CN104163772A - Substituted diaryl ester compound, and preparation method and application thereof - Google Patents

Abstract

The invention discloses a substituted diaryl ether compound, the structural formula of which is as follows: Where Z is -CH- or -N-; W is -CH- or -N-; Y is -O-, -CH ₂ -, -NH- or -NR _y -; R ₁ is hydrogen, halogen, NO ₂ , CN, CF ₃ , OR _a , COR _a , COOR _a , SO ₂ R _a , SO ₂ NR _a R _b , NR _a R _b , NR _a COR _b , unsubstituted\substituted C ₁ -C ₄ alkyl, One of unsubstituted\substituted aryl or unsubstituted\substituted heterocyclic group; R ₂ is hydrogen, unsubstituted\substituted C ₁ -C ₄ alkyl, or halogen; R is unsubstituted\substituted C ₁ -C ₁₂ alkyl, unsubstituted\substituted C ₃ -C ₁₂ cycloalkyl, unsubstituted\substituted C ₂ -C ₁₂ alkenyl, unsubstituted\substituted aryl, substituted alkylamine or unsubstituted One of the substituted\substituted heterocyclic groups. The substituted diaryl ether compound of the invention can obviously inhibit the malignant proliferation phenotype of cancer cells, promote the expression of apoptosis factor protein, and provide a novel potential antitumor drug.

Description

A kind of substituted diaryl ether compound and its preparation method and application

technical field

The invention belongs to the field of chemical medicines, and in particular relates to a substituted diaryl ether compound and its preparation method and application. the

Background technique

Malignant tumor is a common and frequently-occurring disease that seriously threatens human health. It is a major scientific research topic in the field of medical science in all countries in the world. At present, there are no satisfactory prevention and control measures. About 7 million people die of cancer in the world every year, accounting for about a quarter of the total death toll. Among the three major therapies for malignant tumors (surgery, chemotherapy and radiotherapy), chemotherapy is still an important method for clinical treatment. The selectivity of anticancer drugs to cancer cells and normal human cells is not much different, so the adverse reactions in the application process are extensive and serious. In addition, easy drug resistance is also one of the problems in the treatment process. In recent years, with the development of molecular biology, immunology, cell dynamics and people's further understanding of the molecular mechanism of cancer occurrence and development, the research and development of anti-tumor drugs has shifted from traditional cytotoxic drugs to tumor-related drugs. The study of targeted drugs targeting the signaling pathway. the

In 1973, Professor Folkman first proposed tumor angiogenesis factor (tumor angiogenesis factor, TFA), and it has been confirmed that there are many angiogenesis factors in tumor patients, among which vascular endothelial growth factor (vascular endothelial growth factor, VEGF) is the One of the strongest angiogenic factors (Kraizer Y et al. Biochem Biophys Res Commun, 2001, 287:209-215). VEGF and its receptor (vascular endothelial growth factor receptor, VEGFR) are overexpressed in liver cancer tissue compared with normal liver tissue, and are closely related to the growth, metastasis, recurrence and treatment of liver cancer (Suzuki H et al. Am J Physiol, 1999, 276). Therefore, multi-target receptor tyrosine kinase inhibitors targeting VEGF/VEGFR have become an important topic in the research of anticancer drugs. the

In 2005, a multi-target tyrosine kinase inhibitor Sorafenib (shown in the following formula) jointly developed by Bayer AG of Germany and ONYX of the United States was approved by the U.S. FDA for the treatment of advanced cancer. Therapeutics, the first new drug approved for advanced cancer in more than a decade, has made major advances in the treatment of advanced cancer. But sorafenib is often accompanied by a higher incidence of mild and moderate adverse reactions such as rash, diarrhea and oral ulcers, and severe adverse reactions of myocardial infarction (Lin Lin et al., Journal of Clinical Oncology, 2009, 14 (4): 366-368). the

Yang Zhao et al. (Acta Pharm Sin, 2011, 46:1093-1097) based on the structural transformation of bioisosteres, replaced urea with thiourea, and changed the substituents on the A ring to obtain a series of derivatives (as shown in the following formula) , its inhibitory activity to kidney cancer 760-O, liver cancer HepG2, lung cancer A549, breast cancer MDA-MB-435, prostate cancer PC3 and colon cancer cell HT-29 was higher than that of sorafenib. Compound 1b The meta-position of the terminal benzene ring introduces a sulfonamide group, and its inhibitory activity against 6 kinds of tumor cells is better than that of compound 1a. the

The structure of Sorafenib has the advantages of many modification sites and large optimization space. Sorafenib is used as the lead compound to carry out structural modification for specific targets. Through the organic combination of virtual screening and activity screening, independent research and development Compounds similar in structure and pharmacological action to sorafenib can serve as a reference for the development of anti-tumor drugs. the

Contents of the invention

The object of the present invention is to provide a substituted diaryl ether compound. the

Another object of the present invention is to provide a preparation method of the above compound. the

Another object of the present invention is to provide the application of the above compounds. the

Concrete technical scheme of the present invention is as follows:

A substituted diaryl ether compound, its structure is as follows:

Z is -CH- or -N-;

W is -CH- or -N-;

Y is -O-, -CH ₂ -, -NH- or -NR _y -;

R ₁ is hydrogen, halogen, NO ₂ , CN, CF ₃ , OR _a , COR _a , COOR _a , SO ₂ R _a , SO ₂ NR _a R _b , NR _a R _b , NR _a COR _b , unsubstituted\substituted One of the C ₁ -C ₄ alkyl groups, unsubstituted\substituted aryl groups or unsubstituted\substituted heterocyclic groups; wherein, the above-mentioned unsubstituted\substituted C ₁ -C ₄ alkyl groups are preferably methyl , ethyl, propyl, isopropyl or tert-butyl, the above unsubstituted \ substituted aryl includes single or multiple ring compounds, also includes polycyclic compounds containing independent or fused aryl, preferred aryl Containing 6 to about 10 carbon ring atoms, particularly preferred aryl groups include unsubstituted\substituted phenyl, unsubstituted\substituted naphthyl, and the above-mentioned unsubstituted\substituted heterocyclic groups include 1 to 3 independent or Aromatic heterocyclic groups with fused rings and from 5 to about 10 ring atoms, preferably aromatic heterocyclic groups containing one, two or three heteroatoms selected from N, O or S, including, for example: furyl, imidazolyl , thiazolyl, isothiazolyl, quinolinyl (including isoquinolyl), pyridyl, thienyl, pyrrolyl, indolyl, triazolyl, benzimidazolyl, benzofuryl, benzothienyl , benzothiazolyl.

R ₂ is hydrogen, unsubstituted/substituted C ₁ -C ₄ alkyl, or halogen; wherein, the above-mentioned unsubstituted/substituted C ₁ -C ₄ alkyl is preferably methyl, ethyl, propyl, isopropyl or tert-butyl.

R is unsubstituted\substituted C ₁ -C ₁₂ alkyl, unsubstituted\substituted C ₃ -C ₁₂ cycloalkyl, unsubstituted\substituted C ₂ -C ₁₂ alkenyl, unsubstituted\substituted aryl , a substituted alkylamine or an unsubstituted\substituted heterocyclic group; wherein, the above-mentioned unsubstituted\substituted C ₁ -C ₁₂ alkyl groups preferably have 1, 2, 3 or 4 carbon atoms , more preferably methyl, ethyl, propyl, isopropyl or tert-butyl, the above-mentioned unsubstituted \ substituted C ₃ -C ₁₂ cycloalkyl is preferably cyclopentyl, cyclohexyl, cyclopropane Group or cyclobutanyl group, the above-mentioned unsubstituted\substituted aryl group includes single or multiple ring compounds, and also includes polycyclic compounds containing independent or fused aryl groups, the preferred aryl group contains 6 to about 10 carbon rings Atoms, particularly preferred aryl groups include unsubstituted\substituted phenyl, unsubstituted\substituted naphthyl, the above-mentioned unsubstituted\substituted heterocyclic groups include 1 to 3 independent or fused rings and from 5 to about 10 Aromatic heterocyclic group with 3 ring atoms, the preferred aromatic heterocyclic group contains one, two or three heteroatoms selected from N, O or S, for example including: furyl, imidazolyl, thiazolyl, isothiazolyl, Quinolinyl (including isoquinolyl), pyridyl, thienyl, pyrrolyl, indolyl, triazolyl, benzimidazolyl, benzofuryl, benzothienyl, benzothiazolyl.

Preferably, the R _a is hydrogen, unsubstituted\substituted C ₁ -C ₄ alkyl, unsubstituted\substituted C ₂ -C ₄ alkenyl, unsubstituted\substituted aryl or unsubstituted\substituted hetero One of the ring groups; wherein, the above-mentioned unsubstituted\substituted C ₁ -C ₄ alkyl is preferably methyl, ethyl, propyl, isopropyl or tert-butyl, and the above-mentioned unsubstituted\substituted aryl Compounds including single or multiple rings, and also polycyclic compounds containing individual or fused aryl groups, preferably aryl groups containing from 6 to about 10 carbon ring atoms, particularly preferred aryl groups include unsubstituted\substituted phenyl groups , unsubstituted\substituted naphthyl, the above-mentioned unsubstituted\substituted heterocyclic groups include aromatic heterocyclic groups containing 1 to 3 independent or fused rings and from 5 to about 10 ring atoms, preferred aromatic heterocyclic groups Containing one, two or three heteroatoms selected from N, O or S, for example including: furyl, imidazolyl, thiazolyl, isothiazolyl, quinolinyl (including isoquinolyl), pyridyl, thiophene Base, pyrrolyl, indolyl, triazolyl, benzimidazolyl, benzofuryl, benzothienyl, benzothiazolyl.

Further preferably, the R _b is hydrogen, unsubstituted\substituted C ₁ -C ₄ alkyl, unsubstituted\substituted C ₂ -C ₄ alkenyl, unsubstituted\substituted aryl or unsubstituted\substituted One of the heterocyclic groups; wherein, the above-mentioned unsubstituted\substituted C ₁ -C ₄ alkyl is preferably methyl, ethyl, propyl, isopropyl or tert-butyl, and the above-mentioned unsubstituted\substituted aryl The group includes single or multiple ring compounds, and also includes polycyclic compounds containing independent or fused aryl groups. Preferred aryl groups contain 6 to about 10 carbon ring atoms. Particularly preferred aryl groups include unsubstituted\substituted benzene Base, unsubstituted\substituted naphthyl, the above-mentioned unsubstituted\substituted heterocyclic groups include aromatic heterocyclic groups containing 1 to 3 independent or fused rings and from 5 to about 10 ring atoms, preferred aromatic heterocyclic The group contains one, two or three heteroatoms selected from N, O or S, for example including: furyl, imidazolyl, thiazolyl, isothiazolyl, quinolinyl (including isoquinolyl), pyridyl, Thienyl, pyrrolyl, indolyl, triazolyl, benzimidazolyl, benzofuryl, benzothienyl, benzothiazolyl.

Further preferably, the R _y is one of hydrogen or unsubstituted/substituted C ₁ -C ₃ alkyl, preferably hydrogen, methyl, ethyl or propyl, more preferably hydrogen.

In a preferred embodiment of the present invention, both Z and W are -CH-, and Y is -O-, -NH- or -NR _y .

In a preferred embodiment of the present invention, said Z is -CH-, said W is -N-, and said Y is -O-. the

In a preferred embodiment of the present invention, both Z and W are -N-, and Y is -O- or -CH ₂ -.

To further illustrate, the above-mentioned unsubstituted\substituted alkyl, alkenyl, or cycloalkyl can be substituted by one or more of the following groups at the positions that can be substituted: halogen, alkoxy, amino;

To further illustrate, the above-mentioned aryl or heterocyclic group can be substituted by one or more of the following groups: halogen, alkyl, haloalkyl, alkyloxy, cycloalkyl, Cycloalkylaminoacyl, arylalkyl, alkylacyl, arylalkylacyl, heterocyclylacyl. the

The unsubstituted\substituted alkyl, alkenyl, cycloalkyl, aryl or heterocyclic groups mentioned above can also be substituted at one or more available positions by one or more suitable groups, such as halogen , OR _a , =O, SR _a , SOR _a , SO ₂ R _a , NO ₂ , NHR _a , NR _a R _b , =NR _a , NHCOR _a , CN, COR _a , OCOR _a , COOR _a , unsubstituted or Substituted C ₁ -C ₃ alkyl, unsubstituted or substituted C ₂ -C ₄ alkenyl, unsubstituted or substituted aryl, and unsubstituted or substituted heterocyclic group, wherein each of R ₁ and R _{The 2} groups are independently selected from the group consisting of hydrogen, halogen, OH, COH, NO ₂ , NH ₂ , SH, CN, CO ₂ H.

A kind of preparation method of above-mentioned substituted diaryl ether compound, its reaction process is as follows:

Specifically: under the action of a base, the substituted halogenated benzene or substituted aniline shown in the compound (II) is reacted with the substituted ethyl benzoate shown in the compound (III) to generate the compound (IV), and the compound (IV) is carried out Condensation reaction with substituted amine after hydrolysis or acid chlorination to obtain compound (I), namely the substituted diaryl ether compound of the present invention. the

In a preferred embodiment of the present invention, said X is F, Cl, Br or I. the

An application of the above-mentioned substituted diaryl ether compound in the preparation of tumor medicine. the

The beneficial effects of the present invention are: the substituted diaryl ether compound of the present invention can obviously inhibit the malignant proliferation phenotype of cancer cells, promote the expression of apoptosis factor protein, and provide a novel potential antitumor drug. the

Description of drawings

Figure 1 is the experimental results of compound 8 in Example 32 of the present invention inhibiting the proliferation of liver cancer cell HepG2 and the formation of clones, wherein A is the result of compound 8 inhibiting the proliferation of liver cancer cell HepG2, and B is the effect of compound 8 inhibiting the proliferation of liver cancer cell HepG2 at different concentrations Cloning formation and histogram analysis;

Fig. 2 is the Western blot experiment result of compound 8 in Example 32 of the present invention inducing the expression of hepatoma cell HepG2 apoptosis factor protein, wherein A shows the time-course effect of compound 8 inducing the expression of hepatoma cell HepG2 apoptosis factor protein, and B Show the dose-effect relationship of compound 8 inducing the expression of apoptosis factor protein in liver cancer cells HepG2;

Figure 3 is the experimental results of compound 12 inhibiting the proliferation of liver cancer cell HepG2 and the formation of clones in Example 32 of the present invention, wherein A is the experimental result of compound 12 inhibiting the proliferation of liver cancer cell HepG2, and B is the effect of compound 12 on inhibiting the proliferation of liver cancer cell HepG2 at different concentrations Cloning formation and histogram analysis;

Figure 4 is the Western blot experiment result of compound 12 in Example 32 of the present invention inducing the expression of hepatoma cell HepG2 apoptosis factor protein, wherein A shows the time course effect of compound 12 inducing the expression of hepatoma cell HepG2 apoptosis factor protein, B It shows the dose-effect relationship of compound 12 inducing the expression of apoptosis factor protein in liver cancer cell HepG2. the

Detailed ways

The technical solutions of the present invention will be further illustrated and described below through specific embodiments in conjunction with the accompanying drawings. the

The preparation method of substituted diaryl ether compound of the present invention, its reaction process is as follows:

That is, under the action of a base, the substituted halogenated benzene or substituted aniline shown in the compound (II) is reacted with the substituted ethyl benzoate shown in the compound (III) to generate the compound (IV), and the compound (IV) is hydrolyzed or Condensation reaction with substituted amine after acid chlorination to obtain compound (I), namely the substituted diaryl ether compound of the present invention. the

Specifically include the following steps:

(1) The synthetic method of intermediate acid:

(2) The synthesis method of the target compound, including two kinds of A and B, wherein A is as follows:

B is as follows:

The synthesis methods of B1 to B6 in the above process are shown in the following Examples 1 to 6, and the synthesis of compounds 1 to 17 and compounds 18 to 25 are detailed in the following Examples 7 to 31. the

Embodiment 1: the synthesis (B1) of 4-(4-fluorophenoxy)benzoic acid

1.0mL (8.9moL) of 1-fluoro-4-iodobenzene, 1.85g (13.4moL) of p-hydroxybenzoic acid, 170mg (0.9mmoL) of cuprous iodide, and 276mg (2.7mmoL) of N,N-dimethylglycine Dissolve 5.81g (17.8moL) of cesium carbonate in 25mL of 1,4-dioxane, heat and stir at 90°C for 24 hours under the protection of argon. After the reaction was completed, the solvent was evaporated under reduced pressure, water was added, and ethyl acetate was extracted three times. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The solvent was removed and separated by silica gel column chromatography (ethyl acetate:petroleum ether=1:30) to obtain a yellow liquid. 1.0 g (4.1 moL) of the synthesized ethyl 1-fluoro-4-phenoxybenzoate was dissolved in 8 mL of ethanol and 8 mL of water, 1.82 g (4.5 moL) of sodium hydroxide was added, and heated to reflux for 3 hours. After completion of the reaction, cool down, evaporate the solvent under reduced pressure, add 50 mL of water, acidify with 2M hydrochloric acid, extract with dichloromethane, wash the organic layer with saturated brine, and dry over anhydrous magnesium sulfate. The solvent was removed to obtain 1.53 g of white solid, and the yield of the two-step reaction was 76%. the

¹ H NMR (600MHz, DMSO-d ₆ ) δ: 7.00 (d, J=8.8Hz, 2H), 7.17 (d, J=4.4Hz, 1H), 7.18 (d, J=4.6Hz, 1H), 7.28 (t, J = 8.6Hz, 2H), 7.94 (d, J = 8.8Hz, 2H), 12.82 (br s, 1H) ppm.

¹³ C NMR (150 MHz, DMSO-d ₆ ) δ: 116.8, 117.0, 122.0, 125.2, 131.7, 151.1, 161.3, 166.7 ppm.

Embodiment 2: the synthesis (B2) of 4-phenoxybenzoic acid

The synthesis method of Example 1 above gave B2 white solid, and the two-step yield was 87%. the

¹ H NMR (600MHz, DMSO-d ₆ ) δ: 7.02(d, J=8.8Hz, 2H), 7.11(d, J=7.7Hz, 2H), 7.23(t, J=7.3Hz, 1H), 7.45 (t, J = 7.5Hz, 2H), 7.95 (d, J = 8.8Hz, 2H), 12.82 (br s, 1H) ppm;.

¹³ C NMR (150 MHz, DMSO-d ₆ ) δ: 117.2, 119.9, 124.7, 125.2, 130.3, 131.7, 155.1, 161.0, 166.8 ppm.

Embodiment 3: the synthesis (B3) of 4-(4-chlorophenoxy)benzoic acid

The synthesis method of Example 1 above gave B3 white solid, and the two-step yield was 80%. the

¹ H NMR (600MHz, DMSO-d ₆ ) δ: 7.05 (d, J=8.6Hz, 2H), 7.14 (d, J=8.8Hz, 2H), 7.48 (d, J=9.0Hz, 2H), 7.96 (d, J = 8.8Hz, 2H), 12.86 (br s, 1H) ppm.

¹³ C NMR (150 MHz, DMSO-d ₆ ) δ: 117.5, 121.6, 125.7, 128.5, 130.2, 131.7, 154.1, 160.5, 166.7 ppm.

Embodiment 4: the synthesis (B4) of 4-(4-bromophenoxy)benzoic acid

The synthesis method of Example 1 above gave B4 white solid, and the two-step yield was 53%. the

¹ H NMR (600MHz, DMSO-d ₆ ) δ: 7.06 (d, J=8.4Hz, 2H), 7.08 (d, J=8.3Hz, 2H), 7.61 (d, J=8.3Hz, 2H), 7.95 (d, J = 9.0 Hz, 2H), 12.86 (br s, 1H) ppm.

¹³ C NMR (150 MHz, DMSO-d ₆ ) δ: 116.4, 117.6, 122.0, 125.7, 131.7, 133.1, 154.7, 160.4, 166.7 ppm.

Embodiment 5: the synthesis (B5) of 4-(4-methoxyphenoxy group) benzoic acid

The synthesis method of Example 1 above gave B5 white solid, and the two-step yield was 57%. the

¹ H NMR (600MHz, DMSO-d ₆ )δ: 3.77(s, 3H), 6.95(d, J=8.8Hz, 2H), 7.00(d, J=9.0Hz, 2H), 7.08(d, J= 9.0Hz, 2H), 7.92 (d, J = 8.8Hz, 2H), 12.76 (br s, 1H) ppm.

¹³ C NMR (150 MHz, DMSO-d ₆ ) δ: 55.4, 115.3, 116.2, 121.7, 124.6, 131.6, 148.0, 156.3, 162.1, 166.8 ppm.

Embodiment 6: the synthesis (B6) of 4-(4-nitrophenoxy group) benzoic acid

The same synthesis method as in Example 1 above gave B6 yellow solid with a two-step yield of 49%. the

¹ H NMR (600MHz, DMSO-d ₆ ) δ: 7.23-7.25(m, 4H), 8.03(d, J=8.6Hz, 2H), 8.28(d, J=9.2Hz, 2H), 12.99(br s ,1H) ppm.

¹³ C NMR (150 MHz, DMSO-d ₆ ) δ: 118.7, 119.6, 126.3, 127.4, 131.9, 143.0, 158.4, 161.5, 166.6 ppm.

Embodiment 7: the synthesis of compound 1

A 10 mL round bottom flask was charged with N,N-dimethylethylenediamine (44.1 mg, 0.5 mmol), 4-phenoxybenzoic acid (107.1 mg, 0.5 mmol) and 3 mL of freshly distilled dichloromethane. Then N-hydroxybenzotriazole (HOBt) (81.7 mg, 0.6 mmol), 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI) (115.0 mg, 0.6mmol and N-methylmorpholine (NMM) (151.7mg, 1.5mmol).After 30min in ice bath, remove the ice bath.Stir at room temperature overnight, TCL detects that the reaction is complete.The reaction solution is extracted with water/dichloromethane, and the organic phase, the organic phase was successively washed with 10% hydrochloric acid solution (×3), 5% sodium bicarbonate solution (×1), water (×1), saturated sodium chloride solution, dried over anhydrous magnesium sulfate, filtered, and reduced Concentrate under reduced pressure to obtain the crude product. The crude product was separated with a silica gel column (methanol:dichloromethane=1:20) to obtain a yellow oil with a yield of 97%. 

¹ H NMR (400MHz, CDCl ₃ ) δ: 2.31(s, 6H), 2.57(t, J=5.8Hz, 2H), 3.54(q, J=5.0Hz, 2H), 6.95(br s, 1H), 7.01(d, J=8.8Hz, 2H), 7.06(d, J=7.8Hz, 2H), 7.17(t, J=7.5Hz, 1H), 7.38(t, J=8.5Hz, 2H), 7.80( d, J = 8.8 Hz, 2H) ppm.

^{HRMS (ESI, m/z) calcd for C17H21N2O2 [M+H]+} _{285.1603 ; found 285.1598} .

Embodiment 8: the synthesis of compound 2

Compound 2 was obtained by the same synthesis method as in Example 7 as a yellow oil with a yield of 88%. the

¹ H NMR (400MHz, CDCl ₃ ) δ: 1.10(t, J=7.3Hz, 6H), 2.68(q, J=7.3Hz, 4H), 2.76(t, J=5.8Hz, 2H), 3.55(q ,J=5.3Hz,2H),4.89(br s,1H),6.98(d,J=8.8Hz,2H),7.04(d,J=8.5Hz,2H),7.16(t,J=7.5Hz, 1H), 7.37 (t, J=7.5Hz, 3H), 7.80 (d, J=8.8Hz, 2H) ppm.

¹³ C NMR (150 MHz, CDCl ₃ ) δ: 11.1, 36.8, 46.8, 51.5, 117.6, 119.7, 124.1, 128.8, 129.8, 155.9, 160.2, 166.7 ppm.

^{HRMS (ESI, m/z) calcd for C19H25N2O2 [M+H]+} _{313.1916 ; found 313.1911} .

Embodiment 9: the synthesis of compound 3

Compound 3 was obtained as a white solid by the same synthesis method as in Example 7, and the yield was 87%. the

¹ H NMR (400MHz, CDCl ₃ ) δ: 2.49(t, J=4.5Hz, 4H), 2.59(t, J=6.0Hz, 2H), 3.53(q, J=5.5Hz, 2H), 3.71(t ,J=4.8Hz,4H),6.70(br s 1H),7.00(d,J=8.0Hz,2H),7.04(d,J=7.5Hz,2H),7.16(t,J=7.5Hz,1H ), 7.37 (t, J=7.3Hz, 2H), 7.74 (d, J=8.8Hz, 2H) ppm.

¹³ C NMR (150Hz, CDCl ₃ ) δ: 36.0, 53.3, 56.9, 66.9, 117.7, 119.7, 124.2, 128.7, 128.9, 129.9, 155.9, 160.3, 166.6 ppm.

_HRMS (ESI _, m/z) calcd for _C19H23N2O3 [M+H] ⁺ _327.1709 , found 327.1703.

Embodiment 10: the synthesis of compound 4

Compound 4 was obtained as a white solid by the same synthesis method as in Example 7, and the yield was 87%. the

¹ H NMR (400MHz, CDCl ₃ )δ: 1.70–1.79(m,2H),1.92–2.02(m,2H),2.37-2.45(m,2H),4.57(m,1H),6.47(br s, 1H), 6.97(d, J=8.8Hz, 2H), 7.03(d, J=7.5Hz, 2H), 7.16(t, J=7.3Hz, 1H), 7.37(t, J=7.5Hz, 2H) , 7.75 (d, J = 8.8Hz, 2H) ppm.

¹³ C NMR (150 MHz, CDCl ₃ ) δ: 15.1, 31.1, 45.1, 117.6, 119.6, 124.1, 128.8, 129.0, 129.9, 155.9, 160.2, 165.9 ppm.

HRMS (ESI, m/z) calcd for C ₁₇ H ₁₈ NO ₂ [M+H] ⁺ 268.1338, found 268.1332, [M+Na] ⁺ 290.1146, found 290.1151.

Embodiment 11: the synthesis of compound 5

Compound 5 was obtained as a white solid by the same synthesis method as in Example 7, and the yield was 90%. the

¹ H NMR (400MHz, CDCl ₃ )δ: 1.43–1.53(m,2H),1.59-1.76(m,4H),2.04-2.11(m,2H),4.37(m,1H),6.18(br s, 1H), 6.98(d, J=8.8Hz, 2H), 7.03(d, J=7.8Hz, 2H), 7.16(t, J=7.5Hz, 1H), 7.37(t, J=7.5Hz, 2H) , 7.73 (d, J = 8.8Hz, 2H) ppm.

¹³ C NMR (150 MHz, CDCl ₃ ) δ: 23.7, 34.0, 51.6, 117.7, 119.5, 124.0, 128.7, 129.3, 129.8, 156.0, 160.0, 166.5 ppm.

HRMS (ESI, m/z) calcd for C ₁₈ H ₂₀ NO ₂ [M+H] ⁺ 282.1494, found 282.1489, [M+Na] ⁺ 304.1303, found 304.1308.

Embodiment 12: the synthesis of compound 6

Compound 6 was obtained as a white solid by the same synthesis method as in Example 7, and the yield was 89%. the

¹ H NMR (400MHz, CDCl ₃ )δ:0.84-0.91(m,2H),1.38-1.49(m,2H),1.64-1.79(m,4H),2.01-2.05(m,2H),3.97(m ,1H),5.96(br d,1H),7.00(d,J=8.0Hz,2H),7.04(d,J=7.5Hz,2H),7.17(t,J=7.3Hz,2H),7.38( t, J = 7.5Hz, 2H), 7.74 (d, J = 8.8Hz, 2H) ppm.

¹³ C NMR (150 MHz, CDCl ₃ ) δ: 24.9, 25.5, 33.1, 48.6, 117.7, 119.5, 124.0, 128.7, 129.5, 129.8, 156.0, 160.0, 165.9 ppm.

HRMS (ESI, m/z) calcd for C ₁₉ H ₂₂ NO ₂ [M+H] ⁺ 296.1651, found 296.1645, [M+Na] ⁺ 318.1460, found 318.1465.

Embodiment 13: the synthesis of compound 7

Compound 7 was obtained as a white solid by the same synthesis method as in Example 7, and the yield was 90%. the

¹ H NMR (400MHz, CDCl ₃ ) δ: 7.08(t, J=8.8Hz, 4H), 7.16(t, J=7.3Hz, 1H), 7.19(t, J=7.3Hz, 1H), 7.40(q , J=7.3Hz, 4H), 7.63 (d, J=7.5Hz, 2H), 7.77 (br s, 1H), 7.86 (d, J=9.0Hz, 2H).

¹³ C NMR (150 MHz, CDCl ₃ ) δ: 117.3, 119.5, 120.6, 120.7, 124.1, 124.2, 128.5, 128.8, 129.1, 129.2, 129.7, 155.5, 160.5 ppm.

HRMS (ESI, m/z) calculated for C ₁₉ H ₁₆ NO ₂ [M+H] ⁺ 290.1181, found 290.1176, [M+Na] ⁺ 312.0990, found 312.0995.

Embodiment 14: the synthesis of compound 8

Compound 8 was obtained as a white solid by the same synthesis method as in Example 7, and the yield was 47%. the

¹ H NMR (600MHz, DMSO-d ₆ )δ: 7.10(t, J=7.0Hz, 4H), 7.18(t, J=8.9Hz, 2H), 7.22(t, J=7.4Hz, 1H), 7.44 (t, J = 7.5Hz, 2H), 7.81 (q, J = 7.8Hz, 2H), 8.00 (d, J = 8.6Hz, 2H), 10.25 (s, 1H) ppm.

¹³ C NMR (150 MHz, DMSO-d ₆ ) δ: 115.5, 115.7, 117.9, 120.0, 122.6, 122.7, 124.8, 129.8, 130.4, 130.7, 156.0, 160.3, 165.1 ppm.

HRMS (ESI, m/z) calculated for C ₁₉ H ₁₅ FNO ₂ [M+H] ⁺ 308.1087, found 308.1081, [M+Na] ⁺ 330.0896, found 330.0901.

Embodiment 15: the synthesis of compound 9

Compound 9 was obtained as a white solid by the same synthesis method as in Example 7, and the yield was 50%. the

¹ H NMR (600MHz, DMSO-d ₆ )δ: 6.73(d, J=8.6Hz, 2H), 7.07(d, J=8.9Hz, 2H), 7.10(d, J=8.6Hz, 2H), 7.22 (t, J=7.3Hz, 1H), 7.45(t, J=7.6Hz, 2H), 7.50(d, J=8.6Hz, 2H), 7.96(d, J=8.9Hz, 2H), 9.92(s ,1H), 9.95(s,1H)ppm.

¹³ C NMR (150 MHz, DMSO-d ₆ ) δ: 115.0, 117.4, 119.5, 122.3, 124.3, 129.7, 129.8, 130.2, 130.7, 153.7, 155.7, 159.5, 164.2 ppm.

HRMS (ESI, m/z) calcd for C ₁₉ H ₁₆ NO ₃ [M+H] ⁺ 306.1130, found 306.1125, [M+Na] ⁺ 328.0938, found 328.0944.

Embodiment 16: the synthesis of compound 10

Compound 10 was obtained as a white solid by the same synthesis method as in Example 7, and the yield was 91%. the

¹ H NMR (400MHz, CDCl ₃ ) δ: 1.87-1.93 (m, 2H), 1.96 (m, 1H), 2.15 (m, 1H), 2.77-2.91 (m, 2H), 5.40 (q, J=5.3 Hz,1H),6.34(br s,1H),7.01(d,J=8.8Hz,2H),7.05(d,J=8.5Hz,2H),7.14-7.21(m,4H),7.37(q, J=8.5Hz, 3H), 7.77(d, J=9.0Hz, 2H) ppm.

¹³ C NMR (150MHz, CDCl ₃ )δ: 20.1, 29.2, 30.2, 47.9, 117.8, 119.6, 124.1, 126.3, 127.3, 128.7, 128.8, 129.0, 129.2, 129.9, 136.7, 137.7, 156.0, 160.03, 16mpp.

HRMS (ESI, m/z) calcd for C ₂₃ H ₂₂ NO ₂ [M+H] ⁺ 344.1651, found 344.1645, [M+Na] ⁺ 366.1459, found 366.1465.

Embodiment 17: the synthesis of compound 11

Compound 11 was obtained as a white solid by the same synthesis method as in Example 7, and the yield was 90%. the

¹ H NMR (400MHz, CDCl ₃ ) δ: 1.87-1.93 (m, 2H), 1.96 (m, 1H), 2.15 (m, 1H), 2.77-2.91 (m, 2H), 5.40 (q, J=5.3 Hz,1H),6.30(br s,1H),7.01(d,J=8.8Hz,2H),7.05(d,J=8.5Hz,2H),7.14-7.21(m,4H),7.37(q, J=8.3Hz, 3H), 7.77(d, J=8.8Hz, 2H) ppm.

¹³ C NMR (150MHz, CDCl ₃ ) δ: 20.1, 29.2, 30.1, 47.9, 117.7, 119.5, 124.1, 126.2, 127.2, 128.5, 128.9, 129.1, 129.8, 136.7, 136.7, 137.5, 155.9, 160.02, 16mpp.

HRMS (ESI, m/z) calcd for C ₂₃ H ₂₂ NO ₂ [M+H] ⁺ 344.1651, found 344.1645, [M+Na] ⁺ 366.1455, found 366.1465.

Embodiment 18: the synthesis of compound 12

Compound 12 was obtained as a white solid by the same synthesis method as in Example 7, and the yield was 89%. the

¹ H NMR (400MHz, CDCl ₃ ) δ: 1.87-1.93 (m, 2H), 1.96 (m, 1H), 2.15 (m, 1H), 2.77-2.91 (m, 2H), 5.40 (q, J=5.3 Hz,1H),6.29(br s,1H),7.01(d,J=8.8Hz,2H),7.05(d,J=8.5Hz,2H),7.14-7.21(m,4H),7.37(q, J=8.3Hz, 3H), 7.77(d, J=8.8Hz, 2H) ppm.

¹³ C NMR (150MHz, CDCl ₃ )δ: 20.1, 29.2, 30.2, 47.9, 117.8, 119.6, 124.1, 126.3, 127.3, 128.7, 128.8, 129.0, 129.2, 129.9, 136.7, 137.6, 156.0, 160.03, 16mpp.

HRMS (ESI, m/z) calcd for C ₂₃ H ₂₂ NO ₂ [M+H] ⁺ 344.1651, found 344.1645, [M+Na] ⁺ 366.1454, found 366.1465.

Embodiment 19: the synthesis of compound 13

Compound 13 was obtained as a white solid by the same synthesis method as in Example 7, and the yield was 30%. the

¹ H NMR (600MHz, CDCl ₃ ) δ: 1.88-1.92(m, 2H), 1.94(m, 1H), 2.15(m, 1H), 2.78-2.88(m, 2H), 5.38(q, J=5.9 Hz, 1H), 6.41(d, J=8.1Hz, 1H), 6.96(d, J=8.6Hz, 2H), 7.01(d, J=4.4Hz, 1H), 7.02(d, J=4.4Hz, 1H), 7.07(t, J=8.3Hz, 2H), 7.13(d, J=7.3Hz, 1H), 7.12-7.22(m, 2H), 7.33(d, J=7.3Hz, 1H) 7.76(d , J=8.8Hz, 2H)ppm.

¹³ C NMR (150MHz, CDCl ₃ ) δ: 20.1, 29.2, 30.2, 47.9, 116.4, 116.6, 117.3, 121.2, 121.3, 126.3, 127.3, 128.7, 128.9, 129.2, 136.6, 137.7, 151.7, 160.9, 165 mpp.

HRMS (ESI, m/z) calculated for C ₂₃ H ₂₁ FNO ₂ [M+H] ⁺ 362.1556, found 362.1552, [M+Na] ⁺ 384.1369, found 384.1370.

Embodiment 20: the synthesis of compound 14

Compound 14 was obtained as a white solid by the same synthesis method as in Example 7, with a yield of 35%. the

¹ H NMR (600MHz, CDCl ₃ ) δ: 1.87-1.92(m, 2H), 1.94(m, 1H), 2.14(m, 1H), 2.78-2.88(m, 2H), 5.38(q, J=5.9 Hz,1H),6.44(br s,1H),6.98(d,J=8.4Hz,4H),7.14(d,J=7.3Hz,1H),7.12-7.22(m,2H),7.33(d, J = 8.8Hz, 3H) 7.78 (d, J = 8.8Hz, 2H) ppm.

¹³ C NMR (150MHz, CDCl ₃ ) δ: 20.1, 29.2, 30.2, 47.9, 117.9, 120.8, 126.3, 127.3, 128.6, 128.9, 129.2, 129.5, 129.9, 136.6, 137.6, 154.6, 159.8, 165.8 ppm.

HRMS (ESI, m/z) calculated for C ₂₃ H ₂₁ ClNO ₂ [M+H] ⁺ 378.1261, found 378.1255, [M+Na] ⁺ 400.1080, found 400.1075.

Embodiment 21: the synthesis of compound 15

Compound 15 was obtained as a white solid by the same synthesis method as in Example 7, and the yield was 23%. the

¹ H NMR (600MHz, CDCl ₃ ) δ: 1.88-1.93(m, 2H), 1.95(m, 1H), 2.14(m, 1H), 2.78-2.88(m, 2H), 5.38(q, J=5.9 Hz,1H),6.42(br s,1H),6.92(d,J=8.6Hz,2H),6.99(d,J=8.4Hz,2H),7.13(d,J=7.2Hz,1H),7.17 -7.22 (m, 2H), 7.34 (d, J = 7.3Hz, 1H), 7.47 (d, J = 8.6Hz, 2H), 7.78 (d, J = 8.8Hz, 2H) ppm.

¹³ C NMR (150MHz, CDCl ₃ ) δ: 20.0, 29.2, 30.2, 47.9, 116.7, 118.0, 121.2, 126.3, 127.3, 128.7, 128.9, 129.2, 129.5, 132.8, 136.6, 137.6, 155.3, 159.8ppm.

HRMS (ESI, m/z) calcd for C ₂₃ H ₂₁ BrNO ₂ [M+H] ⁺ 422.0756, found 422.0750, [M+Na] ⁺ 444.0571, found 444.0570.

Embodiment 22: the synthesis of compound 16

Compound 16 was obtained as a white solid by the same synthesis method as in Example 7, with a yield of 29%. the

¹ H NMR (600MHz, CDCl ₃ )δ:1.88-1.93(m,2H),1.95(m,1H),2.13-2.17(m,1H),2.78-2.88(m,2H),5.39(q,J =6.1Hz, 1H), 6.50(br s, 1H), 7.05(d, J=9.2Hz, 2H), 7.12(t, J=8.8Hz, 3H), 7.17-7.21(m, 2H), 7.34( d, J = 8.3Hz, 1H), 7.87 (d, J = 8.8Hz, 2H), 8.21 (d, J = 9.2Hz, 2H) ppm.

¹³ C NMR (150MHz, CDCl ₃ )δ: 20.0, 29.2, 30.1, 48.0, 117.8, 119.9, 150.9, 126.3, 127.3, 128.6, 129.2, 129.3, 131.4, 136.4, 137.6, 143.1, 157.5, 162.2, 165m pp MS (ESI, m/z) Calcd. for C ₂₃ H ₂₁ N ₂ O ₄ [M+H] ⁺ 389.1501, found 389.1496, [M+Na] ⁺ 411.1321, found 411.1315.

Embodiment 23: the synthesis of compound 17

Compound 17 was obtained as a white solid by the same synthesis method as in Example 7, with a yield of 29%. the

¹ H NMR (600MHz, CDCl ₃ )δ:1.87-1.91(m,2H),1.93(m,1H),2.13-2.16(m,1H),2.78-2.86(m,2H),3.82(s,3H ), 5.38(q, J=5.9Hz, 1H), 6.40(br s, 1H), 6.92(t, J=9.2Hz, 4H), 7.00(d, J=9.2Hz, 2H), 7.13(d, J = 7.2Hz, 1H), 7.16-7.21 (m, 2H), 7.34 (d, J = 7.2Hz, 1H), 7.74 (d, J = 9.0Hz, 2H) ppm.

¹³ C NMR (150MHz, CDCl ₃ ) δ: 20.1, 29.2, 30.2, 47.8, 114.9, 116.7, 121.2, 126.3, 127.2, 128.3, 128.7, 128.8, 129.2, 136.7, 137.6, 148.9, 156.4, 161.04, 16mpp.

HRMS (ESI, m/z) calcd for C ₂₄ H ₂₄ NO ₃ [M+H] ⁺ 374.1756, found 374.1751, [M+Na] ⁺ 396.1576, found 396.1570.

Embodiment 24: the synthesis of compound 18

Add 4-phenoxybenzoic acid (107.1mg, 0.5mmol) and 3mL ethyl acetate solution into a 10mL round bottom flask, and add two drops of N,N-dimethylformaldehyde dropwise under the protection of argon at 0°C Dimethyl formamide (DMF), followed by slow dropwise addition of thionyl chloride (71.4 mg, 0.6 mmol). Stirring was continued for 1 hour at 0°C, and then changed to reflux for 4 hours, and the solvent was removed under reduced pressure for use. the

Add cyclopropylamine (28.6mg, 34.7μL, 0.5mmol), triethylamine (151.8mg, 1.5mmol) and anhydrous dichloromethane to a 10mL round bottom flask in sequence, and slowly add the acid chloride dichloromethane prepared above solution. Stir at room temperature for 2 hours, and the completion of the reaction was detected by TLC. The reaction solution was extracted with water/dichloromethane, the organic phases were combined, and the organic phase was sequentially washed with 10% hydrochloric acid solution (×3), 5% sodium bicarbonate solution (×1), water (×1), saturated sodium chloride solution Wash, dry over anhydrous magnesium sulfate, filter, and concentrate under reduced pressure to obtain a crude product. The crude product was separated on a silica gel column (ethyl acetate:petroleum ether=1:10) to obtain a white solid with a yield of 85%. the

¹ H NMR (600MHz, CDCl ₃ )δ: 0.60-0.63(m, 2H), 0.80-0.83(m, 2H), 2.87(m, 1H), 6.68(br s, 1H), 6.94(d, J= 8.9Hz, 2H), 7.01(d, J=7.9Hz, 2H), 7.15(t, J=7.3Hz, 1H), 7.35(t, J=8.6Hz, 2H), 7.74(d, J=8.6Hz ,2H)ppm.

¹³ C NMR (150 MHz, CDCl ₃ ) δ: 6.5, 23.1, 117.6, 119.6, 124.1, 128.8, 128.8, 129.9, 155.9, 160.2, 168.3 ppm.

HRMS (ESI, m/z) calcd for C ₁₆ H ₁₆ NO ₂ [M+H] ⁺ 254.1181, found 254.1176; [M+Na] ⁺ 276.0987, found 276.0995.

Embodiment 25: the synthesis of compound 19

Compound 19 was obtained as a yellow solid by the same synthesis method as in Example 24, and the two-step yield was 48%. the

¹ H NMR (600MHz, CDCl ₃ ) δ: 6.89-6.96(m, 2H), 7.08(t, J=7.5Hz, 4H), 7.21(t, J=7.5Hz, 1H), 7.41(t, J= 8.0Hz, 2H), 7.87 (d, J = 8.8Hz, 3H), 8.40 (td, J = 9.2Hz, 1H) ppm.

¹³ C NMR (150 MHz, CDCl ₃ ) δ: 102.4, 102.6, 102.7, 110.2, 110.3, 110.4, 116.8, 119.0, 123.5, 127.4, 128.1, 129.0, 154.6, 160.2, 163.8 ppm.

HRMS (ESI, m/z) calcd for C ₁₉ H ₁₄ F ₂ NO ₂ [M+H] ⁺ 326.0993, found 326.0987, [M+Na] ⁺ 348.0805, found 348.0807.

Embodiment 26: the synthesis of compound 20

Compound 20 was obtained as a yellow solid by the same synthesis method as in Example 24, and the two-step yield was 48%. the

¹ H NMR (600MHz, DMSO-d ₆ )δ: 7.12(d, J=8.9Hz, 2H), 7.13(d, J=8.6Hz, 2H), 7.24(t, J=7.5Hz, 1H), 7.46 (t, J=8.1Hz, 2H), 7.54(t, J=9.2Hz, 1H), 8.00(d, J=8.8Hz, 2H), 8.07(t, J=7.7Hz, 1H), 8.28(dd , J = 5.9, 2.8 Hz, 1H) ppm.

¹³ C NMR (150 MHz, DMSO-d ₆ ) δ: 100.3, 114.5, 117.4, 117.9, 120.1, 124.6, 124.9, 128.1, 129.1, 130.5, 130.8, 136.8, 155.9, 157.9, 160.6, 165.5 ppm.

HRMS (ESI, m/z) calculated for C ₂₀ H ₁₄ FN ₂ O ₂ [M+H] ⁺ 333.1039, found 333.1034, [M+Na] ⁺ 355.0849, found 355.0853.

Embodiment 27: the synthesis of compound 21

Compound 21 was obtained as a yellow solid by the same synthesis method as in Example 24, and the two-step yield was 48%. the

¹ H NMR (600MHz, CDCl ₃ ) δ: 7.05(d, J=8.9Hz, 2H), 7.08(d, J=7.6Hz, 2H), 7.22(t, J=7.3Hz, 1H), 7.37(dd , J=7.3Hz, 2H), 7.41 (t, J=7.6Hz, 2H), 7.79 (br s, 1H), 7.82 (d, J=8.9Hz, 2H) ppm.

¹³ C NMR (150 MHz, CDCl ₃ ) δ: 104.1, 104.3, 117.5, 119.8, 124.5, 128.8, 129.7, 130.2, 155.9, 160.9, 165.1 ppm.

HRMS (ESI, m/z) calcd for C ₁₉ H ₁₃ F ₃ NO ₂ [M+H] ⁺ 344.0898, found 344.0893, [M+Na] ⁺ 366.0708, found 366.0712.

Embodiment 28: the synthesis of compound 22

Compound 22 was obtained as a yellow solid by the same synthesis method as in Example 24, and the two-step yield was 15%. the

¹ H NMR (600MHz, CDCl ₃ ) δ: 7.05(d, J=8.9Hz, 2H), 7.08(d, J=7.6Hz, 2H), 7.22(t, J=7.3Hz, 1H), 7.42(t , J = 7.6Hz, 4H), 7.84 (d, J = 8.6Hz, 2H), 8.06 (br s, 1H) ppm.

¹³ C NMR (150 MHz, CDCl ₃ ) δ: 102.8, 116.7, 119.1, 123.7, 126.9, 128.1, 129.1, 137.8, 154.5, 158.2, 159.7, 160.5, 164.1 ppm.

HRMS (ESI, m/z) calculated for C ₁₉ H ₁₃ F ₂ BrNO ₂ [M+H] ⁺ 404.0098, found 404.0092, [M+Na] ⁺ 425.9912, found 425.9912.

Embodiment 29: the synthesis of compound 23

Compound 22 was obtained as a yellow solid by the same synthesis method as in Example 24, and the two-step yield was 10%. the

¹ H NMR (400MHz, CDCl ₃ ) δ: 7.02(m, 1H), 7.08(t, J=8.8Hz, 4H), 7.22(t, J=7.5Hz, 1H), 7.42(t, J=7.5Hz , 2H), 7.85 (d, J=9.0Hz, 2H), 7.19 (br s, 1H), 8, 46 (m, 1H) ppm.

¹³ C NMR (150MHz, CDCl ₃ ) δ: 104.1, 104.3, 117.5, 119.8, 124.5, 128.8, 129.7, 130.2, 155.9, 161.0, 165.1 ppm.

HRMS (ESI, m/z) calcd for C ₁₉ H ₁₃ F ₃ NO ₂ [M+H] ⁺ 344.0898, found 344.0893, [M+Na] ⁺ 366.0708, found 366.0712.

Embodiment 30: the synthesis of compound 24

Compound 24 was obtained as a yellow solid by the same synthesis method as in Example 24, and the two-step yield was 58%. the

¹ H NMR (600MHz, CDCl ₃ ) δ: 1.61-1.65(m, 2H), 1.68-1.72(m, 2H), 2.52(t, J=6.3Hz, 2H), 2.66(t, J=6.1Hz, 2H), 6.81(d, J=7.5Hz, 1H), 6.89(d, J=8.8Hz, 2H), 6.95(d, J=7.7Hz, 2H), 6.99(t, J=7.8Hz, 2H) , 7.27 (t, J=8.0Hz, 2H), 7.56 (d, J=7.8Hz, 1H), 7.63 (br s, 1H), 7.71 (d, J=8.6Hz, 2H) ppm.

¹³ C NMR (150MHz, CDCl ₃ ) δ: 22.5, 22.8, 24.5, 29.7, 117.8, 119.8, 120.8, 124.3, 125.7, 126.4, 128.8, 128.9, 129.3, 129.9, 135.4, 138.0, 155.8, 160.06, 165mpp.

HRMS (ESI, m/z) calcd for C ₂₃ H ₂₂ NO ₂ [M+H] ⁺ 344.1651, found 344.1645, [M+Na] ⁺ 366.1460, found 366.1465.

Embodiment 31: the synthesis of compound 25

Compound 25 was obtained as a yellow solid by the same synthesis method as in Example 24, and the two-step yield was 41%. the

¹ H NMR (600MHz, CDCl ₃ ) δ: 7.12(d, J=8.6Hz, 4H), 7.22(t, J=7.6Hz, 1H), 7.42(t, J=6.3Hz, 2H), 7.53-7.77 (m,3H),7.76(d,J=8.3Hz,1H),7.90(d,J=8.3Hz,2H),7.98(d,J=8.3Hz,2H),8.04(br s,1H), 8.15(s,1H)ppm.

¹³ C NMR (150MHz, CDCl ₃ ) δ: 117.9, 119.9, 120.9, 121.5, 124.4, 125.8, 126.1, 126.2, 126.4, 127.7, 128.8, 129.0, 129.3, 130.1, 132.5, 134.2, 155.9, 160.9pp

HRMS (ESI, m/z) calculated for C ₂₃ H ₁₈ NO ₂ [M+H] ⁺ 340.1338, found 340.1332, [M+Na] ⁺ 362.1142, found 362.1151.

Example 32: Bioassays for Antitumor Activity Detection

The purpose of this assay is to assess the in vitro cytostatic activity (ability to delay or stop tumor cell growth) or cytotoxic activity (ability to kill tumor cells) of the test sample. The cell lines involved in this embodiment are shown in Table 1 below:

Table 1 Cell lines

name N°ATCC species organize nature HepG2 HB-8065 Humanity the liver liver cancer A549 CCL-185 Humanity lung lung cancer HT-29 HTB-38 Humanity the colon colon cancer Hep3B HB-8064 Humanity the liver liver cancer PLC/PRF/5 CRL-8024 Humanity the liver liver cancer HeLa CCL-2 Humanity cervix cervical cancer

[0235] A375 CRL-1619 Humanity skin skin cancer

Cytotoxic activity was assessed using the MTT colorimetric assay:

Thiazolium blue (MTT) colorimetric method is currently the most widely used method for detecting cell survival and growth. Exogenous MTT is reduced to insoluble blue crystal formazan (Formazan) by succinate dehydrogenase in the mitochondria , which indirectly reflects the number of living cells. All cell lines from different types of human tumors used in this study were obtained from the Shanghai American Type Culture Collection (ATCC). the

The synthesized compounds were formulated into solutions with a concentration of 100mmol/L in 100% DMSO solution, and then diluted tenfold to 10, 1, and 0.1mmol/L solutions, and stored in a -20°C refrigerator. the

Take out the cryopreservation tube of human hepatocellular carcinoma HepG2 cells from the -80°C refrigerator, and thaw quickly in a 37°C water bath. After taking out the cell liquid and centrifuging, discard the supernatant, add the culture medium to blow and mix the cell pellet, add it to a 10mm culture dish already filled with the culture medium, shake well, and culture at 37°C, 5% CO ₂ , and saturated humidity . The medium was changed the next day, the medium was discarded, washed twice with PBS, and new medium was added for culture.

When the cells grow to 60-80% of the culture dish and are in the logarithmic growth cycle, the cells are digested to make a cell suspension. HepG2 (or A549, HT-29, etc.) cell suspensions were evenly inoculated on 96-well plates, and 100 μL was inoculated in each well, that is, 1000 cells/well. After the cells were incubated overnight, the drug-containing media with different concentrations were diluted 1000 times to obtain drug-containing media with concentration units of 0.1, 1, 10, and 100 μmol/L, which were added to 96-well plates. Add 100 μL of drug-containing medium to each well, set 8 auxiliary wells for each concentration as a group, and set another group as a control group with 0.1% DMSO medium. The dressing was changed every 24 hours, and the drug was administered continuously for 72 hours. After 72 hours, add 75 μL of MTT-containing medium (the ratio of MTT solution to medium is 1:4) to each well, incubate in the dark for 4 hours, discard the MTT-containing medium, add 120 μL of 100% DMSO solution to each well, and set a blank The group without cells was directly added to DMSO solution. After fully oscillating and mixing, measure the absorbance OD value of each well at 492 nm (absorption wavelength) on a microplate reader. The cell growth inhibition rate was calculated according to the following formula:

Use GraphPad Prism5.0 software to carry out linear analysis according to the inhibition rate corresponding to each drug concentration, obtain a linear equation, and calculate the corresponding drug concentration (unit: μ mol/L) when the inhibition rate is 50%, which is the effect of the test sample on the tumor. The half-inhibitory concentration (IC ₅₀ value) of the cells. Each drug experiment was repeated three times to take the average value of _IC50 . The specific results are shown in Table 2 and Table 3 below:

Table 2 The compound of the present invention is to HepG2, the biological activity data of A549 and HT-29 cell

Table 3 The biological activity data of compound 8 and compound 12 of the present invention on other different cell lines

The experimental groups of compound 8 and 12 were subjected to western blot analysis of different drug concentrations to observe the time-course effect and dose-effect relationship of promoting the expression of apoptosis factor proteins such as p21, p53 and Cl-caspase3. The results combined with the above MTT ratio The color measurement results are shown in Figures 1 to 4, and the results show that the substituted diaryl ether compounds of the present invention can significantly inhibit the malignant proliferation phenotype of cancer cells, and promote the expression of apoptosis factor proteins such as p21, p53 and Cl-caspase3. Express. the

The above is only a preferred embodiment of the present invention, so the scope of the present invention cannot be limited accordingly, that is, equivalent changes and modifications made according to the patent scope of the present invention and the content of the specification should still be covered by the present invention In the range. the

Claims (10)

1. A substituted diaryl ether compound, characterized in that: its structure is as follows: Z is -CH- or -N-; W is -CH- or -N-; Y is -O-, -CH ₂ -, -NH- or -NR _y -; R ₁ is hydrogen, halogen, NO ₂ , CN, CF ₃ , OR _a , COR _a , COOR _a , SO ₂ R _a , SO ₂ NR _a R _b , NR _a R _b , NR _a COR _b , unsubstituted\substituted One of the C ₁ -C ₄ alkyl, unsubstituted\substituted aryl or unsubstituted\substituted heterocyclic groups; R ₂ is hydrogen, unsubstituted \ substituted C ₁ -C ₄ alkyl, or halogen; R is unsubstituted\substituted C ₁ -C ₁₂ alkyl, unsubstituted\substituted C ₃ -C ₁₂ cycloalkyl, unsubstituted\substituted C ₂ -C ₁₂ alkenyl, unsubstituted\substituted aryl , one of substituted alkylamines or unsubstituted\substituted heterocyclic groups. 2. A substituted diaryl ether compound as claimed in claim 1, characterized in that: said R _a is hydrogen, unsubstituted\substituted C ₁ -C ₄ alkyl, unsubstituted\substituted C ₂ - One of C ₄ alkenyl, unsubstituted\substituted aryl or unsubstituted\substituted heterocyclic group. 3. A substituted diaryl ether compound as claimed in claim 2, characterized in that: said R _b is hydrogen, unsubstituted\substituted C ₁ -C ₄ alkyl, unsubstituted\substituted C ₂ - One of C ₄ alkenyl, unsubstituted\substituted aryl or unsubstituted\substituted heterocyclic group. 4. A substituted diaryl ether compound as claimed in claim 3, characterized in that: said R _y is one of hydrogen and unsubstituted/substituted C ₁ -C ₃ alkyl. 5. A substituted diaryl ether compound as claimed in any one of claims 1 to 4, wherein said Z and W are both -CH-, and said Y is -O-, -NH -or-NR _y . 6. A substituted diaryl ether compound according to any one of claims 1 to 4, wherein said Z is -CH-, said W is -N-, and said Y is - O-. 7. A substituted diaryl ether compound according to any one of claims 1 to 4, characterized in that: said Z and W are both -N-, Y is -O- or -CH ₂ - . 8. A preparation method of the substituted diaryl ether compound according to any one of claims 1 to 7, characterized in that: its reaction process is as follows: Specifically: under the action of a base, the substituted halogenated benzene or substituted aniline shown in the compound (II) is reacted with the substituted ethyl benzoate shown in the compound (III) to generate the compound (IV), and the compound (IV) is carried out Condensation reaction with substituted amine after hydrolysis or acid chlorination to obtain compound (I), namely the substituted diaryl ether compound of the present invention. 9. The preparation method of substituted diaryl ether compounds as claimed in claim 8, characterized in that: said X is F, Cl, Br or I. 10. A use of the substituted diaryl ether compound according to any one of claims 1 to 9 in the preparation of tumor drugs.