z calcd for C ₁₈ H ₁₈ ClF ₄ N ₅ O ₃ ,463.1034; found, 464.3339(M+H ⁺ ).

Compound 42 (100 mg, 0.22 mmol), 2,4-dinitroaniline (47.6 mg, 0.26 mmol), and potassium carbonate (35.9 mg, 0.26 mmol) were dissolved in DMF (2 mL), and the reaction system was placed at 60°C React for 20 hours, add water to dilute the reaction solution, extract with ethyl acetate, collect the organic phase, dry, concentrate, and column chromatography (dichloromethane: methanol = 100:1 to 40:1) to obtain the product (yellow solid, product rate: 26.9%).

Add compound 43 (100 mg, 0.16 mmol), iron powder (13.4 mg, 0.24 mmol), and ammonium chloride (12.7 mg, 0.24 mmol) into the reaction flask, then add methanol (2 mL), water (1 mL), and tetrahydrofuran (2 mL). ), stir the reaction system at 70°C for 6 hours, detect the reaction is complete by TLC, filter while hot, concentrate the filtrate, dissolve it in anhydrous tetrahydrofuran and place it in an ice bath, add triethylamine (32.4mg, 0.32mmol) , add acetyl chloride (25.1 mg, 0.32 mmol) dropwise. After the dropwise addition, stir the reaction system at room temperature for 1 hour. Add water to quench the reaction. Extract with ethyl acetate. Dry the organic phase, concentrate and column chromatography. (Dichloromethane:methanol=50:1 to 25:1) Compound 44 (white solid, two-step yield: 32.8%) was obtained. 1H NMR (600MHz, DMSO-d6) δ10.06(d,J＝22.7Hz,2H),9.48(s,1H),8.89(d,J＝24.0Hz,1H),8.24(s,1H),8.03 (s,1H),7.85(s,2H),7.53(d,J＝30.9Hz,2H),7.40(s,1H),7.29(s,1H),6.74(s,1H),4.74(d, J＝24.0Hz,2H),4.17(d,J＝29.7Hz,2H),3.25(s,2H),2.49(s,2H),2.04(s,3H),2.00(s,3H),1.74( s,3H).MS(ESI ⁺ ):m/z calcd for C ₂₈ H ₃₀ F ₄ N ₈ O ₅ ,634.2275; found,635.2350(M+H ⁺ ).

Example 23

Compound 25 (200 mg, 0.39 mmol), triethylene glycol bis-p-toluenesulfonate (270.5 mg, 0.59 mmol), and potassium carbonate (81.5 mg, 0.59 mmol) were dissolved in DMF (5 mL), and the reaction system was placed React for 6 hours at 60°C. After checking that the raw material reaction is complete, the solvent is concentrated and removed, and compound LY-19 is obtained as a yellow solid through column chromatography (dichloromethane: methanol = 60:1 to 30:1). Yield: 69% . 1H NMR (600MHz, DMSO-d6) δ10.03(s,2H),9.46(s,1H),8.22(d,J＝19.4Hz,2H),8.01(s,1H),7.75(d,J＝ 12.7Hz,3H),7.70(s,1H),7.52(s,1H),7.45(s,2H),7.38(s,1H),7.13(s,1H),4.16(s,2H),4.09( s,2H),3.69(s,2H),3.56(s,2H),3.52(s,2H),3.45(s,2H),2.39(s,3H).13C NMR (101MHz, DMSO-D6) δ170.04,168.86,158.36,157.47,145.92,145.41,139.74,137.48,134.41,132.91,132.43,130.63,128.08,127.04,125.07,116. 81,116.17,115.14,114.79,70.49,70.43,70.20 ,69.82,69.22,68.41,24.51,23.58,21.59.MS(ESI+):m/z calcd for C ₃₃ H ₃₆ BrN ₇ O ₁₀ S,801.1428; found,802.1494(M+

H+).

The synthesis method is the same as that of compound 39c. Compound LY-19 is used as raw material, and the product is obtained as a yellow solid through column chromatography (dichloromethane: methanol = 50:1 to 30:1). Yield: 66.5%. ¹ H NMR (600MHz, DMSO-d6) δ10.01 (s, 2H), 9.42 (s, 1H), 8.17 (d, J = 17.5Hz, 2H), 7.96 (s, 1H), 7.69 (s, 1H) ),7.66(s,1H),7.48(s,1H),7.34(s,1H),7.09(s,1H),4.49(s,1H),4.41(s,1H),4.14(s,2H) ,3.69(s,2H),3.62(s,1H),3.56(s,3H),3.52(s,2H),2.03(s,6H). ¹³ C NMR(151MHz,DMSO-D6)δ170.06,168.89, 158.36,157.51,145.92,139.77,137.48,134.39,132.42,128.02,127.08,125.08,116.84,116.19,115.19,114.79,84.10,83.00,70.56,70.31 ,70.21,69.87,69.24,24.48,23.56.MS(ESI ⁺ ):m/z calcd for C ₂₆ H ₂₉ BrFN ₇ O ₇ ,649.1296; found, 650.1373(M+H ⁺ ).

Example 24

Synthesis of LY-21

Compound 6-fluoronicotinic acid (141.1 mg, 1 mmol) and HATU (456.29 mg, 1.2 mmol) were dissolved in DMF (2 mL), DIPEA (155.1 mg, 1.2 mmol) was added and reacted at room temperature for 40 minutes, and then the compound 15 (500 mg, 1 mmol) was added to the reaction system, and the reaction was continued at room temperature for 10 hours. After the raw materials were completely reacted, 5 mL of water was added, extracted with ethyl acetate, the organic phase was collected, dried, concentrated, and subjected to column chromatography ( Dichloromethane: methanol = 50:1 to 30:1) to obtain the product, yield: 66.3%. ¹ H NMR(600MHz,DMSO-d6)δ10.02(s,1H),9.90(s,1H),9.66(s,1H),9.16(s,1H),8.74(s,1H),8.41(s ,1H),8.13(s,1H),8.11(s,1H),7.70(s,1H),7.59(s,2H),7.42(s,1H),7.35(s,1H),7.30(s, 1H),6.83(s,1H),3.73(s,3H),2.06(s,3H),1.98(s,3H).MS(ESI+):m/z calcd for C ₂₇ H ₂₄ BrFN ₈ O ₄ , 622.1088; found,623.1164(M+H ⁺ ).

Example 25

Add 3-cyano-2-fluoropyridine (10g, 81.9mmol), N-methylmethanesulfonamide (9.83g, 90.07mmol), and potassium carbonate (22.6g, 163.53mmol) into a 500mL round-bottomed flask, and add DMF (200mL), place the reaction system at 70°C for 3 hours, and detect the reaction is complete by TLC. Bring the reaction system to room temperature, slowly add it to 1 liter of water, let it stand, a white solid will precipitate, filter to obtain 13.89 g white solid product, yield: 80.28%. MS (ESI+): m/z calcd for C ₈ H ₉ N ₃ O ₂ S, 211.0415; found, 212.0424 (M+H ⁺ ).

Add compound 45 (8g, 37.87mmol) into the high-pressure reaction kettle, add 10% palladium on carbon (4g, 3.76mmol), dissolve it in methanol (100mL), replace it with nitrogen, add hydrogen to 6MPa, and heat to 50°C for reaction. After 3 hours, it was detected that the reaction was complete, the palladium carbon was filtered off, the filtrate was concentrated, and column chromatography (dichloromethane:methanol=40:1 to 15:1) was performed to obtain the product (7.18g, black solid, yield: 88.06%).

Compound 46 (6g, 27.87mmol), 5-bromo-2,4-dichloropyrimidine (7.62g, 33.44mmol), potassium carbonate (4.74g, 34.31 mmol) into a 250 mL round-bottomed flask, add tetrahydrofuran (100 mL), and stir the reaction system at 60°C. After 4 hours, the reaction is complete by TLC. The reaction system is brought to room temperature, 200 mL of water is added, and the reaction system is stirred with ethyl acetate. The ester was extracted, the organic phase was concentrated, and column chromatography (dichloromethane: methanol = 50:1 to 30:1) was performed to obtain the product (white solid, yield: 87.26%). MS (ESI ⁺ ): m/z calcd for C ₁₂ H ₁₃ BrClN ₅ O ₂ S, 404.9662; found, 443.9273 (M+K ⁺ ).

Dissolve 5-nitrosalicylic acid (10g, 54.61mmol) in anhydrous dichloromethane (70mL) at room temperature, slowly drop sulfoxide dichloride (30mL), and place the reaction system in an oil bath to reflux. After 5 hours, the solvent was concentrated, and the acid chloride was dissolved in anhydrous tetrahydrofuran (50 mL) and placed at 0°C. 1-acetylpiperazine (7g, 54.61 mmol) was dissolved in tetrahydrofuran (50 mL) and slowly added dropwise to the acid chloride, 1 Hours later, after checking that the reaction was complete, the reaction system was slowly added to 200 mL of water, extracted with ethyl acetate, the organic phase was concentrated, and column chromatography (dichloromethane: methanol = 50:1 to 30:1) was performed to obtain the product (14.66g , white solid, yield: 91.54%). MS(ESI+):m/z calcd for C13H15N3O5,293.1012; found,294.1135(M+H+).

Add compound 48b (10g, 34.10mmol), compound 9c (12.45g, 40.92mmol), and potassium carbonate (7.07g, 51.15mmol) into a 500mL round-bottom flask, add DMF (150mL) to dissolve the raw materials, and set the reaction system to Stir at 75°C, check that the reaction is complete after 10 hours, add 300 mL of water, extract with ethyl acetate, concentrate the organic phase, and perform column chromatography (dichloromethane: methanol = 55:1 to 20:1) to obtain the product (8.39g) , light yellow oil, yield: 57.83%). MS(ESI+):m/z calcd for C ₁₉ H ₂₇ N ₃ O ₈ ,425.1798; found, 426.1847(M+H ⁺ ).

The synthesis method is the same as that of compound 35c. Compound 49b (7g, 16.45mmol) is used as raw material, and the product (black solid) is obtained through column chromatography (dichloromethane: methanol = 50:1 to 30:1). Yield: 80% ). MS (ESI ⁺ ): m/z calcd for C ₁₉ H ₂₉ N ₃ O ₆ ,395.2056; found, 418.2138(M+Na ⁺ ).

Add compound 47 (1g, 2.46mmol), 50b (1.07g, 2.71mmol), and p-toluenesulfonic acid monohydrate (186.4mg, 0.98mmol) into a 100mL three-necked flask, replace nitrogen, and add isopropyl alcohol (20mL) , place the reaction system at 90°C for reaction. After 12 hours, TLC detects that the raw material reaction is complete. Bring the reaction system to room temperature, pour the reaction solution into 100 mL of water, extract with ethyl acetate, concentrate the organic phase, and perform column chromatography ( Dichloromethane: methanol = 40:1 to 15:1) to obtain the product (1.15 g, white solid, yield: 60.98%). 1H NMR(600MHz,Methanol-d4)δ8.44(s,1H),7.99(s,1H),7.79(s,1H),7.70(s,1H),7.46(s,1H),7.38(d, J＝19.6Hz,2H),7.22(s,1H),6.95(s,1H),4.18(s,1H),4.10(s,1H),3.80(d,J＝15.4Hz,3H),3.63( m,12H),3.52(s,4H),3.41(s,1H),3.21(s,3H),3.11(s,3H),2.11(d,J＝33.8Hz,3H).MS(ESI+): m/z calcd for C ₃₁ H ₄₁ BrN ₈ O ₈ S,764.1951; found, 418.2138(M+H+).

Dissolve compound LY-48 (500 mg, 0.65 mmol) in anhydrous dichloromethane (10 mL) under an ice bath, add triethylamine (197.3 mg, 1.95 mmol), stir under an ice bath for 0.5 hours, and then drop slowly Add p-toluenesulfonyl chloride (247.8mg, 1.3mmol) Dichloromethane solution (10 mL), after the dropwise addition, the reaction system was placed at room temperature for 20 hours, the solvent was removed by rotary evaporation, and column chromatography (dichloromethane: methanol = 50:1 to 25:1) was performed to obtain the product (358.7 mg, white solid, yield: 60%). MS(ESI+): m/z calcd for C ₃₈ H ₄₇ BrN ₈ O ₁₀ S _2, 918.2040; found _, 919.2125(M+H+).

Dissolve compound LY-50 (100 mg, 0.11 mmol) in 2 mL of tetrahydrofuran, add 1 M tetrabutylammonium fluoride solution (0.17 mL, 0.17 mmol) in tetrahydrofuran, and then reflux. After 2 hours, TLC detects that the reaction is complete. Add 5 mL of water, extracted with ethyl acetate, combined organic phases, dried over anhydrous magnesium sulfate, filtered, concentrated, and column chromatographed (dichloromethane: methanol = 50:1 to 25:1) to 59 mg of white solid, yield: 72.72%. 1H NMR(600MHz,DMSO-d6)δ9.03(s,1H),8.40(s,1H),8.02(s,1H),7.70(s,1H),7.51(s,1H),7.37(s, 1H),7.34(s,1H),7.29(s,1H),6.77(s,1H),4.70(s,2H),4.49(s,1H),4.41(s,1H),3.99(s,2H ),3.66(s,2H),3.60(s,1H),3.52(m,7H),3.41(d,J＝29.7Hz,3H),3.31(d,J＝14.7Hz,1H),3.15(s ,3H),3.12(m,3H),3.01(d,J＝36.9Hz,2H),1.96(d,J＝35.3Hz,3H).13C NMR(151MHz,DMSO-D6)δ168.84,167.01,166.93, 158.96,158.75,156.64,152.83,149.03,147.92,138.17,134.58,125.98,124.62,121.21,119.17,112.94,84.08,82.98,70.41,70.32,70.26, 70.14,69.54,68.36,37.82,36.61,25.56,21.72, 19.91,14.04.MS(ESI+):m/z calcd for C ₃₁ H ₄₀ BrFN ₈ O ₇ S,766.1908; found,767.1995(M+H+).

Example 26

Dissolve 5-nitrosalicylic acid (10g, 54.61mmol) in anhydrous dichloromethane (70mL) at room temperature, slowly drop sulfoxide dichloride (30mL), and place the reaction system in an oil bath to reflux. For 5 hours, the solvent was concentrated, and the acid chloride was dissolved in anhydrous tetrahydrofuran (50 mL) and stirred at 0°C. Morpholine (4.76g, 54.61 mmol) was dissolved in tetrahydrofuran (50 mL) and slowly added dropwise to the acid chloride for 1 hour. After checking that the reaction was complete, the reaction system was slowly added to 200 mL of water, extracted with ethyl acetate, the organic phase was concentrated, and column chromatography (dichloromethane: methanol = 50:1 to 30:1) was performed to obtain the product (white solid, Yield: 90.2%).

The synthesis method was the same as that of compound 49b. Using compound 48b as raw material, the product (yellow solid, yield: 56.2%) was obtained through column chromatography (dichloromethane: methanol = 50:1 to 25:1). MS(ESI ⁺ ):m/z calcd for C ₁₇ H ₂₄ N ₂ O ₈ ,384.1533; found, 385.1478(M+H ⁺ ).

Add compound 49a (1g, 2.6mmol) into a 100 ml round-bottomed flask, add methanol (10 mL), 10% palladium on carbon (0.1g) and replace the hydrogen. The reaction system is stirred at room temperature overnight. The reaction is completed by TLC. , filter, and concentrate the filtrate to obtain compound 50a, which can be directly used in the next reaction without purification. Dissolve compound 50a, compound 47 (1.06g, 2.6mmol), and p-toluenesulfonic acid monohydrate (197.8mg, 1.04mmol) in into isopropyl alcohol (20 mL), and place the reaction system at 90°C to react overnight. When the raw materials react completely, bring the reaction to room temperature, add 50 mL of water, extract with ethyl acetate, collect the organic phase, dry, concentrate, Column chromatography (dichloromethane:methanol=50:1 to 20:1) gave the product (white solid, two-step yield: 62%). 1H NMR(600MHz,Chloroform-d)δ8.32(s,1H),7.88(s,1H),7.78(s,1H),7.73(s,1H),7.44(s,1H),7.37(s, 1H),7.18(s,1H),6.74(s,1H),6.22(s,1H),4.77(s,2H),4.07(s,1H),3.98(s,1H),3.72(d,J ＝21.2Hz,3H),3.62(d,J＝30.7Hz,11H),3.50(s,2H),3.44(s,1H),3.19(s,3H),3.13(s,1H),3.00(s ,3H).13C NMR(151MHz, CHLOROFORM-D) δ167.47,158.45,158.34,155.59,152.82,149.81,148.28,139.68,134.03,133.74,125.75,124.41,122.39,120.39,112.79,93.52,72 .84,70.89,70.47,69.73,68.43,66.98,66.76, 61.63,47.30,42.23,40.48,37.81,35.67.

The synthesis method is the same as that of compound LY-50. Using compound LY-47 as raw material, compound LY-49 (white solid, yield: 50.1 %). ¹ H NMR (600MHz, DMSO-d6) δ9.04(s,1H),8.40(s,1H),8.03(s,1H),7.74(s,2H),7.70(d,J＝7.6Hz,1H ),7.52(s,1H),7.42(s,2H),7.36(dd,J＝6.0,13.5Hz,2H),7.29(s,1H),6.76(s,1H),4.71(s,2H) ,4.07(s,2H),3.97(s,2H),3.63(s,2H),3.50(d,J＝28.6Hz,10H),3.43(s,2H),3.35(s,1H),3.00( s,1H),2.36(s,3H). ¹³ C NMR(151MHz,DMSO-D6)δ166.89,158.96,158.76,156.61,152.84,149.03,147.93,145.40,138.18,134.64,134.57,132.97,1 30.64,128.54, 128.13,125.89,124.61,121.18,119.24,112.93,93.07,70.47,70.38,70.35,70.26,69.53,68.47,68.37,66.69,66.53,47.14,42.09,37.83,3 6.62,21.60.MS(ESI+):m/z calcd for C ₃₆ H ₄₄ BrN ₇ O ₁₀ S ₂ ,877.1774; found, 878.1841(M+H ⁺ ).

The synthesis method is the same as that of compound LY-52. Compound LY-49 is used as raw material and 59 mg of white solid is obtained through column chromatography (dichloromethane: methanol = 50:1 to 25:1). The yield is 72.72%. ¹ H NMR(600MHz,DMSO-d6)δ9.03(s,1H),8.40(s,1H),8.02(s,1H),7.70(s,1H),7.51(s,1H),7.37(s ,1H),7.34(s,1H),7.29(s,1H),6.77(s,1H),4.70(s,2H),4.49(s,1H),4.41(s,1H),3.99(s, 2H),3.66(s,2H),3.60(s,1H),3.52(m,7H),3.41(d,J＝29.7Hz,3H),3.31(d,J＝14.7Hz,1H),3.15( s, 3H), 3.12 (m, 3H), 3.01 (d, J = 36.9Hz, 2H), 1.96 (d, J = 35.3Hz, 3H). ¹³ C NMR (151MHz, DMSO-D6) δ 168.84, 167.01, 166.93,158.96,158.75,156.64,152.83,149.03,147.92,138.17,134.58,125.98,124.62,121.21,119.17,112.94,84.08,82.98,70.41,70.32 ,70.26,70.14,69.54,68.36,37.82,36.61,25.56, 21.72,19.91,14.04.MS(ESI+):m/z calcd for C ₃₁ H ₄₀ BrFN ₈ O ₇ S,766.1908; found,767.1995(M+H ⁺ ).

Test Methods:

This test uses Cisbio's homogeneous time-resolved fluorescence conjugate energy transfer (method) for activity detection. FAK kinase was purchased from CARNA Company; detection kit was purchased from Cisbio Company; detection plate and multifunctional microplate reader were purchased from Perkin Elmer Company. In the detection plate, mix the enzyme, biotin-labeled peptide substrate, ATP, and detection compound, and incubate the reaction. There are 11 concentrations of compounds in total, and the final system concentration ranges from 10 μM to 0.17 nM. Use 10 μL of buffer reaction system (50mM Hepes pH 7.5, 1mM EDTA, 10mM MgCl2, 0.01% Brij-35, 25nM SEB, 1mM DTT, 0.7nM FAK, 1μM biotin-TK peptide, 25μM ATP) and incubate at 23°C for 90 minute. Add 10 μL of stop solution (20 mM EDTA, 0.67 nM TK antibody, 50 nM XL-665), incubate at 23°C for 60 minutes, and read by Envision. Calculate the compound's inhibition rate from the data read by the instrument, and then use mode 205 in IDBS's XLFIT5 to calculate the IC ₅₀ value. The specific results are shown in Table 1.

Example 27

Mark the route:

^{The 18} F ^- trapped on the QMA column was eluted with a mixture of Kryptofix-2.2.2. (13 mg) in anhydrous acetonitrile (0.7 mL) and K ₂ CO ₃ (1 mg) in water (0.3 mL). In the reaction bottle, use nitrogen flow to dry the solvent in the reaction bottle at 100°C. Then Use 0.5 mL anhydrous acetonitrile to azeotropically remove water, repeat 3 times. The anhydrous DMSO solution (0.3 mL) of the labeled precursor compound LY-9 (1.5 mg) was quickly added to the above reaction bottle, sealed, and reacted at 95°C for 20 min. After the reaction, add distilled water (10 mL) to quench the reaction, use a syringe to draw the reaction solution through the pre-activated Sep-Pak C18 solid-phase extraction column, and then use 1 mL acetonitrile to elute the reaction product from the C18 column and collect the eluate. lotion. By radio-HPLC (wavelength 254nm, C18 reversed-phase semi-preparative column Agela Technologies, 5μm, 10×250mm, MeOH:H ₂ O=2.8:1.2, flow rate 4mL/min) liquid phase separation product. The liquid phase retention time of the radiopharmaceutical was 21 minutes, and its accuracy was confirmed by liquid phase co-injection analysis with compound LY-10. The labeling rate was 15% and the radiochemical purity was greater than 95%.

The remaining compounds in Table 1 below of the present invention are also labeled using similar methods. The specific labeling rates are shown in Table 1.

Table 1 In vitro FAK enzyme activity inhibition test results of standard compounds

2. The FAK inhibitor of the present invention has advantages in combined use. Through combination with different drugs, it can be used in the treatment of lung cancer, liver cancer, ovarian cancer, pancreatic cancer, colon cancer, prostate cancer, brain glioma, etc. but is not limited to these types. To achieve better results in cancer, for example, it can be combined with erlotinib, crizotinib, entrectinib, vandetanib, dabrafenib, and larotinib for the treatment of lung cancer; for example, it can be combined with paclitaxel or Carboplatin is used for the treatment of ovarian cancer; for example, it can be combined with gemcitabine, capecitabine, pembrolizumab, paclitaxel, etc. for the treatment of pancreatic cancer; for example, it can be combined with paclitaxel, platinum drugs, etc. for the treatment of esophageal cancer, etc. But it is not limited to these combination methods, which all show better tumor inhibitory effects than single drugs. For example, the tumor inhibitory effect of compound LY-21 in the A549 xenograft model is shown in Figures 1 and 2. It can be seen that compound LY-21 combined with erlotinib performed better than single drug in the lung cancer mouse model. It has a tumor-inhibiting effect, and the mice are also very lively and do not lose weight, indicating that the toxicity is low and the effect of 1+1 is greater than 2 can be achieved. It can be seen that the FAK inhibitor of the present invention has very important significance in combined medication.

The advantages of the FAK inhibitor of the present invention when used alone include the treatment of lung cancer, liver cancer, ovarian cancer, pancreatic cancer, colon cancer, prostate cancer, brain glioma, etc. but are not limited to these types of cancer. For example, the tumor inhibitory effect of compound LY-3 in the A549 xenograft model is shown in Figures 3 and 4. It can be seen that in the A549 xenograft model, the compound LY-3 of the present invention is better than the FAK clinical drug VS when used alone. -6063, and is better than the currently marketed drug erlotinib for the treatment of lung cancer, and its single-use effect is very good.

The FAK inhibitor of the present invention has the advantage of increasing radioactive diagnosis. Taking compound LY-10 as an example, the imaging image in the A549 tumor model is shown in Figure 5. We can clearly see the FAK inhibitor we labeled. It has higher uptake at the tumor site, can clearly distinguish target and non-target sites, and has better retention at the tumor site 1.

The above are only preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions or improvements made within the spirit and principles of the present invention shall be included in the protection of the present invention. within the range.

Claims

A compound targeting focal adhesion kinase, characterized in that the compound targeting focal adhesion kinase has the following structural formula of Formula I:

R 1 is selected from

R 2 is selected from hydrogen or

The R 3 is selected from alkoxy, alkoxy substituted by alkyl, or hydroxyl;

R 7 in R 1 , R 2 or R 3 is the same or different, and each is independently selected from C1-C5 alkyl;

The R 4 is selected from hydrogen, alkoxy, nitro,

The R 8 is selected from a C1-C5 linear alkyl group or a cycloalkyl group; the R 9 or R 10 are the same or different, and each is independently selected from a methyl group or a heteroaryl group containing a halogen substituent; the X Selected from halogen;

The R 5 is selected from halogen or an alkyl group completely substituted by halogen.
The compound targeting focal adhesion kinase according to claim 1, characterized in that, in the compound targeting focal adhesion kinase,

R 7 in R 1 , R 2 or R 3 is the same or different, and each is independently selected from C1-C2 alkyl; and/or,

The alkyl group in the alkoxy group in R 3 or R 4 is selected from C1-C2 alkyl groups;

In the R 3 , the alkoxy group substituted by the alkyl group has the following general formula: The R 11 is selected from halogen, hydroxyl, and OTs group; the n is selected from 1-5;

The R 8 is selected from C1-C3 alkyl or C3-C5 cycloalkyl;

The R 9 and R 10 are independently selected from F-substituted pyrazolyl or pyridinyl.
The compound targeting focal adhesion kinase according to claim 2, characterized in that,

R 7 in R 1 , R 2 or R 3 are all selected from methyl;

The alkyl group in the alkoxy group in R 3 or R 4 is selected from methyl;

The R 5 is selected from bromine or CF 3 ;

The R 11 is selected from F or 18 F; the n is selected from 1-3;

The R 8 is selected from ethyl or cyclopropyl;

The R 9 and R 10 are independently selected from
The compound targeting focal adhesion kinase according to claim 1, characterized in that the compound targeting focal adhesion kinase has the following structural formula:

The n is selected from 1-3.
The compound targeting focal adhesion kinase according to any one of claims 1 or 4, characterized in that,

The R 4 is selected from hydrogen, and preferably the R 2 is also selected from hydrogen.
The compound targeting focal adhesion kinase according to any one of claims 1 or 4, characterized in that the compound targeting focal adhesion kinase includes at least one of the following compounds:
The method for preparing a compound targeting focal adhesion kinase according to any one of claims 1 to 6, characterized in that it includes the following steps:

(1) The hydroxyl group in is replaced by R 3 group;

The R 3 is selected from the group consisting of alkoxy, alkoxy substituted by alkyl, optionally hydroxyl;

(2) Optionally, The nitro group in is further substituted by R 4 group; said R 4 is selected from
The use of the compound according to any one of claims 1 to 6 as a labeling precursor is preferably used as a radioactive labeling precursor.
The use of the compound according to any one of claims 1 to 6 in targeting focal adhesion kinase or in the preparation of tumor therapeutic drugs or in the use of combined drugs to treat tumors or in the preparation of tumor diagnostic imaging agents, Preferably, the tumor is selected from the group consisting of lung cancer, liver cancer, ovarian cancer, pancreatic cancer, colon cancer, prostate cancer, and brain glioma.
A pharmaceutical composition comprising the compound of any one of claims 1-6.