Novel 4-morpholinyl thieno [3, 2-d] pyrimidine compound containing acrylamide structure and application of novel 4-morpholinyl thieno [3, 2-d] pyrimidine compound

Abstract

The invention belongs to the field of medicines, and particularly relates to a novel 4-morpholinyl thieno [3, 2-d] pyrimidine compound containing an acrylamide structure and application of the novel 4-morpholinyl thieno [3, 2-d] pyrimidine compound. The novel 4-morpholinyl thieno [3, 2-d] pyrimidine compound containing the acrylamide structure has a structure as shown in a general formula (I). The invention also relates to a strong PI3K inhibition effect of the compound shown in the general formula (I), and also relates to an application of the compound and the pharmaceutically acceptable salt thereof in preparation of drugs for treating and / or preventing diseases caused by abnormal high expression of PI3K, especially an application in preparation of drugs for treating and / or preventing cancers. The compound provided by the invention is particularly suitable for preparing medicines for treating and / or preventing breast cancer, prostate cancer, colon cancer, lung cancer and brain glioma. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to a novel 4-morpholinothieno[3,2-d]pyrimidine compound containing an acrylamide structure and application thereof. Background Art

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

[0003] With the rapid development of tumor-related sciences such as tumor genetics, the pathogenesis of malignant tumors has been gradually elucidated. Molecular targeted therapy for tumors targeting tumor-related specific molecules and key kinases in signal pathways has attracted more and more attention from scientists. Molecular targeted therapy often has unique advantages such as orientation and positioning. PI3K, or phosphatidylinositol-3 kinase, is an intracellular phosphatidylinositol kinase that has serine / threonine kinase activity and phosphatidylinositol kinase activity. PI3K plays a key role in cell proliferation, growth and survival. When cells are stimulated by external stimuli such as growth factors, receptor tyrosine kinases (RTKs), G protein-coupled receptors and Ras proteins can be activated, thereby activating PI3K kinase. Activated PI3K kinase can phosphorylate phosphatidylinositol bisphosphate (PIP2) to phosphatidylinositol triphosphate (PIP3). PIP3 is a very important second messenger that can activate the phospholipids of downstream serine threonine protein kinase (Akt) effectors, thereby transducing extracellular signals received by cell surface receptors into cells, thereby regulating the biological processes of tumor cells and promoting tumor cell proliferation, differentiation, metastasis, etc. Abnormal PI3K signaling is associated with 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 categories: I, II, and III, among which the most widely studied is class I PI3K. Class I PI3K is divided into four subtypes: PI3Kα, PI3Kβ, PI3Kδ, and PI3Kγ according to their catalytic subunits. Small molecule inhibitors developed with PI3K as the target have become an important direction of anti-tumor drug research in recent years, and have achieved phased results in the treatment of tumors. At present, there are six PI3K inhibitors on the market worldwide, including Idelalisib (Zydelig), Copanlisib (Aliqopa), Duvelisib (Copiktra), Alpelisib (BYL719, Piqray), Umbralisib and Linplisib (Intaride), and multiple PI3K inhibitors have entered the clinical research stage. In recent years, due to safety issues, the indications of many marketed drugs have been withdrawn, and the marketing applications of some drugs have also been rejected. For PI3K inhibitors, reducing toxic side effects and improving safety are the top priorities. Therefore, the development of novel, safe and effective new PI3K inhibitors is still a key area of ​​anti-tumor drug research at home and abroad. GDC-0941 is the first thieno[3,2-d]pyrimidine PI3K inhibitor to enter clinical research. It is a class of orally effective Class I PI3K inhibitors developed by Genentech, with an IC of 1.1 for PI3Kα, β, δ and γ inhibitory activity. 50The values ​​were 3, 33, 3 and 75 nM respectively. GDC-0941 showed significant inhibitory effects on a variety of human tumor cell lines including glioblastoma, breast cancer cells, prostate cancer cells, etc. GDC-0941 has a high plasma protein binding rate in animal models, an appropriate liver clearance rate, good pharmacokinetic properties, and an oral bioavailability of 71%.

[0004] The novel thieno[3,2-d]pyrimidine derivatives designed and synthesized in this application have never been reported. Pharmacological activities have shown that they have significant inhibitory activity against PI3K and a variety of human tumor cells, showing good anti-tumor activity. Summary of the invention

[0005] The inventors designed and synthesized a series of novel 4-morpholinothieno[3,2-d]pyrimidine compounds containing acrylamide structures based on summarizing the structure-activity relationship of PI3K inhibitors and analyzing the three-dimensional structure of PI3K protein. In vitro activity screening showed that the compounds had anti-tumor activity. The present invention relates to novel 4-morpholinothieno[3,2-d]pyrimidine compounds containing acrylamide structures as PI3K inhibitors, which have not been reported in the literature.

[0006] The technical solution adopted by the present invention is: a novel 4-morpholinylthieno[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] in:

[0009] R1 and R2 are the same or different and are independently selected from hydrogen, an alkyl group containing 1 to 6 carbon atoms, a cycloalkyl group containing 3 to 8 carbon atoms, and a phenyl group, wherein the cycloalkyl group and the phenyl group may be optionally substituted by 1 to 3 identical or different R3 groups; or R1 and R2 together with the nitrogen atom to which they are attached form a 4-7 membered heterocyclic group containing 1 to 2 heteroatoms selected from N, O and S, wherein the 4-7 membered heterocyclic group is a heterocyclic group containing sulfur atoms, wherein the sulfur atoms may be oxidized to form sulfones, and the 4-7 membered heterocyclic group containing 1 to 2 heteroatoms selected from N, O and S may be optionally substituted by 1 to 3 identical or different R4 groups; or R1 and R2 are independently 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 hydrogen, halogen, alkyl containing 1-6 carbon atoms, alkoxy containing 1-6 carbon atoms, alkoxyformyl containing 1-6 carbon atoms, alkylsulfonyl containing 1-6 carbon atoms, alkylformyl containing 1-6 carbon atoms, amino substituted by one or two alkyl containing 1-6 carbon atoms, carbamoyl, carbamoyl substituted by one or two alkyl containing 1-6 carbon atoms, alkyl containing 1-6 carbon atoms substituted by hydroxy, alkyl containing 1-6 carbon atoms substituted by alkoxy containing 1-3 carbon atoms;

[0013] R5 and R6 are the same or different and are independently selected from hydrogen and an alkyl group containing 1 to 6 carbon atoms;

[0014] R7 is selected from an alkyl group containing 1 to 6 carbon atoms, an alkoxycarbonyl group containing 1 to 6 carbon atoms;

[0015] R8 is selected from hydrogen, hydroxy, alkyl containing 1-6 carbons, alkoxyformyl containing 1-6 carbons;

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

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

[0018] Q is selected from the following groups:

[0019]

[0020] R9 is selected from hydrogen, amino, hydroxy, alkyl containing 1-6 carbons, alkoxyformyl containing 1-6 carbons, and amino substituted by one or two alkyl containing 1-6 carbons;

[0021] X is selected from C or N atoms;

[0022] Preferably, the novel 4-morpholinothieno[3,2-d]pyrimidine compound containing acrylamide structure and its pharmaceutically acceptable salt,

[0023] In the general formula (I), Selected from the following groups:

[0024]

[0025] Q is selected from the following groups:

[0026]

[0027] Furthermore, the novel 4-morpholinothieno[3,2-d]pyrimidine compounds containing acrylamide structure and pharmaceutically acceptable salts thereof, the compounds of general formula (I) have the following structural formulas, but are not limited to these compounds:

[0028]

[0029]

[0030]

[0031] The following synthetic routes describe the preparation of novel acrylamide-containing 4-morpholinylthieno[3,2-d]pyrimidine compounds of the general formula (I) of the present invention, all of which are prepared by the methods described in these reaction schemes, by methods well known to those skilled in the art of organic chemistry, or are commercially available. All final derivatives of the present invention are prepared by the methods described in the following reaction schemes or by methods analogous thereto, which are well known to those skilled in the art of organic chemistry. All variable factors used in these reaction schemes are as defined below or as defined in the claims.

[0032] According to the novel 4-morpholinothieno[3,2-d]pyrimidine compound containing acrylamide structure of the general formula (I) of the present invention, when Q is When, it can be prepared according to the method of route 1, wherein (I-1) represents a substructure of (I), and other substituents are defined as in the claims;

[0033]

[0034] According to the novel 4-morpholinothieno[3,2-d]pyrimidine compound containing acrylamide structure of the general formula (I) of the present invention, when Q is When, it is prepared according to the method of route 2, wherein (I-2) represents a substructure of (I), the synthesis method of the intermediate e of this route is the same as that of route 1, and the other substituents are defined as in the claims;

[0035]

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

[0037] A pharmaceutical composition comprises any one of the novel 4-morpholinothieno[3,2-d]pyrimidine compounds containing acrylamide structure and a pharmaceutically acceptable salt thereof as an active ingredient and a pharmaceutically acceptable excipient.

[0038] Use of any one of the above-mentioned novel 4-morpholinothieno[3,2-d]pyrimidine compounds containing acrylamide structure and pharmaceutically acceptable salts thereof or the above-mentioned pharmaceutical composition in the preparation of drugs for treating and / or preventing diseases caused by abnormally high expression of PI3K.

[0039] Use of any one of the above novel 4-morpholinothieno[3,2-d]pyrimidine compounds containing acrylamide structure and pharmaceutically acceptable salts thereof or the above pharmaceutical composition in the preparation of drugs for treating and / or preventing proliferative diseases.

[0040] Use of any one of the above novel 4-morpholinothieno[3,2-d]pyrimidine compounds containing acrylamide structure and pharmaceutically acceptable salts thereof or the above pharmaceutical composition in the preparation of drugs for treating and / or preventing cancer.

[0041] Furthermore, any one of the above-mentioned novel 4-morpholinothieno[3,2-d]pyrimidine compounds containing acrylamide structure and pharmaceutically acceptable salts thereof or the above-mentioned pharmaceutical composition is used in the preparation of drugs for treating and / or preventing breast cancer, prostate cancer, colon cancer, lung cancer and brain glioma.

[0042] Moreover, according to some common methods in the field to which the present invention belongs, the novel 4-morpholinylthieno[3,2-d]pyrimidine compound containing acrylamide structure of general formula (I) of the present invention can form pharmaceutically acceptable salts with acids. Pharmaceutically acceptable addition salts include inorganic acid and organic acid addition salts, and 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, naphthalene disulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, benzoic acid, etc.

[0043] In the present invention, "halogen" refers to fluorine, chlorine, bromine or iodine; "alkyl" refers to a straight-chain or branched alkyl; "cycloalkyl" refers to a substituted or unsubstituted cycloalkyl; "aryl" refers to a monocyclic or polycyclic carbon atom aromatic ring system, such as phenyl, naphthyl, etc.; "heteroaryl" refers to a 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, etc.; "heterocyclic group" refers to a monocyclic or polycyclic ring system containing one or more heteroatoms selected from N, O, S, such as pyrrolidinyl, morpholinyl, piperazinyl, piperidinyl, pyrazolidinyl, imidazolidinyl and thiazolinyl, etc. Represents the place where substituents are attached.

[0044] The beneficial effects of the present invention are:

[0045] 1. Through in vitro inhibition of 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 brain glioblastoma cells U87MG activity tests, the compounds of the present invention have a significant inhibitory effect on the above tumor cells, and are particularly useful for preparing drugs for treating and / or preventing breast cancer, prostate cancer, colon cancer, lung cancer and brain glioma.

[0046] 2. Through the PI3K activity test, it was found that the compounds of the present invention have significant PI3K inhibitory activity, and are particularly useful for preparing drugs for treating and / or preventing diseases caused by abnormally high expression of PI3K, and in particular for preparing drugs for treating and / or preventing cancer.

[0047] 3. The 4-morpholinylthieno[3,2-d]pyrimidine compounds containing acrylamide structure of the present invention have novel chemical structures, and most of the compounds have significant inhibitory activity against PI3K in in vitro biological activity studies and have strong anti-proliferative activity against 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 illustrate the compounds of the present invention and their preparation methods. It should be understood that the scope of the following examples and preparation examples does not limit the scope of the present invention in any way. The examples are intended to illustrate rather than limit the scope of the present invention. The H NMR spectra of the compounds were measured using Bruker ARX-400 or ARX-600, and the mass spectra were measured using Agilent 6460QQQ; all reagents used were analytically pure or chemically pure.

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

[0050]

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

[0052]

[0053] 3-Amino-2-thiophenecarboxylic acid methyl ester (124.8 g, 0.79 mmol) and urea (275.4 g, 3.97 mol) were mixed evenly, heated to 190°C and stirred for 2 hours. When the reaction was cooled to 120°C, it was added to 600 mL of 7.5% sodium hydroxide solution while hot, stirred until most of the solid was dissolved, filtered, the insoluble matter was filtered out, the pH was adjusted to 3, a large amount of milky white solid was precipitated, filtered, the filter cake was washed with water until neutral, and dried to obtain 121.0 g of light yellow solid 2,4-dihydroxythieno[3,2-d]pyrimidine. 1 H NMR (300MHz, DMSO-d6) δ 11.58 (s, 1H), 11.24 (s, 1H), 8.06 (d, J = 5.2Hz, 1H), 6.92 (d, J = 5.2Hz, 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 under reflux for 10 hours. The solvent was removed by evaporation under reduced pressure, and the residue was slowly poured into ice water and stirred vigorously to precipitate a large amount of white solid. The filter cake was washed with water until neutral and dried to obtain 12.72 g of light yellow solid 2,4-chlorothieno[3,2-d]pyrimidine. 1 H NMR (300MHz, DMSO-d6) 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] 2,4-Dichlorothieno[3,2-d]pyrimidine (100g, 0.49mol) was placed in 1500mL methanol, and morpholine (93.4g, 1.23mol) was slowly added dropwise under stirring at room temperature. After 2 hours of addition, the reaction was completed. Filter by suction, wash the filter cake with ethanol, and then wash it thoroughly with ethanol. Dry the filter cake to obtain 118.6g of light 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-morpholinothieno[3,2-d]pyrimidine-6-carbaldehyde

[0061]

[0062] Under nitrogen protection, 2-chloro-4-(4-morpholinyl)thieno[3,2-d]pyrimidine (120g, 0.47mol) was dispersed in 1200mL dry tetrahydrofuran, cooled to -10℃, and isopropylmagnesium chloride tetrahydrofuran solution (2mol / L, 118mL) was slowly added dropwise. After the addition was complete, the mixture was kept at -10℃ for 2h. n-Butyllithium hexane solution (2.5mol / L, 226mL) was slowly added dropwise to the above solution. After the addition was complete, the mixture was kept at -10℃ for 2h. Then 91mL of N,N-dimethylformamide was added dropwise to the reaction solution. After the addition was complete, the mixture was kept at -10℃ for 8h. The reaction solution was slowly poured into a five-fold amount of 0.25M hydrochloric acid solution prepared in advance, stirred for 2 hours, then heated to 50°C and stirred for 4 hours, filtered, washed with water until neutral, and dried to obtain 127 g of light yellow solid 2-chloro-4-morpholinothieno[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-morpholinothieno[3,2-d]pyrimidine-6-carbaldehyde

[0064]

[0065] 2-Chloro-4-morpholinylthieno[3,2-d]pyrimidine-6-carboxaldehyde (15g, 52.87mmol) was mixed with 6-methoxypyridine-3-boric acid (12.17g, 79.30mmol) and potassium carbonate (21.30g, 0.158.60mmol), 1,4-dioxane 225mL was added, nitrogen was slowly introduced into the above solution at room temperature for 30min, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (1.17g, 1.60mmol) was added, and the reaction was completed at 100℃ for 4h under nitrogen protection. Filter by suction, wash the filter cake with dichloromethane, add appropriate amount of water to the filtrate, extract with appropriate amount of dichloromethane 3 times, combine the organic phases, wash the organic phase with water 3 times, wash the organic phase with saturated brine 3 times, dry the organic phase with anhydrous sodium sulfate, and evaporate the solvent under reduced pressure to obtain a crude product. The crude product was slurried with anhydrous methanol and dried to obtain 14.96 g of yellow solid 2-(6-methoxy-3-pyridyl)-4-morpholinothieno[3,2-d]pyrimidine-6-carbaldehyde. 1H NMR (600MHz, CDCl3) δ10.19(s,1H),9.23(s,1H),8.58(d,J=8.5Hz,1H),8.08(s,1H),6.82(d ,J=8.6Hz,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: Synthesis of ethyl (E)-3-(2-(6-methoxy-3-pyridinyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acrylate

[0067]

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

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

[0070]

[0071] Ethyl (E)-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 evaporation under reduced pressure, and the residue was added to 50 mL of water. The pH was adjusted to about 3 with concentrated hydrochloric acid solution, and a large amount of yellow solid was precipitated. The mixture was filtered and dried to obtain 2.94 g of yellow solid (E)-3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acrylate. 1H NMR(600MHz,DMSO-d6)δ12.80(s,1H),9.14(s,1H),8.57(d,J=8.5Hz,1H),8.12-7.72(m, 2H),7.09-6.79(m,1H),6.49(d,J=15.9Hz,1H),3.99(br,4H),3.93(s,3H),3.80(br,4H).

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

[0073]

[0074] Add (E)-3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-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'-tetramethyluronium hexafluorophosphate (285.80 mg, 0.75 mmol), triethylamine (152.38 mg, 1.51 mmol) and 5 mL of N,N-dimethylformamide to the reaction bottle, stir at room temperature for 20 hours to complete the reaction. Pour the reaction solution into 30 mL of 10% sodium carbonate aqueous solution, precipitate solid, filter, wash the filter cake with water three times, slurry the filter cake with methanol twice, and dry to obtain 0.18 g of light yellow solid. 1 H NMR (600MHz, DMSO-d6) δ10.43(s,1H),9.16(d,J=2.2Hz,1H),8.59(dd,J=8.6,2.3Hz,1H),7.90(d,J=15.5Hz,1H),7.83(s,1H),7.72(dd,J=8.9,5.0Hz,2 H),7.19(t,J=8.8Hz,2H),6.92(d,J=8.7Hz,1H),6.86(d,J=15.5Hz,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 and following the preparation method of Example 1, Example 2-37 (Compound 2-37) was finally prepared.

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

[0077]

[0078] The 4-fluoroaniline in step 8 of Example 1 was replaced by aniline, and the remaining steps were the same as in Example 1 to obtain compound 2. MS (ESI), m / z: 474.10 [M+H] + .

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

[0080]

[0081] The 4-fluoroaniline in step 8 of Example 1 was replaced by 4-methoxyaniline, and the remaining steps were the same as in Example 1 to obtain compound 3. MS (ESI), m / z: 504.20 [M+H] + .

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

[0083]

[0084] The 4-fluoroaniline in step 8 of Example 1 was replaced by 3,4-dimethoxyaniline, and the remaining steps were the same as those of Example 1 to obtain compound 4. 1 HNMR (600MHz, CDCl3) δ9.23(d,J=2.0Hz,1H),8.57(dd,J=8.7,2.3Hz,1H),7.91(d,J=15.2Hz,1H),7.56(s,1H),7.52(s,1H),7.43(s,1H),6.95(d,J =7.1Hz,1H),6.89-6.76(m,2H),6.55(d,J=15.2Hz,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: Synthesis of (E)-3-(2-(6-methoxy-3-pyridinyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-(4-methylsulfonyl-1-piperazinyl)-2-propen-1-one (Compound 5)

[0086]

[0087] The 4-fluoroaniline in step 8 of Example 1 was replaced by 4-methylsulfonylpiperazine, and the remaining steps were the same as those of Example 1 to obtain compound 5. 1 H NMR (600MHz, DMSO-d6) δ9.15(s,1H),8.57(d,J=8.6Hz,1H),7.88(s,1H),7.82(d,J=15.2Hz,1H),7.30(d,J=15.2Hz,1H),6.92(d,J= 8.6Hz,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: Synthesis of (E)-3-(2-(6-methoxy-3-pyridinyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-morpholinyl-2-propene-1-one (Compound 6)

[0089]

[0090] The 4-fluoroaniline in step 8 of Example 1 was replaced by morpholine, and the remaining steps were the same as in Example 1 to obtain compound 6. MS (ESI), m / z: 468.20 [M+H] + .

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

[0092]

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

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

[0095]

[0096] Replace 4-fluoroaniline in step 8 of Example 1 with diethylamine, and follow the remaining steps as in Example 1 to obtain compound 8. 1 H NMR (600MHz, CDCl3) δ9.23(d,J=2.2Hz,1H),8.58(dd,J=8.7,2.4Hz,1H),7.88(d,J=15.1Hz,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=7.1Hz,3H),1.21(t,J=7.1Hz,3H); MS(ESI),m / z:454.20[M+H] + .

[0097] Example 9: Synthesis of (E)-N-(2-dimethylaminoethyl)-3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-N-methylacrylamide (Compound 9)

[0098]

[0099] The 4-fluoroaniline in step 8 of Example 1 was replaced by N,N,N'-trimethylethylenediamine, and the remaining steps were the same as those of Example 1 to obtain compound 9. 1 HNMR(600MHz,DMSO-d6)δ9.13(d,J=2.0Hz,1H),8.56(dd,J=8.6,2.2Hz,1H),7.84(s,1H), 7.74(t,J=15.5Hz,1H),7.17(t,J=16.0Hz,1H),6.91(d,J=8.7Hz,1H),3.98(br,4H),3.93( s,3H),3.81(br,4H),3.60(t,J=6.2Hz,1H),3.54(t,J=6.5Hz,1H),3.06(d,J=124.6Hz,3H) ,2.54(s,1H),2.45(t,J=6.0Hz,1H),2.28(s,3H),2.22(s,3H); MS(ESI),m / z:483.20[M+H] + .

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

[0101]

[0102] The 4-fluoroaniline in step 8 of Example 1 was replaced by N-methylpiperazine, and the remaining steps were the same as in Example 1 to obtain compound 10. MS (ESI), m / z: 481.20 [M+H] + ,680.20[M+Na] + .

[0103] Example 11: Synthesis of (E)-N-(2-dimethylaminoethyl)-3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acrylamide (Compound 11)

[0104]

[0105] The 4-fluoroaniline in step 8 of Example 1 was replaced by N,N-dimethylethylenediamine, and the remaining steps were the same as those of Example 1 to obtain compound 11. 1 HNMR(600MHz,CD3COOD)δ9.21(d,J=2.4Hz,1H),8.60(dd,J=8.8,2.5Hz,1H),7.87(d,J=15.6Hz,1H),7.79(s,1H),6.96(d,J=8.8Hz,1H),6.83(d,J=1 5.6Hz,1H),4.30-4.17(m,5H),4.04(s,3H),4.02-3.95(m,4H),3.87(t,J= 5.6Hz,2H),3.48(t,J=5.7Hz,2H),3.02(s,7H); MS(ESI),m / z:469.20[M+H] + .

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

[0107]

[0108] The 4-fluoroaniline in step 8 of Example 1 was replaced by 3-dimethylamino-1-propylamine, and the remaining steps were the same as those of Example 1 to obtain compound 12. 1HNMR(600MHz,DMSO-d6)δ9.13(d,J=1.9Hz,1H),8.56(dd,J=8.7,2.2Hz,1H),8.29( t,J=5.4Hz,1H),7.97-7.60(m,2H),6.91(d,J=8.6Hz,1H),6.68(d,J=15.7Hz,1H), 4.04-3.96(m,4H),3.93(s,3H),3.85-3.76(m,4H),3.22(dd,J=12.6,6.6Hz,2H),2 .35(t,J=7.0Hz,2H),2.22(s,6H),1.74-1.52(m,2H);;MS(ESI),m / z:483.20[M+H] + .

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

[0110]

[0111] The 4-fluoroaniline in step 8 of Example 1 was replaced by 2-methoxyethylamine, and the remaining steps were the same as those of Example 1 to obtain compound 13. 1 H NMR (600MHz, DMSO-d6) δ9.15(d,J=2.2Hz,1H),8.58(dd,J=8.7,2.3Hz,1H),8.35(t,J=5.5Hz,1H),7.99-7.61(m,2H),6.92(d,J=8.7Hz,1H),6.76(d,J= 15.8Hz,1H),4.07-3.96(m,4H),3.93(s,3H),3.87-3.75(m,4H),3.41(t,J= 5.4Hz,2H),3.37(t,J=5.3Hz,2H),3.28(s,3H); MS(ESI),m / z:456.20[M+H] + .

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

[0113]

[0114] The 4-fluoroaniline in step 8 of Example 1 was replaced by ethanolamine, and the remaining steps were the same as those of Example 1 to obtain compound 14. 1H NMR(600MHz,DMSO-d6)δ9.14(d,J=1.8Hz,1H),8.57(dd,J=8.6,2.1Hz,1H),8 .32(t,J=5.5Hz,1H),7.73(d,J=13.6Hz,2H),6.91(d,J=8.6Hz,1H),6.75(d,J =15.7Hz,1H),4.77(t,J=5.0Hz,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 15: Synthesis of (E)-N-(3-hydroxy-1-adamantyl)-3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acrylamide (Compound 15)

[0116]

[0117] The 4-fluoroaniline in step 8 of Example 1 was replaced by 3-amino-1-adamantanol, and the remaining steps were the same as in Example 1 to obtain compound 15. 1 HNMR(600MHz,DMSO-d6)δ9.14(s,1H),8.57(d,J=7.6Hz,1H),7.70(dd,J=86.2,47.6Hz,3H),6.91(d,J=8.2Hz,1H),6.74(d,J= 15.5Hz,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: Synthesis of (E)-3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-N-(3-morpholinopropyl)acrylamide (Compound 16)

[0119]

[0120] The 4-fluoroaniline in step 8 of Example 1 was replaced by 3-(4-morpholinyl)-1-propylamine, and the remaining steps were the same as those of Example 1 to obtain compound 16. 1HNMR(600MHz,DMSO-d6)δ9.13(s,1H),8.55(d,J=7.7Hz,1H),8.25(s,1H), 8.05-7.56(m,2H),6.90(d,J=8.3Hz,1H),6.67(d,J=15.5Hz,1H),4.04-3.9 5(m,4H),3.93(s,3H),3.87-3.75(m,4H),3.69-3.54(m,4H),3.23(d,J=4. 9Hz,2H),2.46-2.22(m,6H),1.77-1.53(m,2H); MS(ESI),m / z:525.20[M+H] + .

[0121] Example 17: Synthesis of (E)-1-(3-(2-(6-methoxy-3-pyridinyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acryloyl)piperidine-4-carboxylic acid ethyl ester (Compound 17)

[0122]

[0123] The 4-fluoroaniline in step 8 of Example 1 was replaced by ethyl 4-piperidinate, and the remaining steps were the same as those of Example 1 to obtain compound 17. 1 HNMR(600MHz,DMSO-d6)δ9.15(s,1H),8.70-8.47(m,1H),7.85(s,1H),7.77(d,J=15.2Hz,1H),7.29(d,J= 15.2Hz,1H),6.92(d,J=8.7Hz,1H),4.32(d,J=12.4Hz,1H),4.16(d,J=12.8Hz,1H),4.09(q,J=7.0Hz,2H), 4.04-3.98(m,4H),3.94(s,3H),3.86-3.77(m,4H),3.24(t,J=12.0Hz,1H),2.89(t,J=11.6Hz,1H),2.71-2 .61(m,1H),1.91(s,2H),1.51(dd,J=49.7,10.4Hz,2H),1.20(t,J=7.1Hz,3H); MS(ESI),m / z:538.20[M+H] + .

[0124] Example 18: Synthesis of (E)-(3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acryloyl)glutamic acid diethyl ester (Compound 18)

[0125]

[0126] The 4-fluoroaniline in step 8 of Example 1 was replaced by diethyl glutamate hydrochloride, and the remaining steps were the same as in Example 1 to obtain compound 18. 1 HNMR(600MHz,DMSO-d6)δ9.16(s,1H),8.67(d,J=7.5Hz,1H),8.59(dd,J=8.7,1.7Hz,1H),7.92- 7.66(m,2H),6.93(d,J=8.7Hz,1H),6.79(d,J=15.7Hz,1H),4.49-4.36(m,1H),4.13(q,J=7.1Hz, 2H),4.07(q,J=14.0,7.0Hz,2H),4.04-3.97(m,4H),3.94(s,3H),3.85-3.78(m,4H),2.42(d,J=1 0.0Hz,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: Synthesis of (E)-(3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acryloyl)glycine ethyl ester (Compound 19)

[0128]

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

[0130] Example 20: Synthesis of (E)-(3-(2-(6-methoxy-3-pyridinyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acryloyl)proline methyl ester (Compound 20)

[0131]

[0132] The 4-fluoroaniline in step 8 of Example 1 was replaced with proline methyl ester hydrochloride, and the remaining steps were the same as in Example 1 to obtain compound 20. MS (ESI), m / z: 510.20 [M+H] + .

[0133] Example 21: Synthesis of (E)-(3-(2-(6-methoxy-3-pyridinyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acryloyl)-N-methylglycine ethyl ester (Compound 21)

[0134]

[0135] The 4-fluoroaniline in step 8 of Example 1 was replaced with ethyl 2-(methylamino)acetate hydrochloride, and the remaining steps were the same as in Example 1 to obtain 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)

[0137]

[0138] The 4-fluoroaniline in step 8 of Example 1 was replaced by 4-dimethylaminopiperidine, and the remaining steps were the same as in Example 1 to obtain 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)acrylamide)butyric acid methyl ester (Compound 23)

[0140]

[0141] The 4-fluoroaniline in step 8 of Example 1 was replaced by 4-aminobutyric acid methyl ester hydrochloride, and the remaining steps were the same as those of Example 1 to obtain compound 23. 1 H NMR(600MHz,DMSO-d6)δ9.13(d,J=2.0Hz,1H),8.56(dd,J=8.6,2.2Hz,1H),8.3 1(t,J=5.5Hz,1H),7.86-7.56(m,2H),6.91(d,J=8.6Hz,1H),6.67(d,J=15.7Hz ,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: Synthesis of (E)-1-(1,1-dioxothiomorpholinyl)-3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-2-propene-1-one (Compound 24)

[0143]

[0144] The 4-fluoroaniline in step 8 of Example 1 was replaced with thiomorpholine dioxide, and the remaining steps were the same as in Example 1 to obtain compound 24. MS (ESI), m / z: 516.20 [M+H] + .

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

[0146]

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

[0148] Example 26: Synthesis of (E)-1-((2R,6S)-2,6-dimethylmorpholinyl)-3-(2-(6-methoxy-3-pyridyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-2-propene-1-one (Compound 26)

[0149]

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

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

[0152]

[0153] The 4-fluoroaniline in step 8 of Example 1 was replaced with prolinamide, and the remaining steps were the same as in Example 1 to obtain compound 27. MS (ESI), m / z: 495.10 [M+H] + ,517.20[M+Na] + .

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

[0155]

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

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

[0158]

[0159] The 4-fluoroaniline in step 8 of Example 1 was replaced by 2-aminopropionamide, and the remaining steps were the same as those of Example 1 to obtain compound 29. 1 H NMR (600MHz, DMSO-d6) δ9.14(s,1H),8.57(d,J=8.6Hz,1H),8.41(d,J=7.5Hz,1H),7.89-7.65(m,2H),7.44(s,1H),7.03(s,1H),6.91(d,J=8.7Hz,1 H),6.85(d,J=15.7Hz,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=7.0Hz,3H); MS(ESI),m / z:469.20[M+H] + ,491.10[M+Na] + .

[0160] Example 30: Synthesis of (E)-3-(2-(6-methoxy-3-pyridinyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-N-(1-methylsulfonyl-4-piperidinyl)acrylamide (Compound 30)

[0161]

[0162] The 4-fluoroaniline in step 8 of Example 1 was replaced by 1-methylsulfonyl-4-aminopiperidine, and the remaining steps were the same as those of Example 1 to obtain compound 30.

[0163] 1 H NMR(600MHz,CD3COOD)δ9.26(s,1H),8.64(d,J=6.7Hz,1H),8.05-7.65(m,2H),7.00(d,J=8.7Hz,1H),6.86(d,J=15.6Hz,1H),4.46-4.17 (m,4H),4.14-3.89(m,8H),3.78(d,J=11.4Hz,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] + .

[0164] Example 31: Synthesis of (E)-1-(3-(2-(6-methoxy-3-pyridinyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acryloyl)piperidine-4-carboxamide (Compound 31)

[0165]

[0166] The 4-fluoroaniline in step 8 of Example 1 was replaced by piperidine-4-carboxamide, and the remaining steps were the same as those of Example 1 to obtain compound 31. 1 H NMR(600MHz,CD3COOD)δ9.24(s,1H),8.63(d,J=8.0Hz,1H),7.93(d,J=15.3Hz,1H),7.82(s, 1H),7.24(d,J=15.3Hz,1H),6.98(d,J=8.7Hz,1H),4.69(d,J=12.7Hz,1H),4.31(d,J=13.3Hz ,1H),4.26-4.16(m,4H),4.05(s,3H),4.03-3.91(m,4H),3.32(t,J=12.3Hz,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] + .

[0167] Example 32: Synthesis of (E)-N-(2,6-dioxo-3-piperidinyl)-3-(2-(6-methoxy-3-pyridinyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acrylamide (Compound 32)

[0168]

[0169] The 4-fluoroaniline in step 8 of Example 1 was replaced by 3-aminopiperidine-2,6-dione hydrochloride, and the remaining steps were the same as those of Example 1 to obtain compound 32. 1 H NMR (600MHz, DMSO-d6) δ9.13(s,1H),8.55(d,J=7.7Hz,1H),8.25(s,1H),8.05-7.56(m,2H),6.90(d,J=8.3Hz,1H),6.67(d,J=15.5Hz,1H),4.04-3.9 5(m,4H),3.93(s,3H),3.87-3.75(m,4H),3.69-3.54(m,4H),3.23(d,J=4. 9Hz,2H),2.46-2.22(m,6H),1.77-1.53(m,2H); MS(ESI),m / z:509.10[M+H] + .

[0170] Example 33: Synthesis of (E)-3-(2-(2-amino-5-pyrimidinyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-N-(2-hydroxyethyl)acrylamide (Compound 33)

[0171]

[0172] The 6-methoxypyridine-3-boric acid in step 5 of Example 1 was replaced with 2-aminopyrimidine-5-boric acid or 2-aminopyrimidine-5-boric acid pinacol ester, and the 4-fluoroaniline in step 8 of Example 1 was replaced with ethanolamine. The remaining steps were the same as in Example 1 to obtain compound 33. MS (ESI), m / z: 428.10 [M+H] + ,550.20[M+Na] + .

[0173] Example 34: Synthesis of (E)-(3-(2-(2-amino-5-pyrimidinyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acryloyl)glycine ethyl ester (Compound 34)

[0174]

[0175] The 6-methoxypyridine-3-boric acid in step 5 of Example 1 was replaced with 2-aminopyrimidine-5-boric acid or 2-aminopyrimidine-5-boric acid pinacol ester, and the 4-fluoroaniline in step 8 of Example 1 was replaced with glycine ethyl ester. The remaining steps were the same as in Example 1 to obtain compound 34. MS (ESI), m / z: 470.10 [M+H] + .

[0176] Example 35: Synthesis of (E)-N-(1-adamantyl)-3-(2-(2-amino-5-pyrimidinyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)acrylamide (Compound 35)

[0177]

[0178] The 6-methoxypyridine-3-boric acid in step 5 of Example 1 was replaced with 2-aminopyrimidine-5-boric acid or 2-aminopyrimidine-5-boric acid pinacol ester, and the 4-fluoroaniline in step 8 of Example 1 was replaced with adamantaneamine. The remaining steps were the same as in Example 1 to obtain compound 35. MS (ESI), m / z: 518.20 [M+H] + .

[0179] Example 36: Synthesis of (E)-3-(2-(2-amino-5-pyrimidinyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-N-(2-methoxyethyl)acrylamide (Compound 36)

[0180]

[0181] The 6-methoxypyridine-3-boric acid in step 5 of Example 1 was replaced with 2-aminopyrimidine-5-boric acid or 2-aminopyrimidine-5-boric acid pinacol ester, and the 4-fluoroaniline in step 8 of Example 1 was replaced with 2-methoxyethylamine. The remaining steps were the same as in Example 1 to obtain compound 36. MS (ESI), m / z: 442.20 [M+H] + .

[0182] Example 37: Synthesis of (E)-3-(2-(2-amino-5-pyrimidinyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-N-(1-methylsulfonyl-4-piperidinyl)acrylamide (Compound 37)

[0183]

[0184] The 6-methoxypyridine-3-boric acid in step 5 of Example 1 was replaced with 2-aminopyrimidine-5-boric acid or 2-aminopyrimidine-5-boric acid pinacol ester, and the 4-fluoroaniline in step 8 of Example 1 was replaced with 1-methylsulfonyl-4-aminopiperidine. The remaining steps were the same as in Example 1 to obtain compound 37. MS (ESI), m / z: 545.20 [M+H] + .

[0185] Example 38: Synthesis of (E)-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-(4-methylsulfonyl-1-piperazinyl)-2-propene-1-one (Compound 38)

[0186]

[0187] The 2-chloro-4-morpholinothieno[3,2-d]pyrimidine-6-carbaldehyde used in the synthesis of compound 38 is 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-carbaldehyde

[0189]

[0190] 2-Chloro-4-morpholinylthieno[3,2-d]pyrimidine-6-carboxaldehyde (15g, 52.87mmol) was mixed with 1-THP-4-indazoleboronic acid pinacol ester (26.03g, 79.30mmol) and potassium carbonate (21.30g, 0.158.60mmol), 250mL of 1,4-dioxane and 25mL of water were added, nitrogen was slowly introduced into the above solution at room temperature for 30min, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (1.17g, 1.60mmol) was added, and the reaction was completed at 80℃ for 6h under nitrogen protection. Filtered with suction, the filter cake was washed with dichloromethane, and the filtrate was evaporated under reduced pressure to obtain a crude product. The crude product was slurried with anhydrous methanol and dried to obtain 18.28 g of yellow solid 4-morpholinyl-2-(1-(2-tetrahydropyranyl)-4-indazolyl)thieno[3,2-d]pyrimidine-6-carbaldehyde. 1H NMR (600MHz, CDCl3) δ10.20 (s, 1H), 8.94 (s, 1H), 8.28 (d, J = 7.2Hz, 1H), 8.14 (s, 1H),7.73(d,J=8.3Hz,1H),7.51(t,J=7.7Hz,1H),5.79(d,J=8.9Hz,1H),4.19-4. 09(m,4H),4.05(d,J=11.4Hz,1H),3.97-3.87(m,4H),3.78(t,J=10.3Hz,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: Synthesis of ethyl (E)-3-(4-morpholinyl-2-(1-(2-tetrahydropyranyl)-4-indazolyl)-6-thieno[3,2-d]pyrimidinyl)acrylate

[0192]

[0193] 4-Morpholinyl-2-(1-(2-tetrahydropyranyl)-4-indazolyl)thieno[3,2-d]pyrimidine-6-carboxaldehyde (12.00 g, 26.69 mmol), ethoxycarbonylmethylenetriphenylphosphine (13.95 g, 40.04 mmol) and 120 mL of toluene were added to the reaction bottle and refluxed for about 4 hours to complete the reaction. The reaction system was filtered while hot, and the filter cake was washed with an appropriate amount of toluene. The filter cake was dried to obtain 10.80 g of yellow solid (E)-3-(4-morpholinyl-2-(1-(2-tetrahydropyranyl)-4-indazolyl)-6-thieno[3,2-d]pyrimidinyl) ethyl acrylate. 1 H NMR (600MHz, CDCl3) δ8.93(s,1H),8.27(d,J=7.2Hz,1H),7.87(d,J=15.8Hz,1H),7.71(d,J=8 .3Hz,1H),7.64(s,1H),7.50(t,J=7.8Hz,1H),6.46(d,J=15.8Hz,1H),5.96-5.57(m,1H),4.30 (q,J=7.1Hz,2H),4.16-3.99(m,5H),3.98-3.87(m,4H),3.77(t,J=10.5Hz,1H),2.75-2.53(m ,1H),2.27-2.04(m,2H),1.84-1.64(m,3H),1.36(t,J=7.1Hz,3H); MS(ESI),m / z:520.10[M+H] + .

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

[0195]

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

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

[0198]

[0199] Add (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-methylsulfonylpiperazine (120.30 mg, 0.73 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (347.40 mg, 0.92 mmol), triethylamine (185.30 mg, 1.83 mmol) and 8 mL of N,N-dimethylformamide into the reaction bottle and stir at room temperature for 20 h until the reaction is complete. The reaction solution was poured into 30 mL of 10% sodium carbonate aqueous solution to precipitate solids, which were filtered, and the filter cake was washed three times with water, slurried twice with methanol, and dried to obtain 0.29 g of a light yellow solid (E)-1-(4-methylsulfonyl-1-piperazinyl)-3-(4-morpholinyl-2-(1-(2-tetrahydropyranyl)-4-indazolyl)-6-thieno[3,2-d]pyrimidinyl)-2-propene-1-one. MS (ESI), m / z: 638.20 [M+H] + .

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

[0201]

[0202] (E)-1-(4-methylsulfonyl-1-piperazinyl)-3-(4-morpholinyl-2-(1-(2-tetrahydropyranyl)-4-indazolyl)-6-thieno[3,2-d]pyrimidinyl)-2-propene-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 stirred at 80°C for 2 h until the reaction was completed. Most of the solvent was evaporated under reduced pressure, and the residue was added to 10 mL of saturated sodium carbonate solution to precipitate a yellow solid, which was filtered, and the filter cake was slurried twice with methanol and dried to obtain 0.15 g of a yellow solid (E)-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-(4-methylsulfonyl-1-piperazinyl)-2-propene-1-one. 1 H NMR (600MHz, DMSO-d6) δ13.22(s,1H),8.88(s,1H),8.23(d,J=7.2Hz,1H),7.99(s,1H),7.85(d,J=15.1Hz,1H),7.69(d,J=8.2Hz,1H),7.48(t,J=7 .7Hz,1H),7.34(d,J=15.2Hz,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, Example 39-60 (Compound 39-60) was finally prepared.

[0204] Example 39: Synthesis of (E)-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-N-phenylacrylamide (Compound 39)

[0205]

[0206] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced by aniline, and the remaining steps were the same as those of Example 38 to obtain Compound 39. 1H NMR (600MHz, DMSO-d6) δ13.23(s,1H),10.37(s,1H),8.89(s,1H),8.24(d,J=7.2Hz,1H),8.07-7.86(m,2H),7.70(dd,J=13.4,8.1Hz,3H),7.48(t,J =7.7Hz,1H),7.36(t,J=7.9Hz,2H),7.10(t,J=7.4Hz,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: Synthesis of (E)-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-N-methylacrylamide (Compound 40)

[0208]

[0209] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced with methylamine hydrochloride, and the remaining steps were the same as in Example 38 to obtain compound 40. MS (ESI), m / z: 421.10 [M+H] + .

[0210] Example 41: Synthesis of (E)-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-N-(2-dimethylaminoethyl)acrylamide (Compound 41)

[0211]

[0212] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced by N,N-dimethylethylenediamine, and the remaining steps were the same as those of Example 38 to obtain compound 41. 1 H NMR (600MHz, DMSO-d6) δ13.30(s,1H),8.88(s,1H),8.31-8.16(m,2H),7.86(s,1H),7.74(d,J=15.6Hz,1H),7.69(d,J=8.2Hz,1H),7.47(t,J=7.7Hz, 1H),6.80(d,J=15.6Hz,1H),4.10-3.96(m,4H),3.91-3.79(m,4H),3.32-3 .28(m,2H),2.35(t,J=6.5Hz,2H),2.17(s,6H);MS(ESI),m / z:478.20[M+H] + .

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

[0214]

[0215] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced by 2-methoxyethylamine, and the remaining steps were the same as in Example 38 to obtain compound 42. 1 H NMR (600MHz, DMSO-d6) δ13.24(s,1H),8.89(s,1H),8.39(s,1H),8.23(d,J=7.1Hz,1H),7.85(s,1H),7.76(d,J=15.6Hz,1H),7.69(d,J=8.1Hz ,1H),7.48(t,J=7.7Hz,1H),6.78(d,J=15.6Hz,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 (E)-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-(4-methyl-1-piperazinyl)-2-propen-1-one (Compound 43)

[0217]

[0218] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced by N-methylpiperazine, and the remaining steps were the same as those of Example 38 to obtain compound 43. 1 H NMR (600MHz, DMSO-d6) δ13.22(s,1H),8.88(s,1H),8.23(d,J=7.2Hz,1H),7.97(s,1H),7.80(d,J=15.2Hz,1H),7.68(d,J=8.2Hz,1H),7.48(t,J=7.7 Hz,1H),7.31(d,J=15.2Hz,1H),4.14-3.96(m,4H),3.93-3.79(m,4H),3.7 4-3.53(m,4H),2.42-2.28(m,4H),2.21(s,3H); MS(ESI),m / z:490.20[M+H] + .

[0219] Example 44: Synthesis of (E)-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-thiomorpholin-2-propene-1-one (Compound 44)

[0220]

[0221] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced with thiomorpholine, and the remaining steps were the same as in Example 38 to obtain compound 44. MS (ESI), m / z: 493.10 [M+H] + .

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

[0223]

[0224] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced by tetrahydropyrrole, and the remaining steps were the same as those of Example 38 to obtain compound 45. 1 HNMR(600MHz,DMSO-d6)δ13.21(s,1H),8.88(s,1H),8.23(d,J=7.2Hz,1H), 7.97(s,1H),7.80(d,J=15.4Hz,1H),7.68(d,J=8.2Hz,1H),7.48(t,J=7.7Hz ,1H),7.02(d,J=15.4Hz,1H),4.16-3.97(m,4H),3.91-3.78(m,4H),3.67(t ,J=6.7Hz,2H),3.43(t,J=6.8Hz,2H),2.08-1.89(m,2H),1.87-1.69(m,2H).

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

[0226]

[0227] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced by diethylamine, and the remaining steps were the same as those of Example 38 to obtain compound 46. 1HNMR (600MHz, DMSO-d6) δ13.23(s,1H),8.89(s,1H),8.21(d,J=36.3Hz,2H),7.76(dd,J=56.8,43.6Hz,3H),7.48(s,1H),6.78(d,J =12.9Hz,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: Synthesis of (E)-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-N-(4-methoxyphenyl)acrylamide (Compound 47)

[0229]

[0230] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced by 4-methoxyaniline, and the remaining steps were the same as those of Example 38 to obtain compound 47. 1 H NMR (600MHz, DMSO-d6) δ13.22(s,1H),10.23(s,1H),8.89(s,1H),8.23(d,J=6.5Hz,1H),7.99-7.84(m,2H),7.69(d,J=7.8Hz,1H),7.63(d,J=7.8H z,2H),7.48(t,J=7.0Hz,1H),6.93(d,J=8.0Hz,2H),6.88(d,J=15.5Hz,1H),4.03(br,4H),3.86(br,4H),3.74(s,3H); MS(ESI),m / z:513.20[M+H] + .

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

[0232]

[0233] Replace N-methylsulfonylpiperazine in step 4 of Example 38 with 3-(4-morpholinyl)-1-propylamine, and follow the remaining steps as in Example 38 to obtain Compound 48. 1H NMR(600MHz,DMSO-d6)δ13.24(s,1H),8.88(s,1H),8.43-8.08(m,2H),7.86 (s,1H),7.75(d,J=15.5Hz,1H),7.69(d,J=8.1Hz,1H),7.48(t,J=7.7Hz,1H) ,6.72(d,J=15.6Hz,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: Synthesis of (E)-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-N-(2-hydroxyethyl)acrylamide (Compound 49)

[0235]

[0236] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced by ethanolamine, and the remaining steps were the same as in Example 38 to obtain compound 49. 1 HNMR(600MHz,DMSO-d6)δ13.23(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=15.4Hz,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: Synthesis of (E)-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-N-(2-dimethylaminoethyl)-N-methylacrylamide (Compound 50)

[0238]

[0239] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced by N,N,N′-trimethylethylenediamine, and the remaining steps were the same as those of Example 38 to obtain Compound 50. 1H NMR(600MHz,CD3COOD)δ8.65(s,1H),7.98(d,J=7.2Hz,1H),7.74(d,J=15.4Hz,1H),7.71-7.62(m,2H),7.41(t,J=7.8Hz,1H),6.99(d,J=15.4Hz ,1H),4.12-4.01(m,4H),3.89-3.80(m,4H),3.78(t,J=6.1Hz,2H),3.32 (t,J=6.1Hz,2H),3.14(s,3H),2.86(s,6H); MS(ESI),m / z:492.10[M+H] + .

[0240] Example 51: Synthesis of (E)-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] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced by N-acetylpiperazine, and the remaining steps were the same as those of Example 38 to obtain Compound 51. 1 H NMR (600MHz, DMSO-d6) δ13.25(s,1H),8.89(s,1H),8.23(d,J=7.0Hz,1H),7.98(s,1H),7.83(d,J=15.1Hz,1H),7.69(d,J=7.9Hz,1H),7.48 (t,J=7.5Hz,1H),7.31(d,J=15.1Hz,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: Synthesis of (E)-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] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced by cis-2,6-dimethylmorpholine, and the remaining steps were the same as those of Example 38 to obtain Compound 52. 1H NMR (600MHz, DMSO-d6) δ13.28(s,1H),8.89(s,1H),8.23(d,J=7.1Hz,1H),7.98(s,1H),7.83(d ,J=15.1Hz,1H),7.69(d,J=8.1Hz,1H),7.48(t,J=7.7Hz,1H),7.32(d,J=15.2Hz,1H),4.38(d,J =13.0Hz,1H),4.23(d,J=12.8Hz,1H),4.18-3.97(m,4H),3.95-3.74(m,4H),3.65-3.44(m,2H) ,2.76(t,J=11.9Hz,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: Synthesis of (E)-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-1-(1,1-dioxothiomorpholinyl)-2-propene-1-one (Compound 53)

[0247]

[0248] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced with thiomorpholine dioxide, and the remaining steps were the same as in Example 38 to obtain compound 53. MS (ESI), m / z: 525.10 [M+H] + .

[0249] Example 54: Synthesis of (E)-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] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced by 1-(2-hydroxyethyl)piperazine, and the remaining steps were the same as those of Example 38 to obtain Compound 54. 1H NMR (600MHz, DMSO-d6) δ13.21 (s, 1H), 8.88 (s, 1H), 8.22 (d, J = 6.2Hz, 1H), 7. 95(s,1H),7.79(d,J=14.9Hz,1H),7.68(d,J=7.0Hz,1H),7.59-7.40(m,1H),7 .28(d,J=15.0Hz,1H),4.44(s,1H),4.21-3.97(m,4H),3.95-3.77(m,4H),3. 68(br,2H),3.64-3.43(m,4H),2.50-2.18(m,6H); MS(ESI),m / z:520.20[M+H] + .

[0252] Example 55: Synthesis of (E)-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] Replace N-methylsulfonylpiperazine in step 4 of Example 38 with 1-(2-methoxyethyl)piperazine, and follow the same procedures as in Example 38 to obtain Compound 55. 1 H NMR (600MHz, DMSO-d6) δ13.24(s,1H),8.89(s,1H),8.24(br,1H),7.98(s,1H),7.81(d,J=14.8Hz,1H),7.68(br,1H),7.48(br,1H),7.31(d,J=14. 8Hz,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: Synthesis of (E)-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-N-(1-methylsulfonyl-4-piperidinyl)acrylamide (Compound 56)

[0256]

[0257] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced by 1-methylsulfonyl-4-aminopiperidine, and the remaining steps were the same as those of Example 38 to obtain Compound 56. 1 H NMR (600MHz, DMSO-d6) δ13.26(s,1H),8.86(s,1H),8.38(d,J=7.0Hz,1H),8.22(d,J=7 .0Hz,1H),7.87(s,1H),7.78(d,J=15.6Hz,1H),7.70(d,J=8.0Hz,1H),7.48(t,J=7.5H z,1H),6.73(d,J=15.6Hz,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: Synthesis of (E)-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] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced with 4-fluoropiperidine, and the remaining steps were the same as in Example 38 to obtain compound 57. MS (ESI), m / z: 493.20 [M+H] + .

[0261] Example 58: Synthesis of (E)-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] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced by 4-methylpiperidine, and the remaining steps were the same as in Example 38 to obtain compound 58. MS (ESI), m / z: 489.20 [M+H] + .

[0264] Example 59: Synthesis of (E)-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] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced with 4-methoxypiperidine, and the remaining steps were the same as in Example 38 to obtain compound 59. MS (ESI), m / z: 505.20 [M+H] + .

[0267] Example 60: Synthesis of (E)-3-(2-(4-indazolyl)-4-morpholinyl-6-thieno[3,2-d]pyrimidinyl)-N-(2-methoxyethyl)-N-methylacrylamide (Compound 60)

[0268]

[0269] The N-methylsulfonylpiperazine in step 4 of Example 38 was replaced by 2-methoxy-N-methylethylamine, and the remaining steps were the same as in Example 38 to obtain compound 60. 1 H NMR (600MHz, DMSO-d6) δ13.25(s,1H),8.90(s,1H),8.23(d,J=3.7Hz,1H),7.95(s,1H),7.88-7.61(m,2H),7.48(s,1H),7.21(t,J=14.2Hz,1 H),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=131.4Hz,3H); MS(ESI),m / z:479.20[M+H] + ,501.20[M+Na] + .

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

[0271] Part of the compounds of the novel acrylamide-containing 4-morpholinothieno[3,2-d]pyrimidine compounds of the general formula (I) of the present invention were screened for their in vitro inhibitory activity on 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 brain astroglioma cells U87MG.

[0272] (1) After the cells have recovered and been passaged 2-3 times to stabilize, use trypsin solution (0.25%) to digest them from the bottom of the culture bottle. Pour the cell digestion solution into a centrifuge tube, and then add culture medium to terminate the digestion. Centrifuge the centrifuge tube at 800r / min for 10min, discard the supernatant and add 5mL culture medium. Mix the cells by blowing, and take 10μL of the cell suspension and add it to the cell counting plate for counting. Adjust the cell concentration to 10 4 100 μL of cell suspension was added to all wells except well A1 which was a blank well without cells. The 96-well plate was placed in an incubator and cultured for 24 h.

[0273] (2) Dissolve the test sample in 50 μL of dimethyl sulfoxide, then add an appropriate amount of culture medium to dissolve the sample into a 2 mg / mL solution. Then dilute the sample to 20, 4, 0.8, 0.16, and 0.032 μg / mL in a 24-well plate.

[0274] Each concentration was added to 3 wells, and the growth of cells in the two rows and two columns around was greatly affected by the environment and was used only as blank cell wells. The 96-well plate was placed in an incubator and cultured for 72 hours.

[0275] (3) The drug-containing culture medium in the 96-well plate was discarded, and the cells were rinsed twice with phosphate buffer solution (PBS). 100 μL of MTT (tetrazolium) (0.5 mg / mL) was added to each well and placed in the incubator for 4 hours. The MTT solution was discarded and 100 μL of dimethyl sulfoxide was added. The surviving cells were shaken on a magnetic oscillator to fully dissolve the formazan reaction product of MTT, and the result was measured in an ELISA instrument. The IC value of the drug can be calculated by the Bliss method. 50 The results of the in vitro antiproliferative activity of the compounds 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 brain astrocytoblastoma cells U87MG are shown in Table 1. The IC 50 ≦1.0μM, expressed as A, 1.0μM﹤IC 50 ≦10.0μM, represented by B, 10.0μM﹤IC 50 ≦100.0μM, expressed as C, IC 50 >100.0μM, indicated by D.

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

[0277]

[0278]

[0279]

[0280] Example 62 Compound PI3K Activity Test

[0281] The compounds prepared in the examples were tested for their in vitro PI3Kα enzyme inhibitory activity using the ADP-Glo ​​kinase assay. 1 μL of the test compound was diluted to the desired concentration, and each sample was then added to a 384-well plate as an assay plate. PIP2:3PS lipid kinase substrate working solution was added to each well to which the compound had been added. Then, the kinase solution was added to each well. 1x kinase reaction buffer was added to the control wells. 250 μM ATP solution was added to each of the above wells to start the kinase reaction. After the addition of ATP was complete, the assay plate was covered, mixed on a shaker for 30 to 60 seconds, and incubated at room temperature for 1 hour. ADP-Glo TM The reagent was added to the reaction mixture to stop the enzyme reaction and deplete the unconsumed ATP. After 40 minutes, the 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 before reading on a plate reader for luminescence. Finally, the conversion data was collected on a microplate reader and the RLU (relative light unit) value was converted to an inhibition value using the formula (max-Sample RLU) / (max-min)×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 by the compound at a concentration of 1.0 μM are shown in Table 2. The inhibition rate in Table 2 is ≧50%, represented by A, and 50%﹤inhibition rate ≦0, represented by B.

[0282] Table 2 Inhibitory activity of compounds on PI3Kα

[0283]

[0284]

[0285] It can be clearly seen from the above test results that the novel 4-morpholinylthieno[3,2-d]pyrimidine compounds containing acrylamide structure of general formula (I) to be protected by the present invention have good in vitro anti-tumor activity. Such compounds have good prospects for the development and application of anti-tumor drugs.

[0286] The preferred embodiments of the present invention are described in detail above, but the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, the technical solution of the present invention can be subjected to a variety of equivalent transformations. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations. Any modification, equivalent replacement or improvement made within the technical concept of the present invention is included in the protection scope of the present invention.

Claims

1. A novel 4-morpholinothieno[3,2-d]pyrimidine compound containing an acrylamide structure, characterized in that: The structural formula is shown in general formula (I): in: R1 and R2 are the same or different and are independently selected from hydrogen, an alkyl group containing 1 to 6 carbon atoms, a cycloalkyl group containing 3 to 8 carbon atoms, and a phenyl group, wherein the cycloalkyl group and the phenyl group may be optionally substituted by 1 to 3 identical or different R3 groups; or R1 and R2 together with the nitrogen atom to which they are attached form a 4-7 membered heterocyclic group containing 1 to 2 heteroatoms selected from N, O and S, wherein the 4-7 membered heterocyclic group is a heterocyclic group containing sulfur atoms, wherein the sulfur atoms may be oxidized to form sulfones, and the 4-7 membered heterocyclic group containing 1 to 2 heteroatoms selected from N, O and S may be optionally substituted by 1 to 3 identical or different R4 groups; or R1 and R2 are independently selected from hydrogen and the following groups: R3 is selected from hydrogen, halogen, alkyl containing 1-6 carbons, alkoxy containing 1-6 carbons; R4 is selected from hydrogen, halogen, alkyl containing 1-6 carbon atoms, alkoxy containing 1-6 carbon atoms, alkoxyformyl containing 1-6 carbon atoms, alkylsulfonyl containing 1-6 carbon atoms, alkylformyl containing 1-6 carbon atoms, amino substituted by one or two alkyl containing 1-6 carbon atoms, carbamoyl, carbamoyl substituted by one or two alkyl containing 1-6 carbon atoms, alkyl containing 1-6 carbon atoms substituted by hydroxy, alkyl containing 1-6 carbon atoms substituted by alkoxy containing 1-3 carbon atoms; R5 and R6 are the same or different and are independently selected from hydrogen and an alkyl group containing 1 to 6 carbon atoms; R7 is selected from an alkyl group containing 1 to 6 carbon atoms, an alkoxycarbonyl group containing 1 to 6 carbon atoms; R8 is selected from hydrogen, hydroxy, alkyl containing 1-6 carbons, alkoxyformyl containing 1-6 carbons; n is selected from 1, 2, 3, 4, 5; m is selected from 1, 2, and 3; Q is selected from the following groups: R9 is selected from hydrogen, amino, hydroxy, alkyl containing 1-6 carbons, alkoxyformyl containing 1-6 carbons, and amino substituted by one or two alkyl containing 1-6 carbons; X is selected from C or N atoms.

2. A novel 4-morpholinothieno[3,2-d]pyrimidine compound containing an acrylamide structure according to claim 1, characterized in that: In the general formula (I), Selected from the following groups: Q is selected from the following groups:

3. A novel 4-morpholinothieno[3,2-d]pyrimidine compound containing an acrylamide structure according to claim 1, characterized in that: The compound of general formula (I) is selected from the following compounds:

4. A pharmaceutical composition, characterized in that The invention comprises a novel acrylamide-containing 4-morpholinylthieno[3,2-d]pyrimidine compound and a pharmaceutically acceptable salt thereof as an active ingredient and a pharmaceutically acceptable excipient.

5. Use of the novel 4-morpholinothieno[3,2-d]pyrimidine compound containing an acrylamide structure according to any one of claims 1 to 3 and its pharmaceutically acceptable salt or the pharmaceutical composition according to claim 4 in the preparation of a drug for treating and / or preventing a disease caused by abnormally high expression of PI3K.

6. Use of the novel 4-morpholinothieno[3,2-d]pyrimidine compound containing an acrylamide structure according to any one of claims 1 to 3 and its pharmaceutically acceptable salt or the pharmaceutical composition according to claim 4 in the preparation of a drug for treating and / or preventing a proliferative disease.

7. Use of the novel 4-morpholinothieno[3,2-d]pyrimidine compound containing an acrylamide structure according to any one of claims 1 to 3 and its pharmaceutically acceptable salt or the pharmaceutical composition according to claim 4 in the preparation of a drug for treating and / or preventing cancer.

8. The use according to claim 7, characterized in that: The cancers include breast cancer, prostate cancer, colon cancer, lung cancer and brain glioma.

Images