CN114213416A - Irreversible BTK inhibitor with oxazolo [4,5-b ] pyridine structure and application thereof - Google Patents

Abstract

The inventionA compound with irreversible inhibition on protein tyrosine kinase (BTK) activity, its pharmacologically useful salts, its synthesizing process and its application in preparing the medicines for inhibiting BTK protein are disclosed. In particular, the invention describes compounds of formula a. The compound has good BTK inhibitory activity, improves water solubility, and has pharmacokinetic parameters basically consistent with those of a marketed positive drug, namely ibrutinib, so that the compound can be widely applied to preparation of 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 drugs, and particularly provides a compound with irreversible protein tyrosine kinase (BTK) activity inhibition, and a synthesis method and a use method of the compound.

Background

According to the statistics of the world health organization, the annual growth rate of the incidence rate of the lymphoma is 5-7%, the number of deaths per year exceeds 20 ten thousand, at present, the annual growth rate of the incidence rate of the lymphoma in China is 3-5%, and about 10 ten thousand new cases per year are the eighth most advanced malignant tumor.

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

The main reason for the development of B cell lymphomas is the over-activation of the B cell antigen receptor (BCR) signaling pathway, and BTK kinase is a key process protein in the BCR signaling pathway, widely distributed in the lymphatic, hematopoietic and blood systems, a membrane-bound protein, expressed in immune cells such as B cells, mast cells and macrophages. The large amount of BTK can lead BCR signal channel to be abnormally activated, and influence the proliferation, differentiation and apoptosis of B cells, thereby initiating various B cell malignant tumors; it also causes B cell dysfunction, altered immune tolerance status, and transformation into autoreactive B cells, secreting large amounts of autoantibodies to induce autoimmune diseases. Therefore, BTK is a hot spot in the current clinical research on the treatment of B cell tumors and B cell immune diseases.

The BTK inhibitor can inhibit lymphocyte proliferation by inhibiting over-activation of a BCR signal pathway, and has excellent treatment prospect for treating B cell malignant tumor and autoimmune diseases. In 2013, ibrutinib was approved as the first effective BTK selective inhibitor by FDA, and breakthrough therapy for the treatment of chronic lymphocytic leukemia, mantle cell lymphoma, small lymphocytic lymphoma, fahrenheit macroglobulinemia, marginal zone lymphoma and graft-host disease has epoch-making significance, making BTK a promising therapeutic target. Against this background, the second-generation BTK inhibitors acatinib, temarotinib, zetinib and orbetinib, which improve selectivity for BTK, improve drug resistance and reduce drug toxicity, have all been approved by FDA for marketing. The structure of marketed BTK inhibitors is shown below:

small molecule BTK inhibitors are divided into two classes, reversible inhibitors and irreversible inhibitors, based on their binding pattern to the BTK catalytic domain. Irreversible BTK inhibitors retain terminal electrophilic centers, such as acrylamide groups and 2-butynoamide groups, and can form covalent bonds with the conserved non-catalytic cysteine residue (Cys481) of BTK proteins by Michael addition, nucleophilic addition, addition-elimination, or nucleophilic substitution, resulting in strong irreversible 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 the Tec family while inhibiting BTK, so that side effects such as bleeding, diarrhea, rash and the like are caused, and drug resistance is generated, and therefore, the development of a novel BTK inhibitor is still a problem to be solved urgently.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides a compound with irreversible protein tyrosine kinase (BTK) inhibition activity, a pharmaceutically acceptable salt thereof, a synthetic method thereof and application thereof in drugs for inhibiting BTK proteins.

The purpose of the invention is realized by the following technical scheme: a small molecule compound with oxazolo [4,5-b ] pyridine structure with BTK inhibitory activity, a compound with irreversible BTK inhibitory activity and pharmaceutically acceptable salts thereof, have the structure shown as formula A:

wherein, X, Y, Z has the following structure respectively:

Z＝H、F，

wherein R has the following structure:

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

wherein R has the structure:

alternatively, X, Y, Z has the following structures, respectively:

Z＝H，F，

wherein R has the following structure:

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

or the following structure:

or the following structure:

a compound having irreversible activity to inhibit BTK can be used alone or prepared into pharmaceutically acceptable salts by conventional methods, wherein the pharmaceutically acceptable salts are hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, phosphate, acetate, propionate, butyrate, oxalate, tartrate, methanesulfonate, p-toluenesulfonate, fumarate, taurate, citrate, succinate or their mixture.

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

taking p-bromobenzoyl chloride as an initial raw material, and carrying out nucleophilic substitution on the p-bromobenzoyl chloride and aminopyrazine under the alkaline condition of triethylamine to obtain 4-bromo-N- (pyrazine-2-yl) benzamide; subsequently, 4-bromo-N- (pyrazine-2-yl) benzamide and diboron pinacol ester are coupled through Suzuki to obtain a corresponding borate product; then, reacting the borate product with (R) -phenyl 2- (8-amino-1-bromoimidazo [1,5-A ] pyrazine-3-yl) pyrrolidine-1-carboxylate through Suzuki reaction to obtain a coupling product; removing the carbobenzoxy protecting group of the coupling product under the condition of 33 percent HBr-AcOH solution to obtain a pyrrolidine compound; the pyrrolidine compound, 2-methacrylic acid, 2-butynoic acid and acrylic acid obtain a target product under the action of a condensing agent HATU. The reaction process is as follows:

or the following method is adopted:

p-bromobenzoic acid is taken as a starting material, and is subjected to condensation reaction with 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 pinacol bisboronate through suzuki coupling to obtain a corresponding boric acid ester product; then, carrying out Mitsunobu reaction on the raw material 4-amino-3-iodo-1H-oxazolo [3,4-D ] pyrimidine and the raw material (S) -1-tert-butoxycarbonyl-3-hydroxypiperidine to obtain (3R) -1-Boc-3- (4-amino-3-iodo-1H-pyrazolo [3,4-D ] pyrimidin-1-yl) piperidine; then carrying out Suzuki reaction on the borate product and (3R) -1-Boc-3- (4-amino-3-iodine-1H-pyrazolo [3,4-D ] pyrimidine-1-yl) piperidine to obtain a coupling product; removing Boc protecting group from the coupling product under the condition of 3mol of HCl-EA solution to obtain piperidine compound; the piperidine compound and alpha, beta-unsaturated acid are subjected to the action of a condensing agent HATU to obtain a target product. The reaction process is as follows:

the invention also provides the applications of the oxazolo [4,5-b ] pyridine structure micromolecule compound with BTK inhibitory activity and the pharmaceutically acceptable salt thereof in pharmacy, and the applications are specifically as follows: is used for preparing a medicinal preparation for preventing or treating diseases caused by BTK abnormality. 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 diseases, including rheumatoid arthritis, systemic lupus erythematosus, ankylosing spondylitis, psoriasis, etc.

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

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

Drawings

FIG. 1 is a time-series graph of blood drug concentrations of ibrutinib, IV-2 and VII-2 in SD rats.

Detailed Description

The structure, preparation method and application in preparing pharmaceutical preparations for preventing or treating diseases caused by over-expression of tubulin will be further described in the following without limiting the invention.

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

the thermometer is not corrected; bruker DRX400 nmr; agilent model 5975 mass spectrometer; bruker Vector 22 infrared spectrometer.

Example 1 synthesis of compounds of series i:

1.1 Synthesis of intermediates 1-3:

weighing 1-11.00 g (10.53mmol) of 2-aminopyrazine, placing in a 25mL three-necked flask, and placing N₂Protection, 5mL of dichloromethane is added into a syringe, 1.41g (10.94mmol) of DIPEA is added under ice bath conditions, and finally a dichloromethane solution of 1-22.00 g (9.11mmol) of 4-bromobenzoyl chloride is added dropwise, and the mixture is kept in ice bath for 5min and then placed at room temperature for reaction overnight. The reaction was confirmed to be complete by TLC, the reaction solution was adjusted to neutral with NaOH, and then extracted with ethyl acetate (40 mL. times.3), the organic phases were combined, washed once with 100mL of saturated saline, dried over anhydrous sodium sulfate, spin-dried, and purified by column chromatography to give 1.2g of a pale yellow solid 1-3 with a yield of about 48.2%. MS (ESI) M/z 278.01[ M + H ]]⁺；¹H NMR(400MHz,CDCl₃)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 intermediates 1-4:

weighing 1-31.00 g (3.61mmol) of intermediate, 1.10g (4.33mmol) of pinacol diboron and [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride dichloromethane complex (Pd (dppf) Cl₂)79mg (0.11mmol), 1.06g (10.83mmol) of potassium acetate are placed in a 25mL three-necked flask, N₂After protection, the syringe was filled with tetrahydrofuran solution and refluxed overnight. After the reaction of the dot plate raw materials is finished, cooling the reaction liquid to room temperature, and filtering the reaction liquid by using diatomite to remove Pd (dppf) Cl₂And washing with tetrahydrofuran for 2-3 times, concentrating the filtrate, adding water and adding ethyl acetate for extraction, washing an organic layer with water for 2-3 times, recovering under reduced pressure to obtain a crude product, separating and purifying by column chromatography to obtain 1.04g of light yellow solid 1-4 with the yield of about 89.2%, and transferring nuclear magnetism to the next step.¹H NMR(400MHz,CDCl₃)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 intermediates 1-5:

intermediate 1-4110 mg (0.34mmol), (R) -phenyl 2- (8-amino-1-bromoimidazole [1,5-a ] was weighed in order]Pyrazin-3-yl) pyrrolidine-1-carboxylic acid ester 133mg (0.31mmol), Pd (dppf) Cl₂ 22mg(0.03mmol)，K₂CO₃128mg (0.93mmol) were added to a 10mL three-necked flask, N₂Protection, adding 1, 4-dioxane under ice bath condition: the mixed solution of water (3:1) was transferred to 105 ℃ to start the reaction. After 8h, TLC is used for monitoring the reaction progress, after the reaction is finished, the reaction liquid is cooled to room temperature, and Pd (dppf) Cl is removed by suction filtration through diatomite₂Washing with ethyl acetate for 2-3 times,then adding water (100mL multiplied by 4) and 100mL ethyl acetate into the filtrate for extraction, drying an organic phase by using anhydrous sodium sulfate, concentrating the filtrate, and carrying out column chromatography separation to obtain 110mg of light brown solid 1-5 with the yield of about 61.9%; MS (ESI) M/z 535.24[ M + H ]]⁺；¹H NMR(500MHz,DMSO-d₆)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:

weighing intermediate 1-5300 mg (0.56mmol) and placing in a 10mL single-neck bottle, placing in 0 deg.C environment, slowly adding 33% HBr-AcOH (6.6mmol) dropwise, transferring to room temperature after 5min and reacting for 2 h. After TLC monitoring reaction, adding 50mL of water and dichloromethane (70mL multiplied by 2) into reaction liquid for extraction, removing a dichloromethane layer, adjusting the pH of an aqueous layer to be approximately 10 by NaOH aqueous solution under ice bath conditions, adding dichloromethane (60mL multiplied by 4) for extraction, combining organic layers, drying by anhydrous sodium sulfate, concentrating to obtain a crude product, mixing the sample by a dry method, passing through a column, and carrying out column chromatography gradient elution to obtain light brown powdery solid 206mg, namely an intermediate 1-6, wherein the yield is about 91.6%. MS (ESI) M/z 401.21[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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:

dissolving 2-methacrylic acid 1-78 mg (0.09mmol) in dichloromethane, adding EDC & HCl14mg (0.08mmol) and triethylamine 21 μ L in sequence, and stirringAfter 5min, 1-630 mg (0.08mmol) of intermediate was added and reacted at room temperature. After 3h of reaction, TLC is used for monitoring the reaction, the solvent dichloromethane is removed under reduced pressure, 50mL of ethyl acetate is added for dissolution, 70mL of water is added for extraction, the organic layer is extracted by 1N diluted HCl solution (70mL multiplied by 2), 70mL of saturated NaCl solution is extracted once, the ethyl acetate layer is dried by anhydrous sodium sulfate, the solvent is spun out to obtain a crude product, and the crude product is mixed with a sample by a dry method and is subjected to column chromatography. Gradient elution gave 27mg of a yellow semisolid, compound I-1, in about 77.1% yield; MS (ESI) [ M + H]⁺m/z＝469.21[M+H]⁺；¹H NMR(500MHz,CDCl₃)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).¹³C NMR(126MHz,CDCl₃)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, 1-8 parts of 2-butynoic acid is used for replacing 1-7 parts of 2-methacrylic acid, light brown semisolid I-2 is obtained, and the yield is about 49.6%; MS (ESI) M/z 467.20[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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).¹³C NMR(101MHz,DMSO-d₆)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 from the synthesis of compound I-2, 12mg of a pale yellow semisolid, MS (ESI) M/z 443.20[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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 operation is the same as 1.5, 1-9 of acrylic acid is used for replacing 1-7 of 2-methacrylic acid, and light yellow powdery solid I-4 is obtained, wherein the yield is about 78.3%; MS (ESI) M/z 455.20[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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).¹³CNMR(101MHz,DMSO-d₆)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 compounds of series ii:

2.1 Synthesis of intermediates 2-3:

weighing 2-11.31 g (6.52mmol) of p-bromobenzoic acid, placing in a 25mL single-neck bottle, adding 10mL of dichloromethane, sequentially adding 1.25g (6.52mmol) of EDC & HCl and 2.81mL (16.14mmol) of DIPEA under ice bath condition, stirring for 5min, maintaining 0 deg.C, adding 2-20.50 g (5.38 mmol) of anilinemmol) and left to react at room temperature overnight. The reaction was monitored by TLC for completion, methylene chloride was removed under reduced pressure, saturated brine (200 mL. times.3) and ethyl acetate (150mL) were added and the organic phases were combined, dried over anhydrous sodium sulfate, spun dry and then passed through a column with dry stirring to give 1.1g of 2-3 as a white powdery solid with a yield of about 73.6%. MS (ESI) M/z 275.98[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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 intermediates 2-4:

the same procedure as 1.2 was followed, using intermediate 2-3 instead of intermediate 1-3, to obtain 1.3g of a light brown powdery solid with a yield of about 89.9%;¹H NMR(400MHz,CDCl₃)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 intermediates 2-7:

respectively weighing 4-amino-3-iodo-1H-pyrazolo [3,4-d]Pyrimidine 2-5500 mg (1.92mmol), (R) -1-tert-butoxycarbonyl-3-hydroxypiperidine 2-6774 mg (3.83mmol), triphenylphosphine 755mg (2.88mmol) were added to a three-necked flask, and N was₂Under ice-bath conditions, anhydrous THF was added via syringe, followed by slow dropwise addition of DIAD in anhydrous THF, and the mixture was transferred to room temperature for overnight reaction. After TLC plate material reaction, THF was removed under reduced pressure, and dry-stirred through the column, which was PE-wetted to wash out the yellow DIAD, followed by gradient elution to yield 708mg of a white powdery solid, intermediate 2-7, in about 83.1% yield. MS (ESI) M/z 467.24[ M + Na ]]⁺；¹H NMR(400MHz,DMSO-d₆)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 intermediates 2-8:

intermediate 2-4436 mg (1.35mmol), intermediate 2-7500 mg (1.13mmol), Pd (dppf) Cl were weighed out in order₂ 25mg(0.03mmol)，K₂CO₃468mg (3.39mmol) of the reaction solution are added into a 10mL three-necked flask, nitrogen is used for protection, a mixed solution of 1, 4-dioxane and water (3:1) is added under ice bath condition, and then the mixture is transferred to 105 ℃ to start reflux reaction. After 8h, TLC is used for monitoring the reaction progress, after the reaction is finished, the reaction liquid is cooled to room temperature, and Pd (dppf) Cl is removed by suction filtration through diatomite₂Then, water (100 mL. times.4) and 100mL of ethyl acetate were added to the filtrate for extraction, and the organic phase was extracted with anhydrous Na₂SO₄Drying, concentrating the filtrate, and performing column chromatography to obtain 456mg of light brown solid 2-8 with yield of about 78.6%; MS (ESI) M/z 514.19[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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 intermediates 2-9:

weighing 2-8300 mg (0.58mmol) of the intermediate into a single-neck bottle, adding 2mL of ethyl acetate, dropwise adding 3mL of 3M HCl ethyl acetate solution under an ice bath condition, wherein the reaction solution becomes turbid in the dropwise adding process, and transferring the reaction solution to room temperature for reaction for 2h after the dropwise adding is finished. After the TLC monitoring reaction, the reaction solution is filtered by filter paper, and the filter cake is the product hydrochloride. Then the filter cake is placed in 100mL of K at 0 DEG C₂CO₃Extracting with ethyl acetate (120mL × 4), mixing the organic phases, drying over anhydrous sodium sulfate, and concentrating to obtain 196mg of a light brown powdery solid product 2-9 with a yield of about 81.3%; MS (Mass Spectrometry)(ESI):m/z＝414.27[M+H]⁺；¹H NMR(400MHz,DMSO-d₆)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:

the same procedure as 1.5 was followed, using intermediates 2-9 instead of intermediates 1-6, to give II-1 as a white semi-solid in about 69.8% yield. MS (ESI) M/z 482.23[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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).¹³C NMR(101MHz,DMSO-d₆)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 as 1.5 was followed, using intermediates 2-9 instead of intermediates 1-6 and acrylic acid 2-10 instead of intermediates 1-7, to give II-2 as a white powdery solid in about 67.4% yield. MS (ESI) M/z 467.21[ M + H ]]⁺；¹H NMR(400MHz,CDCl₃)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).¹³C NMR(101MHz,CDCl₃)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 as 1.5 was followed, using intermediates 2-9 instead of intermediates 1-6, 2-fluoroacrylic acid instead of intermediates 1-7, to give a colorless oily liquid with a yield of about 81.6%. MS (ESI) M/z 482.23[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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).¹³C NMR(101MHz,DMSO-d₆)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 same procedure as 1.5 was followed, intermediate 2-9 was used instead of intermediate 1-6, 2-chloroacrylic acid was used instead of intermediate 1-7, and the reaction solution was worked up to give a white semi-solid with a yield of about 71.4%. MS (ESI) M/z 502.31[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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).¹³C NMR(101MHz,DMSO-d₆)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 same procedure as 1.5 was followed, using intermediates 2-9 instead of intermediates 1-6, 2-bromoacrylic acid instead of intermediates 1-7, and the reaction solution was worked up to give a brown solid with a yield of about 75.3%. MS (ESI) M/z 546.12[ M + H ]]⁺；¹H NMR(500MHz,CDCl₃)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).¹³C NMR(126MHz,CDCl₃)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:

4-12.43 g (12.09mol) of p-bromobenzoic acid was placed in a single-necked flask, 10mL of dichloromethane was added, and 4.59g (12.09mmol) of HATU, 5.14mL (30.30mmol) of DIPEA and 4-20.98 mL of cyclohexylamine were added in this order under ice-bath conditions, followed by transfer to room temperature for reaction overnight. The completion of the reaction was confirmed by TLC (EA: PE ═ 1:2), dichloromethane was removed under reduced pressure, saturated brine (200mL × 3) and ethyl acetate (150mL) were added, the organic phases were combined, dried over anhydrous sodium sulfate, and concentratedAfter condensation, the mixture is dried, mixed and put on a column, the column is wetted by PE, and the mixture is eluted by EA and PE (1: 5) to obtain 2.35g of white powdery solid 4-3 with the yield of about 82.9 percent. MS (ESI) M/z 282.15[ M + H ]]⁺；¹H NMR(400MHz,CDCl₃)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 intermediates 3-4:

the operation is the same as 1.2, the intermediate 3-3 is used for replacing the intermediate 1-3, and the white powdery solid 3-4 is obtained, and the yield is about 89.6%;¹H NMR(400MHz,DMSO-d₆)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 intermediates 3-8:

the operation is the same as 2.4, and the intermediate 4-4 is used for replacing the intermediate 2-4 to obtain 3-8 white powdery solid with the yield of about 62.6 percent; MS (ESI) M/z 520.46[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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 intermediates 3-9:

operating as 2.5, using intermediate 3-8 instead of intermediates 2-8, gave 3-9 as a pale yellow powdery solid with a yield of about 87.4%; MS (ESI) M/z 420.37[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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 same procedure as 1.5 was followed, using intermediate 3-9 instead of intermediate 1-6, and the reaction solution was worked up to give solid III-1 as a pale yellow powder with a yield of about 68.8%. MS (ESI) M/z 488.30[ M + H ]]⁺；¹H NMR(400MHz,Acetone-d₆)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).¹³C NMR(101MHz,Acetone-d₆)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 as 1.5 was followed, using intermediates 3 to 9 instead of intermediates 1 to 6 and acrylic acid instead of 2-methacrylic acid 1 to 7, to give III-2 as a white powdery solid in about 77.6% yield. MS (ESI) M/z 496.45[ M + Na ]]⁺；¹H NMR(400MHz,DMSO-d₆)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).¹³C NMR(101MHz,DMSO-d₆)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 as 1.5 was followed, using intermediate 3-9 instead of intermediate 1-6 and 2-fluoroacrylic acid instead of 2-methacrylic acid 1-7, to give compound III-3 as a yellow semi-solid in about 80.3% yield. MS (ESI) M/z 492.25[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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).¹³C NMR(101MHz,DMSO-d₆)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:

the same procedure as 1.5 was followed, using intermediate 3-9 instead of intermediate 1-6, 2-chloroacrylic acid instead of 2-methacrylic acid 1-7, to give solid III-4 as a pale yellow powder with a yield of about 72.4%. MS (ESI) M/z 508.22[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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).¹³C NMR(101MHz,DMSO-d₆)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 as 1.5 was followed, using intermediate 3-9 instead of intermediate 1-6 and 2-bromoacrylic acid instead of 2-methacrylic acid 1-7, to give compound III-5 as a brown solid in about 85.3% yield. MS (ESI) M/z 552.16[ M + H ]]⁺；¹H NMR(500MHz,CDCl₃)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).¹³C NMR(126MHz,CDCl₃)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 intermediates 4-3:

the same procedure as 2.1, 2-aminopyridine was performed, replacing the compound aniline 2-2, to give 4-3 as a brown powdery solid with a yield of about 89.1%. MS (ESI) M/z 276.99[ M + H ]]⁺；¹H NMR(400MHz,CDCl₃)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 intermediates 4-4:

the operation is the same as 2.2, the intermediate 4-3 replaces the intermediate 2-3, and light brown powdery solid 4-4 is obtained, and the yield is about 92.6%;¹H NMR(400MHz,DMSO-d₆)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 intermediates 4-8:

the operation is the same as 2.5, and the intermediate 4-4 replaces the intermediate 2-4 to obtain light brown powdery solid 4-8 with the yield of about 73.4 percent; MS (ESI) M/z 515.25[ M + H ]]⁺；¹H NMR(400MHz,CDCl₃)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 intermediates 4-9:

the operation is the same as 2.4, and the intermediate 4-8 replaces the intermediate 2-8 to obtain light brown powdery solid 4-9 with the yield of about 61.9%; MS (ESI) M/z 437.20[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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, and the intermediate 4-9 replaces the intermediate 1-6 to obtain white powdery solid IV-1 with the yield of about 81.4 percent; MS (ESI) M/z 483.23[ M + H ]]⁺；¹H NMR(500MHz,CDCl₃)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).¹³C NMR(126MHz,CDCl₃)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 intermediates 1 to 6 are replaced by the intermediates 4 to 9, and the 2-methacrylic acid 1 to 7 is replaced by the acrylic acid, so that white powdery solid IV-2 is obtained, and the yield is about 74.3 percent; MS (ESI) M/z 469.21[ M + H ]]⁺；¹H NMR(500MHz,CDCl₃)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, the intermediate 4-9 replaces the intermediate 1-6, and the 2-fluoroacrylic acid replaces the 2-methacrylic acid 1-7, so that the off-white powdery solid IV-3 is obtained, and the yield is about 61.0 percent; MS (ESI) M/z 487.20[ M + H ]]⁺；¹H NMR(400MHz,CDCl₃)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).¹³C NMR(101MHz,CDCl₃)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 operation is the same as 1.5, the intermediate 4-9 replaces the intermediate 1-6, 2-chloroacrylic acid replaces 2-methacrylic acid 1-7, light brown powdery solid IV-4 is obtained, and the yield is about 64.8%; MS (ESI) M/z 503.17[ M + H ]]⁺；¹H NMR(400MHz,CDCl₃)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).¹³C NMR(101MHz,CDCl₃)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, the intermediate 4-9 replaces the intermediate 1-6, the 2-bromoacrylic acid replaces the 2-methacrylic acid 1-7, and light brown powdery solid IV-5 is obtained,the yield is about 67.5%; MS (ESI) M/z 547.12[ M + H ]]⁺；¹H NMR(400MHz,CDCl₃)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).¹³C NMR(101MHz,CDCl₃)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, and the 2-butynoic acid replaces the 2-methacrylic acid 1-7, so that white powdery solid IV-6 is obtained, and the yield is about 45.3%; MS (ESI) M/z 481.21[ M + H ]]⁺；¹H NMR(400MHz,CDCl₃)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).¹³C NMR(101MHz,CDCl₃)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 intermediates 5-3:

placing 5-11.10 g (10mmol) of 2-amino-3-hydroxypyridine and 5-21.10 g (10mmol) of p-bromobenzaldehyde and polyphosphoric acid (30mL) in a reactorIn a 100mL single-neck bottle, heating for 1h at 150 ℃ under the protection of nitrogen. 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 the intermediate 5-3, wherein the yield is about 69.3%; MS (ESI) M/z 274.98[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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 intermediates 5-4:

the operation is the same as 1.2, the intermediate 5-3 replaces the intermediate 1-3 to obtain 5-4 of light yellow semi-solid, and the yield is about 89.8%;¹H NMR(400MHz,DMSO-d₆)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 intermediates 5-8:

the same operation as 2.4 is carried out, and the intermediate 5-4 replaces the intermediate 2-4, so that 5-8 of light brown semisolid is obtained, and the yield is about 76.3%; MS (ESI) M/z 513.42[ M + H ]]⁺；¹H NMR(500MHz,CDCl₃)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 intermediates 5-9:

the operation is the same as 2.5, and the intermediate 5-8 replaces the intermediate 2-8 to obtain light brown powdery solid 5-9 with the yield of about 81.0 percent; MS (ESI) M/z 413.35[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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, and the intermediate 1-6 is replaced by the intermediate 5-9 to obtain white powdery solid V-1 with the yield of about 74.2 percent; MS (ESI) M/z 481.21[ M + H ]]⁺；¹H NMR(400MHz,CDCl₃)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 intermediates 1 to 6 are replaced by the intermediates 5 to 9, and the 2-methacrylic acid 1 to 7 is replaced by the acrylic acid, so that the white powdery solid V-2 is obtained, and the yield is about 79.6 percent; MS (ESI) M/z 467.20[ M + H ]]⁺；¹H NMR(400MHz,CDCl₃)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).¹³C NMR(101MHz,CDCl₃)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, and the 2-fluoroacrylic acid replaces the 2-methylacrylic acid 1-7, so that the white powdery solid V-3 is obtained, and the yield is about 73.8 percent; MS (ESI) M/z 485.19[ M + H ]]⁺；¹H NMR(400MHz,CDCl₃)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).¹³C NMR(101MHz,CDCl₃)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, 2-chloroacrylic acid replaces 2-methacrylic acid 1-7, and light brown powdery solid V-4 is obtained, and the yield is about 61.2%; MS (ESI) M/z 501.16[ M + H ]]⁺；¹H NMR(400MHz,CDCl₃)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).¹³C NMR(101MHz,CDCl₃)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, and the 2-bromoacrylic acid replaces the 2-methacrylic acid 1-7, so that light brown powdery solid V-5 is obtained, and the yield is about 57.4 percent; MS (ESI) M/z 545.10[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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).¹³C NMR(101MHz,DMSO-d₆)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 same procedure as 1.5 was followed, intermediate 5-9 substituted for intermediate 1-6, 2-butynoic acid substituted for 2-methacrylic acid 1-7, to give V-6 as a white powdery solid with a yield of about 45.6%; MS (ESI) M/z 479.19[ M + H ]]⁺；¹H NMR(400MHz,CDCl₃)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).¹³C NMR(101MHz,CDCl₃)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 to 12.43 g (12.09mol) of p-bromobenzoic acid was weighed in a single-necked flask, 10mL of methylene chloride was added, and 4.59g (12.09mmol) of HATU, 5.14mL (30.30mmol) of DIPEA and 6 to 21.13 g (10.08mmol) of 2-amino-4-fluoropyridine were sequentially added thereto at 0 ℃ and then transferred to room temperature for reaction overnight. The end of the reaction was confirmed by TLC (EA: PE ═ 1:5), dichloromethane was removed under reduced pressure, saturated brine (400mL × 3) and ethyl acetate 500mL were added and extracted, the organic phases were combined, dried over anhydrous sodium sulfate, concentrated and then dried over a dry-mix column, and the column was wetted with PE, and 2.27g of a white powdery solid was eluted with EA: PE ═ 1:5 with a yield of about 76.5%. MS (ESI) M/z 294.98[ M + H ]]+；1H 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).

6.2 Synthesis of intermediate 6-4:

the operation is the same as 1.2, and the intermediate 6-3 replaces the intermediate 1-3 to obtain a white solid 6-4. The yield is about 89.6%;¹H NMR(400MHz,DMSO-d₆)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 intermediates 6-8:

the operation is the same as 2.4, the intermediate 6-4 replaces the intermediate 2-4, and the yield is about 78.5 percent; MS (ESI) M/z 551.24[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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 intermediates 6-9:

the operation is the same as 2.5, and the intermediate 6-8 replaces the intermediate 2-8 to obtain light brown semisolid 6-9 with the yield of about 82.7 percent; MS (ESI) M/z 451.20[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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 the faint yellow solid VI-1 is obtained, and the yield is about 77.9 percent; MS (ESI) M/z 505.31[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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).¹³C NMR(101MHz,DMSO-d₆)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, the intermediate 6-9 replaces the intermediate 1-6, 2-chloroacrylic acid replaces 2-methacrylic acid 1-7, and the light brown solid VI-2 is obtained with the yield of about 80.2 percent; MS (ESI) M/z 505.14[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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).¹³C NMR(101MHz,DMSO-d₆)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 as 1.5, the intermediate 6-9 replaces the intermediate 1-6, and the 2-bromoacrylic acid replaces the 2-methacrylic acid 1-7, so that the light brown solid VI-3 is obtained, and the yield is about 76.2 percent; MS (ESI) M/z 583.10[ M + H ]]⁺；¹H 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).¹³C 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 (4.57mol) of 3-fluoro-4-bromobenzoic acid was weighed in a single-necked flask, 5mL of dichloromethane was added, and after the reaction flask was placed in an ice bath, 1.74g (4.57mmol) of HATU, 2.39mL (13.71mmol) of DIPEA and 1.13g (3.81mmol) of 2-aminopyridine were added in this order, followed by transfer to room temperature for overnight reaction. After completion of the reaction, the solvent dichloromethane was removed under reduced pressure by TLC, saturated brine (200mL × 3) and 250mL ethyl acetate were added for extraction, the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and then subjected to dry-method sample-mixing and column-passing, PE column-wetting, and elution with EA: PE ═ 1:4 gave 1.12g of light brown powdery solid with a yield of about 74.6%. MS (ESI) M/z 294.98[ M + H ]]⁺；¹H 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 operation is the same as 1.2, and the intermediate 7-3 replaces the intermediate 1-3 to obtain a white solid with the yield of about 91.6 percent;¹H NMR(400MHz,DMSO-d₆)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 intermediates 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 ]]⁺；¹H NMR(400MHz,DMSO-d₆)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 intermediates 7-9:

the operation is the same as 2.5, and the intermediate 7-8 replaces the intermediate 2-8 to obtain a light brown solid with the yield of about 82.1 percent; MS (ESI) M/z 433.25[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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 procedure as 1.5 was followed, wherein intermediates 1 to 6 were replaced with intermediates 7 to 9 and 1 to 7 of 2-methacrylic acid was replaced with acrylic acid to give VII-1 as a pale brown solid with a yield of about 63.8%; MS (ESI) M/z 487.19[ M + H ]]⁺；¹H NMR(400MHz,Acetone-d₆)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).¹³C NMR(101MHz,Acetone-d₆)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, the intermediate 7-9 replaces the intermediate 1-6, 2-chloroacrylic acid to replace 2-methacrylic acid 1-7, obtaining brown solid VII-2 with the yield of about 63.8%; MS (ESI) M/z 487.19[ M + H ]]⁺；¹H NMR(400MHz,Acetone-d₆)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).¹³C NMR(101MHz,Acetone-d₆)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 operation is the same as 1.5, the intermediate 7-9 replaces the intermediate 1-6, and the 2-bromoacrylic acid replaces the 2-methacrylic acid 1-7, so that brown solid VII-3 is obtained, and the yield is about 63.8%; MS (ESI) M/z 565.11[ M + H ]]⁺；¹H NMR(400MHz,Acetone-d₆)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).¹³C NMR(101MHz,Acetone-d₆)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 as in 1.5Intermediate 7-9 replaces intermediate 1-6, and (E) -4- (dimethylamino) but-2-enoic acid replaces 2-methacrylic acid 1-7 to give white solid VIII-1 in about 63.8% yield; MS (ESI) M/z 565.11[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)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).¹³C NMR(101MHz,DMSO-d₆)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, the intermediate 7-9 replaces the intermediate 1-6, and the 10-hydroxy-2-decenoic acid replaces the 2-methacrylic acid 1-7, so that the faint yellow semisolid VIII-2 is obtained, and the yield is about 63.8%; MS (ESI) M/z 601.30[ M + H ]]⁺；¹H NMR(400MHz,Acetone-d₆)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).¹³C NMR(101MHz,Acetone-d₆)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 preparation of Compounds

The above compound (1 mg) was dissolved in 100% DMSO-d6 to prepare a 10mM stock solution, which was stored under dark conditions in a nitrogen cabinet.

9.2 kinase reaction Processes

(1) A1 XKinase buffer was prepared.

(2) Preparation of compound concentration gradient: test compounds were initially tested at 2400nM, 7 concentrations, and tested in duplicate wells. 100% DMSO-d diluted to 100-fold final concentration in 384source plates₆And (3) solution. 250nl of 100-fold final concentration of compound were transferred to 3575 plates of interest using a dispenser Echo 550.

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

(4) Add 10. mu.l of 2.5 fold final concentration kinase solution to the compound well and positive control well, respectively; mu.l of 1 XKinase buffer was added to the negative control wells.

(5) Centrifuge at 1000rpm for 30 seconds, shake the plate and incubate at room temperature for 10 minutes.

(6) A mixture of ATP and Kinase substrate 2 was made up to a final concentration of 5/3 times using a 1 XKinase buffer.

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

(8) The 384 well plates were centrifuged at 1000rpm for 30 seconds, shaken well and incubated at room temperature for 30 minutes.

(9) Add 30. mu.l of termination detection solution to stop the kinase reaction, centrifuge at 1000rpm for 30 seconds, and mix by shaking.

(10) The conversion was read using a Caliper EZ Reader.

The results are shown in Table 1.

TABLE 1 BTK inhibitory Activity results for Compounds

As can be seen, the small molecular compound provided by the invention has good BTK inhibitory activity, and partial compounds (VII-1, VII-2 and VII-3) have both inhibitory rate and IC₅₀Are superior to the marketed drug, namely the drug Alcaninib, and IC₅₀Similar to ibrutinib.

Example 10 Water solubility test of Compounds

10.1 internal Standard preparation

Accurately weighing 1.00mg of Loratadine (Loratadine) standard, accurately transferring 1mL of dimethyl sulfoxide (DMSO), dissolving, shaking uniformly, preparing a stock solution with the concentration of 1mg/mL, and storing at-20 ℃ for later use. And precisely transferring a proper amount of stock solution, and diluting the stock solution into internal standard working solution with the concentration of 5.0ng/mL by using acetonitrile.

10.2 stock solutions of analytes

Accurately weighing 1.00mg of the substance to be detected, dissolving the substance in 1mL of DMSO to prepare 1.00mg/mL of substance stock solution A to be detected, and storing the substance at-20 ℃ for later use for preparing a standard curve.

Precisely weighing 1.00mg of the substance to be detected, dissolving the substance in 1mL of DMSO to prepare 1.00mg/mL of substance stock solution B for preparing a quality control sample (QC), and storing the substance at-20 ℃ for later use.

10.3 preparation of standard curve:

taking a proper amount of the stock solution A, and carrying out gradient elution on acetonitrile to obtain working solutions with the concentrations of 0.05 mu g/mL,0.1 mu g/mL,0.2 mu g/mL,0.5 mu g/mL,1 mu g/mL,2 mu g/mL,5 mu g/mL,10 mu g/mL,20 mu g/mL,50 mu g/mL,100 mu g/mL and 200 mu g/mL;

precisely transferring 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 concentrations 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 4 ℃ at 12000rpm for 10min, taking supernatant, and performing LC-MS/MS sample injection analysis.

10.4 Quality Control (QC) sample preparation:

taking a proper amount of the stock solution B, and carrying out gradient elution with acetonitrile to obtain working solutions with the concentrations of 0.1 mu g/mL,2.0 mu g/mL and 16 mu g/mL;

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

10.5 preparation of water-soluble sample to be tested

Weighing a proper amount of a substance to be detected, adding 1mL of pure water respectively, preparing a saturated aqueous solution, carrying out vortex 2h, standing at room temperature for 1h, centrifuging at 4 ℃ and 12000rpm for 10min, taking 50 mu L of supernatant, adding 200 mu L of acetonitrile containing internal standard loratadine, carrying out vortex 5min, centrifuging at 4 ℃ and 12000rpm for 10min, taking supernatant, carrying out sample injection analysis by LC-MS/MS (liquid chromatography-mass spectrometry/mass spectrometry), and carrying out 3 parallel experiments on each sample.

The results are shown in Table 2:

TABLE 2.1 Standard Curve equation Table

TABLE 2.2 Water solubility test results for the compounds

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

EXAMPLE 11 Compound PK assay

After the compounds ibrutinib, IV-1 and VIII-2 are respectively administered with single dose through intravenous and oral administration in SD rats, blood samples are collected at different time points, LC-MS/MS is used for measuring the concentrations of the compounds ibrutinib, IV-1 and VIII-2 in rat plasma and calculating related pharmacokinetic parameters, and the bioavailability and the pharmacokinetic properties of the compounds ibrutinib, IV-1 and VIII-2 in the rats are inspected.

Male SD rats 9, about 200g, were randomly divided into 3 groups of 3 rats each. Fasting was performed for 12h before administration, and water was freely available.

Each animal was bled 0.100mL per time through the orbit, and EDTA-disodium was anticoagulated at the following collection time points: group IV: 5min, 15min, 30min, 1h, 2h, 4h, 6h, 8h and 24h after the administration of the test substance; PO group: 15min, 30min, 1h, 2h, 4h, 6h, 8h and 24h after administration of the test substance. Blood samples were collected, placed on ice and plasma was centrifuged within 30min (centrifugation conditions: 3000rpm,10min,4 ℃). And after centrifugation, respectively taking 50 mu L of supernatant, adding 200 mu L of acetonitrile containing internal standard loratadine, vortexing for 5min, centrifuging at 4 ℃ and 12000rpm for 10min, taking the supernatant, carrying out sample injection analysis by LC-MS/MS (liquid chromatography-mass spectrometry/mass spectrometry), and carrying out parallel experiments on 3 parts of each sample.

The experimental test results are as follows:

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

the blood concentration profile of SD rats after intravenous injection (IV.,2.00mg/kg) of Ibrutinib (Ibrutinib), IV-2 and VII-2 over time is shown in figure 1.

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

TABLE 3.2 major pharmacokinetic parameters after gastric gavage in SD rats

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

Claims (8)

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

wherein, X, Y, Z has the following structure respectively:

Z＝H、F，

wherein R has the following structure:

2. the irreversible BTK inhibitor of oxazolo [4,5-b ] pyridine structure according to claim 1, characterized in that X, Y, Z has the following structures respectively:

Z＝H，

wherein R has the structure:

alternatively, X, Y, Z has the following structures, respectively:

Z＝H，F，

wherein R has the following structure:

3. the irreversible BTK inhibitor of oxazolo [4,5-b ] pyridine structure of claim 1, wherein the compound is selected from the following structures:

or the following structure:

or the following structure:

4. use of the irreversible BTK inhibitor of oxazolo [4,5-b ] pyridine structure according to any one of claims 1 to 3 for the preparation of a pharmaceutical preparation for the prevention or treatment of a disease caused by BTK abnormality.

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

6. A pharmaceutically acceptable salt of an irreversible BTK inhibitor of oxazolo [4,5-b ] pyridine structure as claimed in any one of claims 1 to 3, wherein the pharmaceutically acceptable salt is hydrochloride, hydrobromide, hydroiodide, sulphate, bisulphate, phosphate, acetate, propionate, butyrate, oxalate, tartrate, methanesulphonate, p-toluenesulphonate, fumarate, taurate, citrate, succinate or a mixed salt thereof.

7. The use of the pharmaceutically acceptable salt of the irreversible BTK inhibitor of oxazolo [4,5-b ] pyridine structure of claim 6 for the preparation of a pharmaceutical preparation for the prevention or treatment of diseases caused by BTK abnormality.

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

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