CN114213416B - Irreversible BTK inhibitor with oxazolo [4,5-b ] pyridine structure and application thereof - Google Patents

Abstract

The invention discloses a compound with irreversible inhibition of protein tyrosine kinase (BTK) activity, pharmaceutically usable salt thereof, a synthesis method thereof and application thereof in medicines for inhibiting BTK protein. In particular, the present invention describes compounds of formula a. The compound has good BTK inhibition activity, improves water solubility, and has basically consistent pharmacokinetic parameters with the commercially positive drug ibrutinib, so that the compound can be widely applied to preparing pharmaceutical preparations for preventing or treating diseases caused by BTK abnormality.

Description

Irreversible BTK inhibitor with oxazolo [4,5-b ] pyridine structure and application thereof

Technical Field

The invention relates to the field of small molecule medicines, in particular to a compound with irreversible protein tyrosine kinase (BTK) inhibiting activity and a synthesis and use method of the compound.

Background

According to the statistics of world health organization, the annual growth rate of lymphoma incidence is 5% -7%, the annual death number is over 20 ten thousand, at present, the annual growth rate of lymphoma incidence in China is 3% -5%, and the annual new incidence is about 10 ten thousand, so that the lymphoma has become the eighth high-incidence malignant tumor.

Bruton's tyrosine kinase, BTK), a membrane-bound protein, belongs to the Tec family of non-receptor tyrosine kinases, and is found in all hematopoietic cells except T cells and natural killer cells. BTK is an important signal molecule of B cell receptor pathway, expressed at each development stage of B cells, involved in regulating proliferation, differentiation and apoptosis of B cells, and plays an important role in survival and diffusion of malignant B cells.

The main reason for B cell lymphomas is the excessive activation of the B cell antigen receptor (BCR) signaling pathway, and BTK kinase, a key process protein in the BCR signaling pathway, is widely distributed in the lymphatic, hematopoietic and blood systems, is a membrane-bound protein, and is expressed in immune cells such as B cells, mast cells and macrophages. The large-scale expression of BTK can cause abnormal activation of a BCR signal channel and influence proliferation, differentiation and apoptosis of B cells, so that various B cell malignant tumors are initiated; but also can cause B cell dysfunction, immune tolerance state change, and convert into autoreactive B cells, and secrete a large amount of autoantibodies to induce autoimmune diseases. Therefore, BTK is a research hotspot for clinically treating B-cell tumors and B-cell immune diseases at present.

BTK inhibitors can inhibit lymphocyte proliferation by inhibiting the excessive activation of BCR signaling pathways, and show excellent therapeutic prospects for the treatment of B-cell malignancies and autoimmune diseases. In 2013, ibrutinib was approved by the FDA as the first effective BTK selective inhibitor, and breakthrough therapy was used for the treatment of chronic lymphocytic leukemia, mantle cell lymphoma, small lymphocytic lymphoma, fahrenheit macroglobulinemia, marginal zone lymphoma and graft-host disease, which is of epoch-making significance, making BTK a promising therapeutic target. Against this background, the second generation BTK inhibitors acartinib, tiratinib, zebutinib and obutinib, have both improved selectivity to BTK and improved resistance and reduced drug toxicity, all of which have been approved by the FDA for sale. The structure of the marketed BTK inhibitors is shown below:

small molecule BTK inhibitors are classified into two classes, reversible inhibitors and irreversible inhibitors, depending on the mode of binding to the BTK catalytic domain. Irreversible BTK inhibitors retain the electrophilic center of the terminal end, such as acrylamide groups and 2-butynamide groups, and can form covalent bonds with the conserved non-catalytic cysteine residue (Cys 481) of BTK proteins through Michael addition, nucleophilic addition, addition-elimination, nucleophilic substitution or other reactions, thereby achieving irreversible strong binding.

However, the existing irreversible BTK inhibitor still has the defects of poor selectivity, poor water solubility and the like, and can inhibit other kinases of Tec family while inhibiting BTK, which causes side effects such as bleeding, diarrhea, rash and the like and generates drug resistance, so the development of a new BTK inhibition drug is still a problem to be solved urgently.

Disclosure of Invention

The invention aims at overcoming the defects of the prior art and provides a compound with irreversible inhibition of protein tyrosine kinase (BTK) activity and pharmaceutically usable salt thereof, a synthesis method thereof and application thereof in medicines for inhibiting BTK protein.

The aim of the invention is realized by the following technical scheme: a small molecule compound of oxazolo [4,5-b ] pyridine structure with BTK inhibiting activity, a compound with irreversible BTK inhibiting activity and pharmaceutically usable salt thereof, having a structure as shown in formula A:

wherein X, Y, Z has the following structure:

Z＝H、F，

wherein R has the structure:

further, the small molecule compound of oxazolo [4,5-b ] pyridine structure with BTK inhibition activity, X, Y, Z, respectively, has the following structure:

wherein R has the structure:

alternatively, X, Y, Z each have the following structure:

Z＝H，F，

wherein R has the structure:

further, the small molecule compound having the structure of oxazolo [4,5-b ] pyridine with BTK inhibitory activity is selected from the following structures:

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or the structure is as follows:

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or the structure is as follows:

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the compound with irreversible BTK activity can be used singly or prepared into pharmaceutically acceptable salts by a conventional method, wherein the pharmaceutically acceptable salts are hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, phosphate, acetate, propionate, butyrate, oxalate, tartrate, methanesulfonate, p-toluenesulfonate, fumarate, taurine, citrate, succinate or mixed salts thereof.

The invention also provides a preparation method of the compound with irreversible BTK inhibition activity, which comprises the following steps:

the method comprises the steps of taking p-bromobenzoyl chloride as a starting material, and carrying out nucleophilic substitution on the p-bromobenzoyl chloride and aminopyrazine under the alkaline condition of triethylamine to obtain 4-bromo-N- (pyrazin-2-yl) benzamide; subsequently, 4-bromo-N- (pyrazin-2-yl) benzamide and bisboronic acid pinacol ester are coupled by Suzuki to obtain a corresponding borate product; then, the boric acid ester product reacts with (R) -phenyl 2- (8-amino-1-bromoimidazole [1,5-A ] pyrazin-3-yl) pyrrolidine-1-carboxylic acid ester to obtain a coupling product; removing the carbobenzoxy protecting group of the coupling product under the condition of 33% HBr-AcOH solution to obtain pyrrolidine compounds; the target product is obtained by the pyrrolidine compound, 2-methacrylic acid, 2-butynoic acid and acrylic acid under the action of condensing agent HATU. The reaction process is as follows:

or the following method is adopted:

the method comprises the steps of taking p-bromobenzoic acid as an initial raw material, and carrying out condensation reaction on the p-bromobenzoic acid and aniline under the action of condensing agents EDC.HCl/HOBt and N, N-diisopropylethylamine to obtain N-phenyl-4-bromobenzamide; then, coupling N-phenyl-4-bromobenzamide and bisboronic acid pinacol ester to obtain a corresponding borate product through suzuki; then, the raw material 4-amino-3-iodine-1H-azolo [3,4-D ] pyrimidine and the raw material (S) -1-tert-butoxycarbonyl-3-hydroxypiperidine undergo Mitsunobu reaction to obtain (3R) -1-Boc-3- (4-amino-3-iodine-1H-pyrazolo [3,4-D ] pyrimidine-1-yl) piperidine; then the boric acid ester product and (3R) -1-Boc-3- (4-amino-3-iodine-1H-pyrazolo [3,4-D ] pyrimidine-1-yl) piperidine are subjected to Suzuki reaction to obtain a coupling product; removing Boc protecting groups from the coupling product under the condition of 3mol of HCl-EA solution to obtain piperidine compounds; the target product is obtained by the piperidine compound and alpha, beta-unsaturated acid under the action of condensing agent HATU. The reaction process is as follows:

the invention also provides a small molecule compound with a BTK inhibition activity and an application of the small molecule compound with a oxazolo [4,5-b ] pyridine structure and a pharmaceutically acceptable salt thereof in pharmacy, wherein the application is specifically as follows: is used for preparing a pharmaceutical preparation for preventing or treating diseases caused by abnormal BTK. The disease is lymphoma, including chronic lymphocytic leukemia, B-cell lymphoma, mantle cell lymphoma, lymphoplasmacytic lymphoma, diffuse large B-cell lymphoma, non-Hodgkin's lymphoma, follicular central lymphoma, marginal zone B-cell lymphoma, or autoimmune disease, including rheumatoid arthritis, systemic lupus erythematosus, ankylosing spondylitis, psoriasis, etc.

Compared with the prior art, the invention has the following advantages:

the small molecular compound with the oxazolo [4,5-b ] pyridine structure and the pharmaceutically acceptable salt thereof have good BTK inhibitory activity, and simultaneously improve the water solubility, and the pharmacokinetic parameters of the small molecular compound are basically consistent with that of the commercially positive drug ibrutinib, so that the small molecular compound can be widely applied to the preparation of pharmaceutical preparations for preventing or treating diseases caused by BTK abnormality.

Drawings

FIG. 1 is a graph of the plasma concentration of ibrutinib, IV-2 and VII-2 in SD rats.

Detailed Description

The structure, the preparation method and the use of the pharmaceutical preparation for preventing or treating diseases caused by the overexpression of tubulin according to the present invention are further described below with reference to examples, but the present invention is not limited thereto.

Analytical data for the samples were determined by the following instrument:

the thermometer is uncorrected; bruker DRX400 nuclear magnetic resonance apparatus; agilent 5975 mass spectrometer; bruker Vector 22 IR spectrometer.

Example 1 synthesis of series i compounds:

1.1 Synthesis of intermediate 1-3:

2-aminopyrazine 1-1.00 g (10.53 mmol) was weighed into a 25mL three-necked flask, N ₂ 5mL of methylene chloride was added to the syringe, 1.41g (10.94 mmol) of DIPEA was added under ice-bath conditions, and finally a solution of 1-2.00 g (9.11 mmol) of 4-bromobenzoyl chloride in methylene chloride was added dropwise thereto, and the mixture was kept in ice-bath for 5 minutes and then left at room temperature for reaction overnight. The reaction was completed by TLC, the reaction solution was neutralized with NaOH, then extracted with ethyl acetate (40 mL. Times.3), the organic phases were combined, washed once with 100mL of saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography to give 1.2g of pale yellow solid 1-3 in about 48.2% yield. MS (ESI) m/z=278.01 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ9.71(d,J＝1.3Hz,1H),8.57(s,1H),8.41(d,J＝2.5Hz,1H),8.28(dd,J＝2.4,1.6Hz,1H),7.87–7.76(m,2H),7.74–7.62(m,2H).

1.2 Synthesis of intermediate 1-4:

sequentially weighing 1-3.00 g (3.61 mmol) of intermediate, 1.10g (4.33 mmol) of pinacol biborate and [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride dichloromethane Complex (Pd (dppf) Cl) ₂ ) 79mg (0.11 mmol), 1.06g (10.83 mmol) of potassium acetate are placed in a 25mL three-necked flask, N ₂ Protection, after which the syringe was added with tetrahydrofuran solution and refluxed overnight. After the reaction of the spot plate raw materials is finished, cooling the reaction solution to room temperature, and filtering with diatomite to remove Pd (dppf) Cl ₂ Washing with tetrahydrofuran for 2-3 times, concentrating the filtrate, adding water and ethyl acetate for extraction, washing the organic layer with water for 2-3 times, recovering under reduced pressure to obtain crude product, separating and purifying by column chromatography to obtain 1.04g pale yellow solid 1-4, with yield of about 89.2%, and transferring to nuclear magnetism and transferring to the next step. ¹ H NMR(400MHz,CDCl ₃ )δ9.87(d,J＝1.3Hz,1H),8.63(s,1H),8.57(d,J＝2.5Hz,1H),8.39(dd,J＝2.4,1.6Hz,1H),7.83(d,J＝7.8Hz,2H),7.49(d,J＝7.9Hz,2H),1.36(s,12H).

1.3 Synthesis of intermediate 1-5:

sequentially weighing 1-4 mg (0.34 mmol) of the intermediate, (R) -phenyl 2- (8-amino-1-bromoimidazole [1, 5-a)]Pyrazin-3-yl) pyrrolidine-1-carboxylic acid 133mg (0.31 mmol), pd (dppf) Cl ₂ 22mg(0.03mmol)，K ₂ CO ₃ 128mg (0.93 mmol) was added to a 10mL three-necked flask, N ₂ Protection, adding 1, 4-dioxane under ice bath condition: the mixed solution of water (3:1) was then transferred to 105℃to start the reaction. After 8h TLC was used to monitor the progress of the reaction, the reaction mixture was cooled to room temperature and filtered through celite to remove Pd (dppf) Cl ₂ Washing with ethyl acetate for 2-3 times, adding water (100 mL multiplied by 4) and 100mL ethyl acetate into the filtrate, extracting, drying the organic phase with anhydrous sodium sulfate, concentrating the filtrate, and performing column chromatography to obtain 110mg pale brown solid 1-5, wherein the yield is about 61.9%; MS (ESI) m/z=535.24 [ M+H ]] ⁺ ； ¹ H NMR(500MHz,DMSO-d ₆ )δ7.98(s,1H),7.92(d,J＝7.7Hz,2H),7.71(d,J＝4.6Hz,1H),7.64–7.50(m,3H),7.35(s,1H),7.24(s,3H),7.11–7.03(m,1H),7.04–6.95(m,2H),6.67(d,J＝7.3Hz,1H),6.01(s,2H),5.33(d,J＝19.2Hz,1H),4.95(s,2H),3.54–3.47(m,2H),1.91(d,J＝6.6Hz,2H).

1.4 Synthesis of intermediates 1-6:

1-5 mg (0.56 mmol) of intermediate is weighed into a 10mL single-necked flask, placed in an environment of 0 ℃, and 33% HBr-AcOH (6.6 mmol) is slowly added dropwise, after 5min, the mixture is transferred to room temperature for reaction for 2h. After TLC monitoring the reaction, the reaction mixture was extracted with 50mL of water and dichloromethane (70 mL. Times.2), the dichloromethane layer was removed, and the aqueous layer was used in ice-bathThe pH of the aqueous solution of NaOH is regulated to be approximately equal to 10, dichloromethane (60 mL multiplied by 4) is added for extraction, the organic layers are combined, anhydrous sodium sulfate is dried and then concentrated to obtain a crude product, the crude product is subjected to dry sample mixing and column chromatography gradient elution to obtain 206mg of light brown powdery solid, namely an intermediate 1-6, and the yield is about 91.6%. MS (ESI) m/z=401.21 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ11.26(s,1H),9.51(d,J＝1.4Hz,1H),8.60–8.45(m,2H),8.28(d,J＝8.4Hz,2H),7.95(d,J＝5.0Hz,1H),7.87(d,J＝8.4Hz,2H),7.32(d,J＝5.0Hz,1H),6.47(s,2H),5.35(t,J＝7.7Hz,1H),3.14(q,J＝7.3Hz,2H),2.54–2.49(m,2H),2.26(dd,J＝11.9,5.6Hz,2H),2.16–2.10(m,1H).

1.5 Synthesis of Compound I-1:

1-7 mg (0.09 mmol) of 2-methacrylic acid was dissolved in methylene chloride, 14mg (0.08 mmol) of EDC. HCl and 21. Mu.L of triethylamine were sequentially added thereto, followed by stirring for 5 minutes, 1-6 30mg (0.08 mmol) of the intermediate was added thereto, and the mixture was reacted at room temperature. After 3h reaction, TLC monitors the completion of the reaction, removes the solvent dichloromethane under reduced pressure, then adds 50mL of ethyl acetate for dissolution, adds 70mL of water for extraction, then extracts the organic layer by 1N of dilute HCl solution (70 mL multiplied by 2), extracts the organic layer once by 70mL of saturated NaCl solution, dries the ethyl acetate layer by anhydrous sodium sulfate, and then rotates out of the solvent to obtain a crude product, and mixes the crude product with a dry method for passing through a column. Gradient elution gave 27mg of yellow semi-solid, compound I-1, in about 77.1% yield; MS (ESI) [ M+H ]] ⁺ m/z＝469.21[M+H] ⁺ ； ¹ H NMR(500MHz,CDCl ₃ )δ9.74(d,J＝1.2Hz,1H),8.40(d,J＝2.5Hz,1H),8.35–8.26(m,1H),8.05(d,J＝8.0Hz,2H),7.80(d,J＝8.3Hz,3H),7.07(d,J＝5.0Hz,1H),5.50(t,J＝6.6Hz,1H),5.29(s,2H),3.89–3.74(m,2H),2.58(dd,J＝13.2,7.3Hz,2H),2.44–2.30(m,2H),1.24(s,3H). ¹³ C NMR(126MHz,CDCl ₃ )δ171.38,165.20,151.38,148.43,142.06,141.62,140.70,140.42,139.12,137.44,134.15,132.61,130.06,127.95,127.64,118.04,114.66,107.99,58.94,49.49,31.02,25.66,19.68.

1.6 Synthesis of Compound I-2:

the operation is the same as 1.5, 2-butynoic acid 1-8 is used for replacing 2-methacrylic acid 1-7, light brown semi-solid I-2 is obtained, and the yield is about 49.6%; MS (ESI) m/z=467.20 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ11.13(s,1H),8.55–8.37(m,3H),8.18(dd,J＝8.2,2.7Hz,2H),8.11(dd,J＝14.2,7.2Hz,2H),8.03–7.90(d,J＝8.4Hz,1H),7.81–7.64(d,J＝5.6Hz,1H),5.04(t,J＝6.9Hz,1H),3.17–3.10(m,2H),2.01(dd,J＝6.7,3.9Hz,2H),1.96(d,J＝5.1Hz,2H),1.87(s,1H). ¹³ C NMR(101MHz,DMSO-d ₆ )δ164.05,158.26,149.28,148.12,144.77,143.04,141.93,140.63,137.96,136.59,129.32,128.36,127.78,126.19,121.14,110.20,91.89,80.94,58.30,48.55,33.21,24.86.

1.7 Synthesis of Compound I-3:

compound I-3 is a by-product of the synthesis of compound I-2, 12mg of pale yellow semi-solid, MS (ESI): m/z=443.20 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ11.16(s,1H),9.42(s,1H),8.56–8.40(m,2H),8.18(d,J＝8.2Hz,2H),7.94(d,J＝15.3Hz,1H),7.80(dd,J＝29.5,6.3Hz,2H),7.13(d,J＝4.6Hz,1H),5.45(t,J＝6.3Hz,1H),3.73(t,J＝5.3Hz,2H),2.35-2.12(m,2H),2.02-1.92(m,2H),1.81(s,3H).

1.8 Synthesis of Compound I-4:

the same operation as 1.5, acrylic acid 1-9 is used for replacing 2-methacrylic acid 1-7, so as to obtain pale yellow powdery solid I-4, and the yield is about 78.3%; MS (ESI) m/z=455.20 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ11.18(s,1H),9.46(s,1H),8.56–8.40(m,2H),8.18(d,J＝8.2Hz,2H),7.80(dd,J＝29.5,6.3Hz,2H),7.13(d,J＝4.6Hz,1H),6.95(t,J＝15.3Hz,1H),6.05(d,J＝7.9Hz,2H),5.25(t,J＝6.4Hz,1H),4.73(t,J＝5.3Hz,1H),3.23(t,J＝4.8Hz,2H),2.27–2.13(m,2H),1.96(dd,J＝11.9,6.7Hz,2H). ¹³ CNMR(101MHz,DMSO-d ₆ )δ166.39,164.81,156.66,148.62,142.99,141.32,139.98,137.98,136.63,135.33,134.34,131.10,129.69,129.48,127.17,126.63,123.07,121.87,109.56,58.05,48.36,31.09,24.76.

Example 2 synthesis of series ii compounds:

2.1 Synthesis of intermediate 2-3:

2-1.31 g (6.52 mmol) of p-bromobenzoic acid was weighed and placed in a 25mL single-necked flask, 10mL of methylene chloride was added, 1.25g (6.52 mmol) of EDC. HCl was sequentially added under ice bath condition, 2.81mL (16.14 mmol) of DIPEA was stirred for 5min, 2-2.50 g (5.38 mmol) of aniline was added at 0℃and then placed at room temperature for reaction overnight. The reaction was monitored by TLC to completion, methylene chloride was removed under reduced pressure, saturated brine (200 mL. Times.3) and ethyl acetate (150 mL) were added for extraction, the organic phases were combined, dried over anhydrous sodium sulfate, and dried by spin-drying followed by dry-stirring and column-passing to give 1.1g of a white powdery solid, 2-3, in a yield of about 73.6%. MS (ESI) m/z=275.98 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ10.33(s,1H),7.90(s,2H),7.76(dd,J＝8.0,3.7Hz,4H),7.36(t,J＝7.9Hz,2H),7.12(t,J＝7.4Hz,1H).

2.2 Synthesis of intermediate 2-4:

the same procedure as 1.2, intermediate 2-3 was used instead of intermediate 1-3 to give 1.3g of a pale brown powdery solid in about 89.9% yield; ¹ H NMR(400MHz,CDCl ₃ )δ7.92(d,J＝8.2Hz,2H),7.86(d,J＝8.2Hz,3H),7.65(d,J＝7.8Hz,2H),7.38(t,J＝7.9Hz,2H),1.37(s,12H).

2.3 Synthesis of intermediate 2-7:

weighing 4-amino-3-iodine-1H-pyrazolo [3,4-d]Pyrimidine 2-5 500mg (1.92 mmol), 1-t-butoxycarbonyl-3-hydroxypiperidine 2-6 774mg (3.83 mmol), triphenylphosphine 755mg (2.88 mmol) were added to a three-necked flask, N ₂ The reaction was carried out under ice-bath conditions by adding anhydrous THF via syringe, slowly dropping DIAD in anhydrous THF, and transferring to room temperature for overnight reaction. After completion of the TLC plate starting material reaction, THF was removed under reduced pressure, dry-mixed and passed through a column, and the PE wet column was washed out with yellow DIAD, followed by gradient elution to give 708mg of white powdery solid, intermediate 2-7, in a yield of about 83.1%. MS (ESI) m/z=467.24 [ M+Na ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ8.21(s,1H),4.65–4.53(m,1H),3.88(t,J＝61.6Hz,2H),2.98(s,1H),2.19–2.08(m,1H),2.07–1.98(m,1H),1.87(s,1H),1.54(dt,J＝21.8,7.5Hz,2H),1.32(s,9H).

2.4 Synthesis of intermediate 2-8:

sequentially weighing 2-4 mg (1.35 mmol) of intermediate, 2-7 500mg (1.13 mmol) of intermediate, pd (dppf) Cl ₂ 25mg(0.03mmol)，K ₂ CO ₃ 468mg (3.39 mmol) of the mixture was placed in a 10mL three-necked flask, and a mixed solution of 1, 4-dioxane and water (3:1) was added under nitrogen and ice-bath conditions, followed by transfer to 105℃to initiate reflux. After 8h TLC was used to monitor the progress of the reaction, the reaction mixture was cooled to room temperature and filtered through celite to remove Pd (dppf) Cl ₂ The filtrate was then extracted with water (100 mL. Times.4) and 100mL ethyl acetate, and the organic phase was extracted with anhydrous Na ₂ SO ₄ Drying, concentrating filtrate, and separating by column chromatography to obtain 456mg light brown solid 2-8 with yield of about 78.6%; MS (ESI) m/z=514.19 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ10.36(s,1H),9.58–9.24(m,2H),8.46(s,1H),8.19(d,J＝7.5Hz,2H),7.85–7.60(m,2H),7.24(t,J＝7.5Hz,2H),7.09(t,J＝7.2Hz,1H),4.36-4.27(m,1H),3.48(d,J＝11.4Hz,1H),3.53–3.38(m,2H),3.32(d,J＝12.8Hz,2H),3.12–2.95(m,1H),1.47(s,9H),1.89(d,J＝4.6Hz,2H).

2.5 Synthesis of intermediate 2-9:

2-8 mg (0.58 mmol) of intermediate is weighed into a single-necked bottle, 2mL of ethyl acetate is added, 3mL of 3M HCl ethyl acetate solution is dropwise added under the ice bath condition, the reaction solution becomes turbid in the dropwise adding process, and the reaction solution is transferred to room temperature for reaction for 2h after the dropwise adding is completed. After the TLC monitoring reaction is finished, the reaction liquid is filtered by filter paper, and a filter cake is the product hydrochloride. The filter cake was then placed at 100mL K at 0deg.C ₂ CO ₃ In aqueous solution, extracted with ethyl acetate (120 mL. Times.4), the organic phases combined, dried over anhydrous sodium sulfate and concentrated to give 196mg of a pale brown powdered solid product 2-9 in a yield of about 81.3%; MS (ESI) m/z=414.27 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ10.44(s,1H),9.70–9.33(m,2H),8.55(s,1H),8.21(d,J＝8.3Hz,2H),7.90–7.75(m,2H),7.38(t,J＝7.9Hz,2H),7.13(t,J＝7.4Hz,1H),5.23(ddd,J＝14.7,10.2,4.3Hz,1H),3.53–3.38(m,2H),3.32(d,J＝12.6Hz,2H),3.12–2.95(m,1H),2.19(dd,J＝13.3,7.2Hz,2H),1.97(d,J＝3.2Hz,2H).

2.6 Synthesis of Compound II-1:

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the same procedure was followed, except that intermediate 1-6 was replaced with intermediate 2-9, to give white semi-solid II-1 in about 69.8% yield. MS (ESI) m/z=482.23 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ10.36(s,1H),8.30(s,1H),8.16(d,J＝8.1Hz,2H),7.92–7.75(m,4H),7.38(t,J＝7.8Hz,2H),7.13(t,J＝7.4Hz,1H),5.10(d,J＝49.1Hz,2H),3.80(d,J＝9.8Hz,2H),4.11(d,J＝40.3Hz,1H),3.55(d,J＝17.6Hz,1H),3.17(d,J＝54.7Hz,1H),2.29(t,J＝11.8Hz,1H),2.16(d,J＝15.3Hz,1H),1.93(s,3H),1.72–1.54(m,2H). ¹³ C NMR(101MHz,DMSO-d ₆ )δ170.67,165.51,158.66,156.26,154.60,143.50,140.75,139.52,136.22,135.10,129.11,128.93,128.69,124.38,120.88,115.28,97.93,20.56.

2.7 Synthesis of Compound II-2:

the same procedure was followed, except that intermediate 1-6 was replaced with intermediate 2-9 and intermediate 1-7 was replaced with acrylic acid 2-10 to give white powdery solid II-2 in a yield of about 67.4%. MS (ESI) m/z=467.21 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ9.11(s,1H),8.42(d,J＝8.4Hz,1H),8.37–8.27(m,2H),8.13(d,J＝8.3Hz,2H),7.85(d,J＝8.3Hz,2H),7.83–7.77(m,1H),7.12(dd,J＝8.1,5.0Hz,1H),6.06(t,J＝10.2Hz,1H),5.94(d,J＝14.7Hz,1H),5.06(d,J＝4.3Hz,1H),4.12–4.03(m,1H),3.58(d,J＝7.3Hz,1H),2.37(d,J＝11.1Hz,1H),2.29(d,J＝3.6Hz,1H),2.06–1.94(m,2H),1.78(d,J＝11.7Hz,1H),1.26(dd,J＝7.8,6.5Hz,2H). ¹³ C NMR(101MHz,CDCl ₃ )δ165.93,157.92,155.79,155.71,143.42,138.08,136.24,135.36,130.95,129.00,128.56,128.33,127.66,124.65,120.74,58.54,50.04,46.25,42.26,30.55,25.14,19.17.

2.8 Synthesis of Compound II-3:

the same procedure was followed except that 1.5, intermediate 2-9 was used instead of intermediate 1-6, 2-fluoroacrylic acid was used instead of intermediate 1-7 to obtain colorless oily liquid in a yield of about 81.6%. MS (ESI) m/z=482.23 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ10.37(s,1H),8.62(s,1H),8.45(d,J＝6.7Hz,2H),8.07(d,J＝8.0Hz,2H),7.81(s,1H),7.64(dd,J＝16.2,8.3Hz,1H),7.61–7.48(m,1H),7.37(d,J＝14.7Hz,1H),7.12(t,J＝7.0Hz,1H),6.23(d,J＝30.7Hz,1H),6.07(d,J＝15.0Hz,1H),4.61–4.49(m,1H),3.63(d,J＝9.6Hz,2H),3.18–3.05(m,2H),2.21(dd,J＝10.5,5.5Hz,1H),1.99(dd,J＝13.6,7.5Hz,1H),1.80–1.60(m,2H). ¹³ C NMR(101MHz,DMSO-d ₆ )δ165.50,165.08,153.38,151.16,149.62,146.55,139.62,135.26,132.63,131.90,129.17,129.08,127.81,124.38,120.94,120.88,102.04,54.07,46.25,38.69,29.49,18.52,17.16.

2.9 Synthesis of Compound II-4:

the operation is the same as 1.5, intermediate 2-9 is used for replacing intermediate 1-6, 2-chloroacrylic acid is used for replacing intermediate 1-7, and the reaction liquid is subjected to post-treatment to obtain white semi-solid, and the yield is about 71.4%. MS (ESI) m/z=502.31 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ10.35(s,1H),8.30(s,1H),8.16(d,J＝8.0Hz,2H),7.87–7.77(m,2H),7.64(dd,J＝15.6,7.9Hz,1H),7.52(s,1H),7.38(t,J＝7.7Hz,2H),7.13(t,J＝7.3Hz,1H),6.03(d,J＝9.5Hz,1H),5.86(d,J＝5.4Hz,1H),4.97–4.55(m,1H),3.23(t,J＝6.5Hz,2H),2.31(dd,J＝24.0,13.9Hz,2H),2.24–2.13(m,1H),2.06–1.95(m,1H),1.38(dd,J＝15.0,7.4Hz,2H). ¹³ C NMR(101MHz,DMSO-d ₆ )δ167.41,163.85,158.63,143.62,139.55,136.19,135.16,132.17,131.84,129.11,128.70,120.95,65.50,54.06,46.23,38.83,23.04.

2.10 Synthesis of Compound II-5:

the operation is the same as 1.5, intermediate 2-9 is used for replacing intermediate 1-6, 2-bromoacrylic acid is used for replacing intermediate 1-7, the reaction liquid is subjected to post-treatment to obtain brown solid, and the yield is about 75.3%. MS (ESI) m/z=546.12 [ M+H ]] ⁺ ； ¹ H NMR(500MHz,CDCl ₃ )δ8.38(s,1H),8.07(d,J＝8.3Hz,1H),7.82(d,J＝8.1Hz,2H),7.72(dd,J＝5.7,3.3Hz,2H),7.53(dd,J＝5.7,3.3Hz,2H),7.39(t,J＝7.9Hz,1H),7.18(t,J＝7.0Hz,1H),6.63(d,J＝11.5Hz,1H),6.08(d,J＝5.7Hz,1H),3.97–3.85(m,1H),3.74(dd,J＝7.5,1.9Hz,2H),3.56(d,J＝2.1Hz,1H),3.52(d,J＝2.7Hz,1H),2.12–1.87(m,2H),1.73–1.68(m,2H). ¹³ C NMR(126MHz,CDCl ₃ )δ167.78,165.13,136.20,132.43,130.96,129.14,128.85,128.30,124.82,120.43,65.61,47.21,47.16,30.56,29.71,19.19.

Example 3 synthesis of series iii compounds:

3.1 Synthesis of intermediate 3-3:

4-1.43 g (12.09 mol) of p-bromobenzoic acid was placed in a single-necked flask, 10mL of methylene chloride was added, 4.59g (12.09 mmol) of HATU, 5.14mL (30.30 mmol) of DIPEA and 4-2.98 mL of cyclohexylamine were sequentially added under ice-bath conditions, and then the mixture was transferred to room temperature and reacted overnight. The reaction was terminated by TLC (EA: pe=1:2), dichloromethane was removed under reduced pressure, saturated brine (200 ml×3) and ethyl acetate (150 mL) were added for extraction, the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, dry-stirred and run through a column, and PE wet-run, EA: pe=1:5 eluted to give 2.35g of white powdered solid 4-3 in a yield of about 82.9%. MS (ESI) m/z=282.15 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.65–7.59(m,2H),7.58–7.52(m,2H),3.95(tdt,J＝11.7,8.0,3.9Hz,2H),2.07–1.97(m,2H),1.82–1.71(m,2H),1.48–1.35(m,2H),1.23(dd,J＝13.3,2.5Hz,2H).

3.2 Synthesis of intermediate 3-4:

operating in the same way as 1.2, and replacing intermediate 1-3 with intermediate 3-3 to obtain white powdery solid 3-4 with a yield of about 89.6%; ¹ H NMR(400MHz,DMSO-d ₆ )δ7.85(d,J＝8.0Hz,2H),7.73(d,J＝8.0Hz,2H),3.82–3.67(m,2H),2.34–2.21(m,2H),1.76(m,2H),1.61(m,2H),1.37–1.26(dd,J＝30.5,11.9Hz,2H),1.20(s,12H).

3.3 Synthesis of intermediate 3-8:

operating the same as 2.4, and replacing intermediate 2-4 with intermediate 4-4 to obtain white powdery solid 3-8 with a yield of about 62.6%; MS (ESI) m/z=520.46 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ8.20(d,J＝8.0Hz,1H),7.87(d,J＝8.2Hz,2H),7.45(d,J＝8.2Hz,2H),4.20–4.12(m,1H),3.81(t,J＝7.5Hz,2H),3.34(d,J＝10.1Hz,1H),3.14(d,J＝10.7Hz,1H),3.01(d,J＝11.5Hz,1H),3.02–2.54(m,2H),2.18(dd,J＝15.7,7.1Hz,2H),1.95(m,4H),1.83(d,J＝9.3Hz,1H),1.74–1.68(m,2H),1.62(d,J＝12.5Hz,1H),1.46–1.33(m,2H),1.31(s,9H).

3.4 Synthesis of intermediate 3-9:

operating the same as 2.5, and replacing intermediate 2-8 with intermediate 3-8 to obtain pale yellow powdery solid 3-9 with a yield of about 87.4%; MS (ESI) m/z=420.37 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ8.40(d,J＝7.9Hz,1H),8.07(d,J＝7.9Hz,2H),7.75(d,J＝8.0Hz,2H),4.30–4.18(m,1H),3.81(dd,J＝7.5,9.5Hz,2H),3.54(t,J＝9.5Hz,2H),3.44(dd,J＝10.3,10.7Hz,2H),3.09–2.94(m,2H),2.17(dd,J＝15.9,7.3Hz,1H),1.97(d,J＝3.7Hz,1H),1.83(d,J＝9.1Hz,1H),1.78–1.69(m,2H),1.63(d,J＝12.7Hz,1H),1.43–1.23(m,2H),1.22–1.08(m,3H).

3.5 Synthesis of Compound III-1:

the operation is the same as 1.5, intermediate 3-9 is used for replacing intermediate 1-6, and the reaction liquid is post-treated to obtain pale yellow powdery solid III-1, and the yield is about 68.8%. MS (ESI) m/z=488.30 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,Acetone-d ₆ )δ8.28(s,1H),8.05(d,J＝8.2Hz,2H),7.79(d,J＝8.2Hz,2H),7.65(d,J＝7.7Hz,1H),5.76(s,1H),5.56(s,1H),4.41–4.30(m,1H),4.02–3.87(m,2H),2.45–2.32(m,2H),2.27–2.17(m,2H),2.06(dt,J＝4.3,2.2Hz,2H),2.02–1.95(m,3H),1.90(s,3H),1.83–1.71(m,4H),1.66(m,3H),1.21–1.11(m,2H). ¹³ C NMR(101MHz,Acetone-d ₆ )δ170.47,165.13,158.41,155.89,154.67,143.43,141.03,135.91,135.27,128.26,127.97,114.19,98.06,48.88,32.74,25.50,19.73.

3.6 Synthesis of Compound III-2:

the same procedure was followed, except that intermediate 3-9 was used in place of intermediate 1-6 and acrylic acid was used in place of 2-methacrylic acid 1-7 to give III-2 as a white powdery solid in a yield of about 77.6%. MS (ESI) m/z=496.45 [ M+Na ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ8.26(m,1H),8.02(d,J＝8.1Hz,2H),7.74(d,J＝7.7Hz,2H),6.66(d,J＝16.6Hz,1H),6.10(d,J＝5.6Hz,1H),4.83–4.65(m,1H),4.22(dd,J＝14.0,7.6Hz,1H),3.79(dd,J＝12.7,5.4Hz,1H),2.29(dd,J＝11.6,2.8Hz,1H),2.14(dd,J＝10.7,1.8Hz,1H),1.95(dd,J＝13.3,2.6Hz,1H),1.84(d,J＝7.7Hz,2H),1.80–1.70(m,2H),1.64(dd,J＝14.3,7.1Hz,3H),1.33(dd,J＝16.9,9.9Hz,4H),1.25(t,J＝7.9Hz,5H). ¹³ C NMR(101MHz,DMSO-d ₆ )δ167.41,165.87,165.05,158.67,156.31,155.21,149.07,136.62,136.01,135.71,134.10,132.17,131.98,129.28,129.13,128.62,125.63,116.52,98.01,65.49,45.60,30.47,19.12.

3.7 Synthesis of Compound III-3:

the same procedure was followed, except that 1.5, intermediate 3-9 was used instead of intermediate 1-6, 2-fluoroacrylic acid instead of 2-methacrylic acid 1-7, to give a yellow semi-solid compound III-3 in a yield of about 80.3%. MS (ESI) m/z=492.25 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ8.30(s,1H),8.03(d,J＝8.1Hz,2H),7.74(d,J＝8.1Hz,2H),6.25(s,1H),6.09(s,1H),3.84–3.71(m,1H),2.70(m,2H),2.31(dd,J＝22.3,11.9Hz,2H),2.16(d,J＝16.6Hz,2H),1.99(d,J＝13.6Hz,2H),1.85(dt,J＝8.2,10.9Hz,3H),1.75(dt,J＝9.0,11.4Hz,3H),1.62(q,J＝12.2Hz,3H),1.42–1.09(m,2H). ¹³ C NMR(101MHz,DMSO-d ₆ )δ165.34,161.17,160.86,158.63,156.24,154.59,143.69,135.59,135.09,128.52,99.26,99.11,97.98,48.90,38.69,32.88,25.71,25.36.

3.8 Synthesis of Compound III-4:

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the same operation as 1.5, intermediate 3-9 was used instead of intermediate 1-6, 2-chloroacrylic acid was used instead of 2-methacrylic acid 1-7 to give pale yellow powdered solid III-4 in about 72.4% yield. MS (ESI) m/z=508.22 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ8.35–8.27(s,1H),8.03(d,J＝8.0Hz,2H),7.74(d,J＝8.0Hz,2H),5.86(s,1H),5.61(s,1H),3.88–3.71(m,1H),3.62(dq,J＝10.5,6.6Hz,2H),3.21–3.06(m,3H),2.38–2.23(m,2H),2.22–2.09(m,2H),1.98(d,J＝12.6Hz,2H),1.84(m,2H),1.75(q,J＝6.8Hz,2H),1.62(m,2H),1.39–1.31(m,2H). ¹³ C NMR(101MHz,DMSO-d ₆ )δ165.39,163.80,158.62,156.22,154.59,143.73,135.57,135.10,128.50,97.99,54.06,48.89,42.30,32.89,25.72,18.52.

3.9 Synthesis of Compound III-5:

the same procedure was followed, except that 1.5 was used, intermediate 3-9 was used instead of intermediate 1-6, 2-bromoacrylic acid was used instead of 2-methacrylic acid 1-7, to give compound III-5 as a brown solid, with a yield of about 85.3%. MS (ESI) m/z=552.16 [ M+H ]] ⁺ ； ¹ H NMR(500MHz,CDCl ₃ )δ8.37(s,1H),7.93(d,J＝8.3Hz,2H),7.76(d,J＝8.2Hz,2H),6.17(s,1H),6.05(s,1H),4.09–3.95(m,1H),3.22(q,J＝7.3Hz,2H),2.42–2.31(m,2H),2.27(dd,J＝13.0,3.8Hz,2H),2.11–2.00(m,3H),1.84–1.75(m,3H),1.29(dd,J＝11.3,2.8Hz,4H),1.14–1.05(m,2H). ¹³ C NMR(126MHz,CDCl ₃ )δ166.05,165.06,157.60,155.69,155.62,143.67,135.83,135.64,132.29,130.93,128.85,128.59,127.97,65.60,55.65,49.02,33.16,24.95,17.18.

Example 4 synthesis of series iv compounds:

4.1 Synthesis of intermediate 4-3:

the same procedure was used to replace compound aniline 2-2 with 2.1, 2-aminopyridine to give 4-3 as a brown powdered solid in a yield of about 89.1%. MS (ESI) m/z=276.99 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ8.93(s,1H),8.63(d,J＝1.3Hz,1H),8.46(d,J＝2.5Hz,1H),8.35(dd,J＝2.5,1.5Hz,1H),7.60–7.58(m,2H),7.55–7.53(m,2H).

4.2 Synthesis of intermediate 4-4:

operating same as 2.2, intermediate 4-3 replacing intermediate 2-3, obtaining pale brown powdery solid 4-4, yield about 92.6%; ¹ H NMR(400MHz,DMSO-d ₆ )δ10.89(s,1H),8.40(d,J＝4.8Hz,1H),8.19(d,J＝8.4Hz,1H),8.02(d,J＝8.2Hz,2H),7.88–7.82(m,1H),7.79(d,J＝8.2Hz,2H),7.24–7.15(m,1H),1.32(s,12H).

4.3 Synthesis of intermediate 4-8:

operating same as 2.5, intermediate 4-4 replacing intermediate 2-4, obtaining pale brown powdery solid 4-8, yield about 73.4%; MS (ESI) m/z=515.25 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ9.10(s,1H),8.44(d,J＝8.4Hz,1H),8.37(s,1H),8.32(d,J＝4.9Hz,1H),8.14(d,J＝8.3Hz,2H),7.87(d,J＝8.2Hz,2H),7.85–7.77(m,1H),7.13(dd,J＝7.3,5.0Hz,1H),4.87–4.73(m,1H),3.63(dd,J＝13.5,6.4Hz,2H),2.90(t,J＝11.5Hz,2H),2.26–2.17(m,2H),1.84–1.64(m,2H),1.46(s,12H).

4.4 Synthesis of intermediate 4-9:

operating same as 2.4, intermediate 4-8 replacing intermediate 2-8, obtaining pale brown powdery solid 4-9, yield about 61.9%; MS (ESI) m/z=437.20 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ11.66(s,1H),9.75(d,J＝10.1Hz,1H),9.51(d,J＝10.9Hz,1H),8.64(s,1H),8.49(d,J＝4.8Hz,1H),8.36–8.26(m,2H),8.10(t,J＝7.7Hz,1H),7.86(d,J＝8.1Hz,1H),7.47–7.30(m,1H),5.26–5.10(m,1H),3.54(d,J＝10.7Hz,1H),3.50–3.38(m,2H),3.31(d,J＝11.5Hz,1H),3.12–2.93(m,1H),2.15(d,J＝5.2Hz,2H),1.05(t,J＝7.0Hz,1H).

4.5 Synthesis of Compound IV-1:

the operation is the same as 1.5, intermediate 4-9 replaces intermediate 1-6, and white powdery solid IV-1 is obtained, and the yield is about 81.4%; MS (ESI) m/z=483.23 [ M+H ]] ⁺ ； ¹ H NMR(500MHz,CDCl ₃ )δ9.28(s,1H),8.42(d,J＝8.4Hz,1H),8.29(d,J＝4.8Hz,2H),8.12(d,J＝8.0Hz,2H),7.84(d,J＝7.2Hz,2H),7.79(t,J＝7.9Hz,1H),7.14–7.07(m,1H),5.17(s,1H),5.11(s,1H),5.06–4.88(m,1H),3.44–3.29(m,2H),2.86(d,J＝9.8Hz,2H),1.97(s,3H),1.80–1.63(m,2H),1.68-1.58(m,2H). ¹³ C NMR(126MHz,CDCl ₃ )δ171.51,165.26,157.99,155.84,154.41,151.56,147.77,143.23,134.74,128.81,128.40,120.16,115.52,114.53,98.66,98.59,30.28,29.68,20.52.

4.6 Synthesis of Compound IV-2:

the operation is the same as 1.5, the intermediate 4-9 replaces the intermediate 1-6, the acrylic acid replaces the 2-methacrylic acid 1-7, and the white powdery solid IV-2 is obtained, and the yield is about 74.3%; MS (ESI) m/z=469.21 [ M+H ]] ⁺ ； ¹ H NMR(500MHz,CDCl ₃ )δ9.30(s,1H),8.42(d,J＝8.3Hz,1H),8.29(dd,J＝28.2,24.2Hz,2H),8.12(d,J＝5.9Hz,2H),7.84(d,J＝7.9Hz,2H),7.79(t,J＝7.4Hz,1H),7.15–7.05(m,1H),6.69(q,J＝6.6Hz,1H),6.30(d,J＝16.9Hz,1H),5.69(d,J＝4.1Hz,1H),4.86–4.72(m,1H),3.42–3.31(m,1H),2.92(t,J＝11.4Hz,1H),2.38(dd,J＝26.1,12.0Hz,1H),2.26(d,J＝9.3Hz,1H),2.03(dd,J＝20.8,9.0Hz,2H),1.29(dt,J＝31.4,7.1Hz,2H).

4.7 Synthesis of Compound IV-3:

the operation is the same as 1.5, intermediate 4-9 replaces intermediate 1-6, 2-fluoro acrylic acid replaces 2-methacrylic acid 1-7, and off-white powdery solid IV-3 is obtained, and the yield is about 61.0%; MS (ESI) m/z=487.20 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ9.23(s,1H),8.43(d,J＝8.4Hz,2H),8.30(d,J＝4.9Hz,4H),8.13(d,J＝8.3Hz,4H),7.89–7.76(m,6H),7.15–7.07(m,2H),5.12(d,J＝13.3Hz,2H),5.00–4.86(m,2H),2.48–2.17(m,4H),2.13–1.97(m,3H),1.85–1.67(m,2H),1.25(d,J＝5.8Hz,4H). ¹³ C NMR(101MHz,CDCl ₃ )δ165.19,161.62,161.32,157.73,155.47,154.35,151.49,147.62,143.43,138.82,136.82,134.77,128.81,128.45,120.19,114.56,99.81,99.65,98.55,30.20,29.69.

4.8 Synthesis of Compound IV-4:

the procedure is as for 1.5, with intermediate 4-9 replacing intermediate 1-6, 2-chloroacrylic acid replacing 2-methacrylic acid 1-7, to give a pale brown powdery solidIV-4, yield about 64.8%; MS (ESI) m/z=503.17 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ9.29(s,1H),8.43(d,J＝8.4Hz,1H),8.38–8.24(m,2H),8.14(d,J＝8.3Hz,2H),7.82(dd,J＝19.1,8.7Hz,3H),7.17–7.05(m,1H),5.68(s,1H),5.64(s,1H),3.95–3.82(m,1H),2.37(d,J＝10.7Hz,1H),2.31–2.23(m,2H),2.03(d,J＝11.1Hz,1H),1.85–1.72(m,2H),1.26(t,J＝7.1Hz,2H). ¹³ C NMR(101MHz,CDCl ₃ )δ165.22,164.52,157.75,155.44,154.31,151.50,147.53,143.52,138.88,136.76,134.76,128.81,128.48,120.19,117.61,114.62,98.67,53.42,30.16,21.38,13.90.

4.9 Synthesis of Compound IV-5:

the operation is the same as 1.5, intermediate 4-9 replaces intermediate 1-6, 2-bromoacrylic acid replaces 2-methacrylic acid 1-7, light brown powdery solid IV-5 is obtained, and the yield is about 67.5%; MS (ESI) m/z=547.12 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ9.11(s,1H),8.42(d,J＝8.4Hz,1H),8.37–8.27(m,2H),8.13(d,J＝8.3Hz,2H),7.85(d,J＝8.3Hz,2H),7.83–7.77(m,1H),7.12(dd,J＝8.1,5.0Hz,1H),6.43(s,1H),6.06(s,1H),4.12–3.98(m,1H),3.58(d,J＝7.3Hz,1H),2.37(d,J＝11.1Hz,1H),2.29(d,J＝3.6Hz,1H),2.06–1.94(m,2H),1.78(d,J＝11.7Hz,1H),1.26(dd,J＝7.8,6.5Hz,2H). ¹³ C NMR(101MHz,CDCl ₃ )δ165.14,165.00,157.85,155.86,151.48,147.76,143.34,134.75,128.84,128.40,120.21,114.48,98.62,30.18.

4.10 Synthesis of Compound IV-6:

the operation is the same as 1.5, the intermediate 4-9 replaces the intermediate 1-6, the 2-butynoic acid replaces the 2-methacrylic acid 1-7, and white powdery solid IV-6 is obtained, and the yield is about 45.3%; MS (ESI) m/z=481.21 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ8.80(s,1H),8.42(d,J＝9.6Hz,2H),8.36–8.31(m,1H),8.12(dd,J＝8.1,5.7Hz,2H),7.86(t,J＝8.3Hz,2H),7.80(t,J＝7.9Hz,1H),7.12(t,J＝5.9Hz,1H),5.00–4.82(m,1H),4.51(ddd,J＝44.4,22.2,8.9Hz,2H),3.34(dd,J＝14.5,7.3Hz,1H),2.34(dd,J＝12.0,3.6Hz,1H),1.91(s,3H),1.74(dd,J＝25.3,13.3Hz,2H),1.33(t,J＝9.6Hz,2H). ¹³ C NMR(101MHz,CDCl ₃ )δ165.00,157.73,155.94,153.45,153.35,151.38,147.93,138.66,136.94,134.65,128.90,128.88,128.35,128.29,120.27,114.33,89.83,53.35,52.54,46.91,41.30,30.14,25.06,23.76.

Example 5 synthesis of series v compounds:

5.1 Synthesis of intermediate 5-3:

2-amino-3-hydroxypyridine 5-1.10 g (10 mmol) p-bromobenzaldehyde 5-2.10 g (10 mmol) and polyphosphoric acid (30 mL) were placed in a 100mL single-necked flask, nitrogen-protected, and heated at 150℃for 1h. Pouring the reaction solution into 300mL of water, carrying out suction filtration, washing with water, and drying to obtain 1.90g of orange solid, namely intermediate 5-3, with the yield of about 69.3%; MS (ESI) m/z=274.98 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ8.57(dd,J＝4.9,1.4Hz,1H),8.27(dd,J＝8.2,1.4Hz,1H),8.21–8.15(m,2H),7.91–7.84(m,2H),7.50(dd,J＝8.2,4.9Hz,1H).

5.2 Synthesis of intermediate 5-4:

the operation is the same as 1.2, intermediate 5-3 replaces intermediate 1-3, and pale yellow semi-solid 5-4 is obtained, and the yield is about 89.8%; ¹ H NMR(400MHz,DMSO-d ₆ )δ8.58(dd,J＝4.8,1.4Hz,1H),8.27(dd,J＝8.2,1.4Hz,3H),7.93(d,J＝8.3Hz,2H),7.50(dd,J＝8.2,4.9Hz,1H),1.34(s,12H).

5.3 Synthesis of intermediate 5-8:

operating with 2.4, intermediate 5-4 replaces intermediate 2-4 to obtain pale brown semisolid 5-8, and the yield is about 76.3%; MS (ESI) m/z=513.42 [ M+H ]] ⁺ ； ¹ H NMR(500MHz,CDCl ₃ )δ8.63(dd,J＝4.9,1.4Hz,1H),8.50(d,J＝8.3Hz,2H),8.41(s,1H),7.95–7.89(m,3H),7.35(dd,J＝8.1,4.9Hz,1H),4.96–4.83(m,1H),3.51(d,J＝7.1Hz,2H),2.89(t,J＝12.0Hz,1H),2.35–2.17(m,3H),1.79–1.66(m,2H),1.45(s,9H).

5.4 Synthesis of intermediate 5-9:

operating with 2.5, intermediate 5-8 replacing intermediate 2-8, obtaining pale brown powdery solid 5-9, yield about 81.0%; MS (ESI) m/z=413.35 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ8.59(dd,J＝4.8,1.3Hz,1H),8.41(d,J＝8.4Hz,2H),8.34–8.26(m,2H),7.95(d,J＝8.4Hz,2H),7.51(dd,J＝8.2,4.9Hz,1H),3.15–3.06(m,1H),2.95(dd,J＝23.5,11.9Hz,2H),2.12(dd,J＝28.1,18.2,7.4Hz,2H),1.69(d,J＝2.7Hz,1H),1.58(dd,J＝8.8,3.7Hz,2H),1.37(d,J＝7.1Hz,1H).

5.5 Synthesis of Compound V-1:

the operation is the same as 1.5, intermediate 5-9 replaces intermediate 1-6, and white powdery solid V-1 is obtained, and the yield is about 74.2%; MS (ESI) m/z=481.21 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ8.63(dd,J＝4.9,1.2Hz,1H),8.50(d,J＝8.3Hz,2H),8.36(s,1H),7.99–7.85(m,3H),7.36(dd,J＝8.1,4.9Hz,1H),5.19(s,1H),5.10(s,1H),3.91–3.79(m,1H),2.23(t,J＝6.6Hz,2H),2.10–1.98(m,2H),1.85(s,3H),1.73(d,J＝10.5Hz,2H),1.49–1.25(m,2H).

5.6 Synthesis of Compound V-2:

the operation is the same as 1.5, the intermediate 5-9 replaces the intermediate 1-6, the acrylic acid replaces the 2-methacrylic acid 1-7, and the white powdery solid V-2 is obtained, and the yield is about 79.6%; MS (ESI) m/z=467.20 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ8.62(d,J＝4.7Hz,1H),8.49(d,J＝8.0Hz,2H),8.39(s,1H),7.92(dd,J＝7.7,4.6Hz,3H),7.35(dd,J＝8.0,4.9Hz,1H),6.74(t,J＝6.0Hz,1H),6.48(d,J＝13.3Hz,1H),6.19(d,J＝7.6Hz,1H),3.87–3.65(m,1H),3.22(d,J＝16.2Hz,1H),3.03–2.79(m,1H),2.50–2.32(m,2H),2.29(dd,J＝14.8,6.2Hz,1H),2.09–1.93(m,2H),1.75(dd,J＝24.2,11.7Hz,1H). ¹³ C NMR(101MHz,CDCl ₃ )δ165.78,162.29,157.73,156.23,155.84,147.03,143.27,137.90,134.85,134.73,129.03,128.12,127.60,123.84,120.45,118.38,98.59,61.92,48.66,29.70,20.48.

5.7 Synthesis of Compound V-3:

the operation is the same as 1.5, the intermediate 5-9 replaces the intermediate 1-6, the 2-fluoroacrylic acid replaces the 2-methacrylic acid 1-7, and the white powdery solid V-3 is obtained, and the yield is about 73.8%; MS (ESI) m/z=485.19 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ8.62(dd,J＝4.9,1.2Hz,1H),8.49(d,J＝8.3Hz,2H),8.40(s,1H),7.97–7.86(m,3H),7.35(dd,J＝8.1,4.9Hz,1H),5.90(s,1H),5.12(s,1H),3.97–3.82(m,1H),3.38(d,J＝11.2Hz,2H),2.29(dd,J＝12.9,3.7Hz,1H),2.10–2.00(m,2H),1.79(dd,J＝25.7,12.2Hz,1H),1.53–1.37(m,2H). ¹³ C NMR(101MHz,CDCl ₃ )δ164.84,161.62,161.32,157.75,156.20,155.81,154.52,147.02,143.33,143.27,136.97,129.02,126.98,120.46,118.38,29.69.

5.8 Synthesis of Compound V-4:

the operation is the same as 1.5, the intermediate 5-9 replaces the intermediate 1-6, the 2-chloroacrylic acid replaces the 2-methacrylic acid 1-7, and the light brown powdery solid V-4 is obtained, and the yield is about 61.2%; MS (ESI) m/z=501.16 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ8.62(dd,J＝4.9,1.4Hz,1H),8.49(d,J＝8.4Hz,2H),8.37(s,1H),7.96–7.87(m,3H),7.35(dd,J＝8.1,4.9Hz,1H),5.68(s,1H),4.97(s,1H),3.64–3.48(m,1H),3.26(d,J＝5.2Hz,2H),2.46–2.35(m,2H),2.07(t,J＝7.3Hz,1H),1.79(d,J＝13.4Hz,1H),1.33–1.24(m,2H). ¹³ C NMR(101MHz,CDCl ₃ )δ176.14,164.83,164.52,157.84,156.18,155.61,154.46,147.01,143.48,143.27,136.93,129.02,127.01,120.47,118.41,98.58,60.41,30.19,29.69,21.06.

5.9 Synthesis of Compound V-5:

the operation is the same as 1.5, the intermediate 5-9 replaces the intermediate 1-6, the 2-bromoacrylic acid replaces the 2-methacrylic acid 1-7, and the light brown powdery solid V-5 is obtained, and the yield is about 57.4%; MS (ESI) m/z=545.10 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ8.59(d,J＝4.6Hz,1H),8.43(d,J＝8.1Hz,2H),8.33(s,1H),8.29(d,J＝8.1Hz,1H),7.96(d,J＝7.8Hz,2H),7.51(dd,J＝8.1,4.9Hz,1H),6.14(s,1H),5.92(s,1H),4.45–4.35(m,1H),3.25(t,J＝7.3Hz,2H),2.39(d,J＝15.0Hz,1H),2.25–2.12(m,2H),2.01(dd,J＝10.9,4.2Hz,1H),1.33–1.16(m,2H). ¹³ C NMR(101MHz,DMSO-d ₆ )δ165.03,162.87,160.29,158.05,155.93,149.01,147.23,143.64,143.37,138.69,137.00,129.66,128.93,126.28,121.54,119.63,106.77,79.71,62.65,47.15,44.91,27.07,21.53.

5.10 Synthesis of Compound V-6:

the operation is the same as 1.5, the intermediate 5-9 replaces the intermediate 1-6, the 2-butynoic acid replaces the 2-methacrylic acid 1-7, and the white powdery solid V-6 is obtained, and the yield is about 45.6%; MS (ESI) m/z=479.19 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ8.62(d,J＝4.4Hz,1H),8.49(d,J＝8.2Hz,2H),8.43–8.35(m,1H),7.92(t,J＝7.3Hz,3H),7.35(dd,J＝8.0,4.9Hz,1H),5.03–4.83(m,1H),3.96(dd,J＝15.6,8.4Hz,2H),3.08(t,J＝4.2Hz,2H),2.32–2.24(m,2H),1.98(s,3H),1.80–1.66(m,2H). ¹³ C NMR(101MHz,CDCl ₃ )δ171.54,171.54,164.86,164.86,157.76,157.71,156.19,156.19,155.79,155.79,154.47,154.47,147.02,147.02,143.27,143.27,137.02,129.03,127.01,120.47,120.47,118.41,118.41,98.60,77.37,76.73,31.89,29.70,29.30,22.69,20.55,4.06.

Example 6 synthesis of series vi compounds:

6.1 Synthesis of intermediate 6-3:

6-1.43 g (12.09 mmol) of p-bromobenzoic acid was weighed into a single-necked flask, 10mL of methylene chloride was added, 4.59g (12.09 mmol) of HATU, 5.14mL (30.30 mmol) of DIPEA and 6-2.13 g (10.08 mmol) of 2-amino-4-fluoropyridine were sequentially added at 0℃and then transferred to room temperature to react overnight. The reaction was terminated by TLC (EA: pe=1:5), dichloromethane was removed under reduced pressure, saturated brine (400 ml×3) and ethyl acetate were added for extraction 500mL, the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, dry-stirred and run through a column, PE wet-run, EA: pe=1:5 eluted to give 2.27g of white powdery solid in about 76.5% yield. MS (ESI) m/z=294.98 [ M+H ]]+；1H NMR(400MHz,DMSO-d ₆ )δ10.99(s,1H),8.41(ddd,J＝4.8,1.9,0.8Hz,1H),8.17(d,J＝8.4Hz,1H),8.02(dd,J＝9.8,1.9Hz,1H),7.93–7.79(m,3H),7.25–7.14(m,1H).

6.2 Synthesis of intermediate 6-4:

the same procedure as 1.2, intermediate 6-3 was used instead of intermediate 1-3 to give white solid 6-4. The yield was about 89.6%; ¹ H NMR(400MHz,DMSO-d ₆ )δ10.93(d,J＝32.2Hz,1H),8.47–8.35(m,2H),8.18(d,J＝8.4Hz,1H),7.90–7.69(m,3H),7.24–7.14(m,1H),1.33(s,12H).

6.3 Synthesis of intermediate 6-8:

the operation is the same as 2.4, the intermediate 2-4 is replaced by the intermediate 6-4, and the yield is about 78.5%; MS (ESI) m/z=551.24 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ11.05(s,1H),8.44(d,J＝3.1Hz,1H),8.35–8.24(m,2H),8.20(d,J＝8.3Hz,2H),7.91–7.73(m,3H),4.72–4.59(m,1H),3.72(dd,J＝72.6,18.4Hz,2H),3.65(d,J＝44.0Hz,2H),3.05(dd,J＝46.4,21.1Hz,1H),2.24–2.09(m,2H),2.11(dd,J＝12.7,3.7Hz,1H),1.26(s,9H).

6.4 Synthesis of intermediate 6-9:

the operation is the same as 2.5, intermediate 6-8 replaces intermediate 2-8, light brown semisolid 6-9 is obtained, and the yield is about 82.7%; MS (ESI) m/z=451.20 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ11.05(s,1H),8.76(s,1H),8.44(d,J＝2.7Hz,1H),8.31–8.24(m,2H),8.20(d,J＝8.1Hz,1H),7.89–7.76(m,3H),4.83–4.58(m,1H),3.09(dd,J＝10.5,1.5Hz,1H),2.94(dd,J＝22.0,10.9Hz,2H),2.23–2.00(m,2H),1.77(d,J＝12.9Hz,1H),1.65–1.48(m,1H),1.19(dd,J＝17.3,10.2Hz,1H).

6.5 Synthesis of Compound VI-1:

the operation is the same as 1.5, the intermediate 1-6 is replaced by the intermediate 6-9, and the 2-methacrylic acid 1-7 is replaced by the acrylic acid, so that a pale yellow solid VI-1 is obtained, and the yield is about 77.9%; MS (ESI) m/z=505.31 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ11.01(s,1H),8.92(s,1H),8.43(d,J＝3.0Hz,1H),8.32–8.24(m,2H),8.21(d,J＝8.2Hz,1H),7.81(d,J＝8.5Hz,1H),7.71(dt,J＝23.0,6.3Hz,2H),6.52(t,J＝6.5Hz,1H),6.24(d,J＝10.8Hz,1H),5.92(d,J＝5.2Hz,1H),3.13–3.00(m,1H),2.36–2.23(m,2H),2.15(d,J＝12.9Hz,1H),1.95(d,J＝13.6Hz,1H),1.69–1.60(m,2H),1.38(dd,J＝15.0,7.4Hz,1H),1.18(t,J＝7.3Hz,1H). ¹³ C NMR(101MHz,DMSO-d ₆ )δ167.41,165.87,165.05,158.67,156.31,155.21,149.07,136.62,136.01,135.71,134.10,132.17,131.98,129.28,129.13,128.62,125.83,125.63,116.52,98.01,65.49,45.60,30.47,19.12.

6.6 Synthesis of Compound VI-2:

the operation is the same as 1.5, intermediate 6-9 replaces intermediate 1-6, 2-chloroacrylic acid replaces 2-methacrylic acid 1-7, light brown solid VI-2 is obtained, and the yield is about 80.2%; MS (ESI) m/z=505.14 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ11.02(s,1H),8.96(s,1H),8.43(d,J＝3.0Hz,1H),8.33(s,1H),8.26(dd,J＝9.2,4.2Hz,1H),8.21(d,J＝8.2Hz,2H),7.83(dd,J＝14.3,5.7Hz,2H),5.89(s,1H),5.64(s,1H),4.87–4.56(m,1H),3.31(t,J＝10.0Hz,2H),3.18(d,J＝12.6Hz,2H),2.00(d,J＝14.6Hz,1H),1.91(t,J＝7.4Hz,1H),1.77–1.59(m,2H). ¹³ C NMR(101MHz,DMSO-d ₆ )δ165.89,163.87,158.07,149.08,143.82,136.43,136.02,135.68,134.22,132.17,129.30,128.66,125.83,125.64,116.58,116.54,65.49,46.22,30.46,19.11.

6.7 Synthesis of Compound VI-3:

the operation is the same as1.5, intermediate 6-9 replaces intermediate 1-6, 2-bromoacrylic acid replaces 2-methacrylic acid 1-7 to obtain light brown solid VI-3, the yield is about 76.2%; MS (ESI) m/z=583.10 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d6)δ11.01(s,1H),8.43(d,J＝3.0Hz,1H),8.32–8.24(m,2H),8.21(d,J＝8.2Hz,2H),7.88–7.77(m,3H),6.13(s,1H),5.76(s,1H),3.95–3.72(m,1H),2.49(dd,J＝19.6,11.3Hz,1H),2.18(t,J＝12.6Hz,2H),2.00(d,J＝14.6Hz,1H),1.77–1.59(m,2H),1.39(t,J＝7.4Hz,2H). ¹³ C NMR(101MHz,DMSO-d6)δ170.97,165.90,163.42,160.06,158.32,155.35,154.06,142.93,139.05,136.46,131.60,128.06,125.82,122.12,104.17,97.22,93.80,67.49,49.52,45.73,29.44,23.72.

Example 7 synthesis of series vii compounds:

7.1 Synthesis of intermediate 7-3:

1.00g (4.57 mol) of 3-fluoro-4-bromobenzoic acid was weighed into a single-necked flask, 5mL of methylene chloride was added, and after the flask was placed under ice-bath conditions, 1.74g (4.57 mmol) of HATU, 2.39mL (13.71 mmol) of DIPEA and 1.13g (3.81 mmol) of 2-aminopyridine were sequentially added, followed by transfer to room temperature for reaction overnight. After the reaction was completed by TLC, the solvent dichloromethane was removed under reduced pressure, saturated brine (200 ml×3) and ethyl acetate 250mL were added for extraction, the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, dry-mixed and passed through a column, and PE wet-packed, EA: pe=1:4 eluted to give 1.12g of a pale brown powdery solid, the yield was about 74.6%. MS (ESI) m/z=294.98 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d6)δ10.99(s,1H),8.41(ddd,J＝4.8,1.9,0.8Hz,1H),8.17(d,J＝8.4Hz,1H),8.02(dd,J＝9.8,1.9Hz,1H),7.93–7.79(m,3H),7.25–7.14(m,1H).

7.2 Synthesis of intermediate 7-4:

the procedure is as 1.2, intermediate 7-3 is substitutedIntermediate 1-3 to give a white solid with a yield of about 91.6%; ¹ H NMR(400MHz,DMSO-d ₆ )δ10.93(d,J＝32.2Hz,1H),8.47–8.35(m,2H),8.18(d,J＝8.4Hz,1H),7.90–7.69(m,3H),7.24–7.14(m,1H),1.33(s,12H).

7.3 Synthesis of intermediate 7-8:

the operation is the same as 2.4, the intermediate 7-4 replaces the intermediate 2-4, and the yield is about 73.2%; MS (ESI) m/z=583.10 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ10.99(s,1H),9.08(s,1H),8.43(d,J＝3.9Hz,1H),8.28(d,J＝8.8Hz,2H),8.21(t,J＝10.0Hz,2H),7.97–7.82(m,1H),7.70(t,J＝7.8Hz,1H),4.82–4.64(m,1H),2.29–2.15(m,2H),2.15–2.04(m,2H),1.90(dd,J＝9.3,5.7Hz,2H),1.60(dd,J＝24.5,13.2Hz,2H),1.37(s,9H).

7.4 Synthesis of intermediate 7-9:

the same operation as 2.5, intermediate 7-8 replaces intermediate 2-8, light brown solid is obtained, and the yield is about 82.1%; MS (ESI) m/z=433.25 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ8.44–8.41(m,2H),8.24(s,1H),8.05(dd,J＝6.1,1.1Hz,2H),8.04–8.01(m,1H),7.89–7.86(m,2H),3.71–3.68(m,1H),3.17(d,J＝7.0Hz,1H),2.96(t,J＝7.3Hz,2H),2.65(d,J＝3.3Hz,1H),2.39–2.16(m,2H),1.84–1.80(m,2H).

7.5 Synthesis of Compound VII-1:

the same operation as 1.5, intermediate 7-9 replaced intermediate 1-6, acrylic acid replaced 2-methacrylic acid 1-7, to obtain pale brown solid VII-1, yield about 63.8%; MS (ESI) m/z=487.19 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,Acetone-d ₆ )δ9.47(s,1H),8.25–8.17(m,2H),8.09(s,1H),8.01(dd,J＝8.0,1.2Hz,1H),7.92–7.81(m,1H),7.42(ddd,J＝21.1,12.1,4.7Hz,2H),7.15(dd,J＝7.2,4.9Hz,1H),6.62(d,J＝11.2Hz,1H),5.58(t,J＝9.0Hz,2H),4.72(d,J＝4.1Hz,1H),3.42–3.31(m,1H),3.31–3.13(m,2H),2.92(t,J＝11.4Hz,1H),2.38(dd,J＝26.1,12.0Hz,2H),2.26(d,J＝9.3Hz,1H),2.03(dd,J＝20.8,9.0Hz,2H). ¹³ C NMR(101MHz,Acetone-d ₆ )δ168.83,165.56,158.85,156.88,154.27,152.84,146.90,141.84,137.10,135.99,132.27,128.33,124.77,124.07,120.07,114.35,109.51,99.48,96.60,64.11,48.64,28.09,21.08.

7.6 Synthesis of Compound VII-2:

the operation is the same as 1.5, intermediate 7-9 replaces intermediate 1-6, 2-chloroacrylic acid replaces 2-methacrylic acid 1-7, brown solid VII-2 is obtained, and the yield is about 63.8%; MS (ESI) m/z=487.19 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,Acetone-d ₆ )δ9.83(s,1H),8.36(d,J＝8.3Hz,2H),8.29(s,1H),8.13(d,J＝7.9Hz,1H),8.05(d,J＝10.8Hz,1H),7.85(dt,J＝15.5,7.7Hz,2H),7.18(dd,J＝7.1,4.8Hz,1H),6.62(s,1H),6.04(s,1H),3.69–3.49(m,1H),3.31(d,J＝26.2Hz,2H),2.48–2.34(m,1H),2.29(d,J＝3.8Hz,1H),2.10(dd,J＝8.6,4.8Hz,1H),1.80(dd,J＝9.2,4.3Hz,2H),1.30(d,J＝6.0Hz,1H). ¹³ C NMR(101MHz,Acetone-d ₆ )δ163.65,161.06,158.60,158.23,155.82,155.81,154.39,148.15,138.15,137.58,132.09,132.06,124.10,124.07,120.03,116.93,116.06,115.83,114.29,80.13,64.63,44.52,39.44,37.84,21.48.

7.7 Synthesis of Compound VII-3:

the same procedure was followed, with intermediate 7-9 replacing intermediate 1-6, 2-bromoacrylic acid replacing 2-methacrylic acid 1-7, to give brown solid VII-3, yield about 63.8%; MS (ESI) m/z=565.11 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,Acetone-d ₆ )δ9.95(s,1H),8.43–8.32(m,2H),8.28(s,1H),8.12(dd,J＝8.0,1.2Hz,1H),8.08–8.01(m,1H),7.86(ddd,J＝21.1,12.1,4.7Hz,2H),7.19(dd,J＝7.2,4.9Hz,1H),6.62(s,1H),6.35(s,1H),4.46–4.25(m,1H),3.91(dq,J＝13.2,6.6Hz,2H),3.41(q,J＝7.4Hz,2H),2.25(d,J＝11.6Hz,1H),2.03(d,J＝3.4Hz,1H),1.82–1.64(m,2H). ¹³ C NMR(101MHz,Acetone-d ₆ )δ158.37,155.98,152.09,148.19,138.21,132.09,132.06,128.18,124.77,124.11,124.07,120.07,116.05,115.81,114.35,54.82,42.83,37.86,22.32.

Example 8 synthesis of series viii compounds:

8.1 Synthesis of Compound VIII-1:

the operation is the same as 1.5, the intermediate 7-9 replaces the intermediate 1-6, and the (E) -4- (dimethylamino) but-2-enoic acid replaces the 2-methacrylic acid 1-7, so that white solid VIII-1 is obtained, and the yield is about 63.8%; MS (ESI) m/z=565.11 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ10.19(s,1H),9.03(s,1H),8.56(d,J＝4.6Hz,1H),8.19(d,J＝7.9Hz,1H),8.15(t,J＝11.2Hz,1H),8.07–7.96(m,2H),7.85–7.72(m,1H),7.64(t,J＝8.4Hz,1H),6.70(m,1H),6.52(d,J＝10.6Hz,1H),4.43–4.32(m,1H),3.71(d,J＝7.6Hz,2H),2.78(s,6H),3.29–3.15(m,2H),3.02(d,J＝9.4Hz,2H),2.15–2.04(m,2H),1.90(dd,J＝9.3,5.7Hz,1H),1.60(dd,J＝23.7,12.6Hz,1H). ¹³ C NMR(101MHz,DMSO-d ₆ )δ168.24,163.61,160.09,158.85,154.74,151.54,149.02,143.63,136.36,135.28,129.24,128.39,127.67,124.00,120.75,115.44,112.68,98.30,66.05,65.83,46.23,33.92,20.96.

8.2 Synthesis of Compound VIII-2:

the operation is the same as 1.5, inIntermediate 7-9 replaces intermediate 1-6, 10-hydroxy-2-decenoic acid replaces 2-methacrylic acid 1-7, and pale yellow semi-solid VIII-2 is obtained, and the yield is about 63.8%; MS (ESI) m/z=601.30 [ M+H ]] ⁺ ； ¹ H NMR(400MHz,Acetone-d ₆ )δ8.37(d,J＝8.1Hz,2H),8.28(s,1H),8.11(d,J＝7.1Hz,1H),8.05(d,J＝10.9Hz,1H),7.85(d,J＝22.2Hz,2H),7.19(t,J＝6.7Hz,1H),6.81(d,J＝16.2Hz,1H),6.47(m,1H),3.91–3.77(m,1H),3.61–3.44(m,4H),3.30(t,J＝6.5Hz,1H),3.19(t,J＝10.5Hz,1H),2.38(m,2H),2.26–2.19(m,2H),1.96–1.71(m,4H),1.64–1.52(m,2H),1.43–1.35(m,2H),1.33–1.24(m,2H),1.18–1.02(m,2H). ¹³ C NMR(101MHz,Acetone-d ₆ )δ165.18,164.05,156.12,155.74,154.33,152.04,148.34,148.02,138.33,138.06,124.67,123.94,115.90,114.41,114.11,101.90,81.00,78.79,61.43,56.73,54.23,53.67,46.98,42.25,32.77,32.33,29.68,25.27,17.77.

Example 9 in vitro BTK enzyme inhibition assay

9.1 Compound formulation

1mg of the above compound was weighed and dissolved in 100% DMSO-d6 to prepare a 10mM stock solution, which was stored in a nitrogen cabinet in the absence of light.

9.2 kinase reaction procedure

(1) 1 XKinase buffer was prepared.

(2) Preparing a compound concentration gradient: the initial test concentration of test compound was 2400nM,7 concentrations, and multiplex assay. 100% DMSO-d diluted to 100-fold final concentration in 384source plates ₆ A solution. The final concentration of 250nl 100-fold compound was transferred to the plate of interest 3575 using a dispenser Echo 550.

(3) A2.5-fold final concentration of Kinase solution was prepared using a 1 XKinase buffer.

(4) Adding 10 μl of 2.5-fold final concentration kinase solution to each of the compound wells and positive control wells; mu.l of 1 XKinase buffer was added to the negative control wells.

(5) Centrifugation at 1000rpm for 30 seconds, the reaction plate was shaken and mixed well and incubated at room temperature for 10 minutes.

(6) A5/3-fold final concentration of a mixed solution of ATP and Kinase substrate 2 was prepared using a 1 XKinase buffer.

(7) The reaction was initiated by adding 15. Mu.l of a 5/3-fold final concentration of the mixed solution of ATP and substrate.

(8) The 384-well plate was centrifuged at 1000rpm for 30 seconds, and after shaking and mixing, incubated at room temperature for 30 minutes.

(9) The kinase reaction was stopped by adding 30. Mu.l of stop detection solution, centrifuging at 1000rpm for 30 seconds, and shaking and mixing.

(10) The conversion was read with Caliper EZ Reader.

The results are shown in Table 1.

TABLE 1 results of BTK inhibitory Activity of Compounds

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As can be seen, the small molecule compound has good BTK inhibition activity, and partial compounds (VII-1, VII-2 and VII-3) are inhibition ratios or IC ₅₀ All are superior to the acartinib which is a drug on the market, and the IC ₅₀ Similar to ibrutinib.

Example 10 Water solubility test of Compounds

10.1 internal Standard preparation

1.00mg of Loratadine (Loratadine) standard is precisely weighed, 1mL of dimethyl sulfoxide (DMSO) is precisely removed for dissolution, and the solution is uniformly shaken to prepare stock solution with the concentration of 1mg/mL, and the stock solution is stored at the temperature of minus 20 ℃ for standby. The stock solution was precisely removed and diluted with acetonitrile to give an internal standard working solution at a concentration of 5.0 ng/mL.

10.2 stock solution of analyte

1.00mg of the object to be measured is precisely weighed and dissolved in 1mL of DMSO to prepare 1.00mg/mL of object to be measured stock solution A which is used for preparing a standard curve and stored at the temperature of minus 20 ℃ for standby.

1.00mg of the to-be-measured object is precisely weighed and dissolved in 1mL of DMSO to prepare 1.00mg/mL of stock solution B of the to-be-measured object, and the stock solution B is used for preparing a quality control sample (QC) and is stored at-20 ℃ for standby.

10.3 preparation of standard curve:

taking a proper amount of stock solution A, and performing acetonitrile gradient elution to obtain working solutions with the concentration of 0.05 mug/mL, 0.1 mug/mL, 0.2 mug/mL, 0.5 mug/mL, 1 mug/mL, 2 mug/mL, 5 mug/mL, 10 mug/mL, 20 mug/mL, 50 mug/mL, 100 mug/mL and 200 mug/mL;

precisely removing 47.5 mu L of ultrapure water, precisely adding 2.5 mu L of the working solution respectively, swirling for 1min, uniformly mixing, preparing standard curve samples with the concentration of 2.5ng/mL,5ng/mL,10ng/mL,25ng/mL,50ng/mL,100ng/mL,250ng/mL,500ng/mL and 1000ng/mL, precisely adding 200 mu L of acetonitrile containing internal standard loratadine respectively, swirling for 5min, centrifuging at 12000rpm for 10min at 4 ℃, taking supernatant, and carrying out LC-MS/MS sample injection analysis.

10.4 Quality Control (QC) sample preparation:

taking a proper amount of stock solution B, and performing acetonitrile gradient elution to obtain working solutions with the concentration of 0.1 mug/mL, 2.0 mug/mL and 16 mug/mL;

precisely removing 47.5 mu L of ultrapure water, precisely adding 2.5 mu L of the working solution respectively, swirling for 1min, uniformly mixing to prepare QC samples with the concentration of 5ng/mL,100ng/mL and 800ng/mL, precisely adding 200 mu L of acetonitrile containing the internal standard loratadine respectively, swirling for 5min, centrifuging at 12000rpm for 10min at 4 ℃, taking supernatant, and carrying out LC-MS/MS sample injection analysis.

10.5 preparation of Water-soluble sample of analyte

Respectively weighing a proper amount of to-be-measured substances, respectively adding 1mL of pure water to prepare saturated aqueous solution, swirling for 2 hours, standing for 1 hour at room temperature, centrifuging at 12000rpm for 10 minutes at 4 ℃, respectively taking 50 mu L of supernatant, adding 200 mu L of acetonitrile containing internal standard loratadine, centrifuging at 12000rpm for 10 minutes at 4 ℃ for 5 minutes at vortex, taking supernatant, performing LC-MS/MS sample injection analysis, and performing parallel experiments on 3 parts of each sample.

The results are shown in Table 2:

TABLE 2.1 Standard Curve equation form

Table 2.2 results of water solubility test of compounds

Therefore, the compound provided by the invention has better water solubility, and the water solubility of VII series compounds is improved, wherein VII-2 is improved by 15.8% compared with a positive control drug ibrutinib, and VIII-2 is improved by more than 18.4% compared with the positive control drug ibrutinib.

EXAMPLE 11 Compound PK test

After each single dose of the compounds ibrutinib, IV-1 and VIII-2 is respectively administered in the SD rat body by intravenous injection and oral injection, blood samples are collected at different time points, the concentrations of the compounds ibrutinib, IV-1 and VIII-2 in the rat blood plasma are measured by LC-MS/MS, relevant drug generation parameters are calculated, and the bioavailability and drug generation properties of the compounds ibrutinib, IV-1 and VIII-2 in the rat body are examined.

Male SD rats were randomly divided into 3 groups of about 9, about 200g, 3 each. Fasted for 12 hours before administration, and the water is freely drunk.

Each animal was bled 0.100ml per orbital, edta-disodium anticoagulated, and the collection time points were: group IV: 5min, 15min, 30min, 1h, 2h, 4h, 6h, 8h and 24h after administration of the test agent; PO group: 15min, 30min, 1h, 2h, 4h, 6h, 8h and 24h after administration of the test agent. Blood samples were collected and placed on ice and the plasma was centrifuged (centrifugation conditions: 3000rpm,10min,4 ℃) for 30 min. After centrifugation, 50. Mu.L of the supernatant was added with 200. Mu.L of acetonitrile containing the internal standard loratadine, the mixture was vortexed for 5min, centrifuged at 12000rpm at 4℃for 10min, and the supernatant was sampled and analyzed by LC-MS/MS, and 3 samples were run in parallel.

The experimental test results are as follows:

table 3.1 linear range of compounds in plasma, standard curve equation table:

SD rats were given Ibrutinib (Ibrutinib) by intravenous injection (IV, 2.00 mg/kg) and the time-dependent blood concentration profiles of IV-2 and VII-2 were shown in FIG. 1.

After administration of ibrutinib, iv-1, vii-2 (PO., 10 mg/kg) to SD rats, the drug generation parameters after administration were calculated from blood concentration data using the WinNonlin V6.3 non-compartmental model, see table 3.2.

TABLE 3.2 main pharmacokinetic parameters after gastric lavage administration in SD rats

Therefore, the compound provided by the invention has better pharmacokinetic parameters, and VII-2 is basically consistent with or slightly better than that of a positive medicament ibrutinib.

Claims (7)

1. An irreversible BTK inhibitor of the oxazolo [4,5-b ] pyridine structure, characterized by having the structure of formula a:

wherein X, Y, Z has the following structure:

wherein R has the structure:

2. the irreversible BTK inhibitor of the oxazolo [4,5-b ] pyridine structure of claim 1, wherein said compound

The structure is as follows:

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3. use of an irreversible BTK inhibitor of the oxazolo [4,5-b ] pyridine structure according to claim 1 or 2 for the preparation of a pharmaceutical formulation for the prevention or treatment of diseases caused by BTK abnormalities.

4. The use according to claim 3, wherein the disease is lymphoma, autoimmune disease, including rheumatoid arthritis, systemic lupus erythematosus, ankylosing spondylitis or psoriasis, and wherein the lymphoma comprises chronic lymphocytic leukemia, B-cell lymphoma, mantle cell lymphoma, lymphoplasmacytic lymphoma, diffuse large B-cell lymphoma, non-hodgkin's lymphoma, follicular central lymphoma, marginal zone B-cell lymphoma.

5. The irreversible BTK inhibitor of oxazolo [4,5-b ] pyridine structure of claim 1 or 2 in a pharmaceutically acceptable salt, wherein the pharmaceutically acceptable salt is a hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, phosphate, acetate, propionate, butyrate, oxalate, tartrate, methanesulfonate, p-toluenesulfonate, fumarate, taurine, citrate, succinate, or a mixture thereof.

6. Use of an irreversible BTK inhibitor of the oxazolo [4,5-b ] pyridine structure of claim 5 in the manufacture of a pharmaceutical formulation for the prevention or treatment of a disease caused by BTK abnormalities.

7. The use according to claim 6, wherein the disease is lymphoma, autoimmune disease, including rheumatoid arthritis, systemic lupus erythematosus, ankylosing spondylitis or psoriasis, and wherein the lymphoma comprises chronic lymphocytic leukemia, B-cell lymphoma, mantle cell lymphoma, lymphoplasmacytic lymphoma, diffuse large B-cell lymphoma, non-hodgkin's lymphoma, follicular central lymphoma, marginal zone B-cell lymphoma.

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