Tri-substituted quinazoline derivative as well as preparation method and application thereof

Abstract

The invention relates to the technical field of medicinal chemistry, and discloses a tri-substituted quinazoline derivative as well as a preparation method and application thereof. According to the trisubstituted quinazoline derivative provided by the invention, an efficient G4 stable module (polysubstituted quinazoline core) and a powerful HDAC inhibition pharmacophore (hydroximic acid) are integrated into a single molecule through an adjustable connecting chain, so that the bottleneck problems of poor target accessibility, high off-target toxicity and the like caused by a chromatin barrier of a traditional G4 ligand are ingeniously solved. Preferably, the compound a10 shows excellent dual activity in vitro, the stability of the compound a10 to telomere G4 is remarkably superior to that of a lead compound, and the inhibitory activity of the compound a10 to HDAC reaches the nanomole level; the series of compounds show strong inhibitory activity on colorectal cancer cells, the toxicity to normal intestinal epithelial cells is significantly reduced, the selectivity multiple is up to 39 times, and the safety is significantly improved; in-vivo pharmacological experiments further prove that the preferable molecules can effectively inhibit tumor growth, and no obvious toxicity is observed.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pharmaceutical chemistry, in particular to a trisubstituted quinazoline derivative and a preparation method and application thereof. BACKGROUND

[0002] Colorectal cancer (CRC) is the third most common and the second leading cause of cancer-related death worldwide. The clinical treatment, especially the prognosis of advanced and metastatic patients, still faces severe challenges. Current first-line therapy mainly relies on fluorouracil, oxaliplatin or irinotecan and other chemotherapeutic drugs, but the efficacy is limited and often accompanied by drug resistance and side effects. Therefore, there is a great clinical need to develop new, efficient and low-toxicity anti-colorectal cancer drugs.

[0003] In recent years, precise treatment strategies targeting specific molecular targets have shown great potential. Among them, G-quadruplex (G4) and histone deacetylase (HDAC) are two anti-cancer targets that are of great concern and complementary mechanisms. G-quadruplex is a special secondary structure formed by folding nucleic acid sequences rich in guanine (G), which widely exists in telomere ends, proto-oncogene promoters and other key genomic regions. Its formation and opening are closely related to the regulation of gene expression, telomere extension and other important events. Studies have shown that small molecule G4 ligands can induce and stabilize G4 structures, interfere with cancer gene expression and telomere function, thereby exerting anti-tumor effects. However, in the nucleus, genomic DNA is usually highly condensed in the form of chromatin, tightly wrapped with histones, which makes it difficult for many G4 ligands to effectively access their targets, resulting in insufficient efficacy, off-target effects and higher side effects. HDAC is a class of enzymes that catalyze the deacetylation of histone lysine residues, and its overexpression can lead to excessive compression of chromatin (heterochromatinization), thereby inhibiting the transcription of various genes including tumor suppressor genes. In colorectal cancer, abnormal activation of HDAC is a common feature. HDAC inhibitors (HDACi) can increase the acetylation level of histones, causing chromatin structure to change from dense to relaxed (euchromatinization), increasing the accessibility of DNA, not only reactivating silenced tumor suppressor genes, but also creating a more favorable microenvironment for other DNA-targeted drugs (such as G4 ligands). However, single HDAC inhibitors often only achieve stable disease in the treatment of solid tumors, with limited objective response rate.

[0004] Based on the above mechanism, integrating G4 stabilization and HDAC inhibition into a single molecule, i.e. developing G4 / HDAC dual-target ligands, is considered as a promising anticancer strategy. This strategy is expected to achieve multiple benefits through synergistic effects: on the one hand, chromatin relaxation caused by HDAC inhibition can enhance the accessibility and binding efficiency of G4 ligands to their genomic targets; on the other hand, the two may have additive or synergistic effects in inducing DNA damage, regulating tumor immune microenvironment, etc., thereby significantly improving the anti-tumor efficacy and possibly reducing the toxicity to normal cells.

[0005] At present, some quinazoline G4 ligands have been reported, such as CN 120365220 A discloses a class of 2,4-disubstituted quinazoline G4 ligands. However, the prior art still has the following defects: the structure of G4 ligands is relatively simple, the telomere G4 stabilization activity is insufficient, there is no successful precedent for organically integrating high-efficiency G4 stabilization activity and strong HDAC inhibition activity into the same molecule, and the toxicity (selectivity) problem of many molecules to normal cells has not been well solved while improving the activity. Therefore, there is an urgent need in the art to develop a class of dual-target small molecule compounds with novel structure, stronger G4 stabilization ability, high-efficiency HDAC inhibition activity, and high selectivity to tumor cells. SUMMARY

[0006] The present application aims to at least solve one of the above-mentioned technical problems in the prior art. To this end, one of the objects of the present application is to provide a trisubstituted quinazoline derivative.

[0007] A second object of the present application is to provide a preparation method of the trisubstituted quinazoline derivative.

[0008] A third object of the present application is to provide an application of the trisubstituted quinazoline derivative.

[0009] To achieve the above objects, the technical solution adopted by the present application is as follows:

[0010] The first aspect of the present application provides a trisubstituted quinazoline derivative, whose structural formula is shown in formula (I) or formula (II):

[0011] Formula (I); Formula (II);

[0012] In formula (I) and formula (II):

[0013] R1 is independently selected from a nitrogen-containing alkoxy group, a nitrogen-containing alkylamide group, or is connected with a spacer group -L- through an oxygen atom, and the end of the spacer group -L- is a hydroxamic acid group or a bioisostere structure thereof;

[0014] R2is independently selected from H or halogen;

[0015] R3is independently selected from H, a spacer group -L- attached via an oxygen atom and / or a phenyl group, the end of said spacer group -L- being a hydroxamic acid group, a N-hydroxybenzamide group or a structure of a hydroxamic acid group bioisostere, or an amino group substituted with a nitrogen containing heterocyclic group;

[0016] wherein said spacer group -L- is selected from -(CH2) n -(CH2) m -(CH2) p -O-(CH2) a n = 2-6, m = 2-4, p = 2-4.

[0017] In some embodiments of the present application, in said formula (I) and (II), R1is independently selected from -O-(CH2) b -T1, -NH-C(O)-(CH2) c -T2, -O-(CH2) c -C(O)N-T3, -O-(CH2) d -Ph-C(O)N-T3; wherein T1is selected from a 5-6 membered nitrogen containing heterocyclic group, a = 2-4; T2is selected from a pyrrolyl group or a C2-C4dialkylamino group, b = 2-4; T3is selected from a hydroxyl group or an anilino group, c = 3-6.

[0018] In some preferred embodiments of the present application, in said formula (I) and (II), R1, T1is a pyrrolyl group.

[0019] In some embodiments of the present application, in said formula (I) and (II), R2is independently selected from H or F.

[0020] In some embodiments of the present application, in said formula (I) and (II), R3is independently selected from H, -Ph-(CH2) d -C(O)N-T4, -Ph-O-(CH2) d -C(O)N-T4, -O-(CH2) d -Ph-C(O)N-T4, -O-(CH2) e -C(O)N-T4, -O-(CH2) e -O-(CH2) d -C(O)N-T4, C4-C6alkoxy, -O-(CH2) f -T5-Ph-C(O)N-T4, -N-(CH2)- any one of T6, -N-Ph-T6; wherein T4 is selected from hydroxyl, anilino or C4-C6 alkylamino, T5 is selected from 5-6 membered nitrogen-containing heterocyclic group, d = 4-6, e = 2-4; T6 is selected from pyrrolyl, morpholinyl or methylpiperazinyl, f = 2-3.

[0021] In some preferred embodiments of the present application, in the formula (I) and formula (II), in the R3, the T5 is triazolyl.

[0022] In some embodiments of the present application, the tri-substituted quinazoline derivatives are as shown in formula 1 - formula 31 :

[0023]

[0024]

[0025]

[0026]

[0027]

[0028]

[0029]

[0030]

[0031]

[0032]

[0033]

[0034]

[0035]

[0036]

[0037]

[0038] .

[0039] The second aspect of the present application provides a method for preparing the tri-substituted quinazoline derivatives of the first aspect of the present application, comprising the following steps:

[0040] S1, reacting 2, 4-dihalogenated quinazoline compound with arylamine compound and nitrogen-containing heterocyclic amine compound in sequence, or in reverse order, to obtain a 2-(nitrogen-containing heterocyclic group)-4-arylamine quinazoline core intermediate;

[0041] S2, connecting the core intermediate with a side chain module having a protected hydroxamic acid group or a precursor functional group capable of being converted into hydroxamic acid through a bond-forming reaction to obtain a connection product;

[0042] S3, converting the terminal functional group of the connection product into hydroxamic acid or its protected form, and removing the protecting group to obtain a free hydroxamic acid group, to obtain the trisubstituted quinazoline derivative;

[0043] In step S2, the bond-forming reaction is selected from Suzuki-Miyaura coupling reaction, nucleophilic substitution reaction, Buchwald-Hartwig carbon-nitrogen coupling reaction or cycloaddition reaction.

[0044] In some embodiments of the present application, in step S2, the bond-forming reaction is a palladium-catalyzed Suzuki-Miyaura coupling reaction, the terminal functional group of the side chain module is a boronic acid or boronic ester group, and the reaction site on the core intermediate is halogen or halogen-like.

[0045] In some embodiments of the present application, in step S2, the bond-forming reaction is a nucleophilic substitution reaction, the terminal functional group of the side chain module is a halogenated alkyl group, and the core intermediate contains a nucleophilic hydroxyl group or amino group.

[0046] In some embodiments of the present application, in step S2, the bond-forming reaction is a Buchwald-Hartwig carbon-nitrogen coupling reaction, the terminal functional group of the side chain module is an amine group, and the core intermediate contains an aryl halogen or pseudo-halogen that can participate in the coupling reaction.

[0047] In some embodiments of the present application, in step S2, the bond-forming reaction is a copper-catalyzed azide-alkyne cycloaddition reaction, the terminal functional group of the side chain module is an azide group, and the core intermediate contains an alkyne group, or the terminal functional group of the side chain module is an alkyne group, and the core intermediate contains an azide group.

[0048] In some embodiments of the present application, in step S3, the protecting group is tetrahydro-2H-pyran-2-yl, and the removal of the protecting group is carried out under the condition of a protonic acid or a Lewis acid.

[0049] In some embodiments of the present application, the method for preparing the trisubstituted quinazoline derivative comprises at least one of the following:

[0050] Method 1:

[0051] A1, mixing 6-bromo-2, 4-dichloroquinazoline, 4-[2-(pyrrolidin-1-yl)ethoxy]aniline with isopropanol, under the catalysis of concentrated hydrochloric acid, and reacting at 50-70°C for 2-4h to obtain an arylation product; mixing the arylation product, pyrrolidine and N,N-dimethylformamide, in the presence of cesium carbonate and potassium iodide, and reacting at 110-130°C for 6-10h to obtain a core intermediate;

[0052] A2, reacting aryl bromoalkanoic acid with bis(pinacolato)diboron, in the presence of dichloro[l, 1'-bis(diphenylphosphino)ferrocene]palladium(II), potassium acetate and 1,4-dioxane, and reacting at 90-110°C for 6-10h to obtain a side chain boronate carboxylic acid;

[0053] A3, reacting the side chain boronate carboxylic acid with O-(tetrahydro-2H-pyran-2-yl)hydroxylamine, in the presence of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) / 1-hydroxybenzotriazole (HOBt) coupling, triethylamine (TEA), and reacting in dichloromethane at room temperature for 1-3h to obtain a protected side chain boronate; reacting the core intermediate with the protected side chain boronate, in the presence of dichloro[l, 1'-bis(diphenylphosphino)ferrocene]palladium(II), aqueous Na2CO3, and refluxing in a 1,4-dioxane / water mixed solvent for 3-5h; and removing the protecting group by reacting the obtained product with hydrogen chloride / dioxane solution, in a dichloromethane / acetonitrile mixed solvent, at room temperature for 1-4h to obtain the trisubstituted quinazoline derivative;

[0054] Method 2:

[0055] B1, mixing 2, 4-dichloro-6-methoxyquinazoline, 4-[2-(pyrrolidin-1-yl)ethoxy]aniline with isopropanol, under the catalysis of concentrated hydrochloric acid, and reacting at 50-70°C for 2-4h to obtain an arylation product; using hydrogen bromide / acetic acid solution, and reacting at 100-120°C for 10-12h, or using boron tribromide, and reacting in dichloromethane at room temperature for 3-5h to obtain a phenolic hydroxyl intermediate;

[0056] B2, reacting the phenolic hydroxyl intermediate with ω-bromoalkanoate, in the presence of cesium carbonate, and reacting in N,N-dimethylformamide or acetonitrile at 70-90°C for 10-12h to obtain an etherification product;

[0057] B3. hydrolyzing the etherified product in a solution of lithium hydroxide in methanol / water at room temperature for 10-12 h, condensing the resulting carboxylic acid with O-(tetrahydro-2H-pyran-2-yl)hydroxylamine in the presence of N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-l-yl)urea hexafluorophosphate (HATU) and N,N-diisopropylethylamine (DIPEA) in N,N-dimethylformamide at room temperature for 5-7 h; and removing the protecting group from the resulting product using a solution of hydrogen chloride / dioxane in a mixture of dichloromethane / acetonitrile at room temperature for 1-4 h to obtain the trisubstituted quinazoline derivative.

[0058] Method 3:

[0059] C1. mixing 2,4-dichloro-6-methoxyquinazoline, 4-[2-(pyrrolidin-l-yl)ethoxy]aniline and isopropanol, and reacting at 50-70 °C for 2-4 h in the presence of concentrated hydrochloric acid to obtain an arylamine product; using a solution of HBr / HOAc, and reacting at 100-120 °C for 10-12 h to obtain a phenolic intermediate; etherifying the phenolic intermediate with l-azido-6-bromohexane to obtain a terminal azido intermediate;

[0060] C2. reacting the azido intermediate with an alkynyl benzoate in the presence of copper sulfate pentahydrate and sodium ascorbate in a solution of tert-butanol / water at 70-90 °C for 6-7 h to obtain a 1,2,3-triazole ring-containing intermediate;

[0061] C3. hydrolyzing the 1,2,3-triazole ring-containing intermediate in a solution of lithium hydroxide in methanol / water at room temperature for 10-12 h, condensing the resulting carboxylic acid with O-(tetrahydro-2H-pyran-2-yl)hydroxylamine in the presence of N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-l-yl)urea hexafluorophosphate (HATU) and N,N-diisopropylethylamine (DIPEA) in N,N-dimethylformamide at room temperature for 5-7 h; and removing the protecting group from the resulting product using a solution of hydrogen chloride / dioxane in a mixture of dichloromethane / acetonitrile at room temperature for 1-4 h to obtain the trisubstituted quinazoline derivative.

[0062] Method 4:

[0063] D1. reacting 2,4-dichloro-6-bromoquinazoline with 4-aminophenol, etherifying the phenolic hydroxyl group with an ω-bromoalkanoate to obtain 6-bromo-2-chloro-4-(ω-alkyloxyphenoxy)quinazoline; and reacting the 6-bromo-2-chloro-4-(ω-alkyloxyphenoxy)quinazoline with pyrrolidine to obtain a 6-bromo-2-(pyrrolidin-l-yl) key intermediate;

[0064] D2, the 6-bromo intermediate is reacted with various primary or secondary amines (such as 3-(pyrrolidin-1-yl)propylamine) in the presence of methane sulfonic acid (2-dicyclohexylphosphino)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (BrettPhos-Pd-G3) catalyst and sodium tert-butoxide in anhydrous 1,4-dioxane at 110-130 °C for 6-9 h;

[0065] D3, the intermediate obtained in D2 is hydrolyzed in the presence of lithium hydroxide in methanol / water solution at room temperature for 10-12 h, and the resulting carboxylic acid is condensed with O-(tetrahydro-2H-pyran-2-yl)hydroxylamine in the presence of N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)urea hexafluorophosphate (HATU) and N,N-diisopropylethylamine (DIPEA) in N,N-dimethylformamide at room temperature for 5-7 h; the resulting product is deprotected using hydrogen chloride / dioxane solution in dichloromethane / acetonitrile mixed solvent at room temperature for 1-4 h to obtain the trisubstituted quinazoline derivative.

[0066] In some embodiments of the present application, in step A1, the molar ratio of the 6-bromo-2, 4-dichloroquinazoline, 4-[2-(pyrrolidin-1-yl)ethoxy]aniline, isopropyl alcohol is 1:(0.8-1.2).

[0067] In some embodiments of the present application, in step A1, the molar ratio of the arylamination product, pyrrolidine, cesium carbonate, potassium iodide is 1:(1.6-2.4):(2.4-3.6):(0.8-1.2).

[0068] In some embodiments of the present application, in step A2, the molar ratio of the side chain boronate carboxylic acid, O-(tetrahydro-2H-pyran-2-yl)hydroxylamine, dichloro[1, 1'-bis(diphenylphosphino)ferrocene]palladium(II), and potassium acetate is 1:(0.9-1.4):(0.08-0.12):(2.4-3.6).

[0069] In some embodiments of the present application, in step C2, the molar ratio of the azido intermediate, alkynyl benzoate, copper sulfate pentahydrate, and sodium ascorbate is 1:(0.9-1.4):(0.03-0.05):(0.8-1.2).

[0070] In some embodiments of the present application, in step D2, the molar ratio of the 6-bromo intermediate, primary or secondary amine, and BrettPhos-Pd-G3 is 1:(1.2-1.8):(0.12-0.18).

[0071] The third aspect of the present application provides the use of the trisubstituted quinazoline derivative or the pharmaceutically acceptable salt thereof according to the first aspect of the present application in the preparation of a dual-target drug for treating colorectal cancer.

[0072] In some embodiments of the present application, the dual-target drug is a G-quadruplex stabilizer and a histone deacetylase inhibitor.

[0073] Compared with the prior art, the present application has the following beneficial effects:

[0074] The trisubstituted quinazoline derivative provided by the present application integrates the efficient G4 stabilizing module (polysubstituted quinazoline core) and the potent HDAC inhibiting pharmacophore (hydroxamic acid) into a single molecule through an adjustable connecting chain, and ingeniously solves the bottleneck problems of poor target accessibility and high off-target toxicity of traditional G4 ligands caused by the chromatin barrier. The preferred compound a10 exhibits excellent dual activity in vitro, and the stability of telomeric G4 is significantly better than that of the lead compound, and the inhibitory activity on HDAC reaches the nanomolar level; the series of compounds exhibit strong inhibitory activity on colorectal cancer cells, and the toxicity on normal intestinal epithelial cells is significantly reduced, and the selectivity ratio is up to 39 times, and the safety is significantly improved; the in vivo efficacy experiment further confirms that the preferred molecule can effectively inhibit tumor growth, and no obvious toxicity is observed; the present application not only provides a new type of lead compound with synergistic anti-tumor effect, but also provides a new strategy and tool for overcoming the challenge of colorectal cancer treatment. BRIEF DESCRIPTION OF DRAWINGS

[0075] Figure 1 Results of in vivo efficacy evaluation of the trisubstituted quinazoline derivative a10. DETAILED DESCRIPTION

[0076] The content of the present application is further described in detail through specific examples. Unless otherwise specified, the raw materials, reagents or devices used in the examples can be obtained from conventional commercial channels, or can be obtained by existing technical methods. Unless otherwise specified, the test or test method is a conventional method in the art.

[0077] The following examples prepare trisubstituted quinazoline derivatives a1-a12, a15-a22, b1-b3 and b5-b9, and the structures of the compounds are as follows:

[0078]

[0079]

[0080]

[0081]

[0082]

[0083]

[0084] wherein the synthesis routes and related intermediates of the trisubstituted quinazoline derivatives a1-a12, a15-a22, b1-b3 and b5-b9 are shown in the following synthesis route schemes:

[0085] Route 1:

[0086]

[0087] Route 2:

[0088]

[0089] Route 3:

[0090]

[0091]

[0092]

[0093]

[0094] wherein pyrrolydine is a pyrrolyl group; diethylamine is a diethylamine group.

[0095] Route 4:

[0096]

[0097]

[0098] Example 1

[0099] This example prepares the trisubstituted quinazoline derivative a1, with the following steps:

[0100] 1) 6-bromo-2, 4-dichloroquinazoline (4.3 mmol, 1.0 equiv.) and 4-[2-(pyrrolidin-1- yl)ethoxy]aniline (intermediate 1) (4.3 mmol, 1.0 equiv.) were added to 20 mL of isopropanol, stirred, and then 2 drops of concentrated hydrochloric acid were added, and the reaction was allowed to proceed at 60 °C for 2 h, with monitoring of the reaction progress by thin layer chromatography. Upon completion of the reaction, the crude product was precipitated out of solution, collected by suction filtration, and the precipitate was washed with isopropanol and methanol to give intermediate 2 as a light brown solid in 77% yield;

[0101] 2) Intermediate 2 (2.9 mmol, 1.0 equiv.), pyrrolidine (5.8 mmol, 2 equiv.), cesium carbonate (8.7 mmol, 3.0 equiv.) and potassium iodide (2.9 mmol, 1.0 equiv.) were stirred in 8 mL of N,N-dimethylformamide at 110 °C for 12 h, and the reaction progress was monitored by thin layer chromatography. After the reaction was completed, the mixture was cooled to room temperature, 100 mL of water was added, and the aqueous phase was extracted with ethyl acetate (30 mL x 6). The combined organic phase was washed with saturated NaCl solution (30 mL x 3), dried over anhydrous sodium sulfate, and then concentrated, adsorbed on silica gel, and purified by column chromatography to obtain intermediate 3 as a brown solid with a yield of 64%;

[0102] 3) 3-(4-bromophenyl)propanoic acid (4.4 mmol, 1.0 equiv.), bis(pinacolato)diboron (5.3 mmol, 1.2 equiv.), dichloro[l, l'-bis(diphenylphosphino)ferrocene] palladium(II) (0.44 mmol, 0.1 equiv.) and potassium acetate (13.1 mmol, 3.0 equiv.) were added to 10 mL of anhydrous 1, 4-dioxane, and stirred at 100 °C for 8 h under nitrogen protection. After the reaction was completed, the mixture was filtered through diatomite, rinsed with ethyl acetate and methanol, concentrated under vacuum, then adsorbed on silica gel and purified by column chromatography to obtain intermediate 4 as a white liquid with a yield of 63%;

[0103] 4) Intermediate 4 (1.8 mmol, 1.0 equiv.), l-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (2.2 mmol, 1.2 equiv.) and 1-hydroxybenzotriazole (1.8 mmol, 1.0 equiv.) were dissolved in 10 mL of anhydrous dichloromethane and stirred at room temperature, followed by slowly adding triethylamine (5.4 mmol, 3.0 equiv.). After 15 min, O-(tetrahydro-2H-pyran-2-yl)hydroxylamine was added to the reaction system. After the reaction was completed, ice water was added to the system, the aqueous phase was extracted with dichloromethane (30 mL x 6), the combined organic phase was washed with saturated sodium chloride solution (30 mL x 3), dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The residue was adsorbed on silica gel and purified by flash column chromatography to obtain intermediate 8 as a light yellow liquid with a yield of 74%;

[0104] 5) Intermediate 3 (0.6 mmol, 1.0 equiv.), intermediate 8 (0.65 mmol, 1.1 equiv.) and sodium carbonate (1.8 mmol, 3.0 equiv.) were dissolved in 10 mL of 1, 4-dioxane and 3 mL of water, dichlorobis(diphenylphosphino)ferrocenepalladium (0.09 mmol, 0.15 equiv.) was added under nitrogen protection, the reaction was stirred at 100 °C for 8 h, after the reaction was cooled to room temperature, the organic phase was combined and concentrated under reduced pressure, the residue was adsorbed on silica gel and purified by flash column chromatography to obtain intermediate 12 yellow solid product, the yield was 51%;

[0105] 6) Intermediate 12 (100 mg, 0.15 mmol, 1.0 equiv.) was dissolved in 2 mL of dichloromethane and 1 mL of acetonitrile, then 4 mol / L hydrogen chloride / dioxane solution (0.1 mL, 0.75 mmol, 5.0 equiv.) was added, the reaction was stirred at room temperature for 2 h, when thin layer chromatography showed that the reaction was complete, the reaction mixture was filtered, then recrystallized with ethanol and acetonitrile to obtain the trisubstituted quinazoline derivative a1 light yellow solid product, the yield was 34%.

[0106] Example 2

[0107] This example prepared trisubstituted quinazoline derivative a2, the preparation method was the same as example 1, as follows:

[0108] 1) 4-(4-bromophenyl)butyric acid reacted with bis(pinacolato)diboron to obtain intermediate 5 white liquid product, the yield was 63%;

[0109] 2) Intermediate 5 reacted with O-(tetrahydro-2H-pyran-2-yl)hydroxylamine to obtain intermediate 9 light yellow solid product, the yield was 73%;

[0110] 3) Intermediate 3 reacted with intermediate 9 to obtain intermediate 13 yellow solid product, the yield was 55%;

[0111] 4) Intermediate 13 removed the protecting group to obtain trisubstituted quinazoline derivative a2 light yellow solid product, the yield was 33%.

[0112] Example 3

[0113] This example prepared trisubstituted quinazoline derivative a3, the preparation method was the same as example 1, as follows:

[0114] 1) 5-(4-bromophenyl)pentanoic acid reacted with bis(pinacolato)diboron to obtain intermediate 6 white liquid product, the yield was 56%;

[0115] 2) Intermediate 6 reacted with O-(tetrahydro-2H-pyran-2-yl)hydroxylamine to give intermediate 10 as a light yellow solid with a yield of 68%;

[0116] 3) Intermediate 3 reacted with intermediate 10 to give intermediate 14 as a yellow solid with a yield of 58%;

[0117] 4) Intermediate 14 was deprotected to give a trisubstituted quinazoline derivative a3 as a light yellow solid with a yield of 28%.

[0118] Example 4

[0119] In this example, a trisubstituted quinazoline derivative a4 was prepared according to the method of Example 1 as follows:

[0120] 1) 4-(4-bromophenoxy)butyric acid reacted with bis(pinacolato)diboron to give intermediate 7 as a white solid with a yield of 72%;

[0121] 2) Intermediate 7 reacted with O-(tetrahydro-2H-pyran-2-yl)hydroxylamine to give intermediate 11 as a light yellow solid with a yield of 62%;

[0122] 3) Intermediate 3 reacted with intermediate 11 to give intermediate 15 as a yellow solid with a yield of 50%;

[0123] 4) Intermediate 15 was deprotected to give a trisubstituted quinazoline derivative a4 as a light yellow solid with a yield of 47%.

[0124] Example 5

[0125] In this example, a trisubstituted quinazoline derivative a5 was prepared according to the following steps:

[0126] 1) 4-(4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl)phenol (2.2 mmol, 1.0 equiv.), methyl 6-bromohexanoate (4.4 mmol, 2.0 equiv.) and cesium carbonate (6.5 mmol, 3.0 equiv.) were dissolved in 4 mL of N, N-dimethylformamide, and the reaction was stirred at 80°C for 12 h, and the reaction progress was monitored by thin layer chromatography; after the reaction was cooled to room temperature, 100 mL of water was added to the reaction system, the aqueous phase was extracted with ethyl acetate (30 mL x 6), the combined organic phase was washed with saturated sodium chloride solution (30 mL x 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure; the residue was adsorbed on silica gel and purified by flash column chromatography to give intermediate 16 as a white liquid with a yield of 89%;

[0127] 2) Intermediate 3 reacted with intermediate 16 to give intermediate 18 as a yellow solid with a yield of 50% (same as the preparation method of intermediate 12).

[0128] 3) Intermediate 18 (0.34 mmol, 1.0 equiv.) was dissolved in 10 mL of methanol, and lithium hydroxide (3.4 mmol, 10 equiv.) was dissolved in water (2 mol / L) and added to the methanol solution of intermediate 18, which was stirred at room temperature for 12 h, and the reaction progress was monitored by thin layer chromatography. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the pH was adjusted to 7-8 with hydrochloric acid (1 mol / L, aqueous solution). Then, the aqueous phase was concentrated under reduced pressure again to obtain the acidic intermediate, which was directly used in the next step;

[0129] 4) The above acidic intermediate (0.34 mmol, 1.0 equiv.), O-(tetrahydro-2H-pyran-2-yl) hydroxylamine (1.7 mmol, 5.0 equiv.), N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl) urea hexafluorophosphate, N,N-diisopropyl ethylamine (0.5 mmol, 1.5 equiv.) and N,N-diisopropyl ethylamine were added to 4 mL of N,N-dimethylformamide, and the reaction was stirred at room temperature for 2 h, and the reaction progress was monitored by thin layer chromatography. After the reaction was completed, 100 mL of water was added to the reaction system, and the aqueous phase was extracted with ethyl acetate (30 mL x 6), and the combined organic phase was washed with saturated sodium chloride solution (30 mL x 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was adsorbed on silica gel and purified by flash column chromatography to obtain intermediate 20 as a yellow solid product, with a total yield of 25% for the two steps;

[0130] 5) The removal of the protecting group from intermediate 20 obtained a yellow solid product of the trisubstituted quinazoline derivative a5, with a yield of 41% (same preparation method as in Example 1).

[0131] Example 6

[0132] This example prepared a trisubstituted quinazoline derivative a6, and the preparation method was the same as in Example 5, as follows:

[0133] 1) 4-(4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenol was reacted with 7-bromoheptanoic acid methyl ester to obtain intermediate 17 as a white liquid product, with a yield of 90%;

[0134] 2) Intermediate 3 was reacted with intermediate 17 to obtain intermediate 19 as a yellow solid product, with a yield of 71%;

[0135] 3) The hydrolysis of intermediate 19 obtained an acidic intermediate, which was directly used in the next step;

[0136] 4) The above acid intermediate was reacted with O-(tetrahydro-2H-pyran-2-yl) hydroxylamine to give intermediate 21 as a yellow solid with a total yield of 25% for two steps;

[0137] 5) The intermediate 21 was deprotected to give the trisubstituted quinazoline derivative a6 as a yellow solid with a yield of 42%.

[0138] Example 7

[0139] The trisubstituted quinazoline derivative a7 was prepared in this example, and the steps were as follows:

[0140] 1) 2, 4-dichloro-6-methoxyquinazoline was reacted with intermediate 1 to give intermediate 25 as a yellow solid with a yield of 71.8% (same as the preparation method of intermediate 2);

[0141] 2) The intermediate 25 was reacted with pyrrolidine to give intermediate 30 as a brown solid with a yield of 49.1% (same as the preparation method of intermediate 3);

[0142] 3) The intermediate 30 (4.6 mmol, 1.0 equiv.) was dissolved in 5 mL of acetic acid, followed by the addition of hydrogen bromide (23.0 mmol, 5.0 equiv., 33% in acetic acid), and the reaction was stirred at 110°C for 8 h. After the reaction was cooled to room temperature, the mixture was filtered under reduced pressure, ice water was added to the filtrate, the aqueous phase was adjusted to pH 8 with saturated potassium bicarbonate, a precipitate was formed, which was collected and filtered to give intermediate 35 as a dark green solid with a yield of 98.2%;

[0143] 4) The intermediate 35 (0.15 mmol, 1.0 equiv.), methyl 4-(4-bromobutyl)benzoate (0.17 mmol, 1.1 equiv.), and cesium carbonate (0.23 mmol, 1.5 equiv.) were dissolved in 2 mL of N,N-dimethylformamide, and the reaction was stirred at 60°C for 2 h while monitoring the progress of the reaction by thin layer chromatography. After the reaction was cooled to room temperature, 100 mL of water was added to the reaction mixture, the aqueous phase was extracted with ethyl acetate (30 mL x 6), the combined organic phase was washed with saturated NaCl solution (30 mL x 3), and dried over anhydrous sodium sulfate. Then, the organic phase was concentrated under reduced pressure, adsorbed on silica gel, and purified by column chromatography to give intermediate 40 as a brown solid with a yield of 65.6%;

[0144] 5) The intermediate 40 was hydrolyzed and reacted with O-(tetrahydro-2H-pyran-2-yl) hydroxylamine to give intermediate 51 as a dark yellow solid with a yield of 54.1% (same as the preparation method of intermediate 20);

[0145] 6) Deprotection of the protecting group of intermediate 51 to obtain the trisubstituted quinazoline derivative a7 as a yellow solid with a yield of 54.1% (prepared in the same manner as in Example 1).

[0146] Example 8

[0147] In this example, the trisubstituted quinazoline derivative a8 is prepared as follows:

[0148] 1) Reaction of intermediate 35 with methyl 5-bromovalerate to obtain intermediate 41 as a brown solid with a yield of 52.4% (prepared in the same manner as intermediate 40);

[0149] 2) Hydrolysis of intermediate 41 followed by reaction with O-(tetrahydro-2H-pyran-2- yl)hydroxylamine to obtain intermediate 52 as a yellow solid with a yield of 41.4% (prepared in the same manner as intermediate 20);

[0150] 3) Deprotection of the protecting group of intermediate 52 to obtain the trisubstituted quinazoline derivative a8 as a yellow solid with a yield of 54.1% (prepared in the same manner as in Example 1).

[0151] Example 9

[0152] In this example, the trisubstituted quinazoline derivative a9 is prepared as follows:

[0153] Intermediate 52 (0.1 mmol, 1.0 equiv.) was dissolved in 3 mL of N,N- dimethylformamide, N,N-diisopropylethylamine (0.3 mmol, 3.0 equiv.), N,N,N',N'- tetramethyl-O-(7-azabenzotriazol-1-yl)urea hexafluorophosphate (0.15 mmol, 1.5 equiv.) and o-phenylenediamine (0.15 mmol, 1.5 equiv.) were added successively, and the mixture was stirred at room temperature overnight. Then, 30 mL of dichloromethane was added, and the mixture was washed with water and saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate, and then concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography to obtain the trisubstituted quinazoline derivative a9 as a yellow solid with a yield of 75.0%.

[0154] Example 10

[0155] In this example, the trisubstituted quinazoline derivative a10 is prepared as follows:

[0156] 1) Reaction of intermediate 35 with methyl 7-bromoheptanoate to obtain intermediate 42 as a brown solid with a yield of 89.6% (prepared in the same manner as intermediate 40);

[0157] 2) hydrolysis of intermediate 42 followed by reaction with O-(tetrahydro-2H-pyran-2- yl)hydroxylamine to give intermediate 53 as a dark yellow solid in 33.8% yield (same procedure as for intermediate 20);

[0158] 3) deprotection of intermediate 53 to give tri-substituted quinazoline derivative a10 as a yellow solid in 66.2% yield (same procedure as for Example 1).

[0159] Example 11

[0160] In this example, tri-substituted quinazoline derivative a11 is prepared according to the following procedure:

[0161] Intermediate 53 (0.1 mmol, 1.0 equiv.) was dissolved in 3 mL of N,N- dimethylformamide, followed by the addition of N,N-diisopropylethylamine (0.3 mmol, 3.0 equiv.), N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)urea hexafluorophosphate (0.15 mmol, 1.5 equiv.) and o-phenylenediamine (0.15 mmol, 1.5 equiv.). The mixture was stirred at room temperature overnight, then diluted with 30 mL of dichloromethane and washed with water and saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude product, which was purified by column chromatography to give tri-substituted quinazoline derivative a11 as a yellow solid in 76.4% yield.

[0162] Example 12

[0163] In this example, tri-substituted quinazoline derivative a12 is prepared according to the following procedure:

[0164] 1) reaction of intermediate 35 with methyl 3-(4-bromobutyl)benzoate to give intermediate 43 as a brown solid in 57.6% yield (same procedure as for intermediate 40);

[0165] 2) hydrolysis of intermediate 43 followed by reaction with O-(tetrahydro-2H-pyran-2- yl)hydroxylamine to give intermediate 54 as a dark yellow solid in 52.4% yield (same procedure as for intermediate 20);

[0166] 3) deprotection of intermediate 54 to give tri-substituted quinazoline derivative a12 as a yellow solid in 46.2% yield (same procedure as for Example 1).

[0167] Example 13

[0168] In this example, tri-substituted quinazoline derivative a13 is prepared according to the following procedure:

[0169] 1) Intermediate 35 was reacted with tert-butyl 3-(2-bromoethoxy)propanoate to give intermediate 46 as a brown solid in 22.7% yield (same procedure as for intermediate 40);

[0170] 2) Intermediate 46 (0.25 mmol, 1.0 equiv.) was dissolved in 10 mL of dichloromethane, then 1 mL of trifluoroacetic acid was added, the mixture was stirred at room temperature for 4 h, and the reaction progress was monitored by thin layer chromatography, after the reaction was completed, saturated potassium carbonate was added to the reaction mixture to adjust the pH to 8, and the aqueous phase was extracted with ethyl acetate (30 mL x 6), the combined organic phase was washed with saturated NaCl solution (30 mL x 3), and dried over anhydrous sodium sulfate, and the obtained crude product was directly used in the next step reaction;

[0171] 3) The obtained crude product (0.25 mmol, 1.0 equiv.), O-(tetrahydro-2H-pyran-2-yloxy)hydroxylamine (1.25 mmol, 5 equiv.), N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)urea hexafluorophosphate (0.4 mmol, 1.5 equiv.) and N-ethyl-N-isopropylpropylamine (N,N-diisopropylethylamine, 2.5 mmol, 10 equiv.) were dissolved in 2 mL of N,N-dimethylformamide, stirred at room temperature for 2 h, and the reaction progress was monitored by thin layer chromatography, after the reaction was completed, 100 mL of water was added to the reaction mixture, the aqueous phase was extracted with ethyl acetate (30 mL x 6), the combined organic phase was washed with saturated NaCl solution (30 mL x 3), and dried over anhydrous sodium sulfate, and the organic phase was concentrated under reduced pressure, then adsorbed on silica gel, and purified by column chromatography to give intermediate 57 as a yellow solid in 24.7% yield;

[0172] 4) Intermediate 57 was deprotected to give the trisubstituted quinazoline derivative a15 as a light yellow solid in 53.7% yield (same procedure as in Example 1).

[0173] Example 14

[0174] This example prepared the trisubstituted quinazoline derivative a16, and the steps were as follows:

[0175] 1) Intermediate 36 was reacted with methyl 7-bromoheptanoate to give intermediate 47 as a brown solid in 70.1% yield (same procedure as for intermediate 40);

[0176] 2) After intermediate 47 was hydrolyzed and reacted with O-(tetrahydro-2H-pyran-2-yl)hydroxylamine, intermediate 58 was obtained as a dark yellow solid in 26.8% yield (same procedure as for intermediate 20);

[0177] 3) Removal of the protecting group from intermediate 58 to give the trisubstituted quinazoline derivative a16 as a light yellow solid in 72.7% yield (same procedure as in example 1).

[0178] Example 15

[0179] In this example, the trisubstituted quinazoline derivative a17 was prepared as follows:

[0180] 1) Reaction of intermediate 37 with methyl 7-bromoheptanoate to give intermediate 48 as a brown solid in 50.1% yield (same procedure as in intermediate 40);

[0181] 2) Hydrolysis of intermediate 48 followed by reaction with O-(tetrahydro-2H-pyran-2- yl)hydroxylamine to give intermediate 59 as a dark yellow solid in 26.5% yield (same procedure as in intermediate 20);

[0182] 3) Removal of the protecting group from intermediate 59 to give the trisubstituted quinazoline derivative a17 as a yellow solid in 57.2% yield (same procedure as in example 1).

[0183] Example 16

[0184] In this example, the trisubstituted quinazoline derivative a18 was prepared as follows:

[0185] 1) Reaction of intermediate 38 with methyl 7-bromoheptanoate to give intermediate 49 as a brown solid in 22.7% yield (same procedure as in intermediate 40);

[0186] 2) Hydrolysis of intermediate 49 followed by reaction with O-(tetrahydro-2H-pyran-2- yl)hydroxylamine to give intermediate 60 as a dark yellow solid in 79.9% yield (same procedure as in intermediate 20);

[0187] 3) Removal of the protecting group from intermediate 60 to give the trisubstituted quinazoline derivative a18 as a yellow solid in 55.8% yield (same procedure as in example 1).

[0188] Example 17

[0189] In this example, the trisubstituted quinazoline derivative a19 was prepared as follows:

[0190] 1) Reaction of intermediate 39 with methyl 7-bromoheptanoate to give intermediate 50 as a brown solid in 48.0% yield (same procedure as in intermediate 40);

[0191] 2) Hydrolysis of intermediate 50 followed by reaction with O-(tetrahydro-2H-pyran-2- yl)hydroxylamine to give intermediate 61 as a dark yellow solid in 43.4% yield (same procedure as in intermediate 20);

[0192] 3) Deprotection of intermediate 61 to give the trisubstituted quinazoline derivative a19 as a yellow solid in 45.2% yield (same procedure as in example 1).

[0193] Example 18

[0194] In this example, the trisubstituted quinazoline derivative a20 was prepared according to the following procedure:

[0195] 1) Intermediate 40 (0.25 mmol, 1.0 equiv.) was dissolved in 2 mL of ethanol, 80% hydrazine monohydrate (10 mmol, 40 equiv.) was added, the mixture was heated to reflux and stirred for 12 h, the reaction was cooled to room temperature and concentrated under vacuum to give intermediate 62 as a yellow solid in 64% yield;

[0196] 2) Intermediate 62 (0.16 mmol, 1.0 equiv.) was dissolved in 5 mL of tetrahydrofuran, hexanal (1.6 mmol, 10 equiv.) was added, the resulting mixture was stirred at room temperature for 24 h, then the solvent was removed under vacuum, the resulting mixture was then dissolved in 1, 2-dichloroethane, sodium triethoxyborohydride (0.8 mmol, 5 equiv.) was added portionwise under ice bath and nitrogen protection, after the reaction was completed, ice water was added, the aqueous phase was extracted with dichloromethane (30 mL x 6), the organic phases were combined, washed with saturated NaCl solution (30 mL x 3), dried over anhydrous sodium sulfate, then the organic phase was concentrated under vacuum, adsorbed on silica gel and purified by flash column chromatography to give the trisubstituted quinazoline derivative a20 as a yellow solid in 34% yield.

[0197] Example 19

[0198] In this example, the trisubstituted quinazoline derivative a21 was prepared according to the following procedure:

[0199] 1) Intermediate 36 was reacted with 1-azido-6-bromohexane to give intermediate 63 as a brown solid in 65.6% yield (same procedure as for intermediate 40);

[0200] 2) Intermediate 63 (0.15 mmol, 1.0 equiv.) and methyl 3-alkynyl benzoate (0.18 mmol, 1.2 equiv.) were dissolved in a mixed solvent of 6 mL tert-butanol and 2 mL water. The mixture was stirred at room temperature, and then copper sulfate pentahydrate aqueous solution (0.1 mol / L, 0.06 mmol, 0.4 equiv., 0.6 mL) and sodium ascorbate solution (0.1 mol / L, 0.15 mmol, 1.0 equiv., 11.5 mL) were added dropwise. The mixture was heated at 80 °C for 4 h, and the reaction was monitored for completion by thin-layer chromatography. After the reaction was cooled to room temperature, 100 mL of water was added, and the aqueous phase was extracted with ethyl acetate (30 mL × 6). The combined organic phases were washed with saturated NaCl solution (30 mL × 3), dried with anhydrous sodium sulfate, and then the organic phase was concentrated, adsorbed onto silica gel, and purified by column chromatography to obtain intermediate 64, a light brown solid product, with a yield of 53.2%.

[0201] 3) After hydrolyzing intermediate 64, it was reacted with O-(tetrahydro-2H-pyran-2-yl)hydroxylamine to obtain intermediate 65, a yellow solid product, with a yield of 52.1% (same as the preparation method of intermediate 51).

[0202] 4) The protecting group of intermediate 65 was removed to obtain the trisubstituted quinazoline derivative a21, a yellow solid product, with a yield of 47.8% (same as the preparation method in Example 1).

[0203] Example 20

[0204] In this embodiment, the trisubstituted quinazoline derivative a22 is prepared by the following steps:

[0205] Intermediate 35 (0.12 mmol, 1.0 equiv.), 1-bromohexane (0.14 mmol, 1.2 equiv.), and cesium carbonate (0.24 mmol, 2.0 equiv.) were mixed in 2 mL of N,N-dimethylformamide and stirred at 45 °C for 2 h. The reaction progress was monitored by thin-layer chromatography. After the reaction was cooled to room temperature, 100 mL of water was added to the reaction mixture. The aqueous phase was extracted with ethyl acetate (30 mL × 6). The organic phases were combined, washed with saturated NaCl solution (30 mL × 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure, adsorbed onto silica gel, and purified by column chromatography to give the trisubstituted quinazoline derivative a22 as a yellow solid product with a yield of 46.2%.

[0206] Example 21

[0207] In this embodiment, the trisubstituted quinazoline derivative b1 is prepared by the following steps:

[0208] 1) Intermediate 66 (1.6 mmol, 1.0 equiv.), methyl 5-bromopentanoate (2.4 mmol, 1.5 equiv.), cesium carbonate (4.8 mmol, 3 equiv.) were mixed in 8 mL of acetonitrile and stirred at 95 °C for 12 h, and the reaction progress was monitored by thin layer chromatography. After cooling to room temperature, the organic phase was concentrated under reduced pressure and adsorbed on silica gel and purified by column chromatography to give intermediate 68 as a white solid in 45.5% yield.

[0209] 2) Intermediate 68 was reacted with pyrrolidine to give intermediate 73 as a brown solid in 57.3% yield (preparation method as intermediate 3);

[0210] 3) Intermediate 73 was hydrolyzed and reacted with O-(tetrahydro-2H-pyran-2- yl)hydroxylamine to give intermediate 78 as a light yellow solid in 63.7% yield (preparation method as intermediate 20);

[0211] 4) Intermediate 78 was deprotected to give tri-substituted quinazoline derivative b1 as a white solid in 51.2% yield (preparation method as example 1).

[0212] Example 22

[0213] This example prepared tri-substituted quinazoline derivative b2, following steps:

[0214] 1) Intermediate 66 was reacted with methyl 7-bromoheptanoate to give intermediate 69 as a white solid in 45.5% yield (preparation method as intermediate 68);

[0215] 2) Intermediate 69 was reacted with pyrrolidine to give intermediate 74 as a brown solid in 62.0% yield (preparation method as intermediate 3);

[0216] 3) Intermediate 74 was hydrolyzed and reacted with O-(tetrahydro-2H-pyran-2- yl)hydroxylamine to give intermediate 79 as a light yellow solid in 73.1% yield (preparation method as intermediate 20);

[0217] 4) Intermediate 79 was deprotected to give tri-substituted quinazoline derivative b2 as a light yellow solid in 62.1% yield (preparation method as example 1).

[0218] Example 23

[0219] This example prepared tri-substituted quinazoline derivative b3, following steps:

[0220] Intermediate 79 was deprotected to give tri-substituted quinazoline derivative b3 as a white solid in 63.3% yield (preparation method as example 1).

[0221] Example 24

[0222] This example prepares trisubstituted quinazoline derivative b5, following steps:

[0223] 1) Intermediate 66 was reacted with methyl 4-(4-bromobutyl)benzoate to give intermediate 71 as white solid product with a yield of 37.8% (same preparation method as intermediate 68);

[0224] 2) Intermediate 71 was reacted with pyrrolidine to give intermediate 76 as brown solid product with a yield of 38.9% (same preparation method as intermediate 3);

[0225] 3) After hydrolysis of intermediate 76, it was reacted with O-(tetrahydro-2H-pyran-2- yl)hydroxylamine to give intermediate 81 as light yellow solid product with a yield of 31.1% (same preparation method as intermediate 20);

[0226] 4) Intermediate 81 was deprotected to give trisubstituted quinazoline derivative b5 as white solid product with a yield of 53.2% (same preparation method as example 1).

[0227] Example 25

[0228] This example prepares trisubstituted quinazoline derivative b6, following steps:

[0229] 1) Intermediate 72 was reacted with pyrrolidine to give intermediate 77 as brown solid product with a yield of 67.0% (same preparation method as intermediate 3);

[0230] 2) Intermediate 77 (1.1 mmol, 1.0 equiv.), 3-(pyrrolidin-1-yl)propylamine (1.6 mmol, 1.5 equiv.), sodium tert-butoxide (2.1 mmol, 2.0 equiv.) and BrettPhos-Pd-G3 (0.16 mmol, 0.15 equiv.) were added into 15 mL of anhydrous 1, 4-dioxane, stirred at 100 °C under nitrogen protection for 8 h, and the reaction progress was monitored by thin layer chromatography, after cooling to room temperature, the organic phase was concentrated under reduced pressure, adsorbed on silica gel and purified by column chromatography to give dark yellow solid product with a yield of 65.0%;

[0231] 3) The resulting product (0.16 mmol, 1.0 equiv.), O-(tetrahydro-2H-furan-2-yl) hydroxylamine (0.32 mmol, 2 equiv.), N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-l-yl) urea hexafluorophosphate (0.25 mmol, 1.5 equiv.) and N,N-diisopropylethylamine (0.65 mmol, 5 equiv.) were dissolved in 2 mL of N,N-dimethylformamide, stirred at room temperature for 2 h and the reaction progress was monitored by thin layer chromatography, after cooling to room temperature, 100 mL of water was added to the reaction, the aqueous phase was extracted with ethyl acetate (30 mL x 6), the combined organic phase was washed with saturated aqueous NaCl (30 mL x 3), dried over anhydrous sodium sulfate, the organic phase was concentrated under reduced pressure, adsorbed on silica gel and purified by column chromatography to obtain the intermediate 82 yellow solid product with a yield of 43%;

[0232] 4) The intermediate 82 was deprotected to obtain the trisubstituted quinazoline derivative b6 yellow solid product with a yield of 67.0% (same preparation method as in Example 1).

[0233] Example 26

[0234] The trisubstituted quinazoline derivative b7 was prepared in this example, and the steps were as follows:

[0235] 1) The intermediate 83 yellow solid product was synthesized by the same method as the intermediate 82 with a yield of 27.8%;

[0236] 2) The intermediate 83 was deprotected to obtain the trisubstituted quinazoline derivative b7 yellow solid product with a yield of 66.5% (same preparation method as in Example 1).

[0237] Example 27

[0238] The trisubstituted quinazoline derivative b8 was prepared in this example, and the steps were as follows:

[0239] 1) The intermediate 84 yellow solid product was synthesized by the same method as the intermediate 82 with a yield of 32.6%;

[0240] 2) The intermediate 84 was deprotected to obtain the trisubstituted quinazoline derivative b8 yellow solid product with a yield of 62.9% (same preparation method as in Example 1).

[0241] Example 28

[0242] The trisubstituted quinazoline derivative b9 was prepared in this example, and the steps were as follows:

[0243] 1) The intermediate 85 yellow solid product was synthesized by the same method as the intermediate 82 with a yield of 38.1%;

[0244] 2) Removal of the protecting group from intermediate 85 to obtain the trisubstituted quinazoline derivative b9 as a yellow solid in 71.2% yield (same preparation method as in example 1).

[0245] 1) NMR characterization of the intermediates involved in examples 1-28, NMR and high resolution mass spectrometry characterization of the trisubstituted quinazoline derivatives, results as follows:

[0246] Intermediate 2: 1 H NMR (400 MHz, DMSO) δ 9.00 (s, 1H), 7.94 (s, 1H), 7.71 (d, J = 8.8 Hz, 2H), 7.63 (d, J = 8.8 Hz, 1H), 7.12 - 7.06 (m, 2H), 6.84 (dd, J = 8.9, 3.8 Hz, 1H), 4.41 (t, J = 5.0 Hz, 2H), 3.65 - 3.56 (m, 8H), 3.13 (dd, J = 7.2, 3.8 Hz, 2H), 2.03 (t, J = 7.2 Hz, 4H), 1.91 (d, J = 8.4 Hz, 4H).

[0247] Intermediate 3: 1 H NMR (400 MHz, CDCl3) δ 7.75 (d, J = 2.1 Hz, 1H), 7.68 (dd, J = 9.2, 7.4 Hz, 2H), 7.61 - 7.55 (m, 1H), 7.39 (d, J = 8.9 Hz, 1H), 7.01 - 6.92 (m, 2H), 4.15 (t, J = 5.9 Hz, 2H), 3.65 (d, J = 6.6 Hz, 4H), 2.94 (t, J = 5.9 Hz, 2H), 2.68 (d, J = 6.2 Hz, 4H), 2.03 - 1.93 (m, 4H), 1.85 - 1.79 (m, 4H).

[0248] Intermediate 6: 1 H NMR (400 MHz, CDCl3) δ 7.75 - 7.70 (m, 2H), 7.21 - 7.16 (m, 2H), 2.68 - 2.60 (m, 2H), 2.39 - 2.33 (m, 2H), 1.72 - 1.64 (m, 4H), 1.34 (s, 12H).

[0249] Intermediate 8: 1H NMR (400 MHz, CDC13) δ 7.75 (d, J = 7.5 Hz, 2H), 7.22 (t, J = 7.0 Hz, 2H), 4.88 (s, 1H), 3.89 (t, J = 9.8 Hz, 1H), 3.57 (dt, J = 10.0, 4.2 Hz, 1H), 3.00 (t, J = 7.6 Hz, 2H), 2.59 - 2.37 (m, 2H), 1.79 (s, 2H), 1.67 - 1.50 (m, 2H), 1.35 (s, 12H).

[0250] Intermediate 9: 1 H NMR (400 MHz, CDC13) δ 7.76 (d, J = 7.6 Hz, 2H), 7.22 (d, J = 7.4 Hz, 2H), 4.96 (s, 1H), 3.94 (s, 1H), 3.66 (s, 1H), 2.78 - 2.64 (m, 2H), 2.11 (s, 2H), 2.03 (d, J = 7.0 Hz, 2H), 1.82 (s, 4H), 1.36 (s, 12H), 1.34 (s, 2H).

[0251] Intermediate 10: 1 H NMR (400 MHz, CDC13) δ 7.74 (d, J = 7.5 Hz, 2H), 7.20 (d, J = 7.5 Hz, 2H), 4.94 (s, 1H), 3.94 (s, 1H), 3.64 (d, J = 11.3 Hz, 1H), 2.72 - 2.60 (m, 2H), 2.22 - 2.09 (m, 2H), 1.82 (s, 2H), 1.70 (s, 4H), 1.59 (d, J = 15.3 Hz, 4H), 1.36 (s, 12H).

[0252] Intermediate 11: 1 H NMR (400 MHz, CDC13) δ 7.76 (d, J = 8.1 Hz, 2H), 6.94 - 6.85 (m, 2H), 4.96 (s, 1H), 4.06 (t, J = 6.0 Hz, 2H), 3.91 (t, J = 10.5 Hz, 1H), 3.60 (d, J = 11.4 Hz, 1H), 2.45 - 2.28 (m, 2H), 2.18 (q, J = 6.5 Hz, 2H), 1.59 (s, 4H), 1.35 (s, 12H).

[0253] Intermediate 12:1 H NMR (400 MHz, CDC13) δ 9.49 (s, 1H), 9.39 (s, 1H), 8.65 (d, J = 2.6 Hz, 1H), 7.94 (d, J = 8.0 Hz, 1H), 7.80 (dd, J = 8.1, 2.4 Hz, 1H), 7.63 - 7.56 (m, 2H), 7.39 - 7.33 (m, 2H), 7.23 - 7.17 (m, 2H), 6.89 - 6.83 (m, 2H), 4.94 (t, J = 3.2 Hz, 1H), 4.07 (t, J = 5.9 Hz, 2H), 3.86 - 3.75 (m, 5H), 3.66 (ddd, J = 11.1, 5.9, 3.7 Hz, 1H), 3.03 (t, J = 5.9 Hz, 2H), 2.90 - 2.82 (m, 2H), 2.82 (dd, J = 4.3, 3.2 Hz, 4H), 2.66 - 2.59 (m, 2H), 2.04 (d, J = 5.5 Hz, 3H), 1.94 - 1.85 (m, 6H), 1.81 - 1.56 (m, 4H).

[0254] Intermediate 13: 1 H NMR (400 MHz, CDC13) δ 7.91 (s, 1H), 7.76 (dd, J = 8.8, 2.1 Hz, 3H), 7.58 (d, J = 8.7 Hz, 1H), 7.53 (d, J = 8.0 Hz, 2H), 7.22 (d, J = 7.8 Hz, 2H), 6.97 - 6.91 (m, 2H), 4.78 (d, J = 3.5 Hz, 1H), 4.13 (t, J = 6.0 Hz, 2H), 3.90 (d, J = 6.4 Hz, 2H), 3.68 (d, J = 6.5 Hz, 4H), 3.56 (d, J = 10.6 Hz, 2H), 2.91 (t, J = 6.0 Hz, 2H), 2.70 (d, J = 5.8 Hz, 2H), 2.66 - 2.58 (m, 4H), 2.05 (s, 2H), 2.00 - 1.92 (m, 4H), 1.82 (h, J = 3.7 Hz, 6H), 1.73 (d, J = 17.1 Hz, 4H).

[0255] Intermediate 14: 1H NMR (400 MHz, CDC13) δ 7.80-7.71 (m, 4H), 7.58 (d, J = 9.2 Hz, 1H), 7.52 (d, J = 7.8 Hz, 2H), 7.23 (d, J = 7.8 Hz, 2H), 6.99-6.92 (m, 2H), 4.93 (s, 1H), 4.14 (t, J = 6.0 Hz, 2H), 3.92 (d, J = 6.6 Hz, 2H), 3.77-3.65 (m, 4H), 3.61 (d, J = 11.7 Hz, 2H), 2.92 (t, J = 6.0 Hz, 2H), 2.74-2.59 (m, 6H), 2.13 (s, 2H), 2.01-1.92 (m, 6H), 1.82 (p, J = 3.0 Hz, 6H), 1.70 (s, 4H).

[0256] Intermediate 15: 1 H NMR (400 MHz, CDC13) δ 7.80-7.71 (m, 4H), 7.58 (d, J = 9.2 Hz, 1H), 7.52 (d, J = 7.8 Hz, 2H), 7.23 (d, J = 7.8 Hz, 2H), 6.99-6.92 (m, 2H), 4.93 (s, 1H), 4.14 (t, J = 6.0 Hz, 2H), 3.92 (d, J = 6.6 Hz, 2H), 3.77-3.65 (m, 4H), 3.61 (d, J = 11.7 Hz, 2H), 2.92 (t, J = 6.0 Hz, 2H), 2.74-2.59 (m, 6H), 2.13 (s, 2H), 2.01-1.92 (m, 6H), 1.82 (p, J = 3.0 Hz, 6H), 1.70 (s, 4H).

[0257] Intermediate 18: 1H NMR (400 MHz, CDC13) δ 7.84 (s, 1H), 7.80-7.72 (m, 3H), 7.64 (d, J = 8.8 Hz, 1H), 7.62-7.57 (m, 2H), 7.01-6.95 (m, 4H), 4.17 (t, J = 5.9 Hz, 2H), 4.02 (t, J = 6.4 Hz, 2H), 3.70 (s, 3H), 3.67 (s, 4H), 2.95 (t, J = 5.9 Hz, 2H), 2.71-2.64 (m, 4H), 2.38 (t, J = 7.4 Hz, 2H), 2.05-1.92 (m, 5H), 1.89-1.79 (m, 8H), 1.74 (p, J = 7.5 Hz, 2H), 1.55 (tt, J = 9.2, 5.9 Hz, 2H).

[0258] Intermediate 19: 1 H NMR (400 MHz, CDC13) δ 7.84 (s, 1H), 7.80-7.72 (m, 3H), 7.64 (d, J = 8.8 Hz, 1H), 7.62-7.57 (m, 2H), 7.01-6.95 (m, 4H), 4.17 (t, J = 5.9 Hz, 2H), 4.02 (t, J = 6.4 Hz, 2H), 3.70 (s, 3H), 3.67 (s, 4H), 2.95 (t, J = 5.9 Hz, 2H), 2.71-2.64 (m, 4H), 2.38 (t, J = 7.4 Hz, 2H), 2.05-1.92 (m, 5H), 1.89-1.79 (m, 8H), 1.74 (p, J = 7.5 Hz, 2H), 1.55 (tt, J = 9.2, 5.9 Hz, 2H).

[0259] Intermediate 20: 1H NMR (400 MHz, CDC13) δ 8.45 (s, 1H), 7.87 (s, 1H), 7.81-7.73 (m, 3H), 7.66 (d, J = 8.9 Hz, 1H), 7.59 (d, J = 8.6 Hz, 2H), 7.02-6.92 (m, 4H), 4.98 (s, 1H), 4.17 (t, J = 5.9 Hz, 2H), 4.00 (t, J = 6.4 Hz, 2H), 3.95 (s, 1H), 3.68 (s, 4H), 3.64 (s, 1H), 2.97 (t, J = 5.9 Hz, 2H), 2.69 (t, J = 6.0 Hz, 4H), 2.18 (s, 2H), 1.98 (t, J = 6.2 Hz, 4H), 1.89-1.81 (m, 8H), 1.79-1.72 (m, 2H), 1.66-1.49 (m, 6H).

[0260] Intermediate 21: 1 H NMR (400 MHz, CDC13) δ 8.45 (s, 1H), 7.87 (s, 1H), 7.81-7.73 (m, 3H), 7.66 (d, J = 8.9 Hz, 1H), 7.59 (d, J = 8.6 Hz, 2H), 7.02-6.92 (m, 4H), 4.98 (s, 1H), 4.17 (t, J = 5.9 Hz, 2H), 4.00 (t, J = 6.4 Hz, 2H), 3.95 (s, 1H), 3.68 (s, 4H), 3.64 (s, 1H), 2.97 (t, J = 5.9 Hz, 2H), 2.69 (t, J = 6.0 Hz, 4H), 2.18 (s, 2H), 1.98 (t, J = 6.2 Hz, 4H), 1.89-1.81 (m, 8H), 1.79-1.72 (m, 2H), 1.66-1.49 (m, 6H).

[0261] Intermediate 25: 1H NMR (400 MHz, DMSO) δ 8.03 (d, J = 2.7 Hz, 1H), 7.68 (d, J = 6.7 Hz, 2H), 7.64 (d, J = 9.1 Hz, 1H), 7.51 (dd, J = 9.1, 2.7 Hz, 1H), 7.13 - 7.07 (m, 2H), 4.38 (t, J = 5.0 Hz, 2H), 3.95 (s, 3H), 3.60 (t, J = 5.1 Hz, 4H), 3.13 (s, 2H), 2.14 - 1.86 (m, 4H).

[0262] Intermediate 26: 1 H NMR (400 MHz, DMSO) δ 10.27 (s, 1H), 8.08 (d, J = 2.7 Hz, 1H), 7.84 (dd, J = 13.4, 2.5 Hz, 1H), 7.67 (d, J = 9.1 Hz, 1H), 7.60 (dt, J = 9.0, 1.8 Hz, 1H), 7.53 (dd, J = 9.1, 2.6 Hz, 1H), 7.34 (t, J = 9.3 Hz, 1H), 4.47 (t, J = 4.9 Hz, 2H), 3.96 (s, 3H), 3.70 - 3.58 (m, 4H), 3.13 (d, J = 10.4 Hz, 2H), 2.12 - 1.79 (m, 4H).

[0263] Intermediate 27: 1 H NMR (400 MHz, DMSO) δ 10.48 (s, 1H), 10.28 (s, 1H), 8.10 (d, J = 2.7 Hz, 1H), 7.74 - 7.62 (m, 5H), 7.51 (dd, J = 9.2, 2.6 Hz, 1H), 3.95 (s, 3H), 3.51 (h, J = 5.4 Hz, 2H), 3.45 (t, J = 6.9 Hz, 2H), 3.11 - 3.01 (m, 2H), 2.94 (t, J = 7.5 Hz, 2H), 2.10 - 1.96 (m, 2H), 1.89 (dq, J = 12.3, 5.2 Hz, 2H).

[0264] Intermediate 28: 1H NMR (400 MHz, DMSO) δ 10.40 (s, 1H), 10.14 (s, 1H), 8.02 (d, J = 2.7 Hz, 1H), 7.72 - 7.67 (m, 4H), 7.65 (d, J = 9.1 Hz, 1H), 7.52 (dd, J = 9.0, 2.6 Hz, 1H), 3.95 (s, 3H), 3.42 - 3.34 (m, 2H), 3.15 (q, J = 7.1 Hz, 4H), 2.90 (t, J = 7.2 Hz, 2H), 1.25 (t, J = 7.1 Hz, 6H).

[0265] Intermediate 29: 1 H NMR (400 MHz, DMSO) δ 9.71 (s, 1H), 8.56 (t, J = 9.2 Hz, 1H), 7.70 - 7.65 (m, 1H), 7.44 - 7.35 (m, 2H), 7.12 - 7.05 (m, 2H), 6.89 - 6.81 (m, 1H), 4.40 (q, J = 5.6 Hz, 2H), 3.91 (s, 3H), 3.64 - 3.54 (m, 4H), 3.20 - 3.05 (m, 2H), 2.11 - 1.98 (m, 2H), 1.99 - 1.84 (m, 2H).

[0266] Intermediate 30: 1 H NMR (500 MHz, CDCl3) δ 7.74 (d, J = 8.3 Hz, 1H), 7.38 - 7.32 (m, 2H), 7.02 (dd, J = 8.2, 2.7 Hz, 1H), 6.88 - 6.83 (m, 2H), 4.07 (t, J = 5.9 Hz, 2H), 3.84 (t, J = 5.5 Hz, 4H), 3.80 (s, 3H), 3.03 (t, J = 5.9 Hz, 2H), 2.82 (dd, J = 4.2, 3.3 Hz, 4H), 2.03 (d, J = 11.0 Hz, 4H), 1.85 (dd, J = 4.3, 3.2 Hz, 4H).

[0267] Intermediate 31: 1H NMR (400 MHz, CDC13) δ 8.54 (s, 1H), 7.74 (d, J = 8.3 Hz, 1H), 7.45 (d, J = 2.8 Hz, 1H), 7.33 - 7.26 (m, 2H), 7.02 (dd, J = 8.2, 2.7 Hz, 1H), 6.97 - 6.87 (m, 1H), 4.19 (t, J = 5.9 Hz, 2H), 3.84 (t, J = 5.5 Hz, 4H), 3.80 (s, 3H), 2.99 (t, J = 5.9 Hz, 2H), 2.82 (dd, J = 4.2, 3.3 Hz, 4H), 2.03 (d, J = 11.0 Hz, 4H), 1.85 (dd, J = 4.3, 3.2 Hz, 4H).

[0268] Intermediate 32: 1 H NMR (400 MHz, CDC13) δ 11.32 (s, 1H), 7.87 (d, J = 8.5 Hz, 2H), 7.60 (s, 1H), 7.58 - 7.54 (m, 2H), 7.23 - 7.12 (m, 2H), 3.92 (s, 3H), 3.71 (d, J = 6.7 Hz, 4H), 2.85 - 2.78 (m, 2H), 2.71 (q, J = 7.1 Hz, 4H), 2.59 - 2.50 (m, 2H), 2.03 - 1.93 (m, 4H), 1.17 (t, J = 7.1 Hz, 6H).

[0269] Intermediate 33: 1 H NMR (400 MHz, CDC13) δ 11.32 (s, 1H), 7.87 (d, J = 8.5 Hz, 2H), 7.60 (s, 1H), 7.58 - 7.54 (m, 2H), 7.23 - 7.12 (m, 2H), 3.92 (s, 3H), 3.71 (d, J = 6.7 Hz, 4H), 2.85 - 2.78 (m, 2H), 2.71 (q, J = 7.1 Hz, 4H), 2.59 - 2.50 (m, 2H), 2.03 - 1.93 (m, 4H), 1.17 (t, J = 7.1 Hz, 6H).

[0270] Intermediate 34: 1H NMR (400 MHz, CDC13) δ 7.79 (d, J = 8.7 Hz, 2H), 7.16 (s, 1H), 6.98 - 6.91 (m, 2H), 6.84 (t, J = 8.9 Hz, 1H), 6.70 (dd, J = 9.0, 2.5 Hz, 1H), 4.19 (t, J = 5.8 Hz, 2H), 3.80 (s, 3H), 3.71 (s, 4H), 3.00 (t, J = 5.8 Hz, 2H), 2.75 (s, 4H), 1.88 (p, J = 3.2 Hz, 4H).

[0271] Intermediate 35: 1 H NMR (400 MHz, CDC13) δ 7.79 (d, J = 8.7 Hz, 2H), 7.16 (s, 1H), 6.98 - 6.91 (m, 2H), 6.84 (t, J = 8.9 Hz, 1H), 6.70 (dd, J = 9.0, 2.5 Hz, 1H), 4.19 (t, J = 5.8 Hz, 2H), 3.80 (s, 3H), 3.71 (s, 4H), 3.00 (t, J = 5.8 Hz, 2H), 2.75 (s, 4H), 1.88 (p, J = 3.2 Hz, 4H).

[0272] Intermediate 36: 1 H NMR (400 MHz, CDC13) δ 7.79 (d, J = 8.7 Hz, 2H), 7.16 (s, 1H), 6.98 - 6.91 (m, 2H), 6.84 (t, J = 8.9 Hz, 1H), 6.70 (dd, J = 9.0, 2.5 Hz, 1H), 4.19 (t, J = 5.8 Hz, 2H), 3.80 (s, 3H), 3.71 (s, 4H), 3.00 (t, J = 5.8 Hz, 2H), 2.75 (s, 4H), 1.88 (p, J = 3.2 Hz, 4H).

[0273] Intermediate 37: 1H NMR (500 MHz, DMSO) δ 10.02 (d, J = 10.7 Hz, 1H), 9.20 (s, 1H), 7.93-7.87 (m, 2H), 7.60 (d, J = 5.3 Hz, 1H), 7.55 (d, J = 8.6 Hz, 2H), 7.22 (d, J = 8.9 Hz, 1H), 7.16 (dd, J = 8.9, 2.7 Hz, 1H), 3.58-3.49 (m, 6H), 2.71 (t, J = 7.0 Hz, 2H), 2.46 (q, J = 9.0 Hz, 4H), 1.90 (dq, J = 9.6, 4.8 Hz, 4H), 1.73-1.64 (m, 4H).

[0274] Intermediate 38: 1 H NMR (500 MHz, DMSO) δ 10.07 (s, 1H), 9.17 (s, 1H), 7.88 (d, J = 8.6 Hz, 2H), 7.63 (d, J = 2.5 Hz, 1H), 7.58-7.48 (m, 3H), 7.24 (d, J = 8.9 Hz, 1H), 7.20-7.10 (m, 2H), 3.51 (d, J = 6.0 Hz, 4H), 3.46 (s, 2H), 2.73 (t, J = 7.0 Hz, 2H), 2.48 (s, 4H), 2.40 (t, J = 7.0 Hz, 2H), 1.91 (t, J = 3.3 Hz, 4H), 0.98 (t, J = 7.1 Hz, 6H).

[0275] Intermediate 39: 1 H NMR (500 MHz, DMSO) δ 10.02 (d, J = 10.7 Hz, 1H), 9.15 (s, 2H), 7.92-7.86 (m, 2H), 7.60 (d, J = 5.3 Hz, 1H), 7.55 (d, J = 8.6 Hz, 2H), 7.24-7.20 (m, 1H), 7.16 (dd, J = 8.9, 2.7 Hz, 1H), 3.50 (t, J = 5.4 Hz, 6H), 2.71 (t, J = 7.0 Hz, 2H), 2.46 (q, J = 5.4 Hz, 4H), 1.90 (dq, J = 9.6, 4.8 Hz, 4H), 1.71-1.64 (m, 4H).

[0276] Intermediate 40: 1H NMR (500 MHz, CDC13) δ 7.96 (d, J = 7.9 Hz, 2H), 7.69 (d, J = 8.8 Hz, 2H), 7.51 (d, J = 9.1 Hz, 1H), 7.29 (d, J = 7.9 Hz, 2H), 7.23 (dd, J = 9.3, 2.5 Hz, 1H), 7.03 - 6.91 (m, 3H), 4.13 (t, J = 6.0 Hz, 2H), 4.03 (d, J = 5.9 Hz, 2H), 3.90 (s, 3H), 3.71 - 3.59 (m, 4H), 2.92 (t, J = 6.0 Hz, 2H), 2.76 (d, J = 6.6 Hz, 2H), 2.64 (s, 4H), 1.96 (q, J = 4.9 Hz, 4H), 1.90 - 1.84 (m, 4H), 1.83 - 1.78 (m, 4H).

[0277] Intermediate 41: 1 H NMR (500 MHz, DMSO) δ 7.97 (s, 1H), 7.81 (d, J = 8.5 Hz, 2H), 7.62 (s, 1H), 7.44 (s, 1H), 7.09 (d, J = 8.8 Hz, 2H), 4.44 (t, J = 4.8 Hz, 2H), 4.11 (t, J = 6.1 Hz, 2H), 3.77 (t, J = 4.8 Hz, 2H), 3.60 (s, 3H), 3.59 - 3.53 (m, 4H), 3.38 (d, J = 16.8 Hz, 2H), 2.42 (td, J = 7.4, 4.0 Hz, 4H), 2.11 (p, J = 3.6 Hz, 4H), 1.96 (s, 4H), 1.77 - 1.71 (m, 2H), 1.64 - 1.55 (m, 2H).

[0278] Intermediate 42: 1H NMR (400 MHz, CDC13) δ 7.77-7.69 (m, 2H), 7.50 (d, J = 9.1 Hz, 1H), 7.25 (d, J = 2.5 Hz, 1H), 7.21 (d, J = 5.6 Hz, 1H), 7.01 (d, J = 2.7 Hz, 1H), 6.98-6.91 (m, 2H), 4.13 (t, J = 6.0 Hz, 2H), 3.99 (t, J = 6.4 Hz, 2H), 3.66 (s, 3H), 3.63 (d, J = 6.8 Hz, 4H), 2.91 (t, J = 6.0 Hz, 2H), 2.64 (t, J = 4.5 Hz, 4H), 2.34 (t, J = 7.4 Hz, 2H), 1.96 (q, J = 3.6 Hz, 4H), 1.85-1.80 (m, 4H), 1.68 (p, J = 7.5 Hz, 4H), 1.51 (t, J = 7.8 Hz, 2H), 1.42 (q, J = 8.0 Hz, 2H).

[0279] Intermediate 43: 1 H NMR (400 MHz, CDC13) δ 7.91 (d, J = 1.8 Hz, 1H), 7.85 (dt, J = 7.3, 1.7 Hz, 1H), 7.77-7.70 (m, 2H), 7.55 (s, 1H), 7.51 (dd, J = 9.1, 4.5 Hz, 1H), 7.34 (dd, 1H), 7.20 (dd, J = 9.2, 2.6 Hz, 1H), 7.12 (d, J = 2.7 Hz, 1H), 6.97-6.90 (m, 2H), 4.14-4.08 (m, 2H), 3.99-3.87 (m, 5H), 3.70-3.59 (m, 4H), 2.90 (t, J = 6.0 Hz, 2H), 2.71 (q, J = 5.0 Hz, 2H), 2.63 (ddt, J = 6.8, 4.3, 2.5 Hz, 4H), 1.95 (td, J = 5.7, 3.2 Hz, 4H), 1.85-1.75 (m, 8H).

[0280] Intermediate 46: 1H NMR (400 MHz, CDC13) δ 7.80 (d, J = 8.7 Hz, 2H), 7.60 (d, J = 9.1 Hz, 1H), 7.23 (dd, J = 9.0, 2.5 Hz, 1H), 6.95 (d, J = 9.0 Hz, 2H), 4.23 (d, J = 4.7 Hz, 2H), 4.17 (t, J = 5.9 Hz, 2H), 3.86 - 3.77 (m, 4H), 3.67 (d, J = 7.0 Hz, 4H), 2.97 (t, J = 5.9 Hz, 2H), 2.71 (s, 4H), 2.57 (t, J = 6.3 Hz, 2H), 1.98 (q, J = 3.9 Hz, 4H), 1.90 - 1.80 (m, 4H), 1.51 - 1.42 (m, 9H).

[0281] Intermediate 47: 1 H NMR (400 MHz, CDC13) δ 7.80 (d, J = 8.7 Hz, 2H), 7.60 (d, J = 9.1 Hz, 1H), 7.23 (dd, J = 9.0, 2.5 Hz, 1H), 6.95 (d, J = 9.0 Hz, 2H), 4.23 (d, J = 4.7 Hz, 2H), 4.17 (t, J = 5.9 Hz, 2H), 3.86 - 3.77 (m, 4H), 3.67 (d, J = 7.0 Hz, 4H), 2.97 (t, J = 5.9 Hz, 2H), 2.71 (s, 4H), 2.57 (t, J = 6.3 Hz, 2H), 1.98 (q, J = 3.9 Hz, 4H), 1.90 - 1.80 (m, 4H), 1.51 - 1.42 (m, 9H).

[0282] Intermediate 48: 1H NMR (400 MHz, CDC13) δ 11.18 (s, 1H), 8.04 (s, 1H), 7.88 (d, J = 8.6 Hz, 2H), 7.63 (d, J = 9.0 Hz, 1H), 7.56 - 7.50 (m, 2H), 7.19 (d, J = 9.9 Hz, 2H), 4.06 (t, J = 6.4 Hz, 2H), 3.74 - 3.68 (m, 6H), 3.68 (s, 3H), 2.74 (s, 4H), 2.63 - 2.56 (m, 2H), 2.36 (dd, J = 8.6, 6.3 Hz, 2H), 1.99 (q, J = 3.3 Hz, 4H), 1.95 (p, J = 3.1 Hz, 4H), 1.81 (t, J = 7.3 Hz, 2H), 1.73 - 1.67 (m, 2H), 1.59 (s, 2H), 1.51 (d, J = 7.5 Hz, 2H).

[0283] Intermediate 49: 1 H NMR (400 MHz, CDC13) δ 11.23 (s, 1H), 7.79 (d, J = 8.5 Hz, 2H), 7.50 (d, J = 9.2 Hz, 1H), 7.47 (d, J = 8.8 Hz, 2H), 7.14 (s, 1H), 7.09 (dd, J = 9.1, 2.5 Hz, 1H), 3.94 (t, J = 6.4 Hz, 2H), 3.59 (d, J = 2.7 Hz, 7H), 2.71 (dd, J = 6.6, 5.0 Hz, 2H), 2.62 (q, J = 7.1 Hz, 4H), 2.45 (dd, J = 6.6, 5.0 Hz, 2H), 2.26 (t, J = 7.4 Hz, 2H), 1.95 - 1.82 (m, 4H), 1.71 (q, J = 6.9 Hz, 2H), 1.58 (q, J = 7.5 Hz, 2H), 1.41 (t, J = 8.1 Hz, 2H), 1.33 (tt, J = 8.7, 3.5 Hz, 2H), 1.08 (t, J = 7.1 Hz, 6H).

[0284] Intermediate 50: 1H NMR (400 MHz, Chloroform-d) δ 8.22 (d, J = 9.3 Hz, 1H), 7.38-7.34 (m, 2H), 7.23 (s, 1H), 6.89-6.84 (m, 2H), 6.81 (dd, J = 9.2, 2.2 Hz, 1H), 4.07 (t, J = 5.9 Hz, 2H), 4.00 (t, J = 6.3 Hz, 2H), 3.84 (t, J = 5.5 Hz, 4H), 3.62 (s, 3H), 3.03 (t, J = 5.9 Hz, 2H), 2.82 (dd, J = 4.2, 3.3 Hz, 4H), 2.28 (t, J = 8.5 Hz, 2H), 2.03 (t, J = 5.5 Hz, 4H), 1.85 (m, 4H), 1.75-1.68 (m, 4H), 1.50-1.42 (m, 2H), 1.40-1.31 (m, 2H).

[0285] Intermediate 51: 1 H NMR (400 MHz, Chloroform-d) δ 8.22 (d, J = 9.3 Hz, 1H), 7.38-7.34 (m, 2H), 7.23 (s, 1H), 6.89-6.84 (m, 2H), 6.81 (dd, J = 9.2, 2.2 Hz, 1H), 4.07 (t, J = 5.9 Hz, 2H), 4.00 (t, J = 6.3 Hz, 2H), 3.84 (t, J = 5.5 Hz, 4H), 3.62 (s, 3H), 3.03 (t, J = 5.9 Hz, 2H), 2.82 (dd, J = 4.2, 3.3 Hz, 4H), 2.28 (t, J = 8.5 Hz, 2H), 2.03 (t, J = 5.5 Hz, 4H), 1.85 (m, 4H), 1.75-1.68 (m, 4H), 1.50-1.42 (m, 2H), 1.40-1.31 (m, 2H).

[0286] Intermediate 52: 1H NMR (400 MHz, CDC13) δ 7.81 - 7.72 (m, 2H), 7.46 (d, J = 9.0 Hz, 1H), 7.24 (s, 1H), 7.11 (dd, J = 9.3, 2.5 Hz, 1H), 6.94 - 6.84 (m, 2H), 4.92 (s, 1H), 4.10 (t, J = 5.9 Hz, 2H), 4.00 - 3.85 (m, 3H), 3.67 - 3.45 (m, 5H), 2.91 (t, J = 5.9 Hz, 2H), 2.69 - 2.57 (m, 4H), 2.20 (s, 2H), 1.95 (s, 4H), 1.82 - 1.67 (m, 10H), 1.52 (s, 4H).

[0287] Intermediate 53: 1 H NMR (500 MHz, CDC13) δ 7.72 (dd, J = 12.1, 8.3 Hz, 2H), 7.51 (d, J = 8.9 Hz, 1H), 7.26 (s, 1H), 7.22 - 7.16 (m, 1H), 6.95 - 6.89 (m, 2H), 4.90 (s, 1H), 4.12 (t, J = 5.9 Hz, 2H), 3.98 (d, J = 6.9 Hz, 1H), 3.96 - 3.86 (m, 2H), 3.63 (q, J = 8.7 Hz, 5H), 2.92 (t, J = 6.9 Hz, 2H), 2.65 (d, J = 5.9 Hz, 4H), 2.13 (s, 2H), 1.97 (q, J = 13.2 Hz, 4H), 1.83 (q, J = 3.3 Hz, 4H), 1.78 - 1.64 (m, 6H), 1.61 - 1.37 (m, 8H).

[0288] Intermediate 54: 1H NMR (400 MHz, CDC13) δ 8.47 (s, 1H), 7.85-7.77 (m, 2H), 7.61 (d, J = 2.0 Hz, 1H), 7.51 (dq, J = 8.8, 2.2 Hz, 2H), 7.41 (d, J = 2.7 Hz, 1H), 7.27-7.21 (m, 2H), 7.13 (dt, J = 9.1, 2.6 Hz, 1H), 6.92-6.83 (m, 2H), 5.09 (d, J = 3.3 Hz, 1H), 4.10 (t, J = 5.9 Hz, 2H), 4.05-3.94 (m, 1H), 3.84-3.73 (m, 2H), 3.69-3.53 (m, 5H), 2.89 (t, J = 5.9 Hz, 2H), 2.66-2.59 (m, 4H), 2.56 (d, J = 7.3 Hz, 2H), 1.96-1.87 (m, 4H), 1.85-1.73 (m, 6H), 1.72-1.46 (m, 8H).

[0289] Intermediate 57: 1 H NMR (400 MHz, CDC13) δ 8.47 (s, 1H), 7.85-7.77 (m, 2H), 7.61 (d, J = 2.0 Hz, 1H), 7.51 (dq, J = 8.8, 2.2 Hz, 2H), 7.41 (d, J = 2.7 Hz, 1H), 7.27-7.21 (m, 2H), 7.13 (dt, J = 9.1, 2.6 Hz, 1H), 6.92-6.83 (m, 2H), 5.09 (d, J = 3.3 Hz, 1H), 4.10 (t, J = 5.9 Hz, 2H), 4.05-3.94 (m, 1H), 3.84-3.73 (m, 2H), 3.69-3.53 (m, 5H), 2.89 (t, J = 5.9 Hz, 2H), 2.66-2.59 (m, 4H), 2.56 (d, J = 7.3 Hz, 2H), 1.96-1.87 (m, 4H), 1.85-1.73 (m, 6H), 1.72-1.46 (m, 8H).

[0290] Intermediate 58: 1H NMR (400 MHz, CDC13) δ 8.20 (dd, J = 13.7, 2.5 Hz, 1H), 7.90 (d, J = 9.0 Hz, 1H), 7.62 (s, 1H), 7.20 (s, 1H), 7.17 (d, J = 9.1 Hz, 1H), 6.97 (t, J = 9.0 Hz, 1H), 4.93 (s, 1H), 4.19 (t, J = 5.9 Hz, 2H), 4.00 - 3.89 (m, 3H), 3.66 - 3.63 (m, 5H), 2.99 (t, J = 6.9 Hz, 2H), 2.82 (d, J = 5.9 Hz, 4H), 2.28 (s, 2H), 1.97 (q, J = 13.2 Hz, 4H), 1.85 (q, J = 3.3 Hz, 4H), 1.78 - 1.64 (m, 6H), 1.61 - 1.37 (m, 8H).

[0291] Intermediate 59: 1 H NMR (500 MHz, CDC13) δ 10.34 (s, 1H), 9.17 (s, 1H), 9.11 (s, 1H), 7.74 (d, J = 8.8 Hz, 1H), 7.47 - 7.41 (m, 5H), 7.06 (dd, J = 9.0, 2.7 Hz, 1H), 4.94 (t, J = 3.3 Hz, 1H), 4.01 (t, J = 6.3 Hz, 2H), 3.84 (t, J = 5.5 Hz, 4H), 3.80 - 3.73 (m, 1H), 3.66 (ddd, J = 11.0, 5.9, 3.7 Hz, 1H), 2.88 - 2.80 (m, 6H), 2.50 (t, J = 5.8 Hz, 2H), 2.34 - 2.21 (m, 2H), 2.04 (d, J = 5.4 Hz, 4H), 1.91 - 1.85 (m, 5H), 1.81 - 1.66 (m, 6H), 1.65 - 1.55 (m, 3H), 1.50 - 1.41 (m, 2H), 1.41 - 1.29 (m, 2H).

[0292] Intermediate 60: 1H NMR (400 MHz, CDC13) δ 11.29 (s, 1H), 7.89 (d, J = 8.5 Hz, 2H), 7.84 (s, 1H), 7.60-7.52 (m, 3H), 7.34 (s, 1H), 7.19 (dd, J = 9.3, 2.5 Hz, 1H), 4.94 (s, 1H), 4.06-3.89 (m, 3H), 3.67 (d, J = 6.5 Hz, 4H), 3.61 (d, J = 11.6 Hz, 1H), 2.80 (t, J = 5.0 Hz, 2H), 2.69 (q, J = 7.1 Hz, 4H), 2.53 (t, J = 5.0 Hz, 2H), 2.15 (s, 2H), 2.02-1.92 (m, 4H), 1.85-1.64 (m, 8H), 1.56 (s, 2H), 1.48-1.35 (m, 4H), 1.16 (t, J = 7.1 Hz, 6H).

[0293] Intermediate 61: 1 H NMR (400 MHz, CDC13) δ 11.29 (s, 1H), 7.89 (d, J = 8.5 Hz, 2H), 7.84 (s, 1H), 7.60-7.52 (m, 3H), 7.34 (s, 1H), 7.19 (dd, J = 9.3, 2.5 Hz, 1H), 4.94 (s, 1H), 4.06-3.89 (m, 3H), 3.67 (d, J = 6.5 Hz, 4H), 3.61 (d, J = 11.6 Hz, 1H), 2.80 (t, J = 5.0 Hz, 2H), 2.69 (q, J = 7.1 Hz, 4H), 2.53 (t, J = 5.0 Hz, 2H), 2.15 (s, 2H), 2.02-1.92 (m, 4H), 1.85-1.64 (m, 8H), 1.56 (s, 2H), 1.48-1.35 (m, 4H), 1.16 (t, J = 7.1 Hz, 6H).

[0294] Intermediate 62: 11H NMR (500 MHz, DMSO) δ 9.67 (s, 1H), 9.17 (s, 1H), 7.81 (d, J = 8.7 Hz, 2H), 7.74 - 7.69 (m, 3H), 7.28 (t, J = 8.0 Hz, 3H), 7.21 (dd, J = 9.1, 2.5 Hz, 1H), 6.95 (d, J = 8.7 Hz, 2H), 4.44 (s, 2H), 4.05 (dt, J = 13.1, 6.2 Hz, 4H), 3.49 (d, J = 6.3 Hz, 4H), 2.78 (t, J = 5.9 Hz, 2H), 2.63 (d, J = 7.8 Hz, 2H), 2.53 (d, J = 5.7 Hz, 4H), 1.90 (q, J = 4.9 Hz, 4H), 1.77 (d, J = 7.2 Hz, 2H), 1.72 - 1.67 (m, 4H), 1.62 (d, J = 7.7 Hz, 2H), 1.50 (p, J = 7.5 Hz, 2H), 1.39 (q, J = 7.9 Hz, 2H).

[0295] Intermediate 63: 1 1H NMR (400 MHz, DMSO) δ 10.65 (s, 1H), 8.47 (s, 1H), 7.88 (d, J = 8.6 Hz, 2H), 7.71 (s, 1H), 7.67 (d, J = 8.6 Hz, 2H), 7.38 - 7.25 (m, 1H), 4.10 (s, 2H), 3.60 (t, J = 6.3 Hz, 6H), 2.52 (s, 4H), 2.06 - 1.86 (m, 8H), 1.76 - 1.29 (m, 10H).

[0296] Intermediate 64: 1H NMR (400 MHz, CDC13) δ 7.85 (s, 1H), 7.82 (t, J = 2.3 Hz, 1H), 7.72 (s, 1H), 7.69 (d, J = 8.7 Hz, 2H), 7.60 (d, J = 8.4 Hz, 1H), 7.50 (d, J = 9.1 Hz, 1H), 7.18-7.14 (m, 2H), 7.00 (dd, J = 9.1, 2.5 Hz, 1H), 6.80 (d, J = 8.3 Hz, 2H), 4.32 (t, J = 6.7 Hz, 2H), 4.03 (d, J = 6.4 Hz, 2H), 3.90 (s, 3H), 3.61 (s, 4H), 3.56 (t, J = 6.7 Hz, 2H), 2.89 (t, J = 5.7 Hz, 2H), 2.64 (s, 4H), 1.89 (d, J = 3.2 Hz, 4H), 1.75 (s, 4H), 1.63-1.56 (m, 4H), 1.48-1.39 (m, 4H).

[0297] Intermediate 65: 1 H NMR (400 MHz, CDC13) δ 7.85 (s, 1H), 7.82 (t, J = 2.3 Hz, 1H), 7.72 (s, 1H), 7.69 (d, J = 8.7 Hz, 2H), 7.60 (d, J = 8.4 Hz, 1H), 7.50 (d, J = 9.1 Hz, 1H), 7.18-7.14 (m, 2H), 7.00 (dd, J = 9.1, 2.5 Hz, 1H), 6.80 (d, J = 8.3 Hz, 2H), 4.32 (t, J = 6.7 Hz, 2H), 4.03 (d, J = 6.4 Hz, 2H), 3.90 (s, 3H), 3.61 (s, 4H), 3.56 (t, J = 6.7 Hz, 2H), 2.89 (t, J = 5.7 Hz, 2H), 2.64 (s, 4H), 1.89 (d, J = 3.2 Hz, 4H), 1.75 (s, 4H), 1.63-1.56 (m, 4H), 1.48-1.39 (m, 4H).

[0298] Intermediate 67: 1H NMR (500 MHz, CDC13) δ 8.32 (d, J = 2.6 Hz, 1H), 7.90 (d, J = 8.4 Hz, 1H), 7.84 (s, 1H), 7.70 (dd, J = 8.3, 2.5 Hz, 1H), 7.37-7.31 (m, 2H), 6.86-6.77 (m, 2H).

[0299] Intermediate 68: 1 H NMR (400 MHz, CDC13) δ 7.85-7.80 (m, 1H), 7.75-7.66 (m, 3H), 7.56-7.50 (m, 2H), 7.46-7.41 (m, 1H), 6.86-6.82 (m, 2H), 3.93-3.86 (m, 2H), 3.61 (s, 3H), 3.60-3.56 (m, 2H), 1.69 (p, J = 3.4 Hz, 2H), 1.64 (dt, J = 4.7, 3.2 Hz, 2H).

[0300] Intermediate 69: 1 H NMR (400 MHz, DMSO) δ 10.11 (s, 1H), 8.53 (d, J = 8.1 Hz, 1H), 7.87 (ddd, J = 8.3, 6.9, 1.3 Hz, 1H), 7.70 (dd, J = 8.5, 1.3 Hz, 1H), 7.63 (dd, J = 8.1, 6.3 Hz, 3H), 7.04-6.96 (m, 2H), 4.00 (t, J = 6.5 Hz, 2H), 3.59 (s, 3H), 2.32 (t, J = 7.4 Hz, 2H), 1.73 (p, J = 6.7 Hz, 2H), 1.57 (p, J = 7.3 Hz, 2H), 1.50-1.43 (m, 2H), 1.35 (t, J = 7.8 Hz, 2H).

[0301] Intermediate 71: 1H NMR (400 MHz, DMSO) δ 10.10 (s, 1H), 8.52 (d, J = 8.2 Hz, 1H), 7.90 (d, J = 8.0 Hz, 2H), 7.86 (d, J = 7.8 Hz, 1H), 7.69 (d, J = 8.2 Hz, 1H), 7.63 (dd, J = 8.4, 6.3 Hz, 3H), 7.42 - 7.35 (m, 2H), 7.04 - 6.96 (m, 2H), 4.08 - 3.98 (m, 2H), 3.83 (s, 3H), 2.74 (d, J = 7.1 Hz, 2H), 1.81 - 1.71 (m, 4H).

[0302] Intermediate 72: 1 H NMR (400 MHz, DMSO) δ 10.21 (s, 1H), 8.85 (t, J = 2.7 Hz, 1H), 8.00 (dd, J = 8.9, 2.1 Hz, 1H), 7.65 (s, 1H), 7.63 - 7.58 (m, 1H), 7.32 (d, J = 8.8 Hz, 1H), 7.03 - 6.98 (m, 2H), 6.95 (d, J = 8.3 Hz, 1H), 4.00 (t, J = 5.9 Hz, 2H), 3.60 (s, 3H), 2.40 (t, J = 7.0 Hz, 2H), 1.72 (dd, J = 12.8, 6.6 Hz, 8H).

[0303] Intermediate 73: 1 H NMR (400 MHz, CDCl3) δ 7.76 (d, J = 8.8 Hz, 3H), 7.63 (d, J = 8.5 Hz, 1H), 7.51 (t, J = 7.7 Hz, 1H), 7.10 (t, J = 7.6 Hz, 1H), 6.98 - 6.90 (m, 2H), 4.02 (d, J = 5.4 Hz, 2H), 3.70 (s, 7H), 2.43 (dq, J = 5.8, 3.5 Hz, 2H), 1.99 (dd, J = 10.2, 3.3 Hz, 4H), 1.87 (h, J = 3.5 Hz, 4H).

[0304] Intermediate 74: 1H NMR (400 MHz, CDC13) δ 7.86 (s, 1H), 7.80 (d, J = 8.6 Hz, 2H), 7.69 (d, J = 8.4 Hz, 1H), 7.48 (t, J = 7.7 Hz, 1H), 7.10 (t, J = 7.6 Hz, 1H), 6.97-6.91 (m, 2H), 3.99 (t, J = 6.5 Hz, 2H), 3.72 (s, 4H), 3.70 (s, 3H), 2.36 (t, J = 7.5 Hz, 2H), 1.99 (dd, J = 10.3, 3.3 Hz, 4H), 1.82 (p, J = 6.7 Hz, 2H), 1.70 (p, J = 7.5 Hz, 2H), 1.53 (p, J = 6.8 Hz, 2H), 1.48-1.39 (m, 2H).

[0305] Intermediate 76: 1 H NMR (400 MHz, CDC13) δ 7.99 (d, J = 8.0 Hz, 2H), 7.84 (s, 1H), 7.79 (d, J = 8.4 Hz, 2H), 7.68 (d, J = 8.3 Hz, 1H), 7.49 (t, J = 7.7 Hz, 1H), 7.30 (d, J = 7.9 Hz, 2H), 7.10 (t, J = 7.6 Hz, 1H), 6.93 (d, J = 8.7 Hz, 2H), 4.03 (d, J = 5.6 Hz, 3H), 3.93 (s, 3H), 3.72 (s, 4H), 2.88-2.72 (m, 2H), 2.00 (d, J = 6.1 Hz, 5H), 1.86 (p, J = 3.2 Hz, 4H).

[0306] Intermediate 77: 1 H NMR (400 MHz, CDC13) δ 7.97 (s, 1H), 7.80-7.73 (m, 2H), 7.55 (dt, J = 7.0, 2.2 Hz, 1H), 7.52 (s, 1H), 6.95-6.91 (m, 2H), 6.88-6.81 (m, 1H), 4.00 (d, J = 5.5 Hz, 2H), 3.70 (d, J = 3.8 Hz, 7H), 2.44 (dd, J = 7.7, 5.0 Hz, 2H), 2.00 (q, J = 3.4 Hz, 4H), 1.89-1.82 (m, 4H).

[0307] Intermediate 78: 1H NMR (400 MHz, CDC13) δ 8.94 (s, 1H), 7.88 (d, J = 8.1 Hz, 1H), 7.78-7.71 (m, 2H), 7.67 (d, J = 8.4 Hz, 1H), 7.50 (t, J = 7.7 Hz, 1H), 7.10 (t, J = 7.5 Hz, 1H), 6.86 (d, J = 8.5 Hz, 2H), 4.98 (s, 1H), 3.96 (s, 3H), 3.67-3.59 (m, 6H), 2.23 (s, 2H), 2.02-1.92 (m, 4H), 1.90-1.74 (m, 6H).

[0308] Intermediate 79: 1 H NMR (400 MHz, CDC13) δ 8.94 (s, 1H), 7.88 (d, J = 8.1 Hz, 1H), 7.78-7.71 (m, 2H), 7.67 (d, J = 8.4 Hz, 1H), 7.50 (t, J = 7.7 Hz, 1H), 7.10 (t, J = 7.5 Hz, 1H), 6.86 (d, J = 8.5 Hz, 2H), 4.98 (s, 1H), 3.96 (s, 3H), 3.67-3.59 (m, 6H), 2.23 (s, 2H), 2.02-1.92 (m, 4H), 1.90-1.74 (m, 6H).

[0309] Intermediate 81: 1 H NMR (400 MHz, CDC13) δ 8.94 (s, 1H), 7.88 (d, J = 8.1 Hz, 1H), 7.78-7.71 (m, 2H), 7.67 (d, J = 8.4 Hz, 1H), 7.50 (t, J = 7.7 Hz, 1H), 7.10 (t, J = 7.5 Hz, 1H), 6.86 (d, J = 8.5 Hz, 2H), 4.98 (s, 1H), 3.96 (s, 3H), 3.67-3.59 (m, 6H), 2.23 (s, 2H), 2.02-1.92 (m, 4H), 1.90-1.74 (m, 6H).

[0310] Intermediate 82: 1H NMR (400 MHz, DMSO) δ 10.94 (s, 1H), 9.04 (s, 1H), 7.80 (d, J = 8.5 Hz, 2H), 7.17 (d, J = 8.9 Hz, 1H), 7.09 (s, 1H), 7.04 (d, J = 8.9 Hz, 1H), 6.93 (d, J = 8.5 Hz, 2H), 4.82 (s, 1H), 3.96 (s, 3H), 3.47 (s, 5H), 3.14 (s, 2H), 2.48 - 2.31 (m, 10H), 2.16 (s, 3H), 2.07 (s, 2H), 1.90 (d, J = 6.7 Hz, 4H), 1.78 (d, J = 7.4 Hz, 2H), 1.66 (d, J = 12.6 Hz, 6H), 1.51 (s, 4H).

[0311] Intermediate 83: 1 H NMR (400 MHz, DMSO) δ 10.94 (s, 1H), 9.04 (s, 1H), 7.80 (d, J = 8.5 Hz, 2H), 7.17 (d, J = 8.9 Hz, 1H), 7.09 (s, 1H), 7.04 (d, J = 8.9 Hz, 1H), 6.93 (d, J = 8.5 Hz, 2H), 4.82 (s, 1H), 3.96 (s, 3H), 3.47 (s, 5H), 3.14 (s, 2H), 2.48 - 2.31 (m, 10H), 2.16 (s, 3H), 2.07 (s, 2H), 1.90 (d, J = 6.7 Hz, 4H), 1.78 (d, J = 7.4 Hz, 2H), 1.66 (d, J = 12.6 Hz, 6H), 1.51 (s, 4H).

[0312] Intermediate 84: 1H NMR (400 MHz, DMSO) δ 10.94 (s, 1H), 8.98 (s, 1H), 7.84-7.77 (m, 2H), 7.18-7.12 (m, 2H), 7.08 (dd, J = 8.9, 2.3 Hz, 1H), 6.95-6.89 (m, 2H), 5.25 (t, J = 5.4 Hz, 1H), 4.82 (s, 1H), 3.96 (t, J = 5.8 Hz, 2H), 3.92 (s, 1H), 3.61 (t, J = 4.6 Hz, 4H), 3.48 (q, J = 8.0 Hz, 6H), 3.24 (q, J = 6.3 Hz, 2H), 2.59 (t, J = 6.6 Hz, 2H), 2.46 (s, 4H), 2.07 (t, J = 6.7 Hz, 2H), 1.89 (q, J = 4.3 Hz, 4H), 1.67 (d, J = 13.1 Hz, 6H), 1.51 (s, 4H).

[0313] Intermediate 85: 1 H NMR (400 MHz, DMSO) δ 10.94 (s, 1H), 9.16 (s, 1H), 7.83-7.77 (m, 3H), 7.69 (s, 1H), 7.27 (d, J = 1.4 Hz, 2H), 7.02-6.96 (m, 2H), 6.94-6.89 (m, 2H), 6.89-6.83 (m, 2H), 4.82 (s, 1H), 3.99-3.88 (m, 3H), 3.50 (t, J = 5.8 Hz, 5H), 3.17 (d, J = 5.0 Hz, 2H), 3.01 (t, J = 4.9 Hz, 4H), 2.45 (t, J = 5.0 Hz, 4H), 2.22 (s, 3H), 2.07 (t, J = 6.9 Hz, 2H), 1.94-1.88 (m, 4H), 1.67 (d, J = 12.0 Hz, 6H), 1.51 (s, 4H).

[0314] Trisubstituted quinazoline derivative a1: 1H NMR (400 MHz, DMSO) δ 10.45 (s, 1H), 9.70 (s, 1H), 8.74 (s, 1H), 8.63 (s, 1H), 7.90 (dd, J = 8.3, 5.0 Hz, 3H), 7.78-7.71 (m, 3H), 7.68 (dq, J = 7.0, 3.8 Hz, 1H), 7.46 (d, J = 8.6 Hz, 1H), 7.32 (d, J = 8.0 Hz, 2H), 7.04 (d, J = 8.6 Hz, 2H), 4.31 (t, J = 5.2 Hz, 2H), 3.82 (s, 4H), 3.57 (d, J = 6.2 Hz, 4H), 2.87 (t, J = 7.6 Hz, 2H), 2.32 (t, J = 7.6 Hz, 2H), 1.93 (t, J = 7.0 Hz, 8H). 13 C NMR (126 MHz, DMSO) δ 168.70, 167.85, 158.03, 132.14, 131.88, 129.26, 129.17, 126.88, 123.87, 114.83, 110.89, 54.19 (2C), 53.47, 53.09 (2C), 34.31, 30.93, 25.46, 23.16 (2C). ESI-HRMS [M + H] + m / z = 567.3226, calcd for C 33 H 38 N6O3, 567.3078.

[0315] trisubstituted quinazoline derivative a2: 1 H NMR (400 MHz, DMSO) δ 10.45 (s, 1H), 9.70 (s, 1H), 8.74 (s, 1H), 8.63 (s, 1H), 7.90 (dd, J = 8.3, 5.0 Hz, 3H), 7.78-7.71 (m, 3H), 7.68 (dq, J = 7.0, 3.8 Hz, 1H), 7.46 (d, J = 8.6 Hz, 1H), 7.32 (d, J = 8.0 Hz, 2H), 7.04 (d, J = 8.6 Hz, 2H), 4.31 (t, J = 5.2 Hz, 2H), 3.82 (s, 4H), 3.57 (d, J = 6.2 Hz, 4H), 2.87 (t, J = 7.6 Hz, 2H), 2.32 (t, J = 7.6 Hz, 2H), 1.93 (t, J = 7.0 Hz, 8H).13 C NMR (126 MHz, DMSO) d 168.70, 167.85, 158.03, 132.14, 131.88, 129.26, 129.17, 126.88, 123.87, 114.83, 110.89, 54.19 (2C), 53.47, 53.09 (2C), 34.31, 30.93, 25.46, 23.16 (2C). ESI-HRMS [M + H] + m / z = 567.3226, calcd for C 33 H 38 N6O3, 567.3078.

[0316] trisubstituted quinazoline derivative a3: 1 H NMR (500 MHz, DMSO) d 10.16 (s, 1H), 9.44-9.20 (m, 1H), 8.27 (d, J = 8.2 Hz, 1H), 7.95-7.80 (m, 2H), 7.60-7.48 (m, 3H), 7.28 (s, 1H), 7.10 (t, J = 7.6 Hz, 1H), 6.93-6.66 (m, 1H), 3.20 (s, 2H), 2.88-2.79 (m, 2H), 2.77 (t, J = 7.0 Hz, 2H), 2.57-2.51 (m, 4H), 2.44 (t, J = 7.1 Hz, 2H), 2.22 (s, 3H), 1.99 (d, J = 17.4 Hz, 2H), 1.71 (s, 2H), 1.60 (s, 1H), 1.00 (t, J = 7.1 Hz, 6H). 13 C NMR (126 MHz, MeOD) d 171.31, 159.3, 158.87, 151.31, 135.28, 134.56, 132.84, 122.85 (2C), 122.19, 121.20, 119.85 (2C), 54.83, 46.29 (2C), 44.54, 34.92, 32.69, 28.97, 10.12 (2C). ESI-HRMS [M + 2H] 2+ m / z = 298.1838, calcd for C 35 H 42 N6O3,298.1732.

[0317] trisubstituted quinazoline derivative a4: 1H NMR (500 MHz, DMSO) δ 12.40 (s, 1H), 11.20 (s, 1H), 10.94 (s, 1H), 10.51 (s, 1H), 8.92 (s, 1H), 8.74 (s, 1H), 8.12 (d, J = 8.5 Hz, 1H), 8.00 (s, 1H), 7.84 (t, J = 9.1 Hz, 4H), 7.08 (dd, J = 15.6, 8.4 Hz, 4H), 4.40 (s, 2H), 4.03 (t, J = 6.4 Hz, 2H), 3.59 (s, 2H), 3.59 (s, 8H), 3.13 (s, 2H), 2.17 (t, J = 7.4 Hz, 2H), 2.05 - 1.90 (m, 10H). 13 C NMR (126 MHz, DMSO) δ 169.10, 158.97, 157.74, 128.43 (2C), 125.58, 121.40, 115.42 (2C), 114.90 (2C), 110.64, 67.44, 63.96, 54.08 (2C), 53.09, 29.18, 25.27, 23.11 (2C). ESI-HRMS [M + 2H] 2+ m / z = 299.1740, calcd for C 34 H 42 N6O4, 300.1707.

[0318] trisubstituted quinazoline derivative a5: 1 H NMR (400 MHz, DMSO) δ 12.32 (s, 1H), 11.09 (s, 1H), 11.01 (s, 1H), 10.39 (s, 1H), 8.93 (s, 1H), 8.67 (s, 1H), 8.16 (d, J = 8.4 Hz, 1H), 8.02 (d, J = 8.8 Hz, 1H), 7.86 - 7.79 (m, 4H), 7.13 - 7.07 (m, 4H), 4.40 (t, J = 5.0 Hz, 2H), 4.03 (t, J = 6.5 Hz, 2H), 3.70 (s, 2H), 3.60 (s, 8H), 3.12 (s, 2H), 2.03 (s, 4H), 1.98 - 1.88 (m, 6H), 1.73 (t, J = 7.4 Hz, 2H), 1.51 (q, J = 7.5 Hz, 2H), 1.43 (s, 2H), 1.37 - 1.29 (m, 2H). 13C NMR (101 MHz, DMSO) δ 169.54, 159.19, 157.77, 155.69, 130.95, 128.46 (2C), 125.74 (2C), 115.42 (2C), 114.95 (2C), 110.65, 68.00, 63.94, 54.11 (2C), 53.10, 40.66, 32.68, 29.04, 28.83, 25.74, 25.55, 23.09 (2C). ESI-HRMS [M + 2H] 2+ m / z = 320.1974, calcd for C 37 H 46 N6O4, 320.1863.

[0319] trisubstituted quinazoline derivative a6: 1 H NMR (400 MHz, DMSO) δ 12.45 (s, 1H), 11.29 (s, 1H), 11.07 (s, 1H), 10.49 - 10.34 (m, 1H), 8.96 (s, 1H), 8.10 (q, J = 8.9 Hz, 2H), 7.84 (dd, J = 11.3, 8.6 Hz, 4H), 7.08 (dd, J = 17.5, 8.5 Hz, 4H), 4.41 (t, J = 4.9 Hz, 2H), 4.02 (t, J = 6.4 Hz, 2H), 3.76 - 3.67 (m, 4H), 3.63 - 3.59 (m, 4H), 3.16 - 3.09 (m, 2H), 2.09 - 1.96 (m, 6H), 1.95 - 1.83 (m, 4H), 1.78 - 1.71 (m, 2H), 1.58 (q, J = 7.4 Hz, 2H), 1.48 - 1.38 (m, 2H), 1.37 - 1.22 (m, 2H). 13C NMR (126 MHz, DMSO) δ 174.91, 169.49, 159.13, 157.65, 155.62, 150.15, 139.12, 136.34, 133.21, 131.40, 130.85, 128.44 (2C), 125.71 (2C), 121.44, 118.40, 115.33 (2C), 114.86 (2C), 110.58, 67.95, 63.93, 54.06 (2C), 53.70, 53.04, 49.05, 48.76, 47.91, 41.99, 34.13, 32.69, 28.91, 25.65, 25.40, 24.77, 24.56, 23.11 (2C), 18.41, 17.16. ESI-HRMS [M + 2H] 2+ m / z = 313.1902, calcd for C 36 H 44 N6O4, 313.1785.

[0320] trisubstituted quinazoline derivative a7: 1 H NMR (500 MHz, DMSO) δ 12.18 (s, 1H), 11.18 (s, 1H), 11.03 (s, 1H), 10.88 (s, 1H), 8.99 (s, 1H), 8.25 (d, J = 2.6 Hz, 1H), 7.90 (d, J = 9.1 Hz, 1H), 7.81 (d, J = 8.6 Hz, 2H), 7.69 (d, J = 7.9 Hz, 2H), 7.48 (dd, J = 9.2, 2.5 Hz, 1H), 7.31 (d, J = 7.9 Hz, 2H), 7.10 (d, J = 8.6 Hz, 2H), 4.39 (t, J = 4.9 Hz, 2H), 4.18 (d, J = 5.1 Hz, 2H), 3.67 - 3.57 (m, 8H), 3.12 (s, 2H), 2.73 (d, J = 6.8 Hz, 2H), 2.09 - 1.98 (m, 4H), 1.96 - 1.87 (m, 4H), 1.78 (dd, J = 7.3, 3.9 Hz, 4H). 13C NMR (126 MHz, DMSO) δ 164.24, 157.00, 155.40, 155.17, 149.52, 145.43, 130.96, 128.32 (2C), 126.96 (2C), 125.29 (2C), 124.74, 118.89, 114.44 (2C), 110.33, 106.61, 68.48, 63.48, 53.67 (2C), 52.66, 27.19, 22.62 (2C). ESI-HRMS [M + 2H] 2+ m / z = 306.1809, calcd for C 35 H 42 N6O4, 306.1707.

[0321] trisubstituted quinazoline derivative a8: 1 H NMR (400 MHz, MeOD) δ 7.89 (d, J = 2.6 Hz, 1H), 7.76 (dq, J = 9.3, 2.7 Hz, 2H), 7.66 - 7.61 (m, 1H), 7.46 (dt, J = 9.1, 2.9 Hz, 1H), 7.16 - 7.10 (m, 2H), 4.40 (t, J = 4.9 Hz, 2H), 4.15 (q, J = 5.8 Hz, 2H), 3.76 (dd, J = 12.3, 6.8 Hz, 2H), 3.70 (t, J = 4.8 Hz, 2H), 3.64 (s, 4H), 3.25 (q, J = 7.3 Hz, 4H), 2.24 - 2.17 (m, 4H), 2.12 - 2.02 (m, 4H), 1.89 - 1.79 (m, 4H). 13 C NMR (101 MHz, DMSO) δ 174.82, 169.34, 157.47, 155.83, 155.63, 150.07, 131.42, 125.71 (2C), 125.19, 119.45, 114.93 (2C), 113.67, 110.78, 106.97, 68.72, 63.94, 54.15 (2C), 53.14, 40.66, 40.46, 33.77, 32.31, 28.50, 23.07 (2C), 22.28, 21.73, 9.48. ESI-HRMS [M + 2H] 2+ m / z = 268.1598, calcd for C 29 H 38N6O4, 268.1550.

[0322] trisubstituted quinazoline derivative a9: 1 H NMR (500 MHz, CDC13) δ 7.89 (s, 1H), 7.71 (d, J = 8.5 Hz, 2H), 7.47 (d, J = 9.1 Hz, 1H), 7.16-7.08 (m, 2H), 6.98 (t, J = 7.6 Hz, 1H), 6.85 (d, J = 8.4 Hz, 2H), 6.74-6.65 (m, 2H), 4.09 (t, J = 5.8 Hz, 2H), 3.93 (t, J = 6.4 Hz, 2H), 3.59 (d, J = 6.4 Hz, 4H), 2.90 (t, J = 5.8 Hz, 2H), 2.63 (d, J = 5.7 Hz, 4H), 2.43 (t, J = 7.0 Hz, 2H), 1.92 (d, J = 6.1 Hz, 4H), 1.87-1.72 (m, 8H). 13 C NMR (126 MHz, CDC13) δ 172.11, 156.97, 155.01, 153.30, 140.95, 132.52, 127.22, 125.59, 124.09, 123.94 (2C), 123.00, 119.27, 117.91, 114.41 (2C), 110.20, 103.54, 77.24, 68.21, 67.24, 55.18, 54.73 (2C), 46.85, 36.09, 28.21, 25.52, 23.47 (2C), 22.39. ESI-HRMS [M + 2H] 2+ m / z = 305.6865, calcd for C 35 H 43 N7O3, 305.6786.

[0323] trisubstituted quinazoline derivative a10: 1H NMR(400MHz, MeOD)δ7.88(d, J=2.5Hz, 1H),7.80-7.73(m, 2H), 7.63(d, J=9.1Hz, 1H), 7.45(dt, J=9.1, 3.1Hz, 1H), 7.17-7.10(m, 2H), 4.43-4.37(m, 2H), 4.13(t, J=6.4Hz, 2H), 3.79-3.74(m, 2H), 3.70(t, J=4.8Hz, 2H), 3.63(d, J=14.4Hz, 4H), 3.28-3.23(m, 2H), 2.22(dt, J=10.4, 4.8Hz,2H), 2.16-1.99(m, 8H), 1.86(q, J=6.9Hz, 2H), 1.70-1.63(m, 2H), 1.54(dd, J=10.8, 5.0Hz, 2H), 1.44(q, J=8.1Hz, 2H). 13 C NMR(126MHz, CDCl3)(126MHz, DMSO)δ169.57, 157.44, 155.83, 155.58, 131.49, 125.68, 125.16(2C), 114.88(2C),110.77, 106.86, 69.05, 63.94, 54.10(2C), 53.09, 32.67, 28.98, 28.80, 25.76,25.53, 23.08(2C).ESI-HRMS [M + 2H] 2+ m / z=282.1803, calcd for C 31 H 42 N6O4, 282.1707.

[0324] Trisubstituted quinazoline derivative a11: 1H NMR (400 MHz, CDC13) δ 7.75-7.70 (m, 2H), 7.50 (d, J = 9.0 Hz, 1H), 7.42 (s, 1H), 7.19 (dd, J = 9.0, 2.6 Hz, 1H), 7.16 (d, J = 2.9 Hz, 1H), 7.15-7.12 (m, 1H), 7.01 (td, J = 7.6, 1.5 Hz, 1H), 6.93-6.87 (m, 2H), 6.76-6.71 (m, 2H), 5.30 (s, 1H), 4.12 (t, J = 5.9 Hz, 2H), 3.93 (t, J = 6.3 Hz, 2H), 3.83 (s, 2H), 3.64 (d, J = 6.4 Hz, 4H), 3.48 (s, 1H), 2.91 (t, J = 5.9 Hz, 2H), 2.63 (q, J = 5.0 Hz, 4H), 2.40 (t, J = 7.4 Hz, 2H), 1.95 (q, J = 3.4 Hz, 4H), 1.82 (q, J = 3.3 Hz, 4H), 1.77 (q, J = 5.9 Hz, 4H), 1.46 (q, J = 7.9 Hz, 4H). 13 C NMR (126 MHz, CDC13) δ 172.16, 156.91, 155.02, 153.50, 140.88, 127.18, 125.41, 124.24, 123.64, 122.81 (2C), 119.41, 118.05, 114.52 (2C), 110.17, 68.48, 67.32, 55.19 (2C), 54.75 (2C), 46.85, 36.80, 28.82, 28.66, 25.63, 25.56 (2C), 23.49 (2C). ESI-HRMS [M + 2H] 2+ m / z = 319.7043, calcd for C 37 H 47 N7O3, 319.6943.

[0325] trisubstituted quinazoline derivative a12: 1H NMR (500 MHz, DMSO) δ 12.24 (s, 1H), 11.35-11.20 (m, 1H), 11.13 (s, 1H), 10.93-10.80 (m, 1H), 9.12-8.80 (m, 1H), 8.30-8.22 (m, 1H), 7.94 (dd, J = 9.2, 4.2 Hz, 1H), 7.81 (dd, J = 8.6, 5.8 Hz, 2H), 7.65 (s, 1H), 7.58 (d, J = 7.3 Hz, 1H), 7.47 (dd, J = 9.3, 2.7 Hz, 1H), 7.45-7.32 (m, 2H), 7.09 (d, J = 8.5 Hz, 2H), 4.40 (t, J = 5.0 Hz, 2H), 4.18 (d, J = 5.3 Hz, 2H), 3.68-3.55 (m, 8H), 3.13 (q, J = 7.9 Hz, 2H), 2.73 (d, J = 6.8 Hz, 2H), 2.02 (q, J = 8.2 Hz, 4H), 1.96-1.87 (m, 4H), 1.79 (q, J = 4.1 Hz, 4H). 13 C NMR (126 MHz, CDCl3) δ 172.16, 156.91, 155.02, 153.50, 140.88, 127.18, 125.41, 124.24, 123.64, 122.81 (2C), 119.41, 118.05, 114.52 (2C), 110.17, 68.48, 67.32, 55.19 (2C), 54.75 (2C), 46.85, 36.80, 28.82, 28.66, 25.63, 25.56 (2C), 23.49 (2C). ESI-HRMS [M + 2H] 2+ m / z = 307.1791, calcd for C 35 H 42 N6O4, 307.1769.

[0326] trisubstituted quinazoline derivative a15: 1H NMR (500 MHz, DMSO) δ 12.17 (s, 1H), 10.96 (s, 1H), 10.48 (s, 1H), 8.78 (s, 1H), 8.22 (s, 1H), 7.95 (s, 1H), 7.82 (d, J = 8.4 Hz, 2H), 7.49 (d, J = 9.0 Hz, 1H), 7.09 (d, J = 8.5 Hz, 2H), 4.39 (t, J = 5.0 Hz, 2H), 4.28 - 4.22 (m, 2H), 3.77 (t, J = 4.7 Hz, 2H), 3.71 (q, J = 5.1 Hz, 2H), 3.59 (s, 8H), 3.12 (s, 2H), 2.25 (t, J = 6.3 Hz, 2H), 2.01 (s, 4H), 1.93 (s, 4H). 13 C NMR (126 MHz, DMSO) δ 167.43, 157.42, 155.50, 125.54, 114.93 (2C), 110.76, 107.02, 68.99, 68.63, 67.15, 66.87, 63.95, 54.14 (2C), 53.14, 36.27, 35.21, 34.60, 33.65, 31.24, 23.07 (2C). ESI-HRMS [M + 2H] 2+ m / z = 269.6538, calcd for C 29 H 40 N6O4, 269.1628.

[0327] trisubstituted quinazoline derivative a16: 1 H NMR (500 MHz, DMSO) δ 12.17 (s, 1H), 10.96 (s, 1H), 10.48 (s, 1H), 8.78 (s, 1H), 8.22 (s, 1H), 7.95 (s, 1H), 7.82 (d, J = 8.4 Hz, 2H), 7.49 (d, J = 9.0 Hz, 1H), 7.09 (d, J = 8.5 Hz, 2H), 4.39 (t, J = 5.0 Hz, 2H), 4.28 - 4.22 (m, 2H), 3.77 (t, J = 4.7 Hz, 2H), 3.71 (q, J = 5.1 Hz, 2H), 3.59 (s, 8H), 3.12 (s, 2H), 2.25 (t, J = 6.3 Hz, 2H), 2.01 (s, 4H), 1.93 (s, 4H). 13C NMR (126 MHz, DMSO) δ 164.49, 159.58, 157.41, 155.83, 155.55, 144.52, 134.38, 125.69, 125.16 (2C), 119.74, 117.47, 114.83 (2C), 110.92, 110.77, 109.80, 106.90, 106.18, 69.13, 63.92, 55.70 (2C), 54.04 (2C), 53.03, 35.51, 31.13, 29.02, 28.88, 25.87, 23.10 (4C). ESI-HRMS [M + 2H] 2+ m / z = 291.1757, calcd for C 31 H 41 N6O4F, 291.1659.

[0328] trisubstituted quinazoline derivative a17: 1 H NMR (400 MHz, DMSO) δ 12.18 (s, 1H), 10.80 (s, 1H), 10.60 (d, J = 16.5 Hz, 1H), 10.41 (s, 1H), 8.68 (s, 1H), 8.20 - 8.14 (m, 1H), 7.89 (d, J = 9.1 Hz, 1H), 7.77 (d, J = 8.8 Hz, 2H), 7.71 (d, J = 8.7 Hz, 2H), 7.48 (dd, J = 9.1, 2.5 Hz, 1H), 4.11 (t, J = 6.4 Hz, 2H), 3.60 (d, J = 16.0 Hz, 4H), 3.51 (s, 4H), 3.05 (s, 2H), 2.94 (t, J = 7.4 Hz, 2H), 2.79 (s, 2H), 2.03 - 1.88 (m, 8H), 1.75 (q, J = 7.0 Hz, 2H), 1.57 - 1.49 (m, 2H), 1.44 (q, J = 7.6 Hz, 2H), 1.33 (q, J = 8.0 Hz, 2H). 13CNMR (101 MHz, DMSO) δ 169.56, 168.23, 157.47, 155.89, 149.96, 136.92, 133.00, 125.24, 124.81 (2C), 119.40 (2C), 110.75, 106.80, 69.02, 53.50 (2C), 50.16, 42.73, 32.68, 32.61, 28.97, 28.81, 25.77, 25.54, 23.20 (2C). ESI-HRMS [M + 2H] 2+ m / z = 295.6862, calcd for C 32 H 43 N7O4, 295.6761.

[0329] trisubstituted quinazoline derivative a18: 1 H NMR (500 MHz, DMSO) δ 12.12 (s, 1H), 10.79 (s, 1H), 10.60 (s, 1H), 10.39 (s, 1H), 10.26 (s, 1H), 8.67 (s, 1H), 8.17 (d, J = 2.6 Hz, 1H), 7.88 (d, J = 9.1 Hz, 1H), 7.77 (d, J = 8.9 Hz, 2H), 7.72 (d, J = 8.7 Hz, 2H), 7.50 (dd, J = 9.1, 2.4 Hz, 1H), 4.11 (t, J = 6.4 Hz, 2H), 3.67-3.56 (m, 4H), 3.38 (s, 2H), 3.14 (p, J = 6.9 Hz, 4H), 2.93 (t, J = 7.4 Hz, 2H), 2.06-1.91 (m, 6H), 1.76 (q, J = 7.1 Hz, 2H), 1.54 (q, J = 7.4 Hz, 2H), 1.45 (t, J = 7.6 Hz, 2H), 1.34 (q, J = 8.0 Hz, 2H), 1.25 (t, J = 7.2 Hz, 6H). 13C NMR (126 MHz, DMSO) δ 169.54, 168.36, 157.50, 155.92, 149.93, 136.98, 134.67, 132.96, 125.26, 124.86 (2C), 119.95, 119.41 (2C), 110.76, 106.82, 69.00, 60.23, 47.22 (2C), 46.93, 32.68, 30.91, 28.97, 28.81, 25.77, 25.53, 14.56, 8.96 (2C). ESI-HRMS [M + 2H] 2+ m / z = 296.6942, calcd for C 32 H 45 N7O4, 296.6839.

[0330] trisubstituted quinazoline derivative a19: 1 H NMR (500 MHz, DMSO) δ 12.21 (s, 1H), 11.03 (s, 1H), 10.54 (s, 1H), 10.41 (s, 1H), 8.69 (s, 1H), 8.55 (s, 1H), 7.79 (d, J = 8.4 Hz, 2H), 7.46 (s, 1H), 7.07 (d, J = 8.6 Hz, 2H), 7.04 (s, 1H), 4.38 (s, 2H), 4.09 (t, J = 6.4 Hz, 2H), 3.58 (s, 6H), 2.04 - 1.92 (m, 8H), 1.77 (t, J = 7.4 Hz, 2H), 1.53 (q, J = 7.2 Hz, 2H), 1.44 (s, 2H), 1.33 (d, J = 7.5 Hz, 2H), 1.24 (d, J = 3.8 Hz, 4H). 13 C NMR (101 MHz, DMSO) δ 169.55, 163.90, 157.42, 130.13, 125.22, 114.93 (2C), 103.73, 68.72, 63.97, 54.12, 53.13 (2C), 32.65, 31.76, 29.50, 28.73, 27.02, 25.59, 23.09 (2C), 11.45. ESI-HRMS [M + 2H] 2+ m / z = 282.1812, calcd for C31H42N6O4, 282.1707.

[0331] trisubstituted quinazoline derivative a20:1 H NMR (500 MHz, CDC13) δ 7.83 (s, 1H), 7.77 (d, J = 8.5 Hz, 2H), 7.65 (d, J = 7.8 Hz, 2H), 7.55 (d, J = 9.1 Hz, 1H), 7.21 (d, J = 8.1 Hz, 3H), 7.16-7.12 (m, 1H), 6.91 (t, J = 8.5 Hz, 2H), 4.12 (t, J = 5.9 Hz, 2H), 3.97 (t, J = 6.4 Hz, 2H), 3.63 (d, J = 6.5 Hz, 4H), 2.92 (q, J = 7.1 Hz, 4H), 2.65 (q, J = 7.2 Hz, 6H), 1.85-1.80 (m, 4H), 1.73 (q, J = 7.1 Hz, 2H), 1.62 (q, J = 7.6 Hz, 2H), 1.53 (q, J = 7.4 Hz, 2H), 1.46 (d, J = 7.5 Hz, 2H), 1.36 (q, J = 7.9 Hz, 4H), 1.28 (tt, J = 6.2, 3.3 Hz, 4H), 0.88 (t, J = 6.7 Hz, 4H). 13 C NMR (101 MHz, CDC13) δ 167.18, 156.85, 155.53, 154.81, 146.92, 131.71, 130.33, 129.90, 128.66 (2C), 126.96 (2C), 125.64, 124.38, 123.77, 114.36 (2C), 114.21, 110.21, 68.89, 66.81, 54.99 (2C), 54.72, 52.42, 35.70, 31.75, 31.49, 30.98, 29.79, 29.62, 29.33, 29.14, 28.91, 28.02, 27.22, 26.81, 25.89, 25.36, 23.46 (2C), 22.60, 22.41, 14.06, 13.96. ESI-HRMS [M + 2H] 2+ m / z = 361.7490, calcd for C 41 H 55 N7O3, 364.7521.

[0332] trisubstituted quinazoline derivative a21: 1H NMR (500 MHz, DMSO) δ 11.97 (s, 1H), 11.31 (s, 1H), 10.80 - 10.57 (m, 2H), 9.09 (s, 1H), 8.68 (s, 1H), 8.24 (s, 1H), 8.12 (s, 1H), 7.96 (d, J = 7.7 Hz, 1H), 7.80 (s, 1H), 7.76 (d, J = 8.8 Hz, 2H), 7.69 (d, J = 7.7 Hz, 1H), 7.50 (q, J = 8.7 Hz, 2H), 7.10 (d, J = 8.6 Hz, 2H), 4.44 (t, J = 7.0 Hz, 2H), 4.37 (t, J = 5.0 Hz, 2H), 4.11 (t, J = 6.4 Hz, 2H), 3.59 (s, 8H), 3.12 (s, 2H), 2.03 (s, 4H), 1.92 (d, J = 9.4 Hz, 6H), 1.79 (t, J = 7.4 Hz, 2H), 1.52 (dd, J = 11.1, 4.6 Hz, 2H), 1.40 - 1.34 (m, 2H). 13 C NMR (126 MHz, DMSO) δ 157.46, 146.19, 133.92, 131.50, 129.47, 128.04, 126.49, 125.74, 124.23, 122.18, 114.96 (2C), 110.75, 75.24, 68.88, 63.95, 54.23 (2C), 53.22, 50.00, 29.94, 28.82, 26.03, 25.45, 23.04 (2C). ESI-HRMS [M + 2H] 2+ m / z = 353.7055, calcd for C 39 H 47 N9O4, 353.6948.

[0333] trisubstituted quinazoline derivative a22: 1H NMR (500 MHz, CDC13) δ 7.74 (d, J = 8.5 Hz, 2H), 7.53 (d, J = 9.1 Hz, 1H), 7.15 (s, 1H), 7.06 (d, J = 9.1 Hz, 1H), 6.86 (d, J = 8.5 Hz, 2H), 4.08 (t, J = 6.0 Hz, 2H), 3.96 (t, J = 6.6 Hz, 2H), 3.59 (t, J = 7.4 Hz, 4H), 2.88 (t, J = 5.9 Hz, 2H), 2.62 (d, J = 5.9 Hz, 4H), 1.88 (d, J = 6.3 Hz, 4H), 1.72-1.68 (m, 2H), 1.40 (t, J = 7.6 Hz, 2H), 1.29-1.26 (m, 4H), 1.19 (d, J = 5.0 Hz, 4H), 0.83 (t, J = 7.3 Hz, 3H). 13 C NMR (126 MHz, CDC13) δ 156.70, 155.32, 132.10, 129.93, 123.96, 123.05, 114.57 (2C), 109.97, 70.55, 68.87, 67.16, 55.09, 54.74, 47.19, 35.94, 31.92, 31.66, 29.79, 29.71, 29.53, 29.33, 27.22, 25.78, 25.47, 23.49, 22.70, 22.62, 14.13, 14.07. ESI-HRMS [M + 2H] 2+ m / z = 319.7043, calcd for C 37 H 47 N7O3, 319.6943.

[0334] trisubstituted quinazoline derivative b1: 1H NMR (500 MHz, DMSO) δ 10.38 (s, 1H), 9.31 (s, 1H), 8.69 (d, J = 1.8 Hz, 1H), 8.26 (d, J = 8.3 Hz, 1H), 7.86 - 7.82 (m, 2H), 7.54 (t, J = 7.6 Hz, 1H), 7.32 (d, J = 8.4 Hz, 1H), 7.10 (t, J = 7.5 Hz, 1H), 6.96 - 6.92 (m, 2H), 3.96 (t, J = 6.0 Hz, 2H), 3.52 (d, J = 6.7 Hz, 4H), 2.03 (t, J = 7.1 Hz, 2H), 1.92 (dd, J = 6.5, 3.3 Hz, 4H), 1.71 - 1.62 (m, 4H), 1.32 - 1.19 (m, 4H). 13 C NMR (126 MHz, DMSO) δ 169.40, 157.63, 156.68, 150.43, 140.41, 135.34, 130.62, 125.61, 125.13 (2C), 124.70, 117.86, 114.56 (2C), 110.21, 67.74, 48.78, 48.01, 32.37, 28.66 (2C), 25.70, 24.56, 22.27 (2C). ESI-HRMS [M + H] + m / z = 422.2581, calcd for C 23 H 27 N5O3, 422.2187.

[0335] trisubstituted quinazoline derivative b2: 1H NMR (500 MHz, DMSO) δ 9.54 (s, 1H), 9.11 (s, 1H), 8.30 (d, J = 8.2 Hz, 1H), 7.81 (d, J = 8.5 Hz, 2H), 7.59 (t, J = 7.8 Hz, 1H), 7.40 (d, J = 8.4 Hz, 1H), 7.17 (d, J = 8.1 Hz, 2H), 6.94 (d, J = 8.6 Hz, 2H), 6.89 (t, J = 7.9 Hz, 1H), 6.72 (d, J = 7.9 Hz, 1H), 6.54 (t, J = 7.5 Hz, 1H), 4.82 (s, 2H), 3.96 (t, J = 6.5 Hz, 2H), 3.52 (d, J = 6.4 Hz, 4H), 2.34 (t, J = 7.5 Hz, 2H), 1.92 (d, J = 6.2 Hz, 4H), 1.72 (q, J = 7.1 Hz, 2H), 1.64 (p, J = 7.5 Hz, 2H), 1.46 (p, J = 6.8 Hz, 2H), 1.42 - 1.30 (m, 2H). 13 C NMR (126 MHz, DMSO) δ 171.66, 162.78, 157.88, 156.27, 155.30, 142.35, 133.42, 132.69, 126.16, 125.76, 124.06, 123.80, 123.64 (2C), 121.33, 116.66, 116.37, 114.52 (2C), 110.67, 68.01, 46.98, 38.54, 36.21, 31.23, 29.17, 28.94, 25.85, 25.75, 25.43. ESI-HRMS [M + H] + m / z = 525.3098, calcd for C 31 H 36 N6O2, 525.2973.

[0336] trisubstituted quinazoline derivative b3: 1H NMR (400 MHz, DMSO) δ 10.33 (s, 1H), 9.32 (s, 1H), 8.64 (s, 1H), 8.26 (d, J = 8.4 Hz, 1H), 7.87 - 7.80 (m, 2H), 7.54 (t, J = 7.7 Hz, 1H), 7.33 (d, J = 8.3 Hz, 1H), 7.11 (t, J = 7.5 Hz, 1H), 6.97 - 6.90 (m, 2H), 3.95 (t, J = 6.5 Hz, 2H), 3.57 - 3.48 (m, 4H), 1.97 (d, J = 7.3 Hz, 2H), 1.91 (q, J = 3.5 Hz, 4H), 1.70 (t, J = 7.3 Hz, 2H), 1.51 (q, J = 7.5 Hz, 2H), 1.42 (q, J = 7.5 Hz, 2H), 1.35 - 1.29 (m, 2H). 13 C NMR (126 MHz, DMSO) δ 169.58, 157.67, 156.73, 150.46, 140.43, 135.35, 130.59, 125.65, 125.15 (2C), 124.70, 117.87, 114.56 (2C), 110.23, 68.05, 48.77, 48.03, 32.66, 29.06, 28.81, 25.73, 25.55 (2C), 24.56. ESI-HRMS [M + H] + m / z = 450.2821, calcd for C 25 H 31 N5O3, 450.2500.

[0337] trisubstituted quinazoline derivative b5: 1H NMR (500 MHz, DMSO) δ 12.00 (s, 1H), 11.16 (s, 1H), 10.70 (s, 1H), 8.99 (s, 1H), 8.58 (d, J = 8.3 Hz, 1H), 7.86 (q, J = 4.3 Hz, 2H), 7.74 - 7.66 (m, 4H), 7.49 (ddd, J = 8.3, 5.6, 2.7 Hz, 1H), 7.30 (d, J = 7.9 Hz, 2H), 7.04 - 6.97 (m, 2H), 5.76 (s, 1H), 4.03 (d, J = 5.8 Hz, 2H), 3.66 (d, J = 6.9 Hz, 2H), 3.58 (t, J = 7.0 Hz, 2H), 2.71 (d, J = 6.6 Hz, 2H), 2.10 - 2.01 (m, 2H), 1.97 - 1.87 (m, 2H), 1.78 - 1.71 (m, 4H). 13 C NMR (126 MHz, DMSO) δ 164.69, 157.68, 156.81, 150.46, 145.92, 140.27, 135.61, 130.82, 130.46, 128.77 (2C), 127.41, 125.60 (2C), 124.88, 124.80, 117.82, 114.69 (2C), 110.22, 67.91, 55.39, 48.78, 47.53, 35.02, 28.70, 27.64, 25.70, 24.59. ESI-HRMS [M + H]+m / z = 498.2679, calcd for C 29 H 31 N5O3, 498.2500.

[0338] trisubstituted quinazoline derivative b6: 1H NMR (500 MHz, MeOD) δ 7.86 (s, 1H), 7.71 (d, J = 8.5 Hz, 2H), 7.60 (d, J = 8.9 Hz, 1H), 7.48 (dd, J = 8.9, 2.3 Hz, 1H), 6.98 (d, J = 8.6 Hz, 2H), 4.08 - 4.00 (m, 2H), 3.73 - 3.68 (m, 2H), 3.65 (d, J = 15.8 Hz, 4H), 3.50 (t, J = 6.9 Hz, 2H), 3.40 (t, J = 7.8 Hz, 2H), 3.13 (q, J = 9.0 Hz, 2H), 2.25 - 2.15 (m, 6H), 2.05 (dd, J = 7.8, 5.0 Hz, 4H), 1.82 (p, J = 3.0 Hz, 4H). 13 C NMR (126 MHz, MeOD) δ 174.38, 171.38, 157.36, 157.09, 149.50, 130.13, 125.14 (2C), 125.12, 124.93, 118.05, 113.95 (2C), 110.86, 104.83, 67.42, 67.37, 53.88, 52.63 (2C), 42.29, 33.04, 32.00, 28.40, 28.37, 24.55, 22.67, 22.07 (2C), 21.37. ESI-HRMS [M + 2H] 2+ m / z = 274.7067, calcd for C 31 H 41 N7O3, 274.6708.

[0339] trisubstituted quinazoline derivative b7: 1H NMR (500 MHz, DMSO) δ 11.80 (s, 1H), 10.63 (s, 1H), 10.42 (s, 1H), 7.72 (d, J = 8.4 Hz, 2H), 7.65 (d, J = 9.0 Hz, 1H), 7.59 (s, 1H), 7.24 (d, J = 9.0 Hz, 1H), 7.00 (d, J = 8.4 Hz, 2H), 4.06 - 4.00 (m, 2H), 4.00 (d, J = 6.7 Hz, 2H), 3.67 (s, 4H), 3.55 (s, 4H), 3.30 (s, 4H), 2.83 (s, 2H), 2.04 (q, J = 9.5 Hz, 4H), 1.99 (s, 3H), 1.91 (s, 2H), 1.73 - 1.63 (m, 4H), 1.17 (t, J = 7.1 Hz, 2H). 13 C NMR (126 MHz, MeOD) δ 157.30, 156.97, 149.30, 132.50, 130.22, 130.19, 125.10 (2C), 124.43, 117.97, 113.92 (2C), 110.79, 103.14, 67.44, 54.59, 50.71, 49.91, 48.60, 42.07, 41.44, 33.06 (2C), 28.42, 28.37, 22.85, 22.05, 21.42, 21.38. ESI-HRMS [M + 2H] 2+ m / z = 289.1971, calcd for C 31 H 44 N8O3, 289.1841.

[0340] trisubstituted quinazoline derivative b8: 1 H NMR (500 MHz, MeOD) δ 7.77 (s, 2H), 7.51 (s, 2H), 7.22 (s, 1H), 6.93 (s, 2H), 3.99 (s, 2H), 3.90 (s, 2H), 3.57 (s, 6H), 3.31 (s, 4H), 3.28 - 3.20 (m, 2H), 2.21 (s, 2H), 2.02 (s, 4H), 1.81 (s, 4H), 1.38 (d, J = 6.4 Hz, 2H). 13C NMR (126 MHz, DMSO) δ 174.82, 169.40, 157.26, 156.48, 149.29, 145.59, 132.19, 130.79, 125.85 (2C), 124.58, 118.94, 114.36 (2C), 111.05, 101.36, 67.70, 63.91, 63.61, 53.67, 51.64, 49.04, 33.80, 32.37, 28.67, 22.27, 21.70, 18.40, 17.16, 12.59. ESI-HRMS [M + H] + m / z = 550.3238, calcd for C 29 H 39 N7O4, 550.3136.

[0341] trisubstituted quinazoline derivative b9: 1 H NMR (400 MHz, DMSO) δ 12.00 (s, 1H), 11.03 (s, 1H), 10.52 (s, 1H), 10.43 (s, 1H), 8.31 (s, 1H), 8.00 (d, J = 2.3 Hz, 1H), 7.81 (d, J = 9.0 Hz, 1H), 7.64 (d, J = 8.9 Hz, 2H), 7.48 (dd, J = 9.0, 2.3 Hz, 1H), 7.13 (d, J = 8.5 Hz, 2H), 7.06 - 6.93 (m, 4H), 3.99 (t, J = 6.1 Hz, 2H), 3.67 (d, J = 12.3 Hz, 2H), 3.59 (d, J = 8.9 Hz, 4H), 3.48 (d, J = 11.6 Hz, 2H), 3.16 (q, J = 10.4 Hz, 2H), 3.06 (t, J = 12.0 Hz, 2H), 2.81 (d, J = 4.4 Hz, 3H), 2.15 - 1.84 (m, 6H), 1.77 - 1.61 (m, 4H). 13CNMR (126 MHz, DMSO) δ 169.38, 157.35, 156.67, 149.53, 144.83, 142.23, 136.49, 130.64, 126.40, 125.84 (2C), 119.59, 119.13 (2C), 118.21 (2C), 114.55 (2C), 110.98, 106.96, 67.72, 52.77 (2C), 46.93 (2C), 42.41, 33.39, 32.38, 28.68, 22.27, 21.66. ESI-HRMS [M + 2H] 2+ m / z = 306.2167, calcd for C 34 H 42 N8O3, 306.1763.

[0342] 2. In vitro target activity evaluation of the trisubstituted quinazoline derivatives prepared in Examples 1-28:

[0343] The in vitro stability of the trisubstituted quinazoline derivatives to telomeric G4 and double-stranded DNA was evaluated using a fluorescence resonance energy transfer (FRET) experiment, following the steps below:

[0344] 1) Buffer preparation: a buffer solution containing 100 mmol / L KCl and 10 mmol / L Tris-HCl (pH = 7.4) was prepared;

[0345] 2) DNA sample preparation: the fluorescently labeled DNA sequence was diluted to 800 nmol / L with the above buffer, heated at 95°C for 10 min, and then slowly cooled to room temperature to form a specific secondary structure, wherein the fluorescently labeled DNA sequence is:

[0346] Telomeric G4: 5'-FAM-GGGTTAGGGTTAGGGTTAGGG-TAMRA-3';

[0347] Double-stranded DNA: 5' FAM-TATAGCTATA-HEG-TATAGCTATA-TAMRA-3';

[0348] 3) Compound incubation: the DNA sample was mixed with the trisubstituted quinazoline derivative to be tested, so that the final DNA concentration was 400 nmol / L and the trisubstituted quinazoline derivative concentration was 2 μmol / L. The mixture was incubated at 37°C for 1 h;

[0349] 4) Melting curve assay: The assay was performed using Roche LightCycler 2.0 fluorescence quantitative PCR instrument, the excitation wavelength was set at 470 nm, the emission fluorescence intensity at 530 nm was collected, the temperature program was set from 37℃ to 99℃, the temperature interval was 1℃, the signal was collected after 30s equilibrium at each temperature;

[0350] 5) After the experiment, the Prism software was used to fit T m value, the T m value of the compound minus the T m value of the DNA to obtain the ΔT m (℃), the greater the difference, the stronger the stability of the compound to the telomere G4, for double-stranded DNA, the greater the ΔT m (℃) difference, the stronger the stability of the compound to the double-stranded, and the higher the ΔT m to the telomere and the smaller to the double-stranded, wherein the compound A4 prepared in CN 120365220 A Example 1 and the classical HDAC inhibitor SAHA are used as controls.

[0351] Table 1 is the in vitro target activity evaluation results of the tri-substituted quinazoline derivatives in Examples 1-28

[0352]

[0353] wherein, a the T m value of F21T without the compound is 62.4℃, the T m value of F10T is 62.7℃, ΔT m = T m (DNA+ligand)−T m (DNA), the experimental error of ΔT m is about 2.0℃; ΔT m >35℃ indicates that T m has not been reached at 99℃.

[0354] Table 1 is the in vitro target activity evaluation results of the tri-substituted quinazoline derivatives in Examples 1-28, from Table 1, it can be seen that the ΔT m >35℃ of multiple compounds (such as a4, a5, a6, a10, a20, a21, a22 and b7) to the telomere G4 indicates that these compounds have extremely strong G4 stability, can significantly improve the thermal stability of the G4 structure, and make it only dissociate at a higher temperature, which is conducive to maintaining the G4 structure at the physiological temperature of the cell, interfering with the maintenance of the telomere and gene transcription; and the ΔT mGenerally very low (most <1℃), even negative (e.g. a5), proving that these compounds have high selectivity for G4 structure, and will not non-specifically stabilize common double-stranded DNA, which is the key to reduce off-target toxicity, indicating that the compound is a true G4 ligand, rather than an ordinary DNA binder; the ΔT m of telo is 12.9℃, lower than most of the trisubstituted quinazoline derivatives of the application, proving that the structural modification of the application significantly enhances the affinity and stability of the parent quinazoline to G4; the ΔT m of SAHA to G4 and double-stranded DNA are -4.0℃ and 4.0℃, respectively, indicating that the classic HDAC inhibitor SAHA does not have G4 stabilizing activity, and may even destroy G4, which inversely proves the necessity of the dual-target design of the application.

[0355] 3. The trisubstituted quinazoline derivatives prepared in Examples 1-28 were evaluated for histone deacetylase (HDAC) inhibitory activity, with the following steps:

[0356] 1) The gradient-diluted trisubstituted quinazoline derivatives to be tested were mixed with the HDAC-Glo™ I / II kit-provided HeLa cell nucleus extract (containing HDAC enzyme) in the kit buffer;

[0357] 2) The above mixture was incubated at room temperature for 30 min to allow the compound to fully act on the enzyme;

[0358] 3) HDAC-Glo™ I / II detection solution containing fluorescent substrate and developing reagent was added, and the incubation was continued at room temperature for 45 min, and the HDAC activity was proportional to the generated fluorescent signal;

[0359] 4) The chemiluminescence detector was used to read the luminescence intensity, and Prism software was used to calculate the half-inhibitory concentration (IC 50 ) according to the dose-response curve, wherein the compound A4 prepared in CN 120365220 A Example 1 and the classic HDAC inhibitor SAHA were used as controls.

[0360] Table 2 Evaluation results of the histone deacetylase inhibitory activity of the trisubstituted quinazoline derivatives in Examples 1-28

[0361]

[0362] Table 2 is the evaluation results of the histone deacetylase inhibitory activity of the trisubstituted quinazoline derivatives in Examples 1-28. As can be seen from Table 2, the IC 50Dana-mole level, such as a10, a18, a20 and a21, show that the series of compounds have strong HDAC inhibitory activity by introducing hydroxamic acid pharmacophore, among which a10 is particularly prominent (2 nmol / L); IC 50 The compounds a4, a5, b2 and the like, which have a wide range of 2 nmol / L to more than 20 μmol / L, show inhibitory activity, indicating that the HDAC inhibitory activity is adjustable, and the activity can be optimized by structural modification, providing space for structure-activity relationship study and subsequent development; the IC 50 >20 μmol / L, almost no activity, while the compounds of the present application generally have activity, which directly proves that the present application successfully introduces HDAC inhibitory function on the original G4 ligand parent nucleus, realizing the qualitative change from single target to double target; the IC 50 of SAHA is 70 nmol / L, while the activities of a10, a18, a20 and the like in the present application are better than SAHA, indicating that the present application not only successfully designs double targets, but also achieves a high level of HDAC inhibitory single target activity.

[0363] 4. The cytotoxicity and cell safety of the trisubstituted quinazoline derivatives prepared in Examples 1-28 were evaluated, and the steps were as follows:

[0364] 1) Logarithmic growth phase HCT116 (human colorectal cancer cells) and NCM460 (human normal colon epithelial cells) were inoculated in a 96-well plate at a density of 4000 cells per well, and cultured overnight to adhere;

[0365] 2) Remove the old culture medium, add compound solution diluted with fresh culture medium (maximum concentration 50 μmol / L, 8 gradients), each concentration set of duplicate wells, continue to culture for 48 h;

[0366] 3) Remove the drug-containing culture medium, add pre-cooled (-20℃) 70% ethanol, fix overnight at -20℃, remove the ethanol, wash once with PBS, and add 0.5 μg / mL DAPI-containing PBS solution for staining;

[0367] 4) Use high-content imaging system to automatically photograph and count the cell nucleus, use Prism software to fit the dose-response curve, and calculate the half-inhibitory concentration IC 50 , the selectivity index (SI) = IC 50 (NCM460) / IC 50 (HCT116), wherein the compound A4 prepared in CN 120365220 A Example 1 and the classical HDAC inhibitor SAHA are used as controls.

[0368] Table 3: Results of cytotoxicity evaluation of the tri-substituted quinazoline derivatives in Examples 1-28

[0369]

[0370] Table 4: Selectivity fold of cytotoxicity of the tri-substituted quinazoline derivatives in Examples 1-28

[0371]

[0372] Table 3: Results of cytotoxicity evaluation of the tri-substituted quinazoline derivatives in Examples 1-28, and Table 4: Selectivity fold of cytotoxicity of the tri-substituted quinazoline derivatives in Examples 1-28, from which it can be seen that the IC 50 of the tri-substituted quinazoline derivative a10 against HCT116 is the lowest, 0.3 μmol / L, and the activity is the strongest, and a6, a19, b2, b9, etc. also show sub-micromolar activity (IC 50 <2 μmol / L), indicating that the tri-substituted quinazoline derivatives have a significant anti-proliferation effect at the cellular level; the selectivity index of a10 is the highest, reaching 39, and the index of a8, b2, b3, etc. is also greater than 20, indicating that the killing effect of the tri-substituted quinazoline derivatives on cancer cells is much stronger than that on normal cells, indicating a wider therapeutic window and potential low toxic side effects, solving the problem of poor selectivity of traditional chemotherapy and some targeted drugs; the control drugs SAHA and A4 have strong activity (IC 50 about 0.5 μmol / L) against HCT116, but their selectivity indexes are very low (SAHA = 2, A4 = 3), and direct comparison proves that the dual-target design of the present application not only does not lag behind single-target drugs in activity, but also makes significant progress in safety (selectivity).

[0373] 5. Taking the preferred tri-substituted quinazoline derivative a10 as an example, in vivo pharmacodynamic evaluation is carried out, and the steps are as follows:

[0374] 1) Select 5-week-old C57BL / 6J male mice weighing 15-20 g, and cultivate them in a pathogen-free environment for about one week for adaptation;

[0375] 2) MC38 cells in the logarithmic growth phase are inoculated subcutaneously into the front armpit of C57BL / 6J mice at a dose of 5 x 10 5 6 per mouse, and when the tumor volume is about 50 mm 3 3) The mice are randomly divided into 6 groups (6 mice per group) when the tumor volume is about 50 mm

[0376] 3) During the administration period, the tumor volume and the body weight of the mice are measured regularly;

[0377] 4) After the experiment, the tumor weight was weighed and anatomical observation was performed.

[0378] Figure 1 The in vivo efficacy evaluation results of the trisubstituted quinazoline derivative a10, wherein, Figure 1 (A) in the above table is the acute toxicity experiment result of a10 within 7 days, Figure 1 (B) in the above table is the long-term toxicity experiment result of a10 within 26 days, Figure 1 (C) in the above table is the tumor tissue weight after a10 administration for 26 days, Figure 1 (D) in the above table is the tumor volume of a10 within 26 days, Figure 1 (E) in the above table is the tumor tissue dissection diagram after a10 administration for 26 days, and statistics *** represents p<0.05. Figure 1 It can be seen that compared with the control group, the tumor volume growth of the a10 (5 mg / kg) treatment group was significantly slowed down, and the inhibition effect was comparable to that of oxaliplatin, proving that a10 has a clear anti-tumor effect in vivo and can successfully convert the activity at the in vitro and cellular level into disease improvement effect in animals; during the entire 26-day administration period, the body weight of the a10-treated mice did not show a significant decrease, indicating that a10 has good tolerance at an effective dose and does not exhibit obvious acute or subacute toxicity, which is consistent with its high selective cellular data; therefore, a10 exhibits excellent characteristics of high efficiency and low toxicity, and is a double-target anti-colorectal cancer candidate compound with good development prospects.

Claims

1. A trisubstituted quinazoline derivative, characterized in that, Its structural formula is shown in formula (I) or formula (II): Formula (I); Equation (II); In equations (I) and (II): R1 is independently selected from nitrogen-containing alkoxy groups, nitrogen-containing alkamide groups, or groups with a spacer group -L- connected through an oxygen atom, wherein the end of the spacer group -L- is a hydroxamic acid group or its bioelectron isosteric structure; R2 is independently selected from H or halogen; R3 is independently selected from H, spacer group -L- connected by an oxygen atom and / or phenyl group, wherein the end of the spacer group -L- is a hydroxamic acid group, an N-hydroxybenzamide group or a hydroxamic acid group bioelectronic isosteric structure, or an amino group substituted with a nitrogen-containing heterocyclic group; Wherein, the spacer group -L- is selected from -(CH2). n -or-(CH2) m -O-(CH2) p -, n=2-6, m=2-4, p=2-4.

2. The trisubstituted quinazoline derivative according to claim 1, characterized in that, In formulas (I) and (II), R1 is independently selected from -O-(CH2). a -T1、-NH-C(O)-(CH2) b -T2、-O-(CH2) c -C(O)N-T3、-O-(CH2) c Any one of -Ph-C(O)N-T3; wherein, T1 is selected from 5-6 member nitrogen-containing heterocyclic groups, a=2-4; T2 is selected from pyrrole or C2-C4 dialkylamino, b=2-4; T3 is selected from hydroxy or aniline, c=3-6; And / or, the R2 is independently selected from H or F; And / or, the R3 is independently selected from H, -Ph-(CH2), etc. d -C(O)N-T4、-Ph-O-(CH2) d -C(O)N-T4、-O-(CH2) d -Ph-C(O)N-T4、-O-(CH2) d -C(O)N-T4、-O-(CH2) e -O-(CH2) e -C(O)N-T4, C4-C6 alkoxy, -O-(CH2) d -T5-Ph-C(O)N-T4、-N-(CH2) f Any one of -T6 and -N-Ph-T6; wherein T4 is selected from hydroxyl, aniline or C4-C6 alkylamino, T5 is selected from 5-6 member nitrogen-containing heterocyclic groups, d=4-6, e=2-4; T6 is selected from pyrrole, morpholino or methylpiperazinyl, f=2-3.

3. The trisubstituted quinazoline derivative according to claim 1, characterized in that, The trisubstituted quinazoline derivatives are shown in Formulas 1-31: 。 4. The method for preparing the trisubstituted quinazoline derivative according to any one of claims 1-3, characterized in that, Includes the following steps: S1. React a 2,4-dihaloquinazoline compound sequentially with an aromatic amine compound and a nitrogen-containing heterocyclic amine compound, or in the reverse order, to obtain a 2-(nitrogen-containing heterocyclic)-4-arylaminoquinazoline core intermediate. S2. The core intermediate is linked to a side chain module with a protected hydroxamic acid group at the end or a precursor functional group that can be converted into hydroxamic acid through a bonding reaction to obtain the linked product. S3. The terminal functional group of the linking product is converted into hydroxamic acid or its protected form, and the protecting group is removed to obtain a free hydroxamic acid group, thereby obtaining the trisubstituted quinazoline derivative. In step S2, the bonding reaction is selected from the Suzuki-Miyaura coupling reaction, nucleophilic substitution reaction, Buchwald-Hartwig carbon-nitrogen coupling reaction, or cycloaddition reaction.

5. The preparation method according to claim 4, characterized in that, In step S2, the bonding reaction is a palladium-catalyzed Suzuki-Miyaura coupling reaction, the functional group at the end of the side chain module is a boric acid or borate ester group, and the reaction site on the core intermediate is a halogen or halogen-like substance.

6. The preparation method according to claim 4, characterized in that, In step S2, the bonding reaction is a nucleophilic substitution reaction, the functional group at the end of the side chain module is a haloalkyl group, and the core intermediate contains a nucleophilic hydroxyl or amino group.

7. The preparation method according to claim 4, characterized in that, In step S2, the bonding reaction is a Buchwald-Hartwig carbon-nitrogen coupling reaction, the functional group at the end of the side chain module is an amino group, and the core intermediate contains an aryl halogen or pseudohalogen that can participate in the coupling reaction.

8. The preparation method according to claim 4, characterized in that, In step S2, the bonding reaction is a copper-catalyzed azido-alkynyl cycloaddition reaction, the functional group at the end of the side chain module is an azide group, and the core intermediate contains an alkynyl group, or the functional group at the end of the side chain module is an alkynyl group, and the core intermediate contains an azide group.

9. Use of the trisubstituted quinazoline derivative or a pharmaceutically acceptable salt thereof as described in any one of claims 1-3 in the preparation of a dual-target medicament for the treatment of colorectal cancer.

10. The application according to claim 9, characterized in that, The dual-target drug is a G-quadruplex stabilizer and a histone deacetylase inhibitor.

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