CN107522700B

CN107522700B - Quinazolinone and α -unsaturated ketone conjugated derivative and preparation method and application thereof

Info

Publication number: CN107522700B
Application number: CN201710658952.6A
Authority: CN
Inventors: 黄志纾; 谭嘉恒; 王玉青; 王晨曦
Original assignee: National Sun Yat Sen University
Current assignee: National Sun Yat Sen University
Priority date: 2017-08-03
Filing date: 2017-08-03
Publication date: 2020-03-20
Anticipated expiration: 2037-08-03
Also published as: CN107522700A

Abstract

The invention discloses a quinazolinone and α -unsaturated ketone conjugated derivative, wherein the structural formula of the quinazolinone and α -unsaturated ketone conjugated derivative is shown as a formula (I) or a formula (II), wherein R is₁Is hydrogen, amino, substituted amino, five-membered or six-membered heterocyclic group; r₂Is hydrogen, phenyl, substituted phenyl, benzo heterocyclic compound; the substituents being selected from C_1‑5Alkyl radical, C_1‑8The quinazolinone and α -unsaturated ketone conjugated derivative provided by the invention can effectively inhibit the growth of various tumor cells, can obviously inhibit the transcription and expression of protooncogene c-MYC, and has wide application space in preparing antitumor drugs.

Description

Quinazolinone and α -unsaturated ketone conjugated derivative and preparation method and application thereof

Technical Field

The invention belongs to the technical field of medicinal chemistry, and particularly relates to a quinazolinone and α -unsaturated ketone conjugated derivative, and a preparation method and application thereof.

Background

Malignant tumors are a large group of diseases that endanger human health, and according to the World Health Organization (WHO), 800 million people die from cancer every year worldwide, while nearly 200 million people die from cancer every year in china. In recent years, the research and development of anti-malignant tumor drugs have made a great breakthrough, and many new drugs enter clinical research, but still have many challenges. The problems of high toxic and side effects, drug resistance and the like are still main obstacles for clinical tumor drug treatment.

The c-MYC is used as an important protooncogene and regulates the life processes of cell proliferation, differentiation, metabolism, apoptosis and the like. The research shows that c-MYC has abnormal expression in various malignant tumors, such as cervical cancer, breast cancer, colon cancer, small cell lung cancer, malignant glioma, melanoma, osteosarcoma, myelogenous leukemia and the like. High-expression c-MYC in most tumor cells benefits from transcriptional activation of genes, and a C-MYC promoter nucleic acid sensitive element NHE III sequence can form a G-quadruplex structure and can be used as a transcriptional silencer to inhibit transcription of the c-MYC. Therefore, the development of the ligand targeting c-MYCG-quadruplex becomes a hotspot for the research and development of the antitumor drugs.

Quinazoline groups have been extensively studied in the field of medicinal chemistry. Quinazoline alkaloids exist in various plant and animal kingdoms, and quinazoline derivatives have various biological activities, including functions of resisting tumors, bacteria and inflammation, reducing blood pressure and the like. Over the past 15 years, significant progress has been made in the anti-tumor field of quinazoline derivatives, and a large number of quinazoline compounds have been reported, several of which are approved by the FDA for marketing.

Chalcone compounds are widely existed in nature, contain α -unsaturated ketone and aromatic ring in the structure, have hydrogen bond receptor, electrophilicity and lipophilic property, show appropriate flexibility in the whole molecule, can be combined with different receptors, and show various biological activities.

Disclosure of Invention

One of the objectives of the present invention is to provide a quinazolinone and α -unsaturated ketone conjugated derivative.

Another object of the present invention is to provide a process for producing the above derivatives.

The invention also aims to provide application of the derivative in preparing an anti-cancer medicament as a protooncogene c-MYC transcription regulation and control blocker.

The invention is conjugated with α -unsaturated ketone structure on the basis of quinazolinone structure to obtain novel quinazolinone and α -unsaturated ketone conjugated derivatives, and the derivatives have antitumor effect of inhibiting transcription and translation of protooncogene c-myc.

Furthermore, the quinazoline ketone and α -unsaturated ketone conjugated derivative provided by the invention can inhibit the transcription and translation of protooncogene c-myc, inhibit the proliferation of multiple tumor cells and have a wide anti-tumor effect.

The invention realizes the aim through the following technical scheme:

the invention provides a quinazolinone and α -unsaturated ketone conjugated derivative, wherein the structural formula of the quinazolinone and α -unsaturated ketone conjugated derivative is shown as a formula (I) or a formula (II):

wherein R is₁Is hydrogen, amino, substituted amino, five-membered or six-membered heterocyclic group; r₂Is hydrogen, phenyl, substituted phenyl, benzo heterocyclic compound;

the substituents being selected from C_1-5Alkyl radical, C_1-8Alkoxy, halogen radical, phenyl, amino, five-membered or six-membered heterocyclic radical.

Preferably, R₁Is NH (CH)₂)_nR₃，R₃Is amino, substituted amino, five-membered or six-membered heterocyclic group, n is any integer of 1-5, and the substituent in the substituted amino is selected from C_1-5Alkyl radical, C_1-8Alkoxy, five-membered or six-membered heterocyclic group.

Preferably, R₃Is amino, N-diethylmethylamino, N-dimethylmethylamino or morpholinyl.

Preferably, R₂Is phenyl, substituted phenyl, benzo five-membered or benzo six-membered heterocyclic compound, and the substituent in the substituted phenyl is selected from halogen and C_1-5Alkoxy, five-membered or six-membered nitrogen heterocyclic group.

Preferably, R₂Is phenyl, substituted phenyl or 1, 3-benzodioxole, and the substituent in the substituted phenyl is halogeno, morpholinyl, C_1-5Alkyl or C_1-5An alkoxy group.

Preferably, the substituents in the substituted phenyl group are fluoro, morpholinyl, tert-butyl or methoxy.

The invention also provides a preparation method of the quinazolinone and α -unsaturated ketone conjugated derivative, which comprises the following steps:

s1, 2-amino-4, 5-difluorobenzoic acid reacts with 2-pyrrolidone to obtain a compound

S2. Compound

With amine compounds NH-R₁Reacting to obtain an intermediate

S3. Compound

Reacting with ketone compound to obtain intermediate

And S4, reacting the compound obtained in the S2 with the compound obtained in the S3 to obtain the derivative.

The invention also provides application of the quinazolinone and α -unsaturated ketone conjugated derivative in preparation of anti-cancer drugs.

Furthermore, the quinazoline ketone and α -unsaturated ketone conjugated derivative is used as a proto-oncogene c-MYC transcription regulation and control blocking agent to be applied to preparation of anti-cancer drugs.

Further, the anticancer drug is one or more of anti-ovarian cancer, cervical cancer, breast cancer, lung adenocarcinoma, colon cancer, liver cancer, leukemia, small cell lung cancer, skin cancer, epithelial cell cancer, prostate cancer, non-small cell lung cancer, nasopharyngeal carcinoma, glioblastoma, lymphoma or melanoma.

Further, the drug of the present invention also includes a pharmaceutically acceptable salt, carrier, stereoisomer or prodrug molecule thereof.

Further, the medicament of the present invention may be an injection, a tablet, a pill, a capsule, a suspension or an emulsion.

The derivative provided by the invention is applied to cancer cell inhibition experiments, and is found to have a remarkable inhibition effect, particularly a remarkable inhibition effect on cancer cell strain proliferation of the compounds 3a, 3c, 3h and 3 i.

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

the invention relates to a quinazolinone and α -unsaturated ketone conjugated derivative with structural characteristics of general formula (I) or (II), which can effectively inhibit the growth of various tumor cells, can obviously inhibit the transcription and expression of protooncogene c-MYC, and has wide application space in preparing antitumor drugs.

Drawings

FIG. 1 is a graph showing the effect of quinazolinone and α -unsaturated ketone conjugate derivatives on c-MYC transcript levels.

FIG. 2 is a graph showing the effect of quinazolinone conjugated derivatives with α -unsaturated ketone on the translation level of C-MYC according to the present invention.

Detailed Description

The present invention will be described in further detail with reference to the following specific embodiments and the accompanying drawings. Reagents, equipment and methods employed in the present invention are reagents, equipment and methods conventionally commercially available in the art and conventionally used methods, unless otherwise specified.

The synthetic route of the derivatives of the embodiments 14-33 of the invention is as follows:

more specifically, the synthesis routes of the derivatives described in embodiments 14 to 33 of the present invention are as follows:

reacting 2-amino-4, 5-difluorobenzoic acid with 2-pyrrolidone to obtain a compound 2

Reaction of compound 2 with different amines gives intermediate 3

And 4

Compound (I)

Reacting with ketone compound to obtain intermediate a

b

c

d

e

f

g

h

i

And j

And dissolving the intermediates 3 and 4 and the intermediates a, b, c, d, e, f, g, h, i and j in DMF, and performing substitution reaction under the action of trimethylchlorosilane to obtain the final compound. In the presence of an intermediate

The preparation method of the intermediates a, b, c, d, e, f, g, h, i and j comprises the steps of reacting p-benzaldehyde or m-benzaldehyde with acetophenone raw materials under the action of NaOH as a catalyst and ethanol and water as reaction solvents, wherein the molar ratio of the reaction of the p-benzaldehyde or m-benzaldehyde with the acetophenone raw materials is 1: 0.5-1, placing the reaction in an ice bath, stirring and reacting for 2 hours, and then stirring and reacting for 2-6 hours at room temperature.

Further, the final product is prepared by the method that the molar ratio of the intermediate 3 and the intermediate 4 in the step of S4 to the substituted benzaldehyde intermediate a, b, c, d, e, f, g, h, i and j is 1: (1.05-3.0), the reaction temperature is 50-120 ℃, and the reaction time is 12-48 hours.

Example 1: synthesis of Compound 2

2-amino-4, 5-difluorobenzoic acid (5g, 29mmol), 2-pyrrolidone (5mL) was placed in a 100mL single-necked flask, and 20mL of phosphorus oxychloride (POCl) was added dropwise with stirring in an ice bath₃) After 30min, heating and refluxing at 103 ℃ for 24h, dropwise adding the reaction system into 200mL of ice water, adjusting the pH to be alkalescent by using concentrated NaOH solution, performing suction filtration to obtain a crude product, and purifying by silica gel column chromatography (eluent: v (petroleum ether): v (ethyl acetate) ═ 1:1) gave compound m1, 5.4g, 85% yield.

¹H NMR(400MHz,CDCl₃)δ8.20–7.85(m,1H),7.60–7.34(m,1H),4.42–4.06(m,2H),3.19(td,J＝7.9,2.7Hz,2H),2.43–2.16(m,2H).LC-MS m/z:223[M+H]^+.

Example 2: synthesis of Compound 3

Taking the compound 2(2.22g, 10mmol) and diethylaminopropylamine (3ml, 24mmol), heating and stirring overnight at 100 ℃ in a thick-wall pressure-resistant bottle to obtain an orange yellow solution, cooling a reaction system to room temperature, separating out a large amount of light yellow precipitate, performing suction filtration to obtain a filter cake which is a milky white solid, and performing vacuum drying to obtain 2.3g of the milky white solid with the yield of 69%.

¹H NMR(400MHz,CDCl₃)δ7.72(d,J＝11.7Hz,1H),6.72(s,1H),6.67(d,J＝7.7Hz,1H),4.19–4.12(m,2H),3.31(dd,J＝10.6,5.8Hz,2H),3.11(t,J＝7.9Hz,2H),2.63–2.59(m,2H),2.55(q,J＝7.1Hz,4H),2.30–2.19(m,2H),1.89–1.81(m,2H),1.06(t,J＝7.1Hz,6H).

Example 3: synthesis of Compound 4

According to the synthesis of 3, 1.8g of a milky white solid was obtained with a yield of 52%.¹H NMR(400MHz,CDCl₃)δ7.67(d,J＝11.7Hz,1H),6.62(d,J＝7.7Hz,1H),4.13–4.05(m,2H),3.72–3.67(m,2H),3.66–3.61(m,2H),3.25(dd,J＝11.1,5.8Hz,2H),3.04(t,J＝7.9Hz,4H),2.51–2.46(m,3H),2.24–2.12(m,4H),1.86–1.75(m,2H).

Example 4: synthesis of Compound a

P-benzaldehyde (1.34g, 10mol), 3 ', 4' -methylenedioxyacetophenone (1.64g, 10mol) and a round-bottom flask are weighed, 3mL of ethanol, 1mL of water and 0.01g of NaOH are added as catalysts, the mixture is placed in an ice bath and stirred for reaction for 2 hours, and then the mixture is stirred for reaction for 2 hours at room temperature. After TLC monitoring reaction, suction filtration is carried out, filter cakes are taken out and separated and purified by silica gel column chromatography, and light yellow solid 1.21g is obtained with the yield of 43 percent.

¹H NMR(400MHz,DMSO)δ10.05(s,1H),8.12(d,J＝8.1Hz,2H),8.06(d,J＝11.5Hz,1H),7.97(d,J＝6.3Hz,2H),7.91(dd,J＝8.2,1.8Hz,1H),7.77(d,J＝9.3Hz,1H),7.69(s,1H),7.11(d,J＝8.2Hz,1H),6.18(s,2H).

Example 5: synthesis of Compound b

According to the synthesis of a, 1.78g of a yellow solid are obtained, with a yield of 55%.

¹H NMR(400MHz,DMSO)δ10.05(s,1H),8.10(d,J＝7.5Hz,4H),8.07(s,1H),7.97(d,J＝8.1Hz,2H),7.72(d,J＝15.6Hz,1H),7.05(d,J＝8.9Hz,2H),3.77–3.72(m,4H),3.38–3.33(m,4H).

Example 6: synthesis of Compound c

According to the synthesis of a, 1.03g of a milky white solid was obtained with a yield of 35%.

¹H NMR(400MHz,DMSO)δ9.01(s,3H),8.12(d,J＝8.5Hz,1H),8.02(d,J＝15.7Hz,2H),7.99(m,2H),7.76(d,J＝15.6Hz,1H),7.60(d,J＝8.5Hz,1H),1.34(s,9H).

Example 7: synthesis of Compound d

According to the synthesis of a, 2.11g of a yellow solid are obtained with a yield of 83%.

¹H NMR(400MHz,CDCl₃)δ10.08(s,1H),8.10(ddd,J＝5.4,4.5,2.1Hz,2H),7.96(d,J＝8.2Hz,2H),7.83(d,J＝2.5Hz,2H),7.81(s,1H),7.68(d,J＝26.2Hz,1H),7.25–7.18(m,2H).

Example 8: synthesis of Compound e

According to the synthesis of a, 1.64g of a yellow solid are obtained, with a yield of 62%.

¹H NMR(400MHz,CDCl₃)δ10.05(s,1H),8.06(d,J＝8.8Hz,2H),7.93(d,J＝8.1Hz,2H),7.83(d,J＝3.6Hz,1H),7.79(d,J＝7.8Hz,2H),7.65(d,J＝15.7Hz,1H),7.00(d,J＝8.8Hz,2H),3.90(s,3H).

Example 9: synthesis of Compound f

M-benzaldehyde (1.34g, 10mol), 3 ', 4' -methylenedioxyacetophenone (1.64g, 10mol) and a round-bottom flask are weighed, 3mL of ethanol, 1mL of water and 0.01g of NaOH are added as catalysts, the mixture is placed in an ice bath and stirred for reaction for 2 hours, and then the mixture is stirred for reaction for 2 hours at room temperature. After TLC monitoring reaction, suction filtration is carried out, filter cakes are taken out and separated and purified by silica gel column chromatography, and light yellow solid 1.01g is obtained with the yield of 36 percent.

¹H NMR(400MHz,CDCl₃)δ10.08(s,1H),8.15(s,1H),7.91(d,J＝7.6Hz,1H),7.86(d,J＝8.6Hz,1H),7.83(d,J＝15.9Hz,1H),7.68(dd,J＝8.1,1.4Hz,1H),7.63–7.53(m,3H),6.92(d,J＝8.1Hz,1H),6.08(s,2H).

Example 10: synthesis of Compound g

Synthesis according to f, 1.22g of a yellow solid are obtained in 38% yield.

¹H NMR(400MHz,CDCl₃)δ10.10(s,1H),8.18(d,J＝5.5Hz,1H),8.11–8.01(m,2H),7.96–7.79(m,3H),7.73–7.58(m,2H),7.02–6.90(m,2H),3.95–3.85(m,4H),3.42–3.32(m,4H).

Example 11: synthesis of Compound h

According to the synthesis of f, 1.20g of a milky white solid was obtained with a yield of 41%.

¹H NMR(400MHz,CDCl₃)δ10.08(s,1H),8.17(s,1H),8.00(d,J＝8.4Hz,2H),7.94–7.89(m,2H),7.88(d,J＝7.4Hz,1H),7.62(d,J＝5.0Hz,1H),7.54(d,J＝8.3Hz,2H),7.48(dd,J＝8.5,2.3Hz,1H),1.37(s,9H).

Example 12: synthesis of Compound i

Synthesis according to f, 1.68g of a yellow solid are obtained in 66% yield.

¹H NMR(400MHz,CDCl₃)δ10.09(s,1H),8.12–8.05(m,3H),7.86(d,J＝5.6Hz,1H),7.82(s,1H),7.70(dd,J＝7.7,1.5Hz,1H),7.56(d,J＝15.7Hz,1H),7.50(t,J＝7.7Hz,1H),7.23–7.17(m,2H).

Example 13: synthesis of Compound j

Synthesis according to f, 1.47g of a yellow solid are obtained in 55% yield.

¹H NMR(400MHz,CDCl₃)δ10.08(s,1H),8.16(s,1H),8.06(d,J＝8.8Hz,2H),7.89(dd,J＝16.9,7.2Hz,2H),7.85–7.79(m,1H),7.67(d,J＝8.5Hz,1H),7.63–7.60(m,1H),7.00(d,J＝8.6Hz,2H),3.91(s,3H).

Example 14: synthesis of Compound 3a

Adding the compound 3(0.3323g, 1mmol) and the compound a (0.3083g, 1.1mmol) into a rear-wall pressure-resistant bottle, adding 2mL of DMSO and 10mL of trimethylchlorosilane, heating at 100 ℃ overnight, after TLC monitoring reaction, performing suction filtration, and recrystallizing a crude product with methanol/dichloromethane to obtain 0.4180g of light yellow solid with the yield of 70%.

¹H NMR(400MHz,DMSO-d₆)δ8.03(d,J＝6.8Hz,1H),8.02–7.97(m,J＝11.4,5.0Hz,3H),7.92–7.87(m,J＝8.2,3.8,1.7Hz,1H),7.76(d,J＝4.8Hz,1H),7.73–7.71(m,1H),7.70–7.68(m,J＝4.9,2.4Hz,2H),7.63(d,J＝11.8Hz,1H),7.11(d,J＝8.2Hz,1H),6.93(d,J＝7.9Hz,1H),6.18(s,2H),4.20–4.12(m,2H),3.40–3.28(m,4H),3.20–3.06(m,J＝11.9,5.9Hz,6H),2.09–1.95(m,2H),1.22(t,J＝7.2Hz,6H).¹³C NMR(101MHz,DMSO-d₆))δ187.46,159.73,δ153.12(d,J＝194.2Hz),152.07,149.29,148.91,148.52,143.26(d,J＝13.7Hz),142.94,137.82,135.45,135.05,132.84,130.42,129.72,127.98,125.54,123.26,109.54(d,J＝20.6Hz),109.11(d,J＝7.5Hz),108.60,108.41,106.01(d,J＝4.1Hz),102.54,51.09,46.96,44.33,41.72,29.42,25.79,24.96,11.55.ESI-HRMS[M+H]⁺m/z＝595.2642,calcd for C35H35FN4O4,595.2699.Purity:99.6％by HPLC.

Example 15: synthesis of Compound 3b

Compound 3(0.3323g, 1mmol) and compound b (0.3535g, 1.1mmol) were added to a back wall pressure bottle, 2mL DMSO and 10mL trimethylchlorosilane were added, heating was carried out at 100 ℃ overnight, after TLC monitoring reaction, suction filtration was carried out, and the crude product was recrystallized from methanol/dichloromethane to give 0.2961g as a pale yellow solid with a yield of 46%.

¹H NMR(400MHz,CDCl₃)δ8.02(d,J＝8.7Hz,2H),7.84–7.75(m,3H),7.71(d,J＝8.1Hz,2H),7.64–7.56(m,3H),6.93(d,J＝8.8Hz,2H),6.79(d,J＝7.6Hz,1H),4.28(t,J＝7.0Hz,2H),3.91–3.84(m,4H),3.65(d,J＝4.5Hz,1H),3.41–3.27(m,8H),2.69–2.50(m,6H),1.94–1.81(m,2H),1.08(t,J＝6.7Hz,6H).¹³C NMR(101MHz,CDCl₃)δ187.76,160.42,154.96,154.26,150.87(d,J＝246.3Hz),148.71,143.46(d,J＝13.8Hz),142.16,137.38,135.54,133.32,130.66,130.12,128.79,128.69,122.67,113.40,109.74(d,J＝20.8Hz),109.42(d,J＝7.6Hz),105.55(d,J＝4.2Hz),70.59,66.57,52.72,47.49,46.85,43.78,25.81,25.12,11.59.

ESI-HRMS[M+H]⁺m/z＝636.3272,calcd for C38H42FN5O3,636.3321.Purity:97.5％by HPLC.

Example 16: synthesis of Compound 3c

Adding the compound 3(0.3323g, 1mmol) and the compound c (0.3216g, 1.1mmol) into a back-wall pressure-resistant bottle, adding 2mL of DMSO and 10mL of trimethylchlorosilane, heating at 100 ℃ overnight, after TLC monitoring reaction, performing suction filtration, and recrystallizing a crude product with methanol/dichloromethane to obtain 0.3050g of light yellow solid with the yield of 50%.

¹H NMR(400MHz,CDCl₃)δ7.98(d,J＝8.0Hz,1H),7.81(d,J＝16.4Hz,1H),7.77–7.68(m,5H),7.59(d,J＝8.6Hz,2H),7.53(d,J＝7.6Hz,3H),6.78(d,J＝7.5Hz,1H),6.65(s,1H),4.30(t,J＝6.7Hz,2H),3.41–3.34(m,2H),3.33–3.24(m,2H),2.73–2.55(m,6H),1.96–1.85(m,2H),1.37(s,9H),1.10(t,J＝6.6Hz,6H).

¹³C NMR(101MHz,CDCl₃)δ188.75,159.39,156.36(t,J＝273.9Hz),153.94,152.63,151.06,148.62,147.65,142.36(d,J＝14.1Hz),142.20,136.56,134.46,134.22,132.46,131.30,129.88,129.13,127.81,127.79,127.72,127.49,124.63,121.79,108.76(d,J＝21.0Hz),108.47(d,J＝6.7Hz),104.53(d,J＝4.1Hz),64.54,51.46,45.80,42.89,34.14,30.10,28.68,24.79,12.70.

ESI-HRMS[M+H]⁺m/z＝607.3370,calcd for C38H43FN4O2,607.3430.Purity:96.2％by HPLC.

Example 17: synthesis of Compound 3d

Compound 3(0.3323g, 1mmol) and compound d (0.2797g, 1.1mmol) were added to a back wall pressure resistant bottle, 2mL DMSO, 10mL trimethylchlorosilane were added, heating was carried out at 100 ℃ overnight, after TLC monitoring reaction was completed, suction filtration was carried out, and the crude product was recrystallized from methanol/dichloromethane to give 0.2507g as a pale yellow solid with a yield of 44%.

¹H NMR(400MHz,CDCl₃)δ8.03–7.97(m,2H),7.71(m,3H),7.63(d,J＝8.3Hz,2H),7.52(d,J＝8.3Hz,2H),7.47(d,J＝15.6Hz,1H),7.12(t,J＝8.6Hz,2H),6.70(d,J＝7.7Hz,1H),4.24–4.14(m,2H),3.29(dd,J＝10.6,5.6Hz,2H),3.25–3.18(m,2H),2.59(t,J＝5.9Hz,2H),2.53(q,J＝7.0Hz,4H),1.84(dd,J＝11.7,5.8Hz,2H),1.02(t,J＝7.1Hz,6H).¹³C NMR(101MHz,CDCl₃)δ187.50,165.94,163.40,159.40,153.87,149.86(d,J＝245.0Hz),147.64,142.86,142.41(d,J＝14.0Hz),136.84,133.92,133.44,132.66,130.13,130.04,129.14,127.85,127.58,121.16,114.91,114.69,108.74(d,J＝20.7Hz),108.42(d,J＝7.7Hz),104.51(d,J＝3.7Hz),51.60,45.81,42.88,42.48,24.80,23.96,10.44.

ESI-HRMS[M+H]⁺m/z＝569.2650,calcd for C34H34F2N4O2,569.2722.Purity:98.2％by HPLC.

Example 18: synthesis of Compound 3e

Compound 3(0.3323g, 1mmol) and compound e (0.2930g, 1.1mmol) were added to a back wall pressure bottle, 2mL DMSO and 10mL trimethylchlorosilane were added, heating was carried out at 100 ℃ overnight, after TLC monitoring reaction, suction filtration was carried out, and the crude product was recrystallized from methanol/dichloromethane to give 0.3011g as a pale yellow solid with a yield of 52%.

¹H NMR(400MHz,CDCl₃)δ8.05(d,J＝8.7Hz,2H),7.79(t,J＝14.6Hz,3H),7.71(d,J＝7.9Hz,2H),7.64–7.55(m,3H),7.00(d,J＝8.6Hz,2H),6.78(d,J＝7.6Hz,1H),4.27(t,J＝6.8Hz,2H),3.90(s,3H),3.36(m,2H),3.34–3.27(m,2H),2.70–2.53(m,J＝21.5Hz,6H),1.94–1.83(m,2H),1.09(t,J＝6.5Hz,6H).

¹³C NMR(101MHz,CDCl₃)δ188.28,163.53,160.37(d,J＝3.3Hz),153.47(d,J＝285.4Hz),149.62,148.66,143.45(d,J＝14.0Hz),142.77,137.50,135.22,133.44,130.94,130.82,130.12,128.74,128.58,122.38,113.89,109.66(d,J＝20.8Hz),109.32(d,J＝7.7Hz),105.46(d,J＝4.2Hz),55.51,52.86,46.83,43.89,43.76,25.78,25.05,11.67.

ESI-HRMS[M+H]⁺m/z＝581.2850,calcd for C35H37FN4O3,581.2900.Purity:99.1％by HPLC.

Example 19: synthesis of Compound 3f

Adding the compound 3(0.3323g, 1mmol) and the compound f (0.3083g, 1.1mmol) into a rear-wall pressure-resistant bottle, adding 2mL of DMSO and 10mL of trimethylchlorosilane, heating at 100 ℃ overnight, after TLC monitoring reaction, performing suction filtration, and recrystallizing a crude product with methanol/dichloromethane to obtain 0.2106g of light yellow solid with the yield of 35%.

¹H NMR(400MHz,CDCl₃)δ7.82–7.77(m,1H),7.76(d,J＝3.4Hz,1H),7.73(d,J＝2.5Hz,1H),7.66(dd,J＝8.2,1.6Hz,1H),7.62(d,J＝7.5Hz,1H),7.57(d,J＝7.3Hz,1H),7.55–7.53(m,1H),7.51–7.44(m,2H),6.91(dd,J＝8.1,1.5Hz,1H),6.77(d,J＝7.7Hz,2H),6.07(d,J＝1.2Hz,2H),4.25(dd,J＝9.7,4.5Hz,2H),3.38–3.32(m,2H),3.28(dd,J＝8.6,5.0Hz,2H),2.63(t,J＝5.7Hz,2H),2.57(q,J＝7.0Hz,4H),1.91–1.83(m,2H),1.07(t,J＝7.1Hz,6H).¹³C NMR(101MHz,CDCl₃)δ187.99,160.42,154.94,152.10–149.45(d,J＝187.6Hz),149.61,148.40,143.45,143.31(d,J＝13.1Hz),136.44,135.66,133.16,132.83,131.00,129.88,129.42,128.85,128.04,124.77,122.50,109.82(d,J＝20.8Hz),109.57(d,J＝7.6Hz),108.44,107.97,105.61(d,J＝4.3Hz),101.94,52.18,46.75,43.91,43.06,29.70,25.66,24.84,11.10.

ESI-HRMS[M+H]⁺m/z＝595.2642,calcd for C35H35FN4O4,595.2711.Purity:98.1％by HPLC.

Example 20: synthesis of Compound 3g

Adding compound 3(0.3323g, 1mmol) and compound g (0.3535g, 1.1mmol) into a back-wall pressure-resistant bottle, adding 2mL of DMSO and 10mL of trimethylchlorosilane, heating at 100 ℃ overnight, after TLC monitoring reaction, performing suction filtration, and recrystallizing the crude product with methanol/dichloromethane to obtain 0.2360g of light yellow solid with the yield of 37%.

¹H NMR(400MHz,CDCl₃)δ8.03(d,J＝9.0Hz,2H),7.85–7.76(m,4H),7.65(d,J＝7.7Hz,1H),7.63–7.57(m,2H),7.50(t,J＝7.7Hz,1H),6.94(d,J＝9.0Hz,2H),6.81(d,J＝7.7Hz,1H),4.31–4.25(m,2H),3.90–3.86(m,4H),3.65(d,J＝4.5Hz,1H),3.42(t,J＝5.4Hz,2H),3.38–3.34(m,4H),3.34–3.29(m,2H),2.83–2.68(m,6H),2.06–1.95(m,2H),1.17(t,J＝6.7Hz,6H).¹³C NMR(101MHz,CDCl₃)δ187.58,160.34,154.86,154.22,150.70(d,J＝244.5Hz),148.58,148.16,143.30(d,J＝14.2Hz),142.42,136.20,135.75,132.96,130.80,130.64,129.81,129.31,128.68,128.39,128.00,122.37,113.30,109.60(d,J＝20.5Hz),109.28(d,J＝7.6Hz),105.26(d,J＝35.6Hz),66.51,52.53,47.32,46.74,43.90,43.42,25.56,24.85,11.41.

ESI-HRMS[M+H]⁺m/z＝636.3272,calcd for C38H42FN5O3,636.3317.Purity:98.1％by HPLC.

Example 21: synthesis of Compound 3h

Compound 3(0.3323g, 1mmol) and compound h (0.3216g, 1.1mmol) were added to a back wall pressure resistant bottle, 2mL DMSO, 10mL trimethylchlorosilane were added, heated at 100 ℃ overnight, after TLC monitoring reaction, suction filtration was performed, and the crude product was recrystallized from methanol/dichloromethane to give 0.1744g as a pale yellow solid in 29% yield.

¹H NMR(400MHz,CDCl₃)δ8.12–8.03(m,2H),7.83(d,J＝15.7Hz,1H),7.80–7.72(m,3H),7.64(d,J＝7.4Hz,1H),7.60(d,J＝7.5Hz,1H),7.57–7.46(m,2H),7.20(t,J＝8.3Hz,2H),6.85(s,1H),6.77(d,J＝7.6Hz,1H),4.27(t,J＝6.8Hz,2H),3.39–3.25(m,4H),2.62(t,J＝5.3Hz,2H),2.57(q,J＝7.2Hz,4H),1.93–1.81(m,2H),1.25(s,9H),1.07(t,J＝7.1Hz,6H).¹³C NMR(101MHz,CDCl₃)δ187.56,166.14,163.61,159.54,156.73(d,J＝558.3Hz),151.18,148.75,147.79,143.37,142.62(d,J＝14.1Hz),135.57,134.50,133.50,133.47,132.42,130.37,130.27,129.18,128.61,127.63,127.23,121.25,115.12,114.90,108.80(d,J＝20.8Hz),108.46(d,J＝7.6Hz),104.60(d,J＝4.1Hz),52.00,50.24,45.97,42.91,34.41,30.93,28.47,24.80,10.82.

ESI-HRMS[M+H]⁺m/z＝607.3370,calcd for C38H43FN4O2,607.3462.Purity:98.6％by HPLC.

Example 22: synthesis of Compound 3i

Adding the compound 3(0.3323g, 1mmol) and the compound i (0.2797g, 1.1mmol) into a back-wall pressure-resistant bottle, adding 2mL of DMSO and 10mL of trimethylchlorosilane, heating at 100 ℃ overnight, after TLC monitoring reaction, performing suction filtration, and recrystallizing the crude product with methanol/dichloromethane to obtain 0.3622g of light yellow solid with the yield of 64%.

¹H NMR(400MHz,CDCl₃)δ8.12–8.04(m,2H),7.83(d,J＝15.7Hz,1H),7.80–7.72(m,3H),7.65(d,J＝7.6Hz,1H),7.61(d,J＝7.7Hz,1H),7.57–7.47(m,2H),7.20(t,J＝8.3Hz,2H),6.85(s,1H),6.78(d,J＝7.6Hz,1H),4.27(t,J＝7.0Hz,2H),3.39–3.26(m,4H),2.63(t,J＝5.4Hz,2H),2.56(q,J＝7.0Hz,4H),1.93–1.83(m,2H),1.07(t,J＝7.0Hz,6H).¹³CNMR(101MHz,CDCl₃)δ188.36,166.93,164.40,160.33,160.30,154.75,150.77(d,J＝244.5Hz),148.58,144.17,143.41(d,J＝14.1Hz),136.37,135.30,134.30,134.27,133.21,131.16,131.07,129.97,129.41,128.43,128.03,122.05,115.91,115.70,109.59(d,J＝20.8Hz),109.26(d,J＝7.6Hz),105.39(d,J＝4.1Hz),52.79,46.77,43.86,43.71,29.77,25.59,25.00,11.62.

ESI-HRMS[M+H]⁺m/z＝569.2650,calcd for C34H34F2N4O2,569.2705.Purity:97.9％by HPLC.

Example 23: synthesis of Compound 3j

Adding the compound 3(0.3323g, 1mmol) and the compound j (0.2930g, 1.1mmol) into a back-wall pressure-resistant bottle, adding 2mL of DMSO and 10mL of trimethylchlorosilane, heating at 100 ℃ overnight, after TLC monitoring reaction, performing suction filtration, and recrystallizing the crude product with methanol/dichloromethane to obtain 0.3140g of light yellow solid with the yield of 54%.

¹H NMR(400MHz,DMSO-d6)δ8.15(d,J＝8.5Hz,2H),8.03(s,1H),7.96–7.86(m,3H),7.75(d,J＝15.6Hz,1H),7.68(d,J＝7.8Hz,1H),7.61(d,J＝11.8Hz,1H),7.57(t,J＝7.7Hz,1H),7.09(d,J＝8.5Hz,2H),7.01(d,J＝7.3Hz,1H),4.17(t,J＝6.9Hz,2H),3.88(s,3H),3.41–3.32(m,4H),3.19–3.06(m,6H),2.13–2.03(m,2H),1.26(t,J＝7.2Hz,6H).¹³CNMR(101MHz,DMSO-d6)δ188.07,163.85,159.34(d,J＝3.1Hz),153.91(d,J＝427.6Hz),149.36,147.15,146.21,143.17(d,J＝13.9Hz),142.91,136.42,136.13,133.83,131.48,131.36,131.10,130.46,129.93,129.26,123.78,114.59,109.94(d,J＝20.8Hz),109.15(d,J＝7.4Hz),105.18(d,J＝3.9Hz),56.11,49.43,46.96,44.87,34.64,25.61,22.90,8.95.

ESI-HRMS[M+H]⁺m/z＝581.2850,calcd for C35H37FN4O3,581.2904.Purity:97.1％by HPLC.

Example 24: synthesis of Compound 4a

Adding the compound 4(0.3462g, 1mmol) and the compound a (0.3083g, 1.1mmol) into a rear-wall pressure-resistant bottle, adding 2mL of DMSO and 10mL of trimethylchlorosilane, heating at 100 ℃ overnight, after TLC monitoring reaction, performing suction filtration, and recrystallizing a crude product with methanol/dichloromethane to obtain 0.2511g of light yellow solid with the yield of 41%.

¹H NMR(400MHz,CDCl₃)δ7.82(d,J＝5.7Hz,1H),7.80–7.77(m,2H),7.71(d,J＝8.3Hz,2H),7.67(dd,J＝8.1,1.6Hz,1H),7.61(d,J＝8.3Hz,2H),7.55(dd,J＝8.6,7.0Hz,2H),6.92(d,J＝8.1Hz,1H),6.82(d,J＝7.7Hz,1H),6.08(s,2H),4.32–4.26(m,2H),3.80(s,4H),3.39(d,J＝3.9Hz,2H),3.36–3.29(m,2H),2.56(d,J＝20.8Hz,6H),1.92(s,2H).¹³CNMR(101MHz,CDCl₃)δ187.94,160.43(d,J＝3.5Hz),155.04,152.06,150.85(d,J＝244.1Hz),148.68,148.39,143.36(d,J＝13.9Hz),143.08,137.57,135.25,133.44,132.88,130.16,128.80,124.74,122.38,109.83(d,J＝20.9Hz),109.59(d,J＝7.7Hz),108.43,107.97,105.63(d,J＝3.9Hz),101.94,66.89,58.10,53.78,43.93,43.45,25.81,24.02.

ESI-HRMS[M+H]⁺m/z＝608.2435,calcd for C35H33FN4O5,609.2511.Purity:98.9％by HPLC.

Example 25: synthesis of Compound 4b

Adding the compound 4(0.3462g, 1mmol) and the compound b (0.3535g, 1.1mmol) into a back-wall pressure-resistant bottle, adding 2mL of DMSO and 10mL of trimethylchlorosilane, heating at 100 ℃ overnight, after TLC monitoring reaction, performing suction filtration, and recrystallizing a crude product with methanol/dichloromethane to obtain 0.3100g of light yellow solid with the yield of 48%.

¹H NMR(400MHz,CDCl₃)δ8.02(d,J＝8.8Hz,2H),7.80(dd,J＝11.5,4.2Hz,3H),7.71(d,J＝8.2Hz,2H),7.64–7.58(m,3H),6.93(d,J＝8.7Hz,2H),6.82(d,J＝7.7Hz,1H),6.50(s,1H),4.28(t,J＝7.2Hz,2H),3.91–3.84(m,4H),3.79(s,4H),3.41–3.28(m,8H),2.55(d,J＝22.9Hz,6H),1.91(s,2H).¹³C NMR(101MHz,CDCl₃)δ187.74,160.42(d,J＝3.2Hz),154.27,152.33(d,J＝545.4Hz),148.70,143.34(d,J＝13.9Hz),142.13,137.31,135.58,133.25,130.66,130.13,128.85,128.69,122.68,113.40,109.81(d,J＝20.9Hz),109.60(d,J＝7.6Hz),105.66(d,J＝3.9Hz),100.00,66.90,66.56,58.08,53.80,47.48,43.92,43.42,25.79,24.09.

ESI-HRMS[M+H]⁺m/z＝650.3064,calcd forC38H40FN5O4,650.3145.Purity:99.4％by HPLC.

Example 26: synthesis of Compound 4c

Compound 4(0.3462g, 1mmol) and compound c (0.3216g, 1.1mmol) were added to a back wall pressure bottle, 2mL DMSO and 10mL trimethylchlorosilane were added, heated at 100 ℃ overnight, after TLC monitoring reaction was completed, suction filtration was performed, and the crude product was recrystallized from methanol/dichloromethane to obtain 0.1562g as a pale yellow solid with a yield of 25%.

¹H NMR(400MHz,CDCl₃)δ7.99(d,J＝8.3Hz,2H),7.84(d,J＝5.8Hz,1H),7.80(d,J＝11.6Hz,3H),7.65(d,J＝7.5Hz,1H),7.60(t,J＝3.5Hz,2H),7.54(d,J＝8.7Hz,2H),7.53–7.47(m,1H),6.83(d,J＝7.7Hz,1H),6.50(s,1H),4.29(t,J＝7.1Hz,2H),3.87–3.73(m,4H),3.44–3.36(m,2H),3.35–3.27(m,2H),2.68–2.43(m,6H),1.98–1.84(m,2H),1.38(s,9H).¹³C NMR(101MHz,CDCl₃)δ189.69,160.36(d,J＝3.2Hz),156.75,153.42(d,J＝298.4Hz),149.51,148.56,143.50,143.26(d,J＝13.9Hz),136.29,135.56,135.30,133.01,131.03,129.72,129.35,128.80,128.43,128.03,125.59,122.77,109.71(d,J＝20.9Hz),109.46(d,J＝7.8Hz),105.52(d,J＝4.0Hz),66.79,57.99,53.67,43.84,43.34,35.08,31.02,29.60,23.91.

ESI-HRMS[M+H]⁺m/z＝621.3163,calcd for C38H41FN4O3,621.3209.Purity:99.0％by HPLC.

Example 27: synthesis of Compound 4d

Compound 4(0.3462g, 1mmol) and compound d (0.2797g, 1.1mmol) were added to a back wall pressure resistant bottle, 2mL DMSO, 10mL trimethylchlorosilane were added, heating was carried out at 100 ℃ overnight, after TLC monitoring reaction was completed, suction filtration was carried out, and the crude product was recrystallized from methanol/dichloromethane to give 0.2961g as a pale yellow solid in 51% yield.

¹H NMR(400MHz,CDCl₃)δ8.10(dd,J＝7.8,5.8Hz,2H),7.88–7.79(m,3H),7.74(d,J＝8.1Hz,2H),7.63(d,J＝8.0Hz,2H),7.57(d,J＝15.6Hz,1H),7.22(t,J＝8.4Hz,2H),6.84(d,J＝7.6Hz,1H),6.54(s,1H),4.31(t,J＝7.0Hz,2H),3.89–3.76(m,4H),3.46–3.38(m,2H),3.37–3.30(m,2H),2.67–2.46(m,6H),1.99–1.89(m,2H).¹³C NMR(101MHz,CDCl₃)δ188.52,166.96,164.43,160.41(d,J＝3.4Hz),152.32(d,J＝537.2Hz),152.08,148.66,143.87,143.37(d,J＝14.0Hz),137.82,134.43(d,J＝2.9Hz),134.42,133.64,131.16,131.07,130.18,128.90,128.68,122.20,115.94,115.73,109.83(d,J＝21.0Hz),109.59(d,J＝7.8Hz),105.63(d,J＝4.1Hz),66.89,58.11,53.79,53.49,43.93,25.83,24.00.

ESI-HRMS[M+H]⁺m/z＝583.2422,calcd for C34H32F2N4O3,583.2508.Purity:98.9％by HPLC.

Example 28: synthesis of Compound 4e

Adding the compound 4(0.3462g, 1mmol) and the compound e (0.2930g, 1.1mmol) into a rear-wall pressure-resistant bottle, adding 2mL of DMSO and 10mL of trimethylchlorosilane, heating at 100 ℃ overnight, after TLC monitoring reaction, performing suction filtration, and recrystallizing a crude product with methanol/dichloromethane to obtain 0.3594g of light yellow solid with the yield of 60%.

¹H NMR(400MHz,DMSO)δ8.20(d,J＝8.2Hz,2H),8.07–7.99(m,3H),7.79–7.69(m,4H),7.64(d,J＝11.4Hz,1H),7.11(d,J＝7.9Hz,2H),6.90(d,J＝7.8Hz,1H),4.18–4.13(m,2H),4.00–3.93(m,4H),3.88(s,3H),3.47–3.40(m,2H),3.34(dd,J＝14.4,7.1Hz,2H),3.21–3.03(m,6H),2.10–2.02(m,2H).¹³C NMR(101MHz,DMSO)δ187.96,165.97,163.84,159.58,151.97(d,J＝774.2Hz),149.33,143.01(d,J＝14.9Hz),142.60,137.64,135.73,134.70,134.67,132.99,131.36,131.09,130.52,129.68,123.62,116.44,114.59,109.84(d,J＝19.9Hz),105.80(d,J＝6.7Hz),100.01(d,J＝2.6Hz),63.65,56.10,54.57,54.47,51.61,44.61,25.81,22.66.

ESI-HRMS[M+H]⁺m/z＝595.2642,calcd for C35H35FN4O4,595.2690.Purity:99.5％by HPLC.

Example 29: synthesis of Compound 4f

Adding the compound 4(0.3462g, 1mmol) and the compound f (0.3083g, 1.1mmol) into a rear-wall pressure-resistant bottle, adding 2mL of DMSO and 10mL of trimethylchlorosilane, heating at 100 ℃ overnight, after TLC monitoring reaction, performing suction filtration, and recrystallizing a crude product with methanol/dichloromethane to obtain 0.2530g of light yellow solid with the yield of 41%.

¹H NMR(400MHz,MeOD)δ8.13(s,1H),7.82–7.77(m,2H),7.73–7.66(m,3H),7.62(d,J＝3.8Hz,1H),7.60(d,J＝6.1Hz,1H),7.48(t,J＝7.8Hz,1H),7.42(s,1H),7.03(d,J＝7.3Hz,1H),6.86(d,J＝8.1Hz,1H),5.98(s,2H),4.33–4.28(m,2H),3.99(dd,J＝13.4,2.0Hz,2H),3.72(t,J＝12.6Hz,2H),3.53–3.45(m,6H),3.41–3.34(m,4H),2.15–2.07(m,2H).¹³C NMR(101MHz,MeOD)δ189.32,163.47,154.06(d,J＝303.1Hz),151.14,148.56,143.18(d,J＝13.9Hz),142.78,141.42,138.63,131.72,130.86,129.57,128.57,125.13,122.66,120.36,119.17,117.62,115.38,109.56(d,J＝14.6Hz),109.21(d,J＝8.9Hz),107.63,102.15(d,J＝8.5Hz),63.70,56.93,54.92,51.99,30.25,25.32,24.71.

ESI-HRMS[M+H]⁺m/z＝609.2435,calcd for C35H33FN4O5,609.2480.Purity:98.3％by HPLC.

Example 30: synthesis of Compound 4g

Adding compound 4(0.3462g, 1mmol) and compound g (0.3535g, 1.1mmol) into a back-wall pressure-resistant bottle, adding 2mL of DMSO and 10mL of trimethylchlorosilane, heating at 100 ℃ overnight, after TLC monitoring reaction, performing suction filtration, and recrystallizing the crude product with methanol/dichloromethane to obtain 0.3961g of light yellow solid with the yield of 61%.

¹H NMR(400MHz,DMSO)δ8.07(d,J＝8.4Hz,2H),7.92(d,J＝16.1Hz,2H),7.84(d,J＝7.6Hz,1H),7.71(m,3H),7.60(d,J＝12.2Hz,1H),7.58–7.52(m,1H),7.02(d,J＝8.7Hz,2H),6.84(d,J＝7.9Hz,1H),6.59(s,1H),4.17–4.10(m,2H),3.78–3.72(m,4H),3.67–3.60(m,4H),3.37–3.28(m,8H),2.44–2.37(m,6H),1.85–1.77(m,2H).¹³C NMR(101MHz,CDCl₃)δ188.17,160.85(d,J＝3.1Hz),154.70,152.76(d,J＝542.4Hz),149.13,143.77(d,J＝13.9Hz),142.56,137.74,136.01,133.68,131.09,130.56,129.28,129.12,123.11,113.83,110.24(d,J＝20.6Hz),109.99(d,J＝7.4Hz),106.09(d,J＝3.9Hz),67.33,66.99,58.51,54.23,47.91,44.35,43.85,26.22,24.52.

ESI-HRMS[M+H]⁺m/z＝650.3064,calcd for C38H40FN5O4,650.33119.Purity:99.5％by HPLC.

Example 31: synthesis of Compound 4h

Adding compound 4(0.3462g, 1mmol) and compound h (0.3216g, 1.1mmol) into a back-wall pressure-resistant bottle, adding 2mL of DMSO and 10mL of trimethylchlorosilane, heating at 100 ℃ overnight, after TLC monitoring reaction, performing suction filtration, and recrystallizing the crude product with methanol/dichloromethane to obtain 0.1023g of light yellow solid with the yield of 16%.

¹H NMR(400MHz,CDCl₃)δ8.02–7.98(m,2H),7.83–7.80(m,1H),7.78(d,J＝7.3Hz,2H),7.65(d,J＝7.6Hz,1H),7.59(d,J＝5.6Hz,1H),7.57–7.53(m,3H),7.50(t,J＝6.5Hz,1H),6.82(d,J＝7.7Hz,1H),4.31–4.25(m,2H),3.89–3.77(m,4H),3.44–3.36(m,2H),3.32(dd,J＝9.9,4.3Hz,2H),2.69–2.47(m,6H),2.01–1.87(m,2H),1.37(s,9H).¹³C NMR(101MHz,CDCl₃)δ189.80,160.47(d,J＝3.2Hz),156.86,153.53(d,J＝298.4Hz),149.62,148.67,143.61,143.37(d,J＝13.9Hz),136.40,135.67,135.41,133.12,131.14,129.83,129.46,128.91,128.54,128.14,125.70,122.88,109.82(d,J＝20.9Hz),109.57(d,J＝7.8Hz),105.63(d,J＝4.0Hz),66.90,58.10,53.78,43.95,43.45,35.19,31.13,29.71,25.68.

ESI-HRMS[M+H]⁺m/z＝621.3163,calcd for C38H41FN4O3,621.3220.Purity:97.9％by HPLC.

Example 32: synthesis of Compound 4i

Adding compound 4(0.3462g, 1mmol) and compound i (0.2797g, 1.1mmol) into a back-wall pressure-resistant bottle, adding 2mL of DMSO and 10mL of trimethylchlorosilane, heating at 100 ℃ overnight, after TLC monitoring reaction, performing suction filtration, and recrystallizing the crude product with methanol/dichloromethane to obtain 0.4168g of light yellow solid with the yield of 72%.

¹H NMR(400MHz,CDCl₃)δ8.08(t,J＝6.8Hz,2H),7.86(d,J＝12.0Hz,2H),7.80(d,J＝12.2Hz,2H),7.66(d,J＝7.8Hz,1H),7.61(d,J＝7.5Hz,1H),7.55(d,J＝11.5Hz,1H),7.53–7.49(m,1H),7.20(t,J＝8.4Hz,2H),6.83(d,J＝7.7Hz,1H),6.51(s,1H),4.29(t,J＝7.0Hz,2H),3.87–3.73(m,4H),3.44–3.36(m,2H),3.35–3.29(m,2H),2.68–2.47(m,6H),1.91(dt,J＝12.1,6.2Hz,2H).¹³C NMR(101MHz,CDCl₃)δ188.49,166.98,164.45,160.40,154.92,149.55,149.23(d,J＝548.0Hz),148.59,144.23,143.29(d,J＝14.0Hz),136.37,135.35,134.29,133.16,131.20,131.11,130.04,129.49,128.69,128.10,122.16,115.97,115.75,109.76(d,J＝21.0Hz),109.51(d,J＝8.0Hz),105.53(d,J＝3.4Hz),66.71,57.94,53.65,43.94,43.26,25.62,23.85.

ESI-HRMS[M+H]⁺m/z＝583.2442,calcd for C34H32F2N4O3,583.2500.Purity:99.1％by HPLC.

Example 33: synthesis of Compound 4j

Adding the compound 4(0.3462g, 1mmol) and the compound j (0.2930g, 1.1mmol) into a back-wall pressure-resistant bottle, adding 2mL of DMSO and 10mL of trimethylchlorosilane, heating at 100 ℃ overnight, after TLC monitoring reaction, performing suction filtration, and recrystallizing the crude product with methanol/dichloromethane to obtain 0.2010g of light yellow solid with the yield of 34%.

¹H NMR(400MHz,CDCl₃)δ8.08(d,J＝8.8Hz,2H),7.85(d,J＝12.7Hz,1H),7.83–7.79(m,2H),7.68(d,J＝8.1Hz,1H),7.64–7.58(m,2H),7.56–7.50(m,1H),7.03(d,J＝8.8Hz,2H),6.84(d,J＝7.7Hz,1H),4.35–4.27(m,2H),3.93(s,3H),3.90–3.84(m,4H),3.47–3.40(m,2H),3.38–3.31(m,2H),2.80–2.44(m,6H),2.05–1.92(m,2H).¹³C NMR(101MHz,DMSO)δ187.86,163.83,159.29,154.86(d,J＝246.1Hz),151.34,147.84,143.21,143.19(d,J＝3.6Hz),141.13,136.30,136.04,131.60,131.46,130.91,130.70,129.96,129.39,123.42,118.74,114.55,111.77(d,J＝10.1Hz),109.92(d,J＝25.6Hz),104.63(d,J＝7.4Hz),63.61,56.09,54.43,51.51,45.00,34.59,25.61,22.41.

ESI-HRMS[M+H]⁺m/z＝595.2642,calcd for C35H35FN4O4,595.2692.Purity:98.7％by HPLC.

Example 34 inhibition of tumor cell growth by quinazolinone and α -unsaturated ketone conjugate derivatives

The quinazolinone and α -unsaturated ketone conjugated derivatives are selected to perform in vitro cytotoxicity determination on a plurality of tumor cell strains by adopting an MTT method, quinazolinone and α -unsaturated ketone conjugated derivatives with different concentrations are added into cells in a logarithmic phase, after the cells act for 48 hours, MTT is added, the absorbance is determined, the concentration of the compounds for inhibiting the cell growth by 50% is respectively calculated, the value is expressed by an IC50 value, and the result is shown in table 1.

TABLE 1 inhibition of tumor cell line growth by quinazolinone and α -unsaturated ketone conjugated derivatives (IC)₅₀Value/. mu.M)

Example 35: represents the Effect of Compound 3i on the transcription and expression of the intracellular c-MYC Gene

RT-PCR experimental method:

cell culture: inoculating cells into a 6-well plate, 200000 cells/well, adding a compound after 24 hours of cell adherence, culturing for 3-24 hours, collecting cells, extracting RNA, detecting RNA concentration, and performing reverse transcription reaction and PCR (polymerase chain reaction) experiment according to the following system.

The PCR products, primers and preparation system used were as follows:

10 × Dream Taq Green buffer, 2.5 μ L; dNTP mix (2.5mM), 0.5. mu.L; cDNA, 1 uL; primer (sense), 0.5. mu.L; primer (antisense), 0.5 μ L; dream TaqDNA polymerase, 0.25 μ L; ddH20, 20.25 μ L; total, 25. mu.L

Procedure 95 ℃ denaturation for 5 min; denaturation at 95 ℃ for 30 s; annealing at 58 ℃ for 30 s; extension at 72 ℃ for 60s (30 cycles); 72 ℃ for 10 min; finally cooling to 10 ℃.

Electrophoresis is carried out on 1.5% agarose at 120V for 0.5h, and after the electrophoresis is finished, gel imaging is carried out.

As a result, as shown in FIG. 1, quinazolinone conjugated derivative 3i with α -unsaturated ketone was able to reduce the transcription level of c-myc.

Western blot experiment method:

cell culture: counting cells, inoculating, culturing in six-well plate to obtain 500 ten thousand cells, taking out, cracking, collecting cells, adding 50 μ L cell cracking liquid, and extracting supernatant total protein liquid. The BCA method is used for detecting the total protein concentration, protein samples are denatured, the same mass of protein is loaded, and SDS-PAGE gel electrophoresis is used for separating protein bands. And (3) calculating the molecular weight according to the target protein, cutting off the electrophoresis gel band at the corresponding position, and transferring the protein band to the PVDF membrane by a wet transfer method.

Preparing TBST buffer solution: PVDF membrane was blocked with 25mM NaCl, 100mM Tris, 0.2% Tween-20, pH 7.4, and 5% skim milk powder solution (w/v) dissolved in TBST buffer. And respectively applying primary antibody and secondary antibody to incubate the PVDF membrane, rinsing the PVDF membrane for proper times by using TBST buffer solution, and then using a SuperECL Plus hypersensitivity luminescent kit for color development and imaging.

As a result, as shown in FIG. 2, quinazolinone conjugated derivative 3i with α -unsaturated ketone was able to reduce the expression level of C-MYC.

Claims

1. A quinazolinone- α -unsaturated ketone conjugate derivative, wherein the structure formula of the quinazolinone- α -unsaturated ketone conjugate derivative is shown in formula (I):

wherein R1 is NH (CH)₂)nR₃，

R₃Is N, N-diethylmethylamino or morpholinyl; n is any one integer of 1-5;

R₂is substituted phenyl or 1, 3-benzodioxole, and the substituent in the substituted phenyl is fluorine, morpholinyl, tert-butyl or methoxyl.

2. A process for preparing a quinazolinone and α -unsaturated ketone conjugate derivative according to claim 1, comprising the steps of:

S2. Compound

Reacting with amine compound to obtain intermediate

S3. Compound

Reacting with ketone compound to obtain intermediate

And S4, reacting the compound obtained in the S2 with the compound obtained in the S3 to obtain the quinazolinone and α -unsaturated ketone conjugated derivative.

3. Use of the quinazolinone and α -unsaturated ketone conjugate derivative according to claim 1 for the preparation of anticancer drugs.

4. The use as claimed in claim 3, wherein the quinazolinone and α -unsaturated ketone conjugate derivative is used as a blocker of proto-oncogene c-MYC transcription regulation in the preparation of anticancer drugs.

5. The use of claim 3, wherein the anti-cancer is one or more of anti-ovarian cancer, cervical cancer, breast cancer, lung adenocarcinoma, colon cancer, liver cancer, leukemia, small cell lung cancer, skin cancer, epithelial cell cancer, prostate cancer, non-small cell lung cancer, nasopharyngeal cancer, glioblastoma, lymphoma or melanoma.