CN111875601A - Synthetic method and application of indolizine compound - Google Patents

Abstract

A method for synthesizing indolazine compounds, comprising the following steps: mixing a benzopyrone derivative, a bromopyridinium salt derivative, a base and a solvent in a 10ml round-bottomed flask, and stirring at 25°C-100°C for 2 -48 hours; after the reaction is completed, the crude product is purified by silica gel chromatography to obtain the target compound, and its structural formula is shown in formula I,

Description

A kind of synthetic method and application of indolazine compound

technical field

The invention belongs to the technical field of chemistry and medicine, and particularly relates to a preparation method of a novel indolazine compound and its use in treating tumors.

Background technique

Cancer (malignant tumor) is one of the diseases that seriously threaten the global human health, and it is also a major problem faced by the medical field all over the world. According to epidemiological surveys, in 2018, the global cancer incidence rate has reached 2‰. In my country, there are about 3.8 million new cancer cases and 2.29 million deaths each year. The overall cancer incidence rate increases by about 3.9% every year on average, and the incidence and mortality rates show an upward trend year by year. Its treatment and prevention have attracted widespread attention. The current tumor treatment methods mainly include surgical resection, radiotherapy, and chemotherapy, but chemotherapy is still the main treatment. Currently, there are many kinds of chemical drugs used in clinical treatment of cancer, such as platinum, nitrogen mustard, triazole, etc., but most of the drugs are limited due to their high toxicity, many adverse reactions and low bioavailability. Therefore, the development of high-efficiency and low-toxicity anticancer drugs has become a key research topic in the field of medicinal chemistry.

In view of the potential application of indolizine anticancer drugs in tumor treatment, the research on small molecule drugs with indolizine skeleton as the core pharmacophore is a current hotspot. Among the discovered indolazines, the known biological activities include antibacterial, anti-tuberculosis, anticancer, anti-inflammatory, antioxidant, antihistamine and uterine trophic activities, and can also be used as aromatase inhibitors, Phosphodiesterase inhibitors, phosphatase inhibitors, and vasoconstrictor-converting enzyme (ACE) inhibitors. Combined with this research work, it is further discovered that novel indolazine compounds have potential value in the treatment of malignant tumors.

SUMMARY OF THE INVENTION

The present invention aims to solve the technical problem of a new class of substituted indolazine compounds, which can effectively inhibit various types of cancers or tumors.

In order to solve the above-mentioned technical problems, the present invention adopts the following technical solutions:

A method for synthesizing indolazine compounds, comprising the steps of: mixing a benzopyrone derivative, a bromopyridinium salt derivative, a base and a solvent in a 10ml round-bottomed flask, and stirring at 25°C-100°C for 2 -48 hours. After the reaction is completed, the crude product is purified by silica gel chromatography to obtain the target compound, whose structural formula is shown in formula I,

In the above structural formula:

R ¹ is independently substituted with one or more of H, halogen, alkyl, hydroxyl, amino, carboxyl, carbonyl, ester, cyano, alkynyl, alkenyl, phenyl, and aryl;

R ² is independently substituted by one or more of H, hydroxy, alkoxy, phenyl, pyridyl, and aryl;

X ₁ is selected from C and N;

X ₂ is selected from OH and SH;

R ³ is independently selected from one or more of H, halogen, alkyl, hydroxyl, amino, carboxyl, carbonyl, ester, cyano, alkynyl, alkenyl, phenyl, pyridine, thiophene, heterocycle, aryl replaced.

The aromatic group is selected from the group consisting of phenyl, naphthyl, anthracenyl, phenanthryl, pyridyl, thienyl, furanyl, thiazolyl, imidazolyl, oxazolyl, indolizinyl, quinolinyl, indolanyl, benzothiazolyl, benzisothiazolyl, triazolopyridyl, indolizinopyridyl, benzoxazolyl, triazolopyridyl, pyridopyrazinyl, quinazolinyl, benzoxadi azolyl, benzothiadiazolyl, benzoindolizinyl; the alkyl group refers to a C _1-10 straight or branched saturated carbon chain; the halogen is selected from fluorine, chlorine, bromine and iodine.

Synthetic method of indolazine compounds, benzopyrone, bromopyridinium salt, base and solvent are mixed in a 10ml round bottom flask, and stirred at 80°C for 12 hours; after completion of the reaction, the crude product is purified by silica gel chromatography to obtain the target compound.

The base is selected from one of DBU, DIPEA, potassium carbonate, sodium carbonate, potassium acetate, potassium hydroxide, sodium hydroxide; the solvent is selected from 1,4-dioxane, N,N-dimethyl One of formamide, dimethyl sulfoxide, methanol, ethanol, toluene, NMP, THF.

The base is preferably DBU; the solvent is preferably 1,4-dioxane.

The synthesis method of indolazine compounds, benzopyrone, bromopyridinium salt and DBU are mixed in a 10ml round bottom flask, and stirred at 80 ° C for 6 hours; after the reaction is completed, the crude product is purified by silica gel chromatography to obtain the target compound.

A method for synthesizing indolazine compounds, benzopyrone, bromopyridinium salt, metal catalyst, base and 1,4-dioxane are mixed in a 10ml round-bottomed flask, and stirred at 60° C. for 6 hours; After the reaction was completed, the crude product was purified by silica gel chromatography to obtain the title compound. The metal catalyst is selected from one of cuprous iodide, cuprous bromide, cuprous chloride, cupric chloride, and copper sulfate; the base is selected from DBU, DIPEA, potassium carbonate, sodium carbonate, potassium acetate, hydroxide One of potassium and sodium hydroxide.

The metal catalyst is preferably cuprous iodide; the base is preferably DBU.

Indolazine compounds are used for tumors or cancers such as gastric cancer, cervical adenocarcinoma, colon cancer, lung cancer, liver cancer, glioma, esophagus cancer, colon cancer, nasopharyngeal cancer, breast cancer, lymphoma, kidney, pancreas, Bladder cancer, ovarian cancer, uterine cancer, bone cancer, gallbladder cancer, lip cancer, melanoma, tongue cancer, throat cancer, blood cancer, prostate cancer, brain tumor, squamous cell carcinoma, skin cancer, hemangioma, lipoma, cervical cancer and thyroid cancer.

The present invention also provides methods of treating tumors or cancers, limited to in vitro cellular tests.

The present invention relates to the inhibitory activity of said compounds on human hepatoma cells and ovarian cancer cells.

Tumor cell inhibition rate=[1-(experimental well-blank well)/(control well-blank well)]*100%. Terminology Explanation: "Alkyl" is intended to include branched and straight chain saturated hydrocarbon groups having the specified number of carbon atoms. "Halogen" refers to fluorine, chlorine, bromine and iodine atoms. "Indolizine" can also be called indolizine or indolizine.

Abbreviations: DBU refers to 1,8-diazabicycloundec-7-ene; DIPEA refers to N,N-diisopropylethylamine; NMP refers to N-methylpyrrolidone; THF refers to tetrahydrofuran.

The beneficial effects of adopting the above technical scheme are:

The present invention provides three different preparation methods for synthesizing novel indolazine antitumor compounds, and the activity test finds that the compounds have tumor cell inhibitory activity, which further broadens the types of candidate drugs for treating tumors.

Detailed ways

It must be noted that the embodiments of the present invention are used to illustrate rather than limit the present invention. Simple improvements made to the present invention according to the essence of the present invention all belong to the scope of protection of the present invention.

In describing the present invention in the foregoing specification, the purpose of providing the embodiments is to illustrate the actual operation of the present invention and the meaning of the present invention.

The present invention will be further described below in conjunction with specific embodiments. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention. The specific conditions in the examples are not limited to a certain one, and the solvent used in the preparation of compounds 1-10 can be 1,4-dioxane, N,N dimethylformamide, thionyl chloride, methanol, ethanol, etc. , the temperature range can be 25-100 ℃, the alkali can be DBU, triethylamine, potassium carbonate, sodium carbonate, sodium bicarbonate, potassium bicarbonate, potassium hydroxide, sodium hydroxide, wherein DBU is the best. Percentages and parts are weight percentages and parts unless otherwise specified.

The experimental materials and reagents used in the following examples can be obtained from commercial sources unless otherwise specified.

In each example, the nuclear magnetic resonance spectrum was measured by Bruker Ascend III 400M nuclear magnetic resonance apparatus (TMS was used as the internal standard); the high-resolution mass spectrometer was measured by Bruker MicrOTOF Q II; the melting point was measured by X-6 precision micro melting point tester; the silica gel was 200 -300 mesh.

Among them, the Chinese names of reagents represented by chemical formulas or English letter abbreviations are as follows:

Example 1:

Preparation of ethazole:

o-Hydroxyacetophenone (10 mmol) and dimethylformamide dimethyl acetal (10 ml) were heated to reflux at 90°C for 1 hour. The reaction was monitored with a thin layer plate. After completion of the reaction, residual solvent was removed by rotary evaporator. Then, a mixed reagent of concentrated hydrochloric acid and dichloromethane (V:V=1:10) was added to the mixture, and the mixture was heated under reflux at 45°C for 2 hours. After cooling, the mixture was extracted with dichloromethane. The organic phase was washed with saturated sodium bicarbonate solution, then brine, dried over magnesium sulfate, filtered, concentrated, and column chromatographed to give the desired ethazole. Unless otherwise specified, other ethazole derivatives were prepared by this method.

Preparation of bromonium salt:

Ethyl bromoacetate (1.2 equiv) was reacted with pyridine (1.0 equiv) in THF solution for 6 hours at room temperature and the resulting precipitate was filtered and washed twice with petroleum ether. Finally, the prepared pyridinium salt was dried under vacuum for 2 hours. Unless otherwise specified, other bromonium salts are prepared by this method.

Preparation of ethyl-(2-hydroxybenzoyl)indolazine-3-carboxylate (compound 1):

method one:

4H-benzopyran-4-one (0.5 mmol), ethyl bromopyridinium ethyl onium salt (0.55 mmol), 1,8-diazabicycloundec-7-ene (DBU, 1.0 mmol) and 1,4-dioxane (3.0 ml) were dissolved in a 10 ml round bottom flask and stirred at 80°C for 12 hours. Reactions were detected by thin-layer silica gel plates. After completion of the reaction, the reaction mixture was cooled to room temperature and concentrated in vacuo to obtain a crude product, which was further purified by silica gel chromatography (ethyl acetate:petroleum ether=1:5) to finally obtain the target compound 1. Yellow solid (126mg, 81%), Mp107-108°C. 1H NMR (400MHz, CDCl ₃ ) δ 12.03 (s, 1H), 9.58 (d, J=7.0Hz, 1H), 8.46 (dt, J=9.0 ,1.1Hz,1H),7.95–7.83(m,2H),7.52–7.45(m,1H),7.41(m,1H),7.12–7.03(m,2H),7.01–6.92(m,1H), 4.42(q, J=7.1Hz, 2H), 1.42(t, J=7.1Hz, 3H). 13C NMR (101MHz, CDCl ₃ )δ193.02, 162.23, 161.10, 140.17, 134.81, 131.81, 128.05, 127.05, 125.56, 120.94,120.32,118.72,118.22,115.57,115.18,112.10,60.57,14.53.HRMS(ESI+) calculated for C ₁₈ H ₁₆ NO ₄ [M+H] ⁺ :310.1079,found:310.1074.

Method Two

Combine 4H-benzopyran-4-one (0.5 mmol), ethyl bromopyridinium ethyl onium salt (0.55 mmol) and 1,8-diazabicycloundec-7-ene (DBU, 2 ml) In a 10 ml round bottom flask, seal with a rubber stopper and stir at 80°C for 6 hours. Reactions were detected by thin-layer silica gel plates. After the completion of the reaction, the reaction mixture was cooled to room temperature and concentrated in vacuo to obtain a crude product, which was further purified by silica gel chromatography (ethyl acetate:petroleum ether=1:5) to finally obtain the target compound 1 as a yellow solid (138 mg, 89%).

Method three

4H-benzopyran-4-one (0.5 mmol), ethyl bromopyridinium ethyl onium salt (0.55 mmol), cuprous iodide (0.025 mmol), 1,8-diazabicycloundec- 7-ene (DBU, 1.0 mmol) and 1,4-dioxane (3.0 ml) were dissolved in a 10 ml round bottom flask and stirred at 80°C for 12 hours. Reactions were detected by thin-layer silica gel plates. After the reaction was completed, the reaction mixture was cooled to room temperature and concentrated in vacuo to obtain a crude product, which was further purified by silica gel chromatography (ethyl acetate:petroleum ether=1:5) to finally obtain the target compound 1 as a yellow solid (147 mg, 95%).

Embodiment 2

Preparation of ethyl-(4-hydroxy-4'-methoxy-[1,1'-diphenyl]-3-carbonyl)indolazine-3-carboxylate (compound 2):

Referring to method 1 of Example 1, replace 4H-benzopyran-4-one (0.5 mmol) with 6-(4-methoxyphenyl)-4H-benzopyran-4-one (0.5 mmol) ), other conditions remain unchanged, the target compound 2 was prepared. Yellow solid (170mg, 82%), M.p. 169-171°C. 1H NMR (400MHz, CDCl3) δ 11.87 (s, 1H), 9.61 (d, J=7.0Hz, 1H), 8.48 (d, J=9.0 Hz, 1H), 8.05(d, J=2.3Hz, 1H), 7.99(s, 1H), 7.69(dd, J=8.6, 2.3Hz, 1H), 7.51–7.47(m, 2H), 7.44(m ,1H),7.15–7.08(m,2H),6.99–6.94(m,2H),4.41(q,J=7.1Hz,2H),3.85(s,3H),1.41(t,J=7.1Hz, 3H).13C NMR(101MHz,CDCl3)δ213.51,193.14,161.11,158.91,140.16,133.25,132.79,131.60,129.65,128.10,127.67,127.12,125.68,121.15,120.30,118.55,115.62,115.34,114.31,114.00, 112.64,112.20,60.51,55.39,14.50.HRMS(ESI+)calculated for C25H22NO5[M+H]+:416.1498,found:416.1493.

Embodiment 3

Preparation of ethyl-(2-hydroxy-4,5-dimethoxybenzoyl)indolazine-3-carboxylate (compound 3):

Referring to method 1 of Example 1, replace 4H-benzopyran-4-one (0.5 mmol) with 6,7-dimethoxy-4H-benzopyran-4-one (0.5 mmol), other Under the same conditions, the target compound 3 was prepared. Yellow solid (143mg, 78%), Mp142-144°C. 1H NMR (400MHz, CDCl 3 ) δ 12.61 (s, 1H), 9.58 (d, J=7.1Hz, 1H), 8.38 (d, J=8.1 Hz, 1H), 7.95 (s, 1H), 7.51–7.33 (m, 2H), 7.14–6.95 (m, 1H), 6.58 (s, 1H), 4.41 (q, J=7.1Hz, 2H), 3.96 (s, 3H), 3.84 (s, 3H), 1.40 (t, J=7.1Hz, 3H). 13C NMR (101MHz, CDCl 3) δ 191.74, 161.13, 160.05, 155.71, 141.64, 139.71, 127.93, 126.43, 124.73 ,120.20,115.31,114.98,113.66,112.36,112.31,100.86,60.46,56.60,56.20,14.50.HRMS(ESI+)calculated for C ₂₀ H ₂₀ NO ₆ [M+H] ⁺ :370.1291,found:370.1284.

Embodiment 4

Preparation of ethyl-1-(1-hydroxy-2-naphthyl)indolazine-3-carboxylate (compound 4):

Referring to method 1 of Example 1, 4H-benzopyran-4-one (0.5mmol) was replaced with 4H-benzo[h]chromone-4-one (0.5mmol), other conditions were unchanged, and the obtained target compound 4. Yellow solid (135mg, 75%), Mp120-122°C. 1H NMR (400MHz, CDCl 3) δ 14.06 (s, 1H), 9.58 (d, J=7.0Hz, 1H), 8.50 (d, J=8.3 Hz, 1H), 8.47(d, J=9.0Hz, 1H), 7.98(s, 1H), 7.91(d, J=8.8Hz, 1H), 7.79(d, J=8.1Hz, 1H), 7.67– 7.58(m,1H),7.54(ddd,J=8.1,7.0,1.1Hz,1H),7.40(ddd,J=8.9,6.8,1.1Hz,1H),7.33(d,J=8.8Hz,1H) , 7.07 (td, J=6.9, 1.3Hz, 1H), 4.42 (q, J=7.1Hz, 2H), 1.42 (t, J=7.1Hz, 3H). 13C NMR (101MHz, CDCl 3) δ 193.06, 162.61 ,161.18,140.07,136.70,129.64,128.01,127.33,126.76,126.56,125.73,125.48,125.42,124.26,120.39,117.88,115.42,115.20,113.93,112.42,60.55,14.55.HRMS(ESI+)calculated forC ₂₂ H ₁₈ NO ₄ [M+H]+: 360.1236, found: 360.1229.

Embodiment 5

Preparation of ethyl 3-(2-hydroxybenzoyl)pyrro[1,2-a]quinoline-1-carboxylate (compound 5):

Referring to Method 1 of Example 1, the crude compound 5 was prepared by replacing the ethyl bromopyridinium ethyl onium salt (0.55 mmol) with the bromoquinoline ethyl onium salt (0.55 mmol). Further purification by silica gel chromatography (ethyl acetate:petroleum ether=1:3) finally gave the target compound 5. Yellow solid (108mg, 60%), Mp135-137℃. 1HNMR (400MHz, CDCl 3) δ 12.10(s, 1H), 8.41(d, J=8.7Hz, 1H), 8.28(d, J=9.3Hz ,1H),7.90(dd,J＝7.9,1.5Hz,2H),7.85(s,1H),7.82(dd,J＝7.9,1.2Hz,1H),7.66(m,2H),7.56–7.47( m, 2H), 7.08 (dd, J=8.3, 0.7Hz, 1H), 7.01–6.95 (m, 1H), 4.46 (q, J=7.1Hz, 2H), 1.44 (t, J=7.1Hz, 2H) ).13C NMR(101MHz,CDCl 3)δ193.85,162.56,161.72,140.11,135.28,133.19,132.16,128.74,128.69,128.57,127.40,125.74,125.60,120.81,120.55,120.22,118.77,118.33,118.01,114.41, 61.30, 14.44. HRMS(ESI+) calculated for C ₂₂ H ₁₈ NO ₄ [M+H]+: 360.1236, found: 360.1233.

Embodiment 6

Preparation of ethyl 5-(2-hydroxybenzoyl)pyrro[1,2-b]pyridazine-7-carboxylate (compound 6):

Referring to method 1 of Example 1, bromopyridazine ethyl acetate (0.55 mmol) was replaced with bromopyridazine ethyl acetate (0.55 mmol), and other conditions remained unchanged to prepare crude compound 6. Further purification by silica gel chromatography (dichloromethane:methanol=50:1) finally gave the target compound 6. Yellow solid (85mg, 55%), Mp100-102°C. 1HNMR (400MHz, CDCl 3) δ 11.92 (s, 1H), 8.71 (dd, J=9.2, 1.9Hz, 1H), 8.60 (dd, J= 4.5, 1.8Hz, 1H), 7.94 (s, 1H), 7.93 (dd, J=8.0, 1.6Hz, 1H), 7.55–7.49 (m, 1H), 7.18 (dd, J=9.1, 4.5Hz, 1H) ),7.09(dd,J=8.4,0.7Hz,1H),7.03–6.97(m,1H),4.47(q,J=7.1Hz,2H),1.43(t,J=7.1Hz,3H).13C NMR (101MHz, CDCl 3) δ193.18, 162.53, 159.12, 144.92, 135.52, 134.17, 131.90, 128.66, 123.39, _120.36 , 118.97, 118.45, 117.45, 111.91, 99.99, 60.92, 14 calculated for CES. _15N2O4 [M ₊ H] ₊ : 311.1032, found: 311.1026.

Embodiment 7

Preparation of ethyl 1-(2-hydroxybenzoyl)pyrro[2,1-a]phthalazine-7-carboxylate (compound 7):

Referring to method 1 of Example 1, the crude compound 7 was prepared by replacing ethyl bromopyridinium ethyl onium salt (0.55 mmol) with bromophthalazine ethyl onium salt (0.55 mmol), with other conditions unchanged. Further purification by silica gel chromatography (dichloromethane:methanol=60:1) gave the target compound 7 finally. Yellow solid (90mg, 50%), Mp230-232℃. 1HNMR (400MHz, CDCl 3) δ 12.15(s, 1H), 8.83(s, 1H), 8.65(d, J=8.3Hz, 1H), 7.91 (d, J=7.2Hz, 1H), 7.79 (td, J=8.4, 1.4Hz, 2H), 7.74–7.68 (m, 1H), 7.66 (s, 1H), 7.58–7.52 (m, 1H), 7.12(dd,J=8.4,0.8Hz,1H),6.98–6.91(m,1H),4.45(q,J=7.1Hz,2H),1.42(t,J=7.1Hz,3H).13C NMR( 101MHz,CDCl 3)δ196.84,163.27,159.26,146.70,136.34,133.50,132.96,129.57,128.76,127.97,126.60,125.13,122.50,121.60,120.55,119.76,118.96,118.46,114.21,60.82,14.45.HRMS(ESI+ )calculated for C ₂₁ H ₁₇ N ₂ O ₄ [M+H]+:361.1188, found:361.1182.

Embodiment 8

Preparation of (1-(2-hydroxybenzoyl)indolazin-3-yl)(4-methoxyphenyl)methadone (compound 8):

Referring to method 1 of Example 1, replace bromopyridinium ethyl acetate (0.55mmol) with 1-(2-(4-methoxyphenyl)-2-oxoethyl)pyridine-1-bromonium salt (0.55 mmol), other conditions were unchanged, and the crude compound 8 was prepared. The crude product was further purified by silica gel chromatography (ethyl acetate:petroleum ether=1:3) to finally obtain the target compound 8. Yellow solid (136mg, 58%), M.p. 123-125°C. 1H NMR (400MHz, CDCl 3 ) δ 11.98 (s, 1H), 9.92 (d, J=7.0Hz, 1H), 8.47 (d, J= 8.9Hz,1H),7.83(d,J=8.7Hz,2H),7.77(dd,J=7.9,1.4Hz,1H),7.70(s,1H),7.54–7.47(m,1H),7.44( t,J＝7.8Hz,1H),7.15(t,J＝6.9Hz,1H),7.05(d,J＝8.3Hz,1H),6.99(d,J＝8.7Hz,2H),6.88(t, J=7.6Hz, 1H), 3.88(s, 3H). 13C NMR (101MHz, CDCl 3) δ 193.14, 184.62, 162.76, 162.28, 140.66, 134.94, 132.07, 131.62, 131.26, 129.25, 129.09, 128.59, 122. ,120.10,118.74,118.31,116.13,113.82,112.72,55.50.HRMS(ESI+)calculated for C 23H 18NO 4[M+H]+:372.1236,found:372.1234.

Embodiment 9

Preparation of ethyl 1-(2-(acryloyloxy)benzoyl)indolazine-3-carboxylate (compound 9):

Compound 1 (155 mg, 0.5 mmol) was dissolved in 20 ml of chloroform under an ice bath, and then acryloyl chloride (136 mg, 1.5 mmol) and trimethylamine (253 mg, 2.5 mmol) were sequentially added dropwise. The reaction was kept stirring continuously at 0°C for 2 h, then the temperature was returned to room temperature. The reaction was monitored with a thin layer plate. After the reaction was completed, the reaction solution was extracted with ethyl acetate (20 ml×3). The organic phase was dried _{over Na2SO4} and concentrated under reduced pressure. The crude material was purified by column chromatography (petroleum ether/ethyl acetate=5:1) to obtain compound 9 (131 mg, 72%) as a pale yellow solid. Yellow solid (131mg, 72%), Mp112-114°C. 1H NMR (400MHz, CDCl 3) δ 9.55 (d, J=7.0Hz, 1H), 8.57 (d, J=8.9Hz, 1H), 7.66 ( s, 1H), 7.60 (dd, J=7.6, 1.5Hz, 1H), 7.54 (td, J=7.9, 1.6Hz, 1H), 7.43–7.33 (m, 2H), 7.28 (d, J=7.4Hz) ,1H),7.06(td,J＝7.0,1.2Hz,1H),6.43(dd,J＝17.3,1.1Hz,1H),6.15(dd,J＝17.3,10.4Hz,1H),5.85(dd, J=10.5, 1.1Hz, 1H), 4.35 (q, J=7.1Hz, 2H), 1.36 (t, J=7.1Hz, 3H). 13CNMR (101MHz, CDCl 3) δ 187.94, 164.20, 161.13, 147.87, 139.58 ,133.66,132.74,131.18,129.72,127.99,127.46,127.36,126.15,125.73,123.17,120.52,115.59,115.05,113.27,60.41,14.49.HRMS(ESI 6.185 calculated for C 21H ,found:364.1189.

Embodiment ten

Preparation of ethyl-1-(2-((methylsulfonyl)oxy)benzoyl)indolazine-3-carboxylate (compound 10):

Compound 1 (155 mg, 0.5 mmol) was dissolved in CH ₂ Cl ₂ (20 ml) and Et ₃ N (84 μL, 0.6 mmol) was added. To the mixture was added dropwise methanesulfonyl chloride (63 μL, 0.81 mmol) at 0°C. The reaction mixture was stirred at room temperature for 4 hours, then water (20ml) was added and the product was extracted with ethyl acetate (50ml). The separated organic phase was washed with sodium chloride solution and dried _over anhydrous _Na2SO4 . Concentration in vacuo to remove the solvent gave title compound 10 as a pale yellow solid (165 mg, 85%). White solid (165mg, 85%), Mp118-200°C. 1H NMR (400MHz, CDCl ₃ ) δ 9.60 (d, J=7.0Hz, 1H), 8.61 (d, J=8.9Hz, 1H), 7.56 ( m, 4H), 7.52–7.41 (m, 2H), 7.13 (t, J=7.0Hz, 1H), 4.36 (q, J=7.1Hz, 2H), 3.12 (s, 3H), 1.37 (t, J ＝7.1Hz,3H).13CNMR(101MHz,CDCl 3)δ187.48,161.02,146.06,139.57,134.35,131.32,129.71,128.26,127.93,127.00,126.23,123.73,120.41,115.90,115.48,113.24,60.57,37.98, 14.47. HRMS(ESI+) calculated for C ₁₉ H ₁₈ NO ₆ S[M+H]+388.0854, found: 388.0853.

Embodiment 11

Antitumor activity study of compounds 1-10.

Detection method: CCK8 kit;

Materials: Tumor cells: HEPG2 (human hepatoma blast cells), NCI-H460 (human non-small cell lung cancer cells), CCK-8 kit;

Experimental method: with 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfobenzene)-2H-tetrazolium monosodium Salt-based CCK-8 kit to detect the effect of compounds on tumor cell proliferation: tumor cells in logarithmic growth phase HEPG2 or NCI-H460 were digested with 0.25% trypsin, and made into a single medium containing 10% FBS. The cell suspension was seeded in a 96-well plate at 5000 cells/100 μl per well, and incubated at 37°C in a constant temperature incubator containing 5% CO ₂ for 24 h. Cells were treated with 10 μM of various compounds and the same concentration of DMSO for 72 h. After the incubation, CCK8 solution was added, and after 3 hours, the absorbance of each well at 450 nm was detected with a microplate reader, and the inhibition rate (%) was obtained according to the following method.

Cell inhibition rate=[1-(experimental well-blank well)/(control well-blank well)] 100%;

Table 1 Activity test results of compounds 1-10

Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, and substitutions can be made in these embodiments without departing from the principle and spirit of the invention and modifications, the scope of the present invention is defined by the appended claims and their equivalents.

Claims (10)

1. a synthetic method of indolazine compound, is characterized in that: it may further comprise the steps: benzopyrone derivative, bromopyridinium salt derivative, alkali and solvent are mixed in 10ml round-bottomed flask, and in Stir at 25°C-100°C for 2-48 hours; after the reaction is completed, the crude product is purified by silica gel chromatography to obtain the target compound, whose structural formula is shown in formula I,

In the above structural formula: R ¹ is independently substituted with one or more of H, halogen, alkyl, hydroxyl, amino, carboxyl, carbonyl, ester, cyano, alkynyl, alkenyl, phenyl, and aryl; R ² is independently substituted by one or more of H, hydroxy, alkoxy, phenyl, pyridyl, and aryl; X ₁ is selected from C and N; X ₂ is selected from OH and SH; R ³ is independently selected from one or more of H, halogen, alkyl, hydroxyl, amino, carboxyl, carbonyl, ester, cyano, alkynyl, alkenyl, phenyl, pyridine, thiophene, heterocycle, aryl replaced. 2. the synthetic method of indolazine compound according to claim 1 is characterized in that: described aromatic group is selected from phenyl, naphthyl, anthracenyl, phenanthryl, pyridyl, thienyl, furanyl, thiazolyl , imidazolyl, oxazolyl, indolizinyl, quinolinyl, indolanyl, benzothiazolyl, benzisothiazolyl, triazolopyridyl, indolizinopyridyl, benzoxazolyl, Triazolopyridyl, pyridopyrazinyl, quinazolinyl, benzoxadiazolyl, benzothiadiazolyl, benzoindolizinyl; the alkyl refers to C _1-10 straight chain or branched Chain saturated carbon chain; the halogen is selected from fluorine, chlorine, bromine, iodine. 3. the synthetic method of indolazine compound according to claim 1, is characterized in that: benzopyrone derivative, bromopyridinium salt derivative, alkali and solvent are mixed in 10ml round-bottomed flask, and at 80 Stir at °C for 12 hours; after completion of the reaction, the crude product is purified by silica gel chromatography to obtain the title compound. 4. according to the synthetic method of the described indolazine compound of claim 3, it is characterized in that: described alkali is selected from wherein one of DBU, DIPEA, salt of wormwood, sodium carbonate, potassium acetate, potassium hydroxide, sodium hydroxide ; The solvent is selected from one of 1,4-dioxane, N,N-dimethylformamide, dimethyl sulfoxide, methanol, ethanol, toluene, NMP, and THF. 5. The method for synthesizing indolazine compounds according to claim 3 or 4, wherein the base is DBU; and the solvent is 1,4-dioxane. 6. the synthetic method of indolazine compound according to claim 1, is characterized in that: benzopyrone, bromopyridinium salt and DBU are mixed in 10ml round-bottomed flask, and stirred 6 hours at 80 ℃; Reaction After completion, the crude product was purified by silica gel chromatography to give the title compound. 7. the synthetic method of indolazine compound according to claim 1 is characterized in that: benzopyrone, bromopyridinium salt, metal catalyst, alkali and 1,4-dioxane are mixed in 10ml round bottom flask and stirred at 60°C for 6 hours; after completion of the reaction, the crude product was purified by silica gel chromatography to obtain the title compound. 8. the synthetic method of indolazine compound according to claim 7, is characterized in that: described metal catalyst is selected from wherein of cuprous iodide, cuprous bromide, cuprous chloride, cupric chloride, cupric sulfate One; the base is selected from one of DBU, DIPEA, potassium carbonate, sodium carbonate, potassium acetate, potassium hydroxide, and sodium hydroxide. 9. according to the synthetic method of the described indolazine compound of claim 7 or 8, it is characterized in that: described metal catalyst is cuprous iodide; Described alkali is DBU. 10. The purposes of indolazine compound according to claim 1, it is characterized in that: indolazine compound is used for tumor or cancer is gastric cancer, cervical adenocarcinoma, colon cancer, lung cancer, liver cancer, glioma, esophagus Cancer, Intestinal Cancer, Nasopharyngeal Cancer, Breast Cancer, Lymph Cancer, Kidney Cancer, Pancreatic Cancer, Bladder Cancer, Ovarian Cancer, Uterine Cancer, Bone Cancer, Gallbladder Cancer, Lip Cancer, Melanoma, Tongue Cancer, Throat Cancer, Blood Cancer , prostate cancer, brain tumor, squamous cell carcinoma, skin cancer, hemangioma, lipoma, cervical cancer and thyroid cancer.