A novel 4-morpholinyl thieno[3,2-d]pyrimidine compound containing an acrylamide structure and application thereof

Abstract

The application belongs to the field of medicine, and particularly relates to a novel 4-morpholinyl thieno[3,2-d]pyrimidine compound containing an acrylamide structure and application thereof. The novel 4-morpholinyl thieno[3,2-d]pyrimidine compound containing an acrylamide structure has a structure of general formula (I). The application also relates to the compound of general formula (I) having a strong PI3K inhibiting effect, and further relates to the use of the compound and a pharmaceutically acceptable salt thereof in the preparation of a drug for treating and / or preventing a disease caused by abnormal high expression of PI3K, particularly in the preparation of a drug for treating and / or preventing cancer. The compound of the application is particularly suitable for preparing a drug for treating and / or preventing breast cancer, prostate cancer, colon cancer, lung cancer and brain glioma.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and particularly relates to a novel 4-morpholinyl thieno[3,2-d]pyrimidine compound containing an acrylamide structure and application thereof. BACKGROUND

[0002] Cancer, also known as malignant tumor, is a disease that seriously endangers human life and health, and the incidence and mortality of cancer worldwide are extremely high and show a trend of increasing year by year. According to the data of the International Agency for Research on Cancer (IARC) under the World Health Organization, about 19.3 million new cancer cases and about 10 million deaths occurred worldwide in 2020. It is estimated that by 2040, there will be 28.4 million new cancer cases worldwide. Malignant tumor has become a serious public health problem facing the world.

[0003] With the rapid development of tumor-related sciences such as tumor genomics, the mechanism of malignant tumor occurrence has been gradually elucidated. The molecular targeted therapy of tumor with the key kinase in the specific molecules and signal pathways of tumor as the target has been paid more and more attention by scientists. The molecular targeted therapy often has unique advantages such as direction and localization. PI3K, i.e. phosphatidylinositol-3 kinase, is an intracellular phosphatidylinositol kinase which has the activity of serine / threonine kinase and the activity of phosphatidylinositol kinase. PI3K plays a key role in cell proliferation, growth and survival. When the cell is stimulated by external stimuli such as growth factors, the receptor tyrosine kinase (RTKs), G protein-coupled receptor and Ras protein can be activated, and then the PI3K kinase is activated. The activated PI3K kinase can phosphorylate phosphatidylinositol diphosphate (PIP2) to become phosphatidylinositol triphosphate (PIP3), which is a very important second messenger and can activate the phosphatidyl of downstream serine threonine protein kinase (Akt) effectors, so as to transduce the extracellular signal received by the cell surface receptor to the cell, and then regulate the biological processes of tumor cells, promote tumor cell proliferation, differentiation, metastasis, etc. The abnormal PI3K signal is related to the occurrence and development of many cancers (such as prostate cancer, breast cancer, endometrial cancer, lung cancer, liver cancer, colon cancer, etc.). PI3K can be divided into three types of I, II and III, among which the most widely studied is class I PI3K. Class I PI3K is divided into four subtypes of PI3Kα, PI3Kβ, PI3Kδ and PI3Kγ according to their catalytic subunits, etc. The small molecule inhibitors targeting PI3K have become an important direction of the research of anti-tumor drugs in recent years, and have achieved phased results in the treatment of tumors. At present, there are six PI3K inhibitors, Idelalisib (Zydelig), Copanlisib (Aliqopa), Duvelisib (Copiktra), Alpelisib (BYL719, Piqray), Umbralisib and Lumipimycin (Intratumor), which have been marketed worldwide, and many PI3K inhibitors have entered the clinical research stage. In recent years, due to safety problems, the indications of many marketed drugs have been withdrawn, and the marketing applications of some drugs have been rejected. For PI3K inhibitors, reducing the side effects and improving the safety are the primary tasks. Therefore, the development of new PI3K inhibitors with novel structure, safety and effectiveness is still the key research field of anti-tumor drugs at home and abroad. GDC-0941 is the first thieno[3,2-d]pyrimidine PI3K inhibitor entering the clinical research. It is a class of orally effective class I PI3K inhibitors developed by Genentech Company, which has an IC 50The values are 3, 33, 3 and 75 nM respectively. GDC-0941 shows significant inhibitory effect on the proliferation of various human tumor cell lines including glioblastoma, breast cancer cells, prostate cancer cells, etc. GDC-0941 has high plasma protein binding rate, appropriate liver clearance rate, good pharmacokinetic properties in animal models, and oral bioavailability of 71%.

[0004] The structure of the novel thieno[3,2-d]pyrimidine derivatives designed and synthesized by the application has not been reported. It is found through pharmacological activity that the novel thieno[3,2-d]pyrimidine derivatives have significant inhibitory activity on PI3K and significant inhibitory effect on various human tumor cells, and show good antitumor activity. SUMMARY

[0005] Based on the summary of the structure-activity relationship of PI3K inhibitors and the analysis of the three-dimensional spatial structure of PI3K protein, the inventors design and synthesize a series of novel 4-morpholinyl thieno[3,2-d]pyrimidine compounds containing acrylamide structure. Through in vitro activity screening, it is shown that the compounds have antitumor activity. The application relates to novel 4-morpholinyl thieno[3,2-d]pyrimidine compounds containing acrylamide structure as PI3K inhibitors, which have not been reported in the literature.

[0006] The technical scheme adopted by the application is: a novel 4-morpholinyl thieno[3,2-d]pyrimidine compound containing acrylamide structure and a pharmaceutically acceptable salt thereof, the structural formula of which is shown in general formula (I):

[0007] ;

[0008] Among them:

[0009] R1 and R2 are the same or different, and are independently selected from hydrogen, alkyl containing 1-6 carbons, cycloalkyl containing 3-8 carbons, phenyl, wherein the cycloalkyl and phenyl can be optionally substituted by 1-3 R3 which are the same or different; or R1 and R2 together with the nitrogen atom to which they are connected form 4-7 membered heterocyclyl containing 1-2 heteroatoms selected from N, O and S, and when the 4-7 membered heterocyclyl contains a sulfur atom, the sulfur atom can be oxidized to form a sulfone, and the 4-7 membered heterocyclyl containing 1-2 heteroatoms selected from N, O and S can be optionally substituted by 1-3 R4 which are the same or different; or R1 and R2 are selected from hydrogen and the following groups:

[0010]

[0011] R3 is selected from hydrogen, halogen, alkyl containing 1-6 carbons, alkoxy containing 1-6 carbons;

[0012] R4 is selected from the group consisting of hydrogen, halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons, alkoxycarbonyl of 1 to 6 carbons, alkylsulfonyl of 1 to 6 carbons, alkylcarbonyl of 1 to 6 carbons, amino substituted with one or two alkyl groups of 1 to 6 carbons, carbamoyl, carbamoyl substituted with one or two alkyl groups of 1 to 6 carbons, alkyl of 1 to 6 carbons substituted with hydroxy, alkyl of 1 to 6 carbons substituted with alkoxy of 1 to 3 carbons;

[0013] R5 and R6 are the same or different and are each independently selected from the group consisting of hydrogen, alkyl of 1 to 6 carbons;

[0014] R7 is selected from the group consisting of alkyl of 1 to 6 carbons, alkoxycarbonyl of 1 to 6 carbons;

[0015] R8 is selected from the group consisting of hydrogen, hydroxy, alkyl of 1 to 6 carbons, alkoxycarbonyl of 1 to 6 carbons;

[0016] n is selected from 1, 2, 3, 4, 5;

[0017] m is selected from 1, 2, 3;

[0018] Q is selected from the group consisting of:

[0019]

[0020] R9 is selected from the group consisting of hydrogen, amino, hydroxy, alkyl of 1 to 6 carbons, alkoxycarbonyl of 1 to 6 carbons, amino substituted with one or two alkyl groups of 1 to 6 carbons;

[0021] X is selected from a C or N atom;

[0022] Preferably, the above-mentioned novel 4-morpholinylthieno[3,2-d]pyrimidine compounds having an acrylamide structure and pharmaceutically acceptable salts thereof,

[0023] In the general formula (I), is selected from the group consisting of:

[0024]

[0025] Q is selected from the group consisting of:

[0026]

[0027] Further, the above-mentioned novel 4-morpholinylthieno[3,2-d]pyrimidine compounds having an acrylamide structure and pharmaceutically acceptable salts thereof, the compounds of the general formula (I) have the following structural formula, but are not limited to these compounds:

[0028]

[0029] The following synthetic routes describe the preparation of the novel 4-morpholinylthieno[3,2-d]pyrimidine compounds of general formula (I) of the present application containing an acrylamide structure, all starting materials are prepared by the means described in these schemes, by methods well known to one of ordinary skill in the art of organic chemistry or are commercially available. All final derivatives of the present application are prepared by the means described in the following schemes or by methods analogous thereto, which are well known to one of ordinary skill in the art of organic chemistry. All variable factors applied in these schemes are defined below.

[0030] The novel 4-morpholinylthieno[3,2-d]pyrimidine compounds of general formula (I) of the present application containing an acrylamide structure, when Q is can be prepared according to Route 1, wherein (I-1) represents a class of substructure of (I), other substituents are defined as above;

[0031]

[0032] Route 1;

[0033] The novel 4-morpholinylthieno[3,2-d]pyrimidine compounds of general formula (I) of the present application containing an acrylamide structure, when Q is can be prepared according to Route 2, wherein (I-2) represents a class of substructure of (I), the synthesis of intermediate e in this route is the same as Route 1, other substituents are defined as above;

[0034]

[0035] Route 2;

[0036] The substituents R1, R2, Q of all intermediates in the above two routes are defined as above.

[0037] A pharmaceutical composition comprising any one of the novel 4-morpholinylthieno[3,2-d]pyrimidine compounds containing an acrylamide structure and pharmaceutically acceptable salts thereof described above as an active ingredient, together with a pharmaceutically acceptable excipient.

[0038] The use of any one of the novel 4-morpholinylthieno[3,2-d]pyrimidine compounds containing an acrylamide structure and pharmaceutically acceptable salts thereof described above or the pharmaceutical composition described above in the preparation of a medicament for the treatment and / or prevention of diseases caused by the abnormal high expression of PI3K.

[0039] Use of any one of the novel 4-morpholinylthieno[3,2-d]pyrimidines having an acrylamide structure and pharmaceutically acceptable salts thereof or the pharmaceutical composition described above in the manufacture of a medicament for treating and / or preventing a proliferative disease.

[0040] Use of any one of the novel 4-morpholinylthieno[3,2-d]pyrimidines having an acrylamide structure and pharmaceutically acceptable salts thereof or the pharmaceutical composition described above in the manufacture of a medicament for treating and / or preventing a cancer.

[0041] Further, use of any one of the novel 4-morpholinylthieno[3,2-d]pyrimidines having an acrylamide structure and pharmaceutically acceptable salts thereof or the pharmaceutical composition described above in the manufacture of a medicament for treating and / or preventing breast cancer, prostate cancer, colon cancer, lung cancer and brain glioma.

[0042] Also, the novel 4-morpholinylthieno[3,2-d]pyrimidines having an acrylamide structure of the present application can form pharmaceutically acceptable salts with acids according to some of the usual methods in the field to which the present application pertains. The pharmaceutically acceptable addition salts include inorganic acid and organic acid addition salts, and the salts with the following acids are particularly preferred: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalenedisulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, benzoic acid and the like.

[0043] In the present application, "halogen" means fluoro, chloro, bromo or iodo; "alkyl" means straight or branched chain alkyl; "cycloalkyl" means substituted or unsubstituted cycloalkyl; "aryl" means monocyclic or polycyclic aromatic ring system of carbon atoms, such as phenyl, naphthyl and the like; "heteroaryl" means monocyclic or polycyclic ring system containing one or more heteroatoms selected from N, O, S, and the ring system is aromatic, such as imidazolyl, pyridyl, pyrazolyl, (1,2,3)- and (1,2,4)-triazolyl, furanyl, thienyl, pyrrolyl, thiazolyl, benzothiazolyl, oxazolyl, isoxazolyl, naphthyl, quinolyl, isoquinolyl, benzimidazolyl and benzoxazolyl and the like; "heterocyclyl" means monocyclic or polycyclic ring system containing one or more heteroatoms selected from N, O, S, such as pyrrolidinyl, morpholinyl, piperazinyl, piperidinyl, pyrazolidinyl, imidazolidinyl and thiazolidinyl and the like. represents the point of attachment of the substituent.

[0044] The present application has the following advantages:

[0045] 1. The compounds of the present application have significant inhibitory effect on the above-mentioned tumor cells by in vitro inhibition of human breast cancer cells MCF-7, MDA-MB-231, human prostate cancer cells 22RV1, human colon cancer cells HCT116, human lung cancer cells A549 and human brain glioma cells U87MG, and are particularly useful for the preparation of drugs for the treatment and / or prevention of breast cancer, prostate cancer, colon cancer, lung cancer and brain glioma.

[0046] 2. The compounds of the present application have significant inhibitory effect on PI3K activity by PI3K activity test, and are particularly useful for the preparation of drugs for the treatment and / or prevention of diseases caused by abnormal high expression of PI3K, and particularly useful for the preparation of drugs for the treatment and / or prevention of cancer.

[0047] 3. The 4-morpholinyl thieno[3,2-d]pyrimidine compounds of the present application containing acrylamide structure have novel chemical structure, most of the compounds have significant inhibitory activity on PI3K in in vitro biological activity study, and have strong anti-proliferative activity on the tested cells. The compounds can be used for the treatment and prevention of cancer. DETAILED DESCRIPTION

[0048] The examples and preparation examples provided below further illustrate and exemplify the compounds of the present application and methods for preparing them. It is to be understood that the scope of the application is not limited in any way by the following examples and preparation examples. The examples are intended merely to be illustrative and not limiting of the scope of the application. The nuclear magnetic resonance hydrogen spectrum of the compounds was determined by Bruker ARX-400 or ARX-600, and the mass spectrum was determined by Agilent 6460 QQQ; the reagents used were all analytical pure or chemical pure.

[0049] Example 1: E - N Synthesis of (4-fluorophenyl)-3-(2-(6-methoxy-3-pyridinyl)-4-morpholinyl-6- thieno[3,2-d]pyrimidinyl)acrylamide (Compound 1)

[0050] Compound 1

[0051] Step 1: Synthesis of 2,4-dihydroxythieno[3,2-d]pyrimidine

[0052]

[0053] Methyl 3-amino-2-thiophenecarboxylate (124.8 g, 0.79 mmol) was mixed with urea (275.4 g, 3.97 mol) and heated to 190 °C with stirring for 2 hours. The reaction was cooled to 120 °C and added hot to 600 mL of 7.5% sodium hydroxide solution with stirring until most of the solid dissolved. The mixture was filtered, the pH was adjusted to 3 and a large amount of white solid precipitated. The mixture was filtered, the filter cake was washed with water until neutral and dried to give 2,4-dihydroxythieno[3,2-d]pyrimidine as a light yellow solid, 121.0 g. 1 H NMR (300 MHz, DMSO- d 6 ) δ 11.58 (s, 1H), 11.24 (s, 1H), 8.06 (d, J = 5.2Hz, 1H), 6.92 (d, J = 5.2 Hz, 1H)。

[0054] Step 2: Synthesis of 2,4-dichlorothieno[3,2-d]pyrimidine

[0055]

[0056] 2,4-Dihydroxythieno[3,2-d]pyrimidine (14.8 g, 88.0 mmol) was slowly added to 80 mL of phosphorus oxychloride and heated to reflux for 10 hours. The solvent was evaporated under reduced pressure and the residue was slowly poured into ice water with vigorous stirring. A large amount of white solid precipitated and was filtered, the filter cake was washed with water until neutral and dried to give 2,4-dichlorothieno[3,2-d]pyrimidine as a light yellow solid, 12.72 g. 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.71 (d, J = 5.4 Hz, 1H), 7.74 (d, J =5.4 Hz, 1H)。

[0057] Step 3: Synthesis of 2-chloro-4-(4-morpholinyl)thieno[3,2-d]pyrimidine

[0058]

[0059] 100 g (0.49 mol) of 2,4-dichlorothieno[3,2-d]pyrimidine was placed in 1500 mL of methanol, and morpholine (93.4 g, 1.23 mol) was slowly added dropwise with stirring at room temperature. The reaction was complete 2 hours later. The mixture was filtered, and the filter cake was washed with ethanol, followed by thorough washing with ethanol. The filter cake was dried to obtain 118.6 g of a pale yellow solid, 2-chloro-4-(4-morpholinyl)thieno[3,2-d]pyrimidine. MS (ESI), m / z (%): 256.1 [M+H] + .

[0060] Step 4: Synthesis of 2-chloro-4-morpholinylthiopheno[3,2-d]pyrimidine-6-carbaldehyde

[0061]

[0062] Under nitrogen protection, 120 g (0.47 mol) of 2-chloro-4-(4-morpholinyl)thieno[3,2-d]pyrimidine was dispersed in 1200 mL of dry tetrahydrofuran. The solution was cooled to -10 °C, and a tetrahydrofuran solution of isopropyl magnesium chloride (2 mol / L, 118 mL) was slowly added dropwise. After the addition was complete, the reaction was maintained at -10 °C for 2 h. A hexane solution of n-butyllithium (2.5 mol / L, 226 mL) was then slowly added dropwise to the above solution. After the addition was complete, the reaction was maintained at -10 °C for 2 h. Finally, the reaction solution was further treated with... N, N- 91 mL of dimethylformamide was added dropwise, and the mixture was kept at -10 °C for 8 h. The reaction solution was then slowly poured into five times the volume of a pre-prepared 0.25 M hydrochloric acid solution, stirred for 2 h, and then heated to 50 °C and stirred for 4 h. The mixture was filtered, washed with water until neutral, and dried to obtain 127 g of a pale yellow solid, 2-chloro-4-morpholinylthiopheno[3,2-d]pyrimidine-6-carboxaldehyde. MS (ESI), m / z (%): 284.1 [M+H] + .

[0063] Step 5: Synthesis of 2-(6-methoxy-3-pyridyl)-4-morpholinylthiopheno[3,2-d]pyrimidine-6-carboxaldehyde

[0064]

[0065] 2-Chloro-4-morpholinylthiopheno[3,2-d]pyrimidine-6-carboxaldehyde (15 g, 52.87 mmol) was mixed with 6-methoxypyridine-3-boronic acid (12.17 g, 79.30 mmol) and potassium carbonate (21.30 g, 0.158.60 mmol). 225 mL of 1,4-dioxane was added, and nitrogen gas was slowly passed through the solution at room temperature for 30 min. Then, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (1.17 g, 1.60 mmol) was added, and the reaction was carried out at 100°C for 4 h under nitrogen protection. The mixture was filtered, and the filter cake was washed with dichloromethane. The filtrate was added with an appropriate amount of water, and the mixture was extracted three times with dichloromethane. The organic phases were combined, washed three times with water, and three times with saturated brine. The organic phase was dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain the crude product. The crude product was pulped with anhydrous methanol and dried to obtain 14.96 g of yellow solid 2-(6-methoxy-3-pyridyl)-4-morpholinylthiopheno[3,2-d]pyrimidine-6-carboxaldehyde. 1 H NMR (600 MHz, CDCl3) δ 10.19 (s, 1H), 9.23 (s, 1H), 8.58 (d, J = 8.5 Hz,1H), 8.08 (s, 1H), 6.82 (d, J = 8.6 Hz, 1H), 4.20–4.07 (m, 4H), 4.02 (s, 3H), 3.96–3.80 (m, 4H); MS (ESI) m / z: 357.00 [M+H] + .

[0066] Step 6: ( E Synthesis of ethyl acrylate 3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acrylate

[0067]

[0068] 2-(6-methoxy-3-pyridyl)-4-morpholinylthiopheno[3,2-d]pyrimidine-6-carboxaldehyde (5.00 g, 14.03 mmol), ethoxyformylmethylenetriphenylphosphine (7.33 g, 21.04 mmol), and 40 mL of toluene were added to a reaction flask, and the mixture was refluxed for approximately 3 h until the reaction was complete. The reaction mixture was filtered while hot, and the filter cake was washed with an appropriate amount of toluene. The filter cake was then dried to obtain a yellow solid. EEthyl acrylate 4.41 g of 3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acrylate. MS (ESI) m / z: 427.10 [M+H] + .

[0069] Step 7: ( E Synthesis of 3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acrylic acid

[0070]

[0071] Will( E Ethyl 3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acrylate (4.40 g, 10.32 mmol), sodium hydroxide (1.24 g, 30.95 mmol), 40 mL of ethanol, and 10 mL of water were added to a reaction flask and stirred at 60°C for 5 h until the reaction was complete. Most of the solvent was removed by vacuum evaporation, and the residue was added to 50 mL of water. The pH was adjusted to approximately 3 with concentrated hydrochloric acid solution, resulting in the precipitation of a large amount of yellow solid. The solid was filtered, and the filter cake was dried to obtain a yellow solid. E 2.94 g of 3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acrylic acid. 1 H NMR (600MHz, DMSO- d 6 ) δ 12.80 (s, 1H), 9.14 (s, 1H), 8.57 (d, J = 8.5 Hz, 1H), 8.12 –7.72 (m, 2H), 7.09 – 6.79 (m, 1H), 6.49 (d, J = 15.9 Hz, 1H), 3.99 (br, 4H), 3.93 (s, 3H), 3.80 (br, 4H).

[0072] Step 8: ( E )- N Synthesis of 3-(4-fluorophenyl)-3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acrylamide (compound 1)

[0073] Compound 1

[0074] Add ( to the reaction flask) E)-3-(2-(6-methoxy-3-pyridinyl)-4-morpholino-6-thieno[3,2-d]pyrimidinyl)acrylic acid (200.00 mg, 0.50 mmol), 4-fluoroaniline (67.00 mg, 0.60 mmol), 2-(7-azabenzotriazole)- N,N,N',N' - Tetramethyl-uranium hexafluorophosphate (285.80 mg, 0.75 mmol), triethylamine (152.38 mg, 1.51 mmol) and N,N - Dimethylformamide 5 mL, stirring at room temperature for 20 h reaction completed. The reaction liquid was poured into 30 mL 10% sodium carbonate aqueous solution, a solid was precipitated, filtered, the filter cake was washed with water three times, the filter cake was slurried with methanol twice, and dried to obtain a light yellow solid 0.18 g. 1 H NMR (600 MHz, DMSO- d 6 ) δ 10.43 (s, 1H), 9.16 (d, J = 2.2 Hz, 1H), 8.59 (dd, J = 8.6, 2.3 Hz, 1H), 7.90 (d, J = 15.5 Hz, 1H), 7.83 (s, 1H), 7.72(dd, J = 8.9, 5.0 Hz, 2H), 7.19 (t, J = 8.8 Hz, 2H), 6.92 (d, J = 8.7 Hz,1H), 6.86 (d, J = 15.5 Hz, 1H), 4.07 – 3.99 (m, 4H), 3.93 (s, 3H), 3.86 –3.78 (m, 4H); MS (ESI), m / z: 492.10 [M+H] + .

[0075] By replacing appropriate raw materials and reagents, the preparation method of Example 1 was followed to finally obtain Examples 2-37 (Compound 2-37).

[0076] Example 2: ( E ) N - Synthesis of phenyl-3-(2-(6-methoxy-3-pyridinyl)-4-morpholino-6-thieno[3,2-d]pyrimidinyl)acrylamide (Compound 2)

[0077]

[0078] Replacing 4-fluoroaniline in step 8 of Example 1 with aniline, and following the same steps as in Example 1, compound 2 was obtained. MS (ESI), m / z: 474.10 [M+H] + .

[0079] Example 3: ( E )- N Synthesis of -(4-methoxyphenyl)-3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acrylamide (compound 3)

[0080]

[0081] Replacing 4-fluoroaniline in step 8 of Example 1 with 4-methoxyaniline, and following the same steps as in Example 1, compound 3 was obtained. MS (ESI), m / z: 504.20 [M+H] + .

[0082] Example 4: ( E )- N Synthesis of -(3,4-dimethoxyphenyl)-3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acrylamide (compound 4)

[0083]

[0084] In Example 1, step 8, 4-fluoroaniline was replaced with 3,4-dimethoxyaniline, and the remaining steps were the same as in Example 1, to obtain compound 4. 1 H NMR (600 MHz, CDCl3) δ 9.23 (d, J = 2.0 Hz, 1H), 8.57 (dd, J =8.7, 2.3 Hz, 1H), 7.91 (d, J = 15.2 Hz, 1H), 7.56 (s, 1H), 7.52 (s, 1H), 7.43(s, 1H), 6.95 (d, J = 7.1 Hz, 1H), 6.89 – 6.76 (m, 2H), 6.55 (d, J = 15.2 Hz,1H), 4.08 – 4.02 (m, 4H), 4.01 (s, 3H), 3.91 (s, 3H), 3.90 – 3.85 (m, 7H); MS(ESI), m / z: 534.20 [M+H] + .

[0085] Example 5: ( E Synthesis of 3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-(4-methanesulfonyl-1-piperazinyl)-2-propen-1-one (compound 5)

[0086]

[0087] In Example 1, step 8, 4-fluoroaniline was replaced with 4-methanesulfonylpiperazine, and the remaining steps were the same as in Example 1, to obtain compound 5. 1 H NMR (600 MHz, DMSO- d 6 ) δ 9.15 (s, 1H), 8.57 (d, J = 8.6 Hz, 1H),7.88 (s, 1H), 7.82 (d, J = 15.2 Hz, 1H), 7.30 (d, J = 15.2 Hz, 1H), 6.92 (d, J = 8.6 Hz, 1H), 4.07 – 3.97 (m, 4H), 3.93 (s, 3H), 3.87 – 3.76 (m, 6H), 3.70(br, 2H), 3.17 (br, 4H), 2.91 (s, 3H); MS (ESI), m / z: 545.10 [M+H] + .

[0088] Example 6: ( E Synthesis of )-3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-morpholinyl-2-propen-1-one (compound 6)

[0089]

[0090] Replacing 4-fluoroaniline in step 8 of Example 1 with morpholine, and following the same steps as in Example 1, compound 6 was obtained. MS (ESI), m / z: 468.20 [M+H] + .

[0091] Example 7: ( E Synthesis of 3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-(1-pyrrolidinyl)-2-propen-1-one (compound 7)

[0092]

[0093] Compound 7 was obtained by replacing 4-fluoroaniline in step 8 of Example 1 with tetrahydropyrrole, and the remaining steps were the same as Example 1. MS (ESI), m / z: 452.20 [M+H] + .

[0094] Example 8: ( E )- N, N Synthesis of N,N-diethyl-3-(2-(6-methoxy-3-pyridinyl)-4-morpholino-6- thieno[3,2-d]pyrimidinyl)acrylamide (Compound 8)

[0095]

[0096] Compound 8 was obtained by replacing 4-fluoroaniline in step 8 of Example 1 with diethylamine, and the remaining steps were the same as Example 1. 1 H NMR (600 MHz, CDCl3) δ 9.23 (d, J = 2.2 Hz, 1H), 8.58 (dd, J = 8.7, 2.4Hz, 1H), 7.88 (d, J = 15.1 Hz, 1H), 7.54 (s, 1H), 6.84 (dd, J = 21.7, 11.9Hz, 2H), 4.10 – 4.03 (m, 4H), 4.01 (s, 3H), 3.94 – 3.84 (m, 5H), 3.59 – 3.41(m, 4H), 1.30 (t, J = 7.1 Hz, 3H), 1.21 (t, J = 7.1 Hz, 3H); MS (ESI), m / z:454.20 [M+H] + .

[0097] Example 9: ( E )- N Synthesis of N-(2-dimethylaminoethyl)-3-(2-(6-methoxy-3-pyridinyl)-4-morpholino-6- thieno[3,2-d]pyrimidinyl)- N methyl acrylamide (Compound 9)

[0098]

[0099] Compound 9 was obtained by replacing 4-fluoroaniline in step 8 of Example 1 with N,N,N'- trimethylethylenediamine, the remaining steps are the same as Example 1 to give compound 9. 1 H NMR (600 MHz, DMSO- d 6 ) δ 9.13 (d, J = 2.0 Hz, 1H), 8.56 (dd, J = 8.6, 2.2 Hz, 1H), 7.84 (s, 1H), 7.74 (t, J = 15.5 Hz, 1H), 7.17 (t, J =16.0 Hz, 1H), 6.91 (d, J = 8.7 Hz, 1H), 3.98 (br, 4H), 3.93 (s, 3H), 3.81(br, 4H), 3.60 (t, J = 6.2 Hz, 1H), 3.54 (t, J = 6.5 Hz, 1H), 3.06 (d, J =124.6 Hz, 3H), 2.54 (s, 1H), 2.45 (t, J = 6.0 Hz, 1H), 2.28 (s, 3H), 2.22 (s,3H); MS (ESI), m / z: 483.20 [M+H] + .

[0100] Example 10: Synthesis of (S)-3-(2-(6-methoxy-3-pyridinyl)-4-morpholino-6- thieno[3,2-d]pyrimidinyl)-1-(4-methyl-1-piperazinyl)-2-propen-1-one (Compound 10) E

[0101]

[0102] Replace 4-fluoroaniline in step 8 of Example 1 with N - methylpiperazine, the remaining steps are the same as Example 1 to give compound 10. MS (ESI), m / z: 481.20 [M+H] + , 680.20 [M+Na] + .

[0103] Example 11: Synthesis of (S)-3-(2-(6-methoxy-3-pyridinyl)-4-morpholino-6- thieno[3,2-d]pyrimidinyl)-1-(4-methyl-1-piperazinyl)-2-propen-1-one (Compound 10) E N ​​Synthesis of 11-(2-dimethylaminoethyl)-3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acrylamide (compound 11)

[0104]

[0105] Replace 4-fluoroaniline in step 8 of Example 1 with N,N -Dimethylethylenediamine, and the remaining steps are the same as in Example 1, to obtain compound 11. 1 H NMR (600 MHz, CD3COOD) δ 9.21 (d, J = 2.4 Hz, 1H), 8.60 (dd, J =8.8, 2.5 Hz, 1H), 7.87 (d, J = 15.6 Hz, 1H), 7.79 (s, 1H), 6.96 (d, J = 8.8Hz, 1H), 6.83 (d, J = 15.6 Hz, 1H), 4.30 – 4.17 (m, 5H), 4.04 (s, 3H), 4.02 –3.95 (m, 4H), 3.87 (t, J = 5.6 Hz, 2H), 3.48 (t, J = 5.7 Hz, 2H), 3.02 (s,7H); MS (ESI), m / z: 469.20[M+H] + .

[0106] Example 12: ( E )- N Synthesis of 3-(3-dimethylaminopropyl)-3-(2-(6-methoxy-3-pyridyl)-4-morpholino-6-thieno[3,2-d]pyrimidinyl)acrylamide (compound 12)

[0107]

[0108] In Example 1, step 8, 4-fluoroaniline was replaced with 3-dimethylamino-1-propylamine, and the remaining steps were the same as in Example 1, to obtain compound 12. 1 H NMR (600 MHz, DMSO- d 6 ) δ 9.13 (d, J = 1.9 Hz, 1H), 8.56 (dd, J= 8.7, 2.2 Hz, 1H), 8.29 (t, J = 5.4 Hz, 1H), 7.97 – 7.60 (m, 2H), 6.91 (d, J = 8.6 Hz, 1H), 6.68 (d, J = 15.7 Hz, 1H), 4.04 – 3.96 (m, 4H), 3.93 (s, 3H),3.85 – 3.76 (m, 4H), 3.22 (dd, J = 12.6, 6.6 Hz, 2H), 2.35 (t, J = 7.0 Hz,2H), 2.22 (s, 6H), 1.74 – 1.52 (m, 2H);; MS (ESI), m / z: 483.20[M+H] + .

[0109] Example 13: Synthesis of (2-methoxyethyl)-3-(2-(6-methoxy-3-pyridinyl)-4- morpholino-6-thieno[3,2-d]pyrimidinyl)acrylamide (Compound 13) E N

[0110] Example 1, step 8, replacing 4-fluoroaniline with 2-methoxyethylamine. Compound 13 was obtained. 1 H NMR (600 MHz, DMSO- 6 ) δ 9.15 (d,

[0111] = 2.2 Hz, 1H), 8.58 (dd, d =8.7, 2.3 Hz, 1H), 8.35 (t, J = 5.5 Hz, 1H), 7.99 – 7.61 (m, 2H), 6.92 (d, J =8.7 Hz, 1H), 6.76 (d, J = 15.8 Hz, 1H), 4.07 – 3.96 (m, 4H), 3.93 (s, 3H),3.87 – 3.75 (m, 4H), 3.41 (t, J = 5.4 Hz, 2H), 3.37 (t, J J J ​​= 5.3 Hz, 2H), 3.28(s, 3H); MS (ESI), m / z: 456.20[M+H] + .

[0112] Example 14: Synthesis of (2-hydroxyethyl)-3-(2-(6-methoxy-3-pyridinyl)-4- morpholino-6-thieno[3,2-d]pyrimidinyl)acrylamide (Compound 14) E )- N Example 14: Synthesis of (2-hydroxyethyl)-3-(2-(6-methoxy-3-pyridinyl)-4- morpholino-6-thieno[3,2-d]pyrimidinyl)acrylamide (Compound 14)

[0113]

[0114] Example 14: Synthesis of (2-hydroxyethyl)-3-(2-(6-methoxy-3-pyridinyl)-4- morpholino-6-thieno[3,2-d]pyrimidinyl)acrylamide (Compound 14) 1 H NMR (600 MHz, DMSO- d 6 ) δ 9.14 (d, J = 1.8 Hz, 1H), 8.57 (dd, J = 8.6,2.1 Hz, 1H), 8.32 (t, J = 5.5 Hz, 1H), 7.73 (d, J = 13.6 Hz, 2H), 6.91 (d, J = 8.6 Hz, 1H), 6.75 (d, J = 15.7 Hz, 1H), 4.77 (t, J = 5.0 Hz, 1H), 4.14 –3.96 (m, 4H), 3.93 (s, 3H), 3.86 – 3.73 (m, 4H), 3.58 – 3.42 (m, 2H), 3.30 –3.20 (m, 2H); MS (ESI), m / z: 442.20[M+H] + .

[0115] Example 14: Synthesis of (2-hydroxyethyl)-3-(2-(6-methoxy-3-pyridinyl)-4- morpholino-6-thieno[3,2-d]pyrimidinyl)acrylamide (Compound 14) E )- N Example 14: Synthesis of (2-hydroxyethyl)-3-(2-(6-methoxy-3-pyridinyl)-4- morpholino-6-thieno[3,2-d]pyrimidinyl)acrylamide (Compound 14)

[0116]

[0117] In Example 1, step 8, 4-fluoroaniline was replaced with 3-amino-1-adamantanol, and the remaining steps were the same as in Example 1, to obtain compound 15. 1 H NMR (600 MHz, DMSO- d 6 ) δ 9.14 (s, 1H), 8.57 (d, J = 7.6 Hz, 1H), 7.70 (dd, J = 86.2, 47.6 Hz, 3H), 6.91 (d, J = 8.2 Hz, 1H), 6.74 (d, J =15.5 Hz, 1H), 4.51 (s, 1H), 4.14 – 3.71 (m, 11H), 2.14 (s, 2H), 2.02 – 1.72(m, 6H), 1.62 – 1.37 (m, 6H); MS (ESI), m / z: 548.20[M+H] + .

[0118] Example 16: ( E )-3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)- N Synthesis of 3-(3-morpholinopropyl)acrylamide (compound 16)

[0119]

[0120] In Example 1, step 8, 4-fluoroaniline was replaced with 3-(4-morpholino)-1-propylamine, and the remaining steps were the same as in Example 1, to obtain compound 16. 1 H NMR (600 MHz, DMSO- d 6 ) δ 9.13 (s, 1H), 8.55 (d, J = 7.7 Hz,1H), 8.25 (s, 1H), 8.05 – 7.56 (m, 2H), 6.90 (d, J = 8.3 Hz, 1H), 6.67 (d, J = 15.5 Hz, 1H), 4.04 – 3.95 (m, 4H), 3.93 (s, 3H), 3.87 – 3.75 (m, 4H), 3.69– 3.54 (m, 4H), 3.23 (d, J= 4.9 Hz, 2H), 2.46 – 2.22 (m, 6H), 1.77 – 1.53(m, 2H); MS (ESI), m / z: 525.20[M+H] + .

[0121] Example 17: ( E Synthesis of ethyl piperidine-4-carboxylate (compound 17)

[0122]

[0123] By replacing 4-fluoroaniline in step 8 of Example 1 with ethyl 4-piperidinecarboxylate, and following the same steps as in Example 1, compound 17 was obtained. 1 H NMR (600 MHz, DMSO- d 6 ) δ 9.15 (s, 1H), 8.70 – 8.47 (m, 1H), 7.85 (s, 1H), 7.77 (d, J = 15.2 Hz, 1H), 7.29 (d, J = 15.2 Hz, 1H), 6.92 (d, J =8.7 Hz, 1H), 4.32 (d, J = 12.4 Hz, 1H), 4.16 (d, J = 12.8 Hz, 1H), 4.09 (q, J = 7.0 Hz, 2H), 4.04 – 3.98 (m, 4H), 3.94 (s, 3H), 3.86 – 3.77 (m, 4H), 3.24(t, J = 12.0 Hz, 1H), 2.89 (t, J = 11.6 Hz, 1H), 2.71 – 2.61 (m, 1H), 1.91(s, 2H), 1.51 (dd, J = 49.7, 10.4 Hz, 2H), 1.20 (t, J = 7.1 Hz, 3H); MS(ESI), m / z: 538.20[M+H] + .

[0124] Example 18: ( ESynthesis of 3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acryloyl)glutamate diethyl ester (compound 18)

[0125]

[0126] By replacing 4-fluoroaniline in step 8 of Example 1 with diethyl glutamate hydrochloride, and following the same steps as in Example 1, compound 18 was obtained. 1 H NMR (600 MHz, DMSO- d 6 ) δ 9.16 (s, 1H), 8.67 (d, J = 7.5 Hz, 1H), 8.59 (dd, J = 8.7, 1.7 Hz, 1H), 7.92 – 7.66 (m, 2H), 6.93 (d, J = 8.7Hz, 1H), 6.79 (d, J = 15.7 Hz, 1H), 4.49 – 4.36 (m, 1H), 4.13 (q, J = 7.1 Hz, 2H), 4.07 (q, J = 14.0, 7.0 Hz, 2H), 4.04 – 3.97 (m, 4H), 3.94 (s, 3H), 3.85 – 3.78 (m, 4H), 2.42 (d, J = 10.0 Hz, 2H), 2.12 – 2.00 (m, 1H), 1.97 – 1.85(m, 1H), 1.29 – 1.13 (m, 6H); MS (ESI), m / z: 584.20[M+H] + .

[0127] Example 19: ( E Synthesis of ethyl glycine ester (compound 19) of 3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acryloyl)glycine

[0128]

[0129] Replacing 4-fluoroaniline in step 8 of Example 1 with glycine ethyl ester, and following the same steps as in Example 1, compound 19 was obtained. MS (ESI), m / z: 484.20 [M+H] + .

[0130] Example 20: Synthesis of (E)-4-(3-(2-(6-methoxy-3-pyridinyl)-4-morpholinyl-6- thieno[3,2-d]pyrimidinyl)acrylamidyl)-1-piperidinyl (Compound 20) E

[0131]

[0132] Replace 4-fluoroaniline in step 8 of Example 1 with proline methyl ester hydrochloride, and the remaining steps are the same as Example 1 to give Compound 20. MS (ESI), m / z: 510.20 [M+H] + .

[0133] Example 21: Synthesis of (E)-4-(3-(2-(6-methoxy-3-pyridinyl)-4-morpholinyl-6- thieno[3,2-d]pyrimidinyl)acrylamidyl)-1-piperidinyl (Compound 21) E N - methyl glycine ethyl ester (Compound 21)

[0134]

[0135] Replace 4-fluoroaniline in step 8 of Example 1 with 2-(methylamino)acetic acid ethyl ester hydrochloride, and the remaining steps are the same as Example 1 to give Compound 21. MS (ESI), m / z: 498.20 [M+H] + .

[0136] Example 22: Synthesis of (E)-1-(4-dimethylamino-1-piperidinyl)-3-(2-(6-methoxy-3- pyridinyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-2-propen-1-one (Compound 22) E

[0137]

[0138] Replace 4-fluoroaniline in step 8 of Example 1 with 4-dimethylamino-piperidine, and the remaining steps are the same as Example 1 to give Compound 22. MS (ESI), m / z: 509.20 [M+H] + .

[0139] Example 23: Synthesis of (E)-4-(3-(2-(6-methoxy-3-pyridinyl)-4-morpholinyl-6- thieno[3,2-d]pyrimidinyl)acrylamidyl)butyric acid methyl ester (Compound 23)

[0140]

[0141] ​​​In Example 1, step 8, 4-fluoroaniline was replaced with methyl 4-aminobutyrate hydrochloride, and the remaining steps were the same as in Example 1, to obtain compound 23. 1 H NMR (600 MHz, DMSO- d 6 ) δ 9.13 (d, J = 2.0 Hz, 1H), 8.56 (dd, J = 8.6, 2.2 Hz, 1H), 8.31 (t, J = 5.5 Hz, 1H), 7.86 – 7.56 (m, 2H), 6.91 (d, J = 8.6 Hz, 1H), 6.67 (d, J = 15.7 Hz, 1H), 4.03 – 3.96 (m, 4H), 3.93 (s,3H), 3.86 – 3.76 (m, 4H), 3.60 (s, 3H), 3.26 – 3.17 (m, 2H), 2.37 (t, J = 7.4Hz, 2H), 1.82 – 1.66 (m, 2H); MS (ESI), m / z: 498.20[M+H] + .

[0142] Example 24: ( E Synthesis of 1-(1,1-dioxothiomorpholine)-3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-2-propen-1-one (compound 24)

[0143]

[0144] Replacing 4-fluoroaniline in step 8 of Example 1 with thiomorphon dioxide, and following the same steps as in Example 1, compound 24 was obtained. MS (ESI), m / z: 516.20 [M+H] + .

[0145] Example 25: ( E Synthesis of 1-(3-isopropoxy-1-azacyclobutane)-3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-2-propen-1-one (compound 25)

[0146]

[0147] In Example 1, step 8, 4-fluoroaniline was replaced with 3-isopropoxyazinobutane hydrochloride, and the remaining steps were the same as in Example 1, yielding compound 25. MS (ESI), m / z: 496.20 [M+H] + .

[0148] Example 26: ( E Synthesis of 2-(2R,6S)-2,6-dimethylmorpholino)-3-(2-(6-methoxy-3-pyridyl)-4-morpholino-6-thieno[3,2-d]pyrimidinyl)-2-propen-1-one (compound 26)

[0149]

[0150] Replacing 4-fluoroaniline in step 8 of Example 1 with cis-2,6-dimethylmorpholine, and following the same steps as in Example 1, compound 26 was obtained. MS (ESI), m / z: 496.20 [M+H] + .

[0151] Example 27: ( E Synthesis of 1-(3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acryloyl)pyrrolidine-2-carboxamide (compound 27)

[0152]

[0153] Replacing 4-fluoroaniline in step 8 of Example 1 with proline amide, and following the same steps as in Example 1, compound 27 was obtained. MS (ESI), m / z: 495.10 [M+H] + , 517.20 [M+Na] + .

[0154] Example 28: ( E )- N Synthesis of 2-(2-amino-2-oxoethyl)-3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acrylamide (compound 28)

[0155]

[0156] Replacing 4-fluoroaniline in step 8 of Example 1 with 2-aminoacetamide, and following the same steps as in Example 1, compound 28 was obtained. MS (ESI), m / z: 455.20 [M+H] + .

[0157] Example 29: ( E )-N Synthesis of (1-amino-l-oxo-2-propyl)-3-(2-(6-methoxy-3-pyridinyl)-4- morpholino-6-thieno[3,2-d]pyrimidinyl)acrylamide (Compound 29)

[0158]

[0159] Replace 4-fluoroaniline in Example 1, Step 8 with 2-aminopropanamide and follow the remaining steps of Example 1 to obtain Compound 29. 1 H NMR (600 MHz, DMSO- d 6 ) δ 9.14 (s, 1H), 8.57 (d, J = 8.6 Hz, 1H),8.41 (d, J = 7.5 Hz, 1H), 7.89 – 7.65 (m, 2H), 7.44 (s, 1H), 7.03 (s, 1H),6.91 (d, J = 8.7 Hz, 1H), 6.85 (d, J = 15.7 Hz, 1H), 4.71 – 4.24 (m, 1H),4.08 – 3.97 (m, 4H), 3.93 (s, 3H), 3.87 – 3.76 (m, 4H), 1.27 (d, J = 7.0 Hz,3H); MS (ESI), m / z: 469.20 [M+H] + , 491.10 [M+Na] + .

[0160] Example 30: (S)-3-(2-(6-methoxy-3-pyridinyl)-4-morpholino-6- thieno[3,2-d]pyrimidinyl)-(1-methylsulfonyl-4-piperidinyl)acrylamide (Compound 30) E N Synthesis of (1-amino-l-oxo-2-propyl)-3-(2-(6-methoxy-3-pyridinyl)-4- morpholino-6-thieno[3,2-d]pyrimidinyl)acrylamide (Compound 29)

[0161]

[0162] Replace 4-fluoroaniline in Example 1, Step 8 with 1-methylsulfonyl-4- aminopiperidine and follow the remaining steps of Example 1 to obtain Compound 30. 1 H NMR (600 MHz, CD3COOD) δ 9.26 (s, 1H), 8.64 (d, J ​= 6.7 Hz,1H), 8.05 – 7.65 (m, 2H), 7.00 (d, J = 8.7 Hz, 1H), 6.86 (d, J = 15.6 Hz,1H), 4.46 – 4.17 (m, 4H), 4.14 – 3.89 (m, 8H), 3.78 (d, J = 11.4 Hz, 2H),3.15 – 2.69 (m, 5H), 2.15 – 2.09 (m, 2H), 1.81 – 1.55 (m, 2H); MS (ESI), m / z:559.20[M+H] + .

[0163] Example 31: ( E Synthesis of 1-(3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acryloyl)piperidine-4-carboxamide (compound 31)

[0164]

[0165] By replacing 4-fluoroaniline in step 8 of Example 1 with piperidine-4-carboxamide, and following the same steps as in Example 1, compound 31 was obtained. 1 H NMR (600 MHz, CD3COOD) δ 9.24 (s, 1H), 8.63 (d, J = 8.0 Hz, 1H), 7.93 (d, J = 15.3 Hz, 1H), 7.82 (s, 1H), 7.24 (d, J = 15.3 Hz, 1H), 6.98 (d, J = 8.7 Hz, 1H), 4.69 (d, J = 12.7 Hz, 1H), 4.31 (d, J = 13.3 Hz, 1H), 4.26 –4.16 (m, 4H), 4.05 (s, 3H), 4.03 – 3.91 (m, 4H), 3.32 (t, J= 12.3 Hz, 1H), 3.00 – 2.85 (m, 1H), 2.78 – 2.56 (m, 1H), 2.05 – 1.97 (m, 2H), 1.89 – 1.63(m, 2H); MS (ESI), m / z: 509.20 [M+H] + , 531.10 [M+Na] + .

[0166] Example 32: Synthesis of (2,6-dioxo-3-piperidyl)-3-(2-(6-methoxy-3-pyridyl)-4- morpholinyl-6-thieno[3,2-d]pyrimidinyl)acrylamide (Compound 32) E N

[0167]

[0168] Replace 4-fluoroaniline in Example 1, Step 8 with 3-aminopiperidine-2,6-dione hydrochloride and follow the rest of the procedure as in Example 1 to obtain Compound 32. 1 H NMR (600 MHz, DMSO- d 6 ) δ 9.13 (s, 1H), 8.55 (d, J =7.7 Hz, 1H), 8.25 (s, 1H), 8.05 – 7.56 (m, 2H), 6.90 (d, J = 8.3 Hz, 1H),6.67 (d, J = 15.5 Hz, 1H), 4.04 – 3.95 (m, 4H), 3.93 (s, 3H), 3.87 – 3.75 (m,4H), 3.69 – 3.54 (m, 4H), 3.23 (d, J = 4.9 Hz, 2H), 2.46 – 2.22 (m, 6H), 1.77– 1.53 (m, 2H); MS (ESI), m / z: 509.10[M+H] + .

[0169] Example 33: Synthesis of (2-hydroxyethyl)-3-(2-(2-amino-5-pyrimidinyl)-4- morpholinyl-6-thieno[3,2-d]pyrimidinyl)acrylamide (Compound 33) E N

[0170] ​​​​

[0171] In Example 1, step 5, 6-methoxypyridine-3-boronic acid was replaced with 2-aminopyrimidine-5-boronic acid or pinacol ester of 2-aminopyrimidine-5-boronic acid; in Example 1, step 8, 4-fluoroaniline was replaced with ethanolamine; and the remaining steps were the same as in Example 1, yielding compound 33. MS (ESI), m / z: 428.10 [M+H] + 550.20 [M+Na] + .

[0172] Example 34: ( E Synthesis of ethyl glycine ester (compound 34) of 3-(2-(2-amino-5-pyrimidinyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acryloyl)glycine

[0173]

[0174] In Example 1, step 5, 6-methoxypyridine-3-boronic acid was replaced with 2-aminopyrimidine-5-boronic acid or pinacol ester of 2-aminopyrimidine-5-boronic acid; in Example 1, step 8, 4-fluoroaniline was replaced with glycine ethyl ester; the remaining steps were the same as in Example 1, yielding compound 34. MS (ESI), m / z: 470.10 [M+H] + .

[0175] Example 35: ( E )- N Synthesis of -(1-adamantyl)-3-(2-(2-amino-5-pyrimidinyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acrylamide (compound 35)

[0176]

[0177] In Example 1, step 5, 6-methoxypyridine-3-boronic acid was replaced with 2-aminopyrimidine-5-boronic acid or pinacol ester of 2-aminopyrimidine-5-boronic acid; in Example 1, step 8, 4-fluoroaniline was replaced with adamantaneamine; and the remaining steps were the same as in Example 1, yielding compound 35. MS (ESI), m / z: 518.20 [M+H] + .

[0178] Example 36: ( E )-3-(2-(2-amino-5-pyrimidinyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)- N Synthesis of 2-(methoxyethyl)acrylamide (compound 36)

[0179]

[0180] Replace 6-methoxypyridine-3-boronic acid in step 5 of example 1 with 2- aminopyrimidine-5-boronic acid or 2-aminopyrimidine-5-boronic acid pinacol ester, replace 4-fluoroaniline in step 8 of example 1 with 2-methoxyethylamine, and follow the remaining steps of example 1 to obtain compound 36. MS (ESI), m / z: 442.20 [M+H] + .

[0181] Example 37: E )-3-(2-(2-amino-5-pyrimidinyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-(4- methylsulfonyl-1-piperazinyl)-2-propen-1-one N Synthesis of (1-methylsulfonyl-4-piperidinyl)acrylamide (compound 37)

[0182]

[0183] Replace 6-methoxypyridine-3-boronic acid in step 5 of example 1 with 2- aminopyrimidine-5-boronic acid or 2-aminopyrimidine-5-boronic acid pinacol ester, replace 4-fluoroaniline in step 8 of example 1 with 1-methylsulfonyl-4- aminopiperidine, and follow the remaining steps of example 1 to obtain compound 37. MS (ESI), m / z: 545.20 [M+H] + .

[0184] Example 38: E )-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-(4- methylsulfonyl-1-piperazinyl)-2-propen-1-one

[0185] Synthesis of (1-methylsulfonyl-4-piperidinyl)acrylamide (compound 37)

[0186]

[0187] The 2-chloro-4-morpholinylthieno[3,2-d]pyrimidine-6-carboxaldehyde used in the synthesis of compound 38 was derived from the preparation method of example 1 and is therefore not described again.

[0188] Step 1: Synthesis of 4-morpholinyl-2-(1-(2-tetrahydropyranyl)-4-indazolyl)thieno[3,2- d]pyrimidine-6-carboxaldehyde

[0189]

[0190] 2-Chloro-4-morpholinylthiopheno[3,2-d]pyrimidine-6-carboxaldehyde (15 g, 52.87 mmol) was mixed with 1-THP-4-indazoleboronic acid pinacol ester (26.03 g, 79.30 mmol) and potassium carbonate (21.30 g, 0.158.60 mmol). 250 mL of 1,4-dioxane and 25 mL of water were added. The solution was slowly purged with nitrogen at room temperature for 30 min. Then, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (1.17 g, 1.60 mmol) was added. The reaction was carried out at 80°C for 6 h under nitrogen protection until completion. The mixture was filtered, the filter cake was washed with dichloromethane, and the filtrate was evaporated to dryness under reduced pressure to obtain the crude product. The crude product was pulped with anhydrous methanol and dried to obtain 18.28 g of a yellow solid 4-morpholino-2-(1-(2-tetrahydropyrano)-4-indazolyl)thieno[3,2-d]pyrimidine-6-carboxaldehyde. 1 H NMR (600 MHz, CDCl3) δ 10.20 (s, 1H), 8.94 (s, 1H), 8.28 (d, J = 7.2 Hz, 1H),8.14 (s, 1H), 7.73 (d, J = 8.3 Hz, 1H), 7.51 (t, J = 7.7 Hz, 1H), 5.79 (d, J = 8.9 Hz, 1H), 4.19 – 4.09 (m, 4H), 4.05 (d, J = 11.4 Hz, 1H), 3.97 – 3.87(m, 4H), 3.78 (t, J = 10.3 Hz, 1H), 2.76 – 2.53 (m, 1H), 2.34 – 2.05 (m, 2H), 1.91 – 1.62 (m, 3H); MS (ESI) m / z: 450.10 [M+H] + .

[0191] Step 2: ( E Synthesis of ethyl 3-(4-morpholino-2-(1-(2-tetrahydropyrano)-4-indazolyl)-6-thieno[3,2-d]pyrimidinyl)acrylate

[0192]

[0193] 12.00 g (26.69 mmol) of 4-morpholino-2-(1-(2-tetrahydropyrano)-4-inzolyl)thieno[3,2-d]pyrimidin-6-carboxaldehyde, ethoxyformylmethylenetriphenylphosphine (13.95 g, 40.04 mmol), and 120 mL of toluene were added to a reaction flask, and the mixture was refluxed for approximately 4 h until the reaction was complete. The reaction system was filtered while hot, and the filter cake was washed with an appropriate amount of toluene. The filter cake was then dried to obtain a yellow solid. E 10.80 g of ethyl acrylate of 3-(4-morpholino-2-(1-(2-tetrahydropyrano)-4-indazolyl)-6-thieno[3,2-d]pyrimidinyl) acrylate. 1 H NMR (600 MHz, CDCl3) δ 8.93 (s, 1H), 8.27 (d, J = 7.2 Hz, 1H), 7.87 (d, J = 15.8 Hz, 1H), 7.71 (d, J = 8.3 Hz, 1H), 7.64 (s,1H), 7.50 (t, J = 7.8 Hz, 1H), 6.46 (d, J = 15.8 Hz, 1H), 5.96 – 5.57 (m,1H), 4.30 (q, J = 7.1 Hz, 2H), 4.16 – 3.99 (m, 5H), 3.98 – 3.87 (m, 4H), 3.77(t, J = 10.5 Hz, 1H), 2.75 – 2.53 (m, 1H), 2.27 – 2.04 (m, 2H), 1.84 – 1.64(m, 3H), 1.36 (t, J = 7.1 Hz, 3H); MS (ESI), m / z: 520.10 [M+H] + .

[0194] Step 3: ( E Synthesis of 3-(4-morpholino-2-(1-(2-tetrahydropyrano)-4-indazolyl)-6-thieno[3,2-d]pyrimidinyl)acrylic acid

[0195]

[0196] Will( E) -3-(4-morpholinyl-2-(1-(2-tetrahydropyranyl)-4-indazolyl)-6- thieno[3,2-d]pyrimidinyl)acrylic acid ethyl ester (10.00 g, 19.24 mmol), sodium hydroxide (2.31 g, 57.73 mmol), ethanol 80 mL and water 20 mL were added into a reaction flask, stirred at 60 °C for 5 h. Most of the solvent was removed by evaporation under reduced pressure, the residue was added into 80 mL water, pH was adjusted to about 3 with concentrated hydrochloric acid solution, a large amount of yellow solid was precipitated, suction filtration, the filter cake was dried to give yellow solid (7.38 g). MS (ESI), m / z: 492.20 [M+H] E ) -3-(4-morpholinyl-2-(1-(2-tetrahydropyranyl)-4-indazolyl)-6- thieno[3,2-d]pyrimidinyl)acrylic acid ethyl ester (10.00 g, 19.24 mmol), sodium hydroxide (2.31 g, 57.73 mmol), ethanol 80 mL and water 20 mL were added into a reaction flask, stirred at 60 °C for 5 h. Most of the solvent was removed by evaporation under reduced pressure, the residue was added into 80 mL water, pH was adjusted to about 3 with concentrated hydrochloric acid solution, a large amount of yellow solid was precipitated, suction filtration, the filter cake was dried to give yellow solid (7.38 g). MS (ESI), m / z: 492.20 [M+H] + .

[0197] Step 4: E Synthesis of -1-(4-methanesulfonyl-1-piperazinyl)-3-(4-morpholinyl-2-(1-(2- tetrahydropyranyl)-4-indazolyl)-6-thieno[3,2-d]pyrimidinyl)-2-propen-1-one

[0198]

[0199] Into a reaction flask was added E ) -3-(4-morpholinyl-2-(1-(2-tetrahydropyranyl)-4-indazolyl)-6- thieno[3,2-d]pyrimidinyl)acrylic acid (300.00 mg, 0.61 mmol), N - methanesulfonylpiperazine (120.30 mg, 0.73 mmol), 2-(7-azabenzotriazole)- N,N,N',N' - hexafluorophosphate (347.40 mg, 0.92 mmol), triethylamine (185.30 mg, 1.83 mmol) and N,N - dimethylformamide 8 mL, stirred at room temperature for 20 h. The reaction was completed. The reaction solution was poured into 30 mL of 10% aqueous sodium carbonate solution, and a solid was precipitated, filtered, the filter cake was washed with water three times, the filter cake was slurried with methanol twice, and dried to give a yellow solid (0.29 g). MS (ESI), m / z: 638.20 [M+H] E ) -3-(4-morpholinyl-2-(1-(2-tetrahydropyranyl)-4-indazolyl)-6- thieno[3,2-d]pyrimidinyl)acrylic acid ethyl ester (10.00 g, 19.24 mmol), sodium hydroxide (2.31 g, 57.73 mmol), ethanol 80 mL and water 20 mL were added into a reaction flask, stirred at 60 °C for 5 h. Most of the solvent was removed by evaporation under reduced pressure, the residue was added into 80 mL water, pH was adjusted to about 3 with concentrated hydrochloric acid solution, a large amount of yellow solid was precipitated, suction filtration, the filter cake was dried to give yellow solid (7.38 g). MS (ESI), m / z: 492.20 [M+H] + .

[0200] Step 5: ESynthesis of 38-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-(4-methanesulfonyl-1-piperazinyl)-2-propen-1-one

[0201]

[0202] Will( E 1-(4-methanesulfonyl-1-piperazinyl)-3-(4-morpholinyl-2-(1-(2-tetrahydropyranyl)-4-indazolyl)-6-thieno[3,2-d]pyrimidinyl)-2-propen-1-one (0.25 g, 0.39 mmol) was added to 5 mL of 1,4-dioxane-saturated hydrogen chloride solution, and the reaction was carried out by stirring at 80°C for 2 h. After the reaction was completed, most of the solvent was removed by vacuum evaporation, and the residue was added to 10 mL of saturated sodium carbonate solution, precipitating a yellow solid. The solid was filtered, and the filter cake was slurried twice with methanol and dried to obtain a yellow solid. E )-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-(4-methanesulfonyl-1-piperazinyl)-2-propen-1-one 0.15 g. 1 H NMR (600 MHz, DMSO- d 6 ) δ 13.22 (s, 1H), 8.88 (s, 1H),8.23 (d, J = 7.2 Hz, 1H), 7.99 (s, 1H), 7.85 (d, J = 15.1 Hz, 1H), 7.69 (d, J = 8.2 Hz, 1H), 7.48 (t, J = 7.7 Hz, 1H), 7.34 (d, J = 15.2 Hz, 1H), 4.11 –3.97 (m, 4H), 3.91 – 3.79 (m, 6H), 3.71 (br, 2H), 3.25 – 3.11 (m, 4H), 2.92(s, 3H); MS (ESI) m / z: 554.20 [M+H] + .

[0203] By replacing appropriate raw materials and reagents, and following the preparation method of Example 38, Examples 39-60 (compounds 39-60) were finally obtained.

[0204] Example 39: ( E)-3-(2-(4-azaindol-1-yl)-4-morpholin-6-yl-thieno[3,2-d]pyrimidin-5-yl)-N- methyl-acrylamide N Synthesis of phenylacrylamide (compound 39)

[0205]

[0206] Replace morpholin-4-yl with piperidin-4-yl in step 4 of example 38 N - Replace morpholin-4-yl with piperidin-4-yl in step 4 of example 38 1 H NMR (600 MHz, DMSO- d 6 ) δ 13.23 (s, 1H), 10.37 (s, 1H), 8.89 (s, 1H),8.24 (d, J = 7.2 Hz, 1H), 8.07 – 7.86 (m, 2H), 7.70 (dd, J = 13.4, 8.1 Hz,3H), 7.48 (t, J = 7.7 Hz, 1H), 7.36 (t, J = 7.9 Hz, 2H), 7.10 (t, J = 7.4 Hz,1H), 6.93 (d, J = 15.6 Hz, 1H), 4.11 – 4.01 (m, 4H), 3.94 – 3.84 (m, 4H); MS(ESI), m / z: 483.10[M+H] + .

[0207] Example 40: ( E )-3-(2-(4-azaindol-1-yl)-4-morpholin-6-yl-thieno[3,2-d]pyrimidin-5-yl)-N- methyl-acrylamide N Synthesis of methylacrylamide (compound 40)

[0208]

[0209] Replace morpholin-4-yl with piperidin-4-yl in step 4 of example 38 N - Replace morpholin-4-yl with piperidin-4-yl in step 4 of example 38 + .

[0210] Example 41: ( E )-3-(2-(4-azaindol-1-yl)-4-morpholin-6-yl-thieno[3,2-d]pyrimidin-5-yl)-N- methyl-acrylamideN Synthesis of 2-(2-dimethylaminoethyl)acrylamide (compound 41)

[0211]

[0212] In step 4 of embodiment 38 N -Methanesulfonylpiperazine is replaced with N,N -Dimethylethylenediamine, and the remaining steps are the same as in Example 38, to obtain compound 41. 1 H NMR (600 MHz, DMSO- d 6 ) δ 13.30 (s, 1H), 8.88 (s, 1H),8.31 – 8.16 (m, 2H), 7.86 (s, 1H), 7.74 (d, J = 15.6 Hz, 1H), 7.69 (d, J =8.2 Hz, 1H), 7.47 (t, J = 7.7 Hz, 1H), 6.80 (d, J = 15.6 Hz, 1H), 4.10 – 3.96(m, 4H), 3.91 – 3.79 (m, 4H), 3.32 – 3.28 (m, 2H), 2.35 (t, J = 6.5 Hz, 2H),2.17 (s, 6H); MS (ESI), m / z: 478.20[M+H] + .

[0213] Example 42: ( E )-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)- N Synthesis of 2-(methoxyethyl)acrylamide (compound 42)

[0214]

[0215] In step 4 of embodiment 38 N The methanesulfonylpiperazine was replaced with 2-methoxyethylamine, and the remaining steps were the same as in Example 38, to obtain compound 42. 1 H NMR (600 MHz, DMSO- d 6 ) δ 13.24 (s, 1H), 8.89 (s, 1H), 8.39 (s, 1H), 8.23 ​​(d, J= 7.1 Hz, 1H), 7.85 (s, 1H), 7.76 (d, J = 15.6 Hz, 1H),7.69 (d, J = 8.1 Hz, 1H), 7.48 (t, J = 7.7 Hz, 1H), 6.78 (d, J = 15.6 Hz,1H), 4.02 (br, 4H), 3.85 (br, 4H), 3.49 – 3.37 (m, 4H), 3.29 (s, 3H); MS(ESI), m / z: 465.20[M+H] + .

[0216] Example 43: Synthesis of (S)-3-(2-(4-indazolyl)-4-morpholino-6- thieno[3,2-d]pyrimidinyl)-1-(4-methyl-1-piperazinyl)-2-propen-1-one (Compound 43) E

[0217]

[0218] Example 38, step 4 was replaced with N -methanesulfonylpiperazine was replaced with N -methylpiperazine and the remaining steps were the same as Example 38 to give Compound 43. 1 H NMR (600 MHz, DMSO- d 6 ) δ 13.22 (s, 1H), 8.88 (s, 1H), 8.23 (d, J = 7.2 Hz, 1H), 7.97 (s, 1H), 7.80 (d, J = 15.2 Hz, 1H), 7.68 (d, J = 8.2Hz, 1H), 7.48 (t, J = 7.7 Hz, 1H), 7.31 (d, J = 15.2 Hz, 1H), 4.14 – 3.96 (m,4H), 3.93 – 3.79 (m, 4H), 3.74 – 3.53 (m, 4H), 2.42 – 2.28 (m, 4H), 2.21 (s,3H); MS (ESI), m / z: 490.20[M+H] + .

[0219] Example 44:​E ) -3- (2- (4-indazolyl) -4-morpholinyl-6- thieno [3, 2-d] pyrimidinyl) -1- (1-pyrrolidinyl) -2-propen-1-one (Compound 44).

[0220]

[0221] The procedure of Example 38, Step 4 was followed using N - methanesulfonylpiperazine was replaced by thiomorpholine, and the remaining steps were the same as Example 38 to give Compound 44. MS (ESI), m / z: 493.10 [M+H] + .

[0222] Example 45: ( E ) -3- (2- (4-indazolyl) -4-morpholinyl-6- thieno [3, 2-d] pyrimidinyl) -1- (1-pyrrolidinyl) -2-propen-1-one (Compound 44).

[0223]

[0224] The procedure of Example 38, Step 4 was followed using N - methanesulfonylpiperazine was replaced by thiomorpholine, and the remaining steps were the same as Example 38 to give Compound 44. MS (ESI), m / z: 493.10 [M+H] 1 H NMR (600 MHz, DMSO- d 6 ) δ 13.21 (s, 1H), 8.88 (s, 1H), 8.23 (d, J = 7.2 Hz, 1H), 7.97 (s, 1H), 7.80 (d, J = 15.4 Hz, 1H), 7.68 (d, J = 8.2 Hz,1H), 7.48 (t, J = 7.7 Hz, 1H), 7.02 (d, J = 15.4 Hz, 1H), 4.16 – 3.97 (m,4H), 3.91 – 3.78 (m, 4H), 3.67 (t, J = 6.7 Hz, 2H), 3.43 (t, J = 6.8 Hz, 2H),2.08 – 1.89 (m, 2H), 1.87 – 1.69 (m, 2H).

[0225] Example 46: ( E)-3-(2-(4-Indazolyl)-4-morpholino-6-thieno[3,2-d]pyrimidinyl)- N Synthesis of (2-diethylaminoethyl)acrylamide (Compound 46)

[0226]

[0227] The compound of Example 38, Step 4 was used in place of the compound of Example 38, Step 1. N - methanesulfonylpiperazine was replaced with diethylamine and the remaining steps were the same as Example 38 to give Compound 46. 1 H NMR (600 MHz, DMSO- d 6 ) δ 13.23 (s, 1H), 8.89 (s, 1H), 8.21 (d, J = 36.3 Hz, 2H), 7.76 (dd, J = 56.8, 43.6 Hz, 3H), 7.48 (s, 1H), 6.78 (d, J =12.9 Hz, 1H), 4.30 – 3.96 (m, 4H), 3.96 – 3.67 (m, 4H), 3.60 – 3.09 (m, 6H),2.49 – 2.32 (m, 2H), 0.98 (s, 6H); MS (ESI), m / z: 506.20[M+H] + .

[0228] Example 47: ( E )-3-(2-(4-Indazolyl)-4-morpholino-6-thieno[3,2-d]pyrimidinyl)- N Synthesis of (4-methoxyphenyl)acrylamide (Compound 47)

[0229]

[0230] The compound of Example 38, Step 4 was used in place of the compound of Example 38, Step 1. N - methanesulfonylpiperazine was replaced with diethylamine and the remaining steps were the same as Example 38 to give Compound 46. 1 H NMR (600 MHz, DMSO- d 6 ) δ 13.22 (s, 1H), 10.23 (s, 1H), 8.89(s, 1H), 8.23 (d, J = 6.5 Hz, 1H), 7.99 – 7.84 (m, 2H), 7.69 (d, J= 7.8 Hz, 1H), 7.63 (d, J = 7.8 Hz, 2H), 7.48 (t, J = 7.0 Hz, 1H), 6.93 (d, J = 8.0 Hz, 2H), 6.88 (d, J = 15.5 Hz, 1H), 4.03 (br, 4H), 3.86 (br, 4H), 3.74 (s, 3H);MS (ESI), m / z: 513.20[M+H] + .

[0231] Example 48: ( E )-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)- N Synthesis of 3-(3-morpholinopropyl)acrylamide (compound 48)

[0232]

[0233] In step 4 of embodiment 38 N The methanesulfonylpiperazine was replaced with 3-(4-morpholino)-1-propane, and the remaining steps were the same as in Example 38, to obtain compound 48. 1 H NMR (600 MHz, DMSO- d 6 ) δ13.24 (s, 1H), 8.88 (s, 1H),8.43 – 8.08 (m, 2H), 7.86 (s, 1H), 7.75 (d, J = 15.5 Hz, 1H), 7.69 (d, J =8.1 Hz, 1H), 7.48 (t, J = 7.7 Hz, 1H), 6.72 (d, J = 15.6 Hz, 1H), 4.03 (br,4H), 3.85 (br, 4H), 3.58 (br, 4H), 3.26 – 3.18 (m, 2H), 2.42 – 2.26 (m, 6H),1.68 – 1.56 (m, 2H); MS (ESI), m / z: 534.10[M+H] + .

[0234] Example 49: ( E)-3-(2-(4-azaindolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)- N Synthesis of (2-hydroxyethyl)acrylamide (Compound 49)

[0235]

[0236] The compound from Example 38, Step 4 was used in place of N - methanesulfonylpiperazine was replaced with ethanolamine and the remaining steps were the same as Example 38 to give Compound 49. 1 H NMR (600 MHz, DMSO- d 6 ) δ 13.23 (s, 1H), 8.88 (s, 1H), 8.52 – 8.15(m, 2H), 8.10 – 7.62 (m, 3H), 7.48 (s, 1H), 6.79 (d, J = 15.4 Hz, 1H), 4.78(s, 1H), 4.02 (s, 4H), 3.85 (s, 4H), 3.49 (s, 2H), 3.28 (s, 2H); MS (ESI), m / z: 4451.10[M+H] + .

[0237] Example 50: ( E )-3-(2-(4-azaindolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)- N -(2-dimethylaminoethyl)- N Synthesis of (2-hydroxyethyl)acrylamide (Compound 49)

[0238]

[0239] The compound from Example 38, Step 4 was used in place of N - methanesulfonylpiperazine was replaced with N,N,N' - trimethylethylenediamine and the remaining steps were the same as Example 38 to give Compound 50. 1 H NMR (600 MHz, CD3COOD) δ 8.65 (s, 1H), 7.98 (d, J =7.2 Hz, 1H), 7.74 (d, J = 15.4 Hz, 1H), 7.71 – 7.62 (m, 2H), 7.41 (t, J = 7.8Hz, 1H), 6.99 (d, J= 15.4 Hz, 1H), 4.12 – 4.01 (m, 4H), 3.89 – 3.80 (m, 4H), 3.78 (t, J = 6.1 Hz, 2H), 3.32 (t, J = 6.1 Hz, 2H), 3.14 (s, 3H), 2.86 (s,6H); MS (ESI), m / z: 492.10[M+H] + .

[0240] Example 51: ( E Synthesis of 3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-(4-acetyl-1-piperazinyl)-2-propen-1-one (compound 51)

[0241]

[0242] In step 4 of embodiment 38 N -Methanesulfonylpiperazine is replaced with N - Acetylpiperazine, and the remaining steps are the same as in Example 38, to obtain compound 51. 1 H NMR (600 MHz, DMSO- d 6 ) δ 13.25 (s, 1H), 8.89 (s, 1H), 8.23(d, J = 7.0 Hz, 1H), 7.98 (s, 1H), 7.83 (d, J = 15.1 Hz, 1H), 7.69 (d, J =7.9 Hz, 1H), 7.48 (t, J = 7.5 Hz, 1H), 7.31 (d, J = 15.1 Hz, 1H), 4.20 – 3.96(m, 4H), 3.93 – 3.80 (m, 4H), 3.79 – 3.43 (m, 8H), 2.05 (s, 3H); MS (ESI), m / z: 518.20 [M+H] + .

[0243] Example 52: ( E Synthesis of )-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-((2R,6S)-2,6-dimethylmorpholinyl)-2-propen-1-one (compound 52)

[0244]

[0245] In step 4 of embodiment 38 N The methanesulfonylpiperazine was replaced with cis-2,6-dimethylmorpholine, and the remaining steps were the same as in Example 38, to obtain compound 52. 1 H NMR (600 MHz, DMSO- d 6 ) δ 13.28 (s, 1H), 8.89 (s, 1H),8.23 (d, J = 7.1 Hz, 1H), 7.98 (s, 1H), 7.83 (d, J = 15.1 Hz, 1H), 7.69 (d, J = 8.1 Hz, 1H), 7.48 (t, J = 7.7 Hz, 1H), 7.32 (d, J = 15.2 Hz, 1H), 4.38 (d, J = 13.0 Hz, 1H), 4.23 (d, J = 12.8 Hz, 1H), 4.18 – 3.97 (m, 4H), 3.95 – 3.74(m, 4H), 3.65 – 3.44 (m, 2H), 2.76 (t, J = 11.9 Hz, 1H), 2.37 (t, J = 11.9Hz, 1H), 1.33 – 1.00 (m, 6H); MS (ESI), m / z: 505.20[M+H] + .

[0246] Example 53: ( E Synthesis of )-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-(1,1-dioxothiomorpholinyl)-2-propen-1-one (compound 53)

[0247]

[0248] In step 4 of embodiment 38 N The methanesulfonylpiperazine was replaced with thiomorphon dioxide, and the remaining steps were the same as in Example 38, yielding compound 53. MS (ESI), m / z: 525.10 [M+H] + .

[0249] Example 54: ( E Synthesis of 3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-(4-(2-hydroxyethyl)-1-piperazinyl)-2-propen-1-one (compound 54)

[0250]

[0251] In step 4 of embodiment 38 N -Replace 1-(2-hydroxyethyl)piperazine with methanesulfonylpiperazine, and follow the same steps as in Example 38 to obtain compound 54. 1 H NMR (600 MHz, DMSO- d 6 ) δ 13.21 (s, 1H), 8.88 (s, 1H),8.22 (d, J = 6.2 Hz, 1H), 7.95 (s, 1H), 7.79 (d, J = 14.9 Hz, 1H), 7.68 (d, J = 7.0 Hz, 1H), 7.59 – 7.40 (m, 1H), 7.28 (d, J MS (ESI), m / z: 520.20[M+H] + .

[0252] Example 55: ( E Synthesis of 3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-(4-(2-methoxyethyl)-1-piperazinyl)-2-propen-1-one (compound 55)

[0253]

[0254] In step 4 of embodiment 38 N The methanesulfonylpiperazine was replaced with 1-(2-methoxyethyl)piperazine, and the remaining steps were the same as in Example 38, to obtain compound 55. 1 H NMR (600 MHz, DMSO- d 6) δ 13.24 (s, 1H), 8.89 (s,1H), 8.24 (br, 1H), 7.98 (s, 1H), 7.81 (d, J = 14.8 Hz, 1H), 7.68 (br, 1H),7.48 (br, 1H), 7.31 (d, J = 14.8 Hz, 1H), 4.18 – 3.95 (m, 4H), 3.94 – 3.79(m, 4H), 3.77 – 3.53 (m, 4H), 3.51 – 3.42 (m, 2H), 3.25 (s, 3H), 2.50 – 2.22(m, 6H); MS (ESI), m / z:534.20[M+H] + , 556.20 [M+Na] + .

[0255] Example 56: ( E )-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)- N Synthesis of 1-(1-methylsulfonyl-4-piperidinyl)acrylamide (compound 56)

[0256]

[0257] In step 4 of embodiment 38 N -Replace 1-methylsulfonylpiperazine with 1-methylsulfonyl-4-aminopiperidine, and follow the same steps as in Example 38 to obtain compound 56. 1 H NMR (600 MHz, DMSO- d 6 ) δ 13.26 (s, 1H), 8.86 (s, 1H),8.38 (d, J = 7.0 Hz, 1H), 8.22 (d, J = 7.0 Hz, 1H), 7.87 (s, 1H), 7.78 (d, J = 15.6 Hz, 1H), 7.70 (d, J = 8.0 Hz, 1H), 7.48 (t, J = 7.5 Hz, 1H), 6.73 (d, J= 15.6 Hz, 1H), 4.23 – 3.99 (m, 4H), 3.96 – 3.76 (m, 5H), 3.70 – 3.55 (m,2H), 3.13 – 2.81 (m, 5H), 2.15 – 1.79 (m, 2H), 1.68 – 1.31 (m, 2H); MS (ESI),m / z: 568.20[M+H] + 590.20 [M+Na] + .

[0258] Example 57: ( E Synthesis of 3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-(4-fluoro-1-piperidinyl)-2-propen-1-one (compound 57)

[0259]

[0260] In step 4 of embodiment 38 N The methanesulfonylpiperazine was replaced with 4-fluoropiperidine, and the remaining steps were the same as in Example 38, yielding compound 57. MS (ESI), m / z: 493.20 [M+H] + .

[0261] Example 58: ( E Synthesis of 3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-(4-methyl-1-piperidinyl)-2-propen-1-one (compound 58)

[0262]

[0263] In step 4 of embodiment 38 N The methanesulfonylpiperazine was replaced with 4-methylpiperidine, and the remaining steps were the same as in Example 38, yielding compound 58. MS (ESI), m / z: 489.20 [M+H] + .

[0264] Example 59: ( E Synthesis of 3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-(4-methoxy-1-piperidinyl)-2-propen-1-one (compound 59)

[0265]

[0266] In step 4 of embodiment 38 NThe methanesulfonylpiperazine was replaced with 4-methoxypiperidine, and the remaining steps were the same as in Example 38, yielding compound 59. MS (ESI), m / z: 505.20 [M+H] + .

[0267] Example 60: ( E )-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)- N -(2-Methoxyethyl)- N Synthesis of -methacrylamide (compound 60)

[0268]

[0269] In step 4 of embodiment 38 N -Methanesulfonylpiperazine is replaced with 2-methoxy- N -Methylethylamine, and the remaining steps are the same as in Example 38, to obtain compound 60. 1 H NMR (600 MHz, DMSO- d 6 ) δ 13.25 (s, 1H), 8.90 (s, 1H),8.23 (d, J = 3.7 Hz, 1H), 7.95 (s, 1H), 7.88 – 7.61 (m, 2H), 7.48 (s, 1H),7.21 (t, J = 14.2 Hz, 1H), 4.30 – 3.94 (m, 4H), 3.94 – 3.77 (m, 4H), 3.71(br, 1H), 3.65 – 3.44 (m, 3H), 3.28 (s, 3H), 3.08 (d, J = 131.4 Hz, 3H); MS(ESI), m / z: 479.20[M+H] + , 501.20 [M+Na] + .

[0270] Example 61: In vitro inhibition of tumor cell activity

[0271] Partial synthesis of novel 4-morpholinylthiopheno[3,2-d]pyrimidine compounds containing acrylamide structures according to the general formula (I) of this invention was screened in vitro for their inhibitory activity against human breast cancer cells MCF-7 and MDA-MB-231, human prostate cancer cells 22RV1, human colon cancer cells HCT116, human lung cancer cells A549, and human astrocytoma cells U87MG.

[0272] (1) After the cells were recovered and subcultured 2-3 times stably, they were digested from the bottom of the culture bottle with trypsin solution (0.25%). After the cell digestion solution was poured into a centrifuge tube, culture solution was added to terminate the digestion. The centrifuge tube was centrifuged at 800 r / min for 10 min, and after the supernatant was discarded, 5 mL of culture solution was added, the cells were mixed by blowing, 10 μL of the cell suspension was taken and added to a cell counting plate for counting, and the cell concentration was adjusted to 10 4 cells / well. Except that 100 μL of cell suspension was added to the 96-well plate except for the blank well in A1, the rest were added. The 96-well plate was placed in an incubator for 24 h.

[0273] (2) The test sample was dissolved with 50 μL of dimethyl sulfoxide, and then an appropriate amount of culture solution was added to dissolve the sample into a 2 mg / mL drug solution, and then the sample was diluted into 20, 4, 0.8, 0.16, 0.032 μg / mL in a 24-well plate.

[0274] Three wells were added for each concentration, and the cell growth in the surrounding two rows and two columns was greatly affected by the environment, and only blank cell wells were used. The 96-well plate was placed in an incubator for 72 h.

[0275] (3) The drug-containing culture solution in the 96-well plate was discarded, and the cells were washed twice with phosphate buffer solution (PBS), and 100 μL of MTT (tetrazolium) (0.5 mg / mL) was added to each well. After being placed in an incubator for 4 h, the MTT solution was discarded, and 100 μL of dimethyl sulfoxide was added. The survival cells were fully dissolved with MTT reaction product formazan on a magnetic shaker, and the results were measured in an enzyme marker. The IC 50 values of the drugs were calculated by Bliss method. The activity results of the in vitro anti-proliferative activity of the compounds on human breast cancer cells MCF-7, MDA-MB-231, human prostate cancer cells 22RV1, human colon cancer cells HCT116, human lung cancer cells A549 and human brain astrocytoma cells U87MG are shown in Table 1, and in Table 1, IC 50 ≦1.0 μM is represented by A, 1.0 μM﹤IC 50 ≦10.0 μM is represented by B, 10.0 μM﹤IC 50 ≦100.0 μM is represented by C, IC 50 >100.0 μM is represented by D.

[0276] Table 1 Inhibitory activity of compounds on MCF-7, MDA-MB-231, 22RV1, HCT116, A549 and U87MG tumor cell lines

[0277]

[0278] Example 62 Compound PI3K activity test

[0279] The in vitro PI3Kα enzyme inhibitory activity of the compounds prepared in the examples was tested by using ADP-Glo kinase assay. 1 μL of the test compound was diluted to the desired concentration, and then each sample was added to a 384-well plate as an assay plate. PIP2:3PS liposome kinase substrate working solution was added to each well to which the compound had been added. Then, kinase solution was added to each well. The control well was added with 1x kinase reaction buffer. 250 μM ATP solution was added to each well to start the kinase reaction. After the addition of ATP was completed, the assay plate was covered, mixed on a shaker for 30 to 60 seconds, and incubated at room temperature for 1 h. ADP-Glo TM reagent was added to the reaction mixture to stop the enzyme reaction and deplete the unused ATP. After 40 minutes, kinase detection reagent was added to the reaction mixture to convert ADP to ATP. The mixture was shaken for 60 seconds and equilibrated for 40 minutes, and then read on a plate reader to emit light. Finally, conversion data was collected on the enzyme reader, and RLU (relative light unit) values were converted to inhibition values using the formula (max-sample RLU) / (max-min) x 100%. "max" represents the RLU of the DMSO control, and "min" represents the RLU of the no enzyme control. The results of the inhibition of PI3Kα activity of the compounds at a concentration of 1.0 μM are shown in Table 2. In Table 2, the inhibition rate ≧ 50% is represented by A, and 50% < inhibition rate ≦ 0 is represented by B.

[0280] Table 2 Inhibition activity of compounds on PI3Kα

[0281]

[0282] From the above test results, it can be clearly seen that the novel 4-morpholinyl thieno[3,2-d]pyrimidine compounds with acrylamide structure of general formula (I) to be protected by the present application have good in vitro anti-tumor activity. The compounds have good application prospects for the development of anti-tumor drugs.

[0283] The preferred embodiments of the present application are described in detail above, but the present application is not limited to the specific details in the above-described embodiments. Within the technical concept of the present application, various equivalent transformations of the technical solutions of the present application can be made. In order to avoid unnecessary repetition, the present application will not be described again for various possible combinations. Any modification, equivalent replacement or improvement made within the technical concept of the present application is included in the protection scope of the present application.

Claims

1. A 4-morpholinylthiopheno[3,2-d]pyrimidine compound containing an acrylamide structure, characterized in that, The structural formula is shown in general formula (I): In general formula (I), Selected from the following groups: Q is selected from the following groups: 。 2. The 4-morpholinylthiopheno[3,2-d]pyrimidine compound containing an acrylamide structure according to claim 1, characterized in that, The compounds of general formula (I) are selected from the following compounds:

3. A pharmaceutical composition, characterized in that, The active ingredient comprises a 4-morpholinothiopheno[3,2-d]pyrimidine compound containing an acrylamide structure as described in any one of claims 1-2, and a pharmaceutically acceptable salt thereof, as well as a pharmaceutically acceptable excipient.

4. The use of the 4-morpholinylthiopheno[3,2-d]pyrimidine compound containing an acrylamide structure as described in any one of claims 1-2, and its pharmaceutically acceptable salt, or the pharmaceutical composition of claim 3, in the preparation of a medicament for treating and / or preventing diseases caused by abnormally high expression of PI3K.

5. The use of the 4-morpholinylthiopheno[3,2-d]pyrimidine compound containing an acrylamide structure as described in any one of claims 1-2, and its pharmaceutically acceptable salt, or the pharmaceutical composition of claim 3, in the preparation of a medicament for the treatment and / or prevention of proliferative diseases.

6. The use of the 4-morpholinylthiopheno[3,2-d]pyrimidine compound containing an acrylamide structure as described in any one of claims 1-2, and its pharmaceutically acceptable salt, or the pharmaceutical composition of claim 3, in the preparation of a medicament for treating and / or preventing cancer.

7. The application according to claim 6, characterized in that, The cancer in question is breast cancer, prostate cancer, colon cancer, lung cancer, or glioma.

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