Benzimidazolium-containing myricetin derivative, preparation method and application
阅读说明：本技术 一种含苯并咪唑的杨梅素衍生物、制备方法及用途 (Benzimidazolium-containing myricetin derivative, preparation method and application ) 是由 薛伟 陈梅 尹诗涛 詹文亮 胡蝶 周然 贺鸣 卢平 于 2020-12-02 设计创作，主要内容包括：本发明公开了一种含苯并咪唑的杨梅素衍生物、其制备方法及用途,其结构通式如下所示：其中,R为苯基、取代苯基、取代芳杂环基；n为碳链中碳的个数分别为2－5；本发明(中间体B)的合成中,实验过程中分别使用了无水碳酸钾和结晶碳酸钾作为催化剂,实验证明：使用结晶碳酸钾为催化剂得到的产率(62.5％)比使用无水碳酸钾为催化剂的产率(53.8％)的高。且结晶碳酸钾比无水碳酸钾廉价,其合成工艺简单,与前技术相比,具有明显的有益效果。(The invention discloses a myricetin derivative containing benzimidazole, a preparation method and application thereof, wherein the structural general formula is as follows:)
1. An application of a myricetin derivative containing benzimidazole in preparing a medical cancer cell inhibiting medicament is disclosed, wherein the derivative has a structural general formula as follows:
wherein R is phenyl, substituted aromatic heterocyclic radical; n is the number of carbons in the carbon chain of 2-5.
2. Use according to claim 1, characterized in that: the substituted phenyl is p-methylphenyl, p-chlorophenyl, m-bromophenyl, m-fluorophenyl, p-nitrophenyl or m-nitrophenyl.
3. Use according to claim 1, characterized in that: the aromatic heterocyclic group in the substituted aromatic heterocyclic group is thienyl, furyl or pyridyl.
4. The method for preparing a benzimidazole-containing myricetin derivative according to claim 1, wherein: comprises the following steps:
(1) the method comprises the following steps of taking o-phenylenediamine and substituted aromatic aldehyde or heterocyclic aldehyde as raw materials, taking methanol as a solvent, and refluxing to prepare an intermediate A: substituted 1H-benzimidazoles, as shown below:
(2) taking myricitrin and methyl iodide as raw materials, taking crystallized potassium carbonate as a catalyst, and preparing an intermediate B by acid regulation: 3-hydroxy-5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one, as follows:
(3) taking the intermediate B and dibromoalkane with different chain lengths as raw materials, potassium carbonate as a catalyst, N, N-dimethylformamide as a solvent, and preparing an intermediate C, namely 3-bromo-5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one under ice bath, wherein the intermediate C is shown as follows:
(4) and (3) refluxing the intermediate C and the intermediate A as raw materials, potassium carbonate as a catalyst and N, N-dimethylformamide as a solvent to prepare a target compound I: the myricetin derivative containing benzimidazole is shown as follows:
Technical Field
The invention relates to the technical field of chemical industry, in particular to a benzimidazole-containing myricetin derivative, a preparation method of the benzimidazole-containing myricetin derivative, and application of the benzimidazole-containing myricetin derivative in the aspect of cancer cell inhibition medicaments.
Background
Myricetin (Myricetin), also known as Myricetin, Myricetin belong to flavonols, have extensive biological activity, the appearance is yellow needle-shaped crystal, can be dissolved in methanol, ethanol, acetone, ethyl acetate, slightly soluble in water, slightly insoluble in chloroform, petroleum ether, widely exist in various plants, such as: fruits, vegetables and beverages which are eaten daily have rich sources. Pharmacological research shows that myricetin has biological activities of oxidation resistance, virus resistance, cancer resistance, bacteriostasis and the like, has certain research and application values, and is increasingly researched by people in recent years.
In 2005, Lyu et al (Lyu, S.Y.; RHIm, J.Y.; et al. ArcH. PHarm. Res.2005,28, 1293-one 1301.) found that myricetin at 5, 10 and 50. mu. mol/L could inhibit the plaque of I, II type herpes simplex virus by 50% -80% by using plaque reduction experiment.
In 2014, Zhao et al (Zhao hong Ju. Guizhou university, 2014) reported a series of derivatives containing heterocycloalkyl myricetin, and tested the in vitro proliferation inhibitory activity of all compounds on breast cancer cells MDA-MB-231 by using MTT method, wherein, at the concentration of 1 μmol/L, the inhibitory activity of part of the compounds is higher than that of the control drug gefitinib (9.73 +/-8.04%).
A series of acylhydrazone-containing myricetin derivatives were reported in 2015 by Xue et al (Xue, W.; Song, B.A.; ZHao, H.J.; et al, Eur.J.Med.chem.2015,97,155 163.). The MTT method is utilized to carry out in-vitro proliferation inhibition activity test of human breast cancer cells MDA-MB-231 on the synthesized compound, and research results show that: the myricetin acylhydrazone derivatives have good inhibition rate on human breast cancer cells MDA-MB-231.
In 2016, plum et al (plum is rich, Lihui, Hantao, et al. J. pharmaceutical of northwest, 2016,3,270-274.) used in vitro culture of HeLa cells from human cervical cancer, and the cell proliferation inhibition rate was determined by MTT and SRB methods; detecting apoptosis by a flow cytometer, and analyzing a cell cycle by a computer; the Western Blot method detects the contents of caspase-3, caspase-9 and survivin. The results show that myricetin has inhibition effect on the in vitro proliferation of HeLa cells and has concentration and time dependent tolerance; inducing HeLa cell apoptosis in vitro by myricetin, and blocking cell cycle in S phase; myricetin promotes the expression of caspase-3 and caspase-9 proteins and inhibits the expression of survivin proteins. Conclusion myricetin can inhibit the proliferation of HeLa cells and induce the apoptosis of the HeLa cells, and the apoptosis pathway of myricetin is related to an intracellular pathway and an extracellular pathway.
In 2020, Tang et al (Tang, X.; Zhang, C.; Chen, M.; et al. New J. chem.2020,44, 2374-: part of the compounds have protective activity EC on tobacco mosaic virus at the concentration of 500 mu g/mL50Values of 196.1, 425.3, and 386.7 μ g/mL were superior to the control drug ningnanmycin (447.92 μ g/mL).
In 2020 Jiang et al (Jiang, S.C.; Su, S.J.; Chen, M.; et al.J. Agric.food chem.2020,68, 5641-5647.) A series of dithiocarbamate myricetin derivatives were reported and the antibacterial activity of the target compound was tested by the turbidity method, and the results showed that: EC of partial compound on citrus canker50The values are 0.01 mu g/mL and 0.83 mu g/mL, which are superior to those of control drugs of thiediazole copper (59.97 mu g/mL) and bismerthiazol (48.93 mu g/mL), and the EC of partial compounds on rice bacterial blight bacteria50The values are 1.58. mu.g/mL and 3.28. mu.g/mL respectively, which are superior to the control drugs, namely, Thiobiazole (83.04. mu.g/mL) and bismerthiazol (56).05μg/mL)。
In conclusion, the myricetin has wide sources and resource advantages in research and application of medicines, but no reports about the synthesis of a benzimidazole-containing myricetin derivative by introducing a benzimidazole-containing active group into a myricetin structure are found, and no reports about the cancer cell inhibiting medicament of the benzimidazole-containing myricetin derivative are found.
Disclosure of Invention
The present invention aims to overcome the above disadvantages and to provide a benzimidazole-containing myricetin compound and a derivative thereof.
One of the objects of the present invention is to provide a myricetin compound containing benzimidazole.
The invention also aims to provide the intermediate compound and a preparation method thereof.
It is a further object of the present invention to provide a composition comprising the above compound.
It is a further object of the present invention to provide the use of the above compounds.
The invention further aims to provide the application of the benzimidazole-containing myricetin derivative in the aspect of cancer cell inhibition medicaments.
In order to realize the purpose, the invention adopts the following technical scheme:
a myricetin derivative containing benzimidazole has the following structural general formula:
wherein R is phenyl, substituted aromatic heterocyclic radical; n is the number of carbons in the carbon chain of 2-5 respectively.
The benzimidazole-containing myricetin derivative is characterized in that: the substituted phenyl is p-methylphenyl, p-chlorophenyl, m-bromophenyl, m-fluorophenyl, p-nitrophenyl or m-nitrophenyl.
The benzimidazole-containing myricetin derivative is characterized in that: the substituted aromatic heterocyclic group is thienyl, furyl or pyridyl.
The preparation method of the myricetin derivative containing benzimidazole of the invention comprises the following synthetic route:
(1) using o-phenylenediamine and substituted aromatic aldehyde or heterocyclic aldehyde as raw materials, methanol (CH)3OH) as a solvent, refluxing at 60 ℃ to prepare an intermediate A: substituted 1H-benzimidazoles, as shown below:
(2) taking myricitrin and methyl iodide as raw materials, taking crystallized potassium carbonate as a catalyst, and preparing an intermediate B by acid regulation: 3-hydroxy-5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one, as follows:
(3) taking the intermediate B and dibromoalkane with different chain lengths as raw materials, potassium carbonate as a catalyst, and N, N-dimethylformamide as a solvent, and preparing an intermediate C under ice bath: 3-bromo-5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one, as follows:
(4) taking the intermediate C and the intermediate A as raw materials, potassium carbonate as a catalyst, N, N-dimethylformamide as a solvent, and refluxing at 105 ℃ to prepare a target compound I: the myricetin derivative containing benzimidazole is shown as follows:
the invention relates to application of a myricetin derivative containing benzimidazole in preparation of a medicament for inhibiting cancer cells.
In the synthesis of the intermediate B, anhydrous potassium carbonate and crystalline potassium carbonate are respectively used as catalysts in the experimental process, and experiments prove that: the use of crystalline potassium carbonate as catalyst gave a higher yield (62.5%) than the anhydrous potassium carbonate (53.8%). And the crystallized potassium carbonate is cheaper than anhydrous potassium carbonate, the synthesis process is simple, and compared with the prior art, the method has obvious beneficial effects. The invention mainly takes dibromoalkane as a bridge, and the natural products myricetin and benzimidazole are subjected to active splicing to obtain the compound with higher anticancer activity.
Detailed Description
Example 1
The preparation method of 5, 7-dimethoxy-3- (3- (2- (p-tolyl) -1H-benzimidazole) propoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I1) is as follows:
(1) preparation of 2- (p-tolyl) -1H-benzimidazole (intermediate a):
a50 mL round bottom flask was charged with 0.50g (4.62mmol) o-phenylenediamine and 30mL CH3After the temperature of OH was raised to 60 ℃, 0.55g (4.62mmol) of p-tolualdehyde was slowly added, and the reaction was followed by TLC (petroleum ether: ethyl acetate: 3:1, V/V). After the reaction is stopped, the mixture is decompressed and concentrated to obtain yellow solid, and the yellow solid (intermediate A) is obtained by ethanol recrystallization for standby. Yield: and (4) 64.4%.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
into a 250mL round bottom flask were successively added 5.00g (10.77mmol) of myricitrin and 19.34g (140mmol) of crystalline K2CO3And 120mL of DMF, and after stirring at room temperature for 0.5 to 1h, 7.50mL (120mmol) of iodomethane was slowly added dropwise, and the mixture was stirred at room temperature for 48h, followed by TLC (methanol: ethyl acetate ═ 1:4, V/V). After the reaction is stopped, filtering and precipitating, washing filter residues by dichloromethane, combining the filter residues, diluting the filter residues by 100mL of water, extracting the filter residues for three times by dichloromethane, combining organic layers, concentrating the organic layers under reduced pressure, then dissolving the concentrate in 100mL of absolute ethyl alcohol, heating to reflux, adding 12mL of concentrated hydrochloric acid under reflux after the solution is clarified, then separating out yellow solid, continuing the reaction for 2 hours, cooling to room temperature, and filtering to obtain a crude product, namely 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxyl myricetin (an intermediate B), wherein the yield is as follows: 62.5 percent。
(3) Preparation of 3- (3-bromopropoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
1.23g (3.17mmol) of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B) and 1.31g K are sequentially added into a 100mL single-neck round-bottom flask2CO3(9.50mmol) and 30mL DMF, stirring in ice for 0.5-1 h, adding 1.92g (9.50mmol)1, 3-dibromopropane, continuing the reaction at this temperature for 12h, and monitoring the reaction by TLC (ethyl acetate). After the reaction is stopped, the reaction solution is dispersed by 100mL of water, a white solid is separated out, the mixture is filtered and dried, then the solid is added into a round-bottom flask filled with 30mL of solution (ethyl acetate: n-hexane ═ 3:1) and stirred for 4-5 h at normal temperature, filtered and purified by reduced pressure column chromatography (petroleum ether: ethyl acetate ═ 2:1, V/V) to obtain a white solid (intermediate C), and the yield: 74.6 percent.
(4) Preparation of 5, 7-dimethoxy-3- (3- (2- (p-tolyl) -1H-benzimidazole) propoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I1): in a 100mL one-necked flask were charged 0.24g (1.18mmol) of 2- (p-tolyl) -1H-benzimidazole (intermediate A), and 0.41g (2.94mmol) of K2CO3And 30mL of DMF, stirring at 80 ℃ for 0.5 to 1H, adding 0.5g (0.98mmol) of 3- (3-bromopropoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C), heating to 105 ℃, tracing the reaction by TLC (ethyl acetate: petroleum ether: 3:1, V/V), stopping the reaction, cooling, pouring into 200mL of water, adjusting pH to 4 to 5 with 5% HCl solution, precipitating a white precipitate, clarifying the solution, performing suction filtration to obtain a crude product, and purifying by column chromatography (ethyl acetate: petroleum ether: 3:1, V/V) to obtain a white solid (target compound I1) with a yield: 46.7 percent.
Example 2
The preparation method of 3- (3- (2- (4-chlorophenyl) -1H-benzimidazole) propoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I2) is as follows:
(1) preparation of 2- (4-chlorophenyl) -1H-benzimidazole (intermediate A):
the procedure is as in (1) in example 1, except that 4-chlorobenzaldehyde is used as the starting material.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (3-bromopropoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) of example 1.
(4) Preparation of 3- (3- (2- (4-chlorophenyl) -1H-benzimidazole) propoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (title compound I2):
the procedure is as in (4) of example 1, except that 2- (4-chlorophenyl) -1H-benzimidazole (intermediate A) is used as the starting material. Yield: 52.7 percent.
Example 3
The preparation method of 3- (3- (2- (3-chlorophenyl) -1H-benzimidazole) propoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I3) is as follows:
(1) preparation of 2- (3-chlorophenyl) -1H-benzimidazole (intermediate A):
the procedure is as in (1) in example 1, except that 3-chlorobenzaldehyde is used as the starting material.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (3-bromopropoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) of example 1.
(4) Preparation of 3- (3- (2- (3-chlorophenyl) -1H-benzimidazole) propoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (title compound I3):
the procedure is as in (4) of example 1, except that 2- (3-chlorophenyl) -1H-benzimidazole (intermediate A) is used as the starting material. Yield: 31.0 percent.
Example 4
The preparation method of 3- (3- (2- (furyl) -1H-benzimidazole) propoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I4) is as follows:
(1) preparation of 2- (furyl) -1H-benzimidazole (intermediate a):
the procedure is as in (1) in example 1, except that furan-2-carbaldehyde is used as the starting material.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (3-bromopropoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) of example 1.
(4) Preparation of 3- (3- (2- (furyl) -1H-benzimidazole) propoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I4):
the procedure is as in (4) of example 1, except that 2- (furyl) -1H-benzimidazole (intermediate A) is used as the starting material. Yield: 45.7 percent.
Example 5
The preparation method of 5, 7-dimethoxy-3- (3- (2- (thienyl) -1H-benzimidazole) propoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I5) is as follows:
(1) preparation of 2- (thienyl) -1H-benzimidazole (intermediate a):
the procedure is as in (1) of example 1, except that thiophene-2-carbaldehyde is used as the starting material.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (3-bromopropoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) of example 1.
(4) Preparation of 5, 7-dimethoxy-3- (3- (2- (thienyl) -1H-benzimidazole) propoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I5):
the procedure is as in step (4) of example 1, except that 2- (thienyl) -1H-benzimidazole (intermediate A) is used as the starting material. Yield: 36.8 percent.
Example 6
The preparation method of 3- (3- (2- (3-fluorophenyl) -1H-benzimidazole) propoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I6) is as follows:
(1) preparation of 2- (3-fluorophenyl) -1H-benzimidazole (intermediate a):
the procedure is as in (1) in example 1, except that 3-fluorobenzaldehyde is used as a starting material.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (3-bromopropoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) of example 1.
(4) Preparation of 3- (3- (2- (3-fluorophenyl) -1H-benzimidazole) propoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I6):
the procedure is as in (4) in example 1, except that 2- (3-fluorophenyl) -1H-benzimidazole (intermediate A) is used as a starting material. Yield: 68.1 percent.
Example 7
The preparation method of 3- (3- (2- (3-bromophenyl) -1H-benzimidazole) propoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I7) is as follows:
(1) preparation of 2- (3-bromophenyl) -1H-benzimidazole (intermediate a):
the procedure is as in (1) in example 1, except that 3-bromobenzaldehyde is used as a starting material.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (3-bromopropoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) of example 1.
(4) Preparation of 3- (3- (2- (3-bromophenyl) -1H-benzimidazole) propoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I7):
the procedure is as in (4) of example 1, except that 2- (3-bromophenyl) -1H-benzimidazole (intermediate A) is used as the starting material. Yield: 46.3 percent.
Example 8
The preparation method of 5, 7-dimethoxy-3- (3- (2- (pyridyl) -1H-benzimidazole) propoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I8) is as follows:
(1) preparation of 2- (pyridyl) -1H-benzimidazole (intermediate a):
the procedure is as in step (1) of example 1, except that a pyridylaldehyde is used as a starting material.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (3-bromopropoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) of example 1.
(4) Preparation of 5, 7-dimethoxy-3- (3- (2- (pyridyl) -1H-benzimidazole) propoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I8):
the procedure is as in (4) in example 1, except that 2- (pyridyl) -1H-benzimidazole (intermediate A) is used as a starting material. Yield: 26.1 percent.
Example 9
The preparation method of 5, 7-dimethoxy-3- (3- (2-phenyl-1H-benzimidazole) propoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I9) is as follows:
(1) preparation of 2-phenyl-1H-benzimidazole (intermediate a):
the procedure is as in step (1) of example 1, except that benzaldehyde is used as a starting material.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (3-bromopropoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) of example 1.
(4) Preparation of 5, 7-dimethoxy-3- (3- (2-phenyl-1H-benzimidazole) propoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I9):
the procedure is as in (4) of example 1, except that 2-phenyl-1H-benzimidazole (intermediate A) is used as the starting material. Yield: 37.1 percent.
Example 10
The preparation method of 5, 7-dimethoxy-3- (3- (2- (3-nitrophenyl) -1H-benzimidazole) propoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I10) is as follows:
(1) preparation of 2- (3-nitrophenyl) -1H-benzimidazole (intermediate A):
the procedure is as in step (1) of example 1, except that 3-nitrobenzaldehyde is used as a starting material.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (3-bromopropoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) of example 1.
(4) Preparation of 5, 7-dimethoxy-3- (3- (2- (3-nitrophenyl) -1H-benzimidazole) propoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (title compound I10):
as in step (4) of example 1, except that starting from the preparation of 2- (3-nitrophenyl) -1H-benzimidazole (intermediate a), the yield: 36.4 percent.
Example 11
The preparation method of 5, 7-dimethoxy-3- (3- (2- (4-nitrophenyl) -1H-benzimidazole) propoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I11) is as follows:
(1) preparation of 2- (4-nitrophenyl) -1H-benzimidazole (intermediate A):
the procedure is as in step (1) of example 1, except that 4-nitrobenzaldehyde is used as a starting material.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (3-bromopropoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) of example 1.
(4) Preparation of 5, 7-dimethoxy-3- (3- (2- (3-nitrophenyl) -1H-benzimidazole) propoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (title compound I11):
the procedure is as in (4) of example 1, except that the preparation of 2- (4-nitrophenyl) -1H-benzimidazole (intermediate A) is used as starting material. Yield: 37.1 percent.
Example 12
The preparation method of 5, 7-dimethoxy-3- (4- (2- (p-tolyl) -1H-benzimidazole) butoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I12) is as follows:
(1) preparation of 2- (p-tolyl) -1H-benzimidazole (intermediate a):
as in step (1) of example 1.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (4-bromobutoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
the procedure is as in (3) in example 1, except that 1, 4-dibromobutane is used as the starting material.
(4) Preparation of 5, 7-dimethoxy-3- (4- (2- (p-tolyl) -1H-benzimidazole) butoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (title compound I12):
as in step (4) in example 1, yield: 45.0 percent.
Example 13
The preparation method of 3- (4- (2- (4-chlorophenyl) -1H-benzimidazole) butoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I13) is as follows:
(1) preparation of 2- (4-chlorophenyl) -1H-benzimidazole (intermediate A):
as in step (2) of example 2.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (4-bromobutoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) of example 12.
(4) Preparation of 3- (4- (2- (4-chlorophenyl) -1H-benzimidazole) butoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (title compound I13):
as in step (4) of example 2. Yield: 62.9 percent.
Example 14
The preparation method of 3- (4- (2- (3-chlorophenyl) -1H-benzimidazole) butoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I14) is as follows:
(1) preparation of 2- (3-chlorophenyl) -1H-benzimidazole (intermediate A):
as in step (1) of example 3.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (4-bromobutoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) of example 12.
(4) Preparation of 3- (4- (2- (3-chlorophenyl) -1H-benzimidazole) butoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (title compound I14):
as in step (4) of example 3. Yield: 41.3 percent.
Example 15
The preparation of 3- (4- (2- (furyl) -1H-benzimidazole-) butoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (title compound I15) is as follows:
(1) preparation of 2- (furyl) -1H-benzimidazole (intermediate a):
as in step (1) of example 4.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (4-bromobutoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) of example 12.
(4) Preparation of 3- (4- (2- (furyl) -1H-benzimidazole) butoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-4-one (target compound I15):
as in step (4) of example 4. Yield: 56.8 percent.
Example 16
The preparation method of 5, 7-dimethoxy-3- (4- (2- (thienyl) -1H-benzimidazole) butoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I16) is as follows:
(1) preparation of 2- (thienyl) -1H-benzimidazole (intermediate a):
as in step (1) of example 5.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (4-bromobutoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) of example 12.
(4) Preparation of 5, 7-dimethoxy-3- (4- (2- (thienyl) -1H-benzimidazole) butoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I16):
as in step (4) of example 5. Yield: 32.6 percent.
Example 17
The preparation method of 3- (4- (2- (3-fluorophenyl) -1H-benzimidazole) butoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I17) is as follows:
(1) preparation of 2- (3-fluorophenyl) -1H-benzimidazole (intermediate a):
as in step (1) of example 6.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (4-bromobutoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) of example 12.
(4) Preparation of 3- (4- (2- (3-fluorophenyl) -1H-benzimidazole) butoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (title compound I17):
as in step (4) in example 6. Yield: 46.6 percent.
Example 18
The preparation method of 3- (4- (2- (3-bromophenyl) -1H-benzimidazole) butoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I18) is as follows:
(1) preparation of 2- (3-bromophenyl) -1H-benzimidazole (intermediate a):
as in step (1) of example 7.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (4-bromobutoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) in example 12
(4) Preparation of 3- (4- (2- (3-bromophenyl) -1H-benzimidazole) butoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (title compound I18):
the procedure was as in step (4) of example 7. Yield: 37.8 percent.
Example 19
The preparation method of 5, 7-dimethoxy-3- (4- (2- (pyridyl) -1H-benzimidazole) butoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I19) is as follows:
(1) preparation of 2- (pyridyl) -1H-benzimidazole (intermediate a):
as in step (1) of example 8.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (4-bromobutoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) of example 12.
(4) Preparation of 5, 7-dimethoxy-3- (4- (2- (pyridyl) -1H-benzimidazole) butoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I19):
the procedure was as in step (4) of example 8. Yield: 36.0 percent.
Example 20
The preparation method of 5, 7-dimethoxy-3- (4- (2-phenyl-1H-benzimidazole) butoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I20) is as follows:
(1) preparation of 2-phenyl-1H-benzimidazole (intermediate a):
as in step (1) of example 9.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (4-bromobutoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) of example 12.
(4) Preparation of 5, 7-dimethoxy-3- (4- (2-phenyl-1H-benzimidazole) butoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I20):
as in step (4) in example 9. Yield: 40.5 percent.
Example 21
The preparation method of 5, 7-dimethoxy-3- (4- (2- (3-nitrophenyl) -1H-benzimidazole) butoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I21) is as follows:
(1) preparation of 2- (3-nitrophenyl) -1H-benzimidazole (intermediate A):
as in step (1) of example 10.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (4-bromobutoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) of example 12.
(4) Preparation of 5, 7-dimethoxy-3- (4- (2- (3-nitrophenyl) -1H-benzimidazole) butoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (title compound I21):
as in step (4) in example 10. Yield: 51.0 percent.
Example 22
The preparation method of 5, 7-dimethoxy-3- (4- (2- (4-nitrophenyl) -1H-benzimidazole) butoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (target compound I22) is as follows:
(1) preparation of 2- (4-nitrophenyl) -1H-benzimidazole (intermediate A):
as in step (1) of example 11.
(2) Preparation of 3-hydroxy-3 ', 4 ', 5 ', 5, 7-pentamethoxy myricetin (intermediate B):
as in step (2) of example 1.
(3) Preparation of 3- (4-bromobutoxy) -5, 7-dimethoxy-2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (intermediate C):
as in step (3) of example 12.
(4) Preparation of 5, 7-dimethoxy-3- (4- (2- (4-nitrophenyl) -1H-benzimidazole) butoxy) -2- (3,4, 5-trimethoxyphenyl) -4H-chromen-4-one (title compound I22):
as in step (4) in example 11. Yield: 44.2 percent
The physicochemical properties and mass spectrum data of the synthesized benzimidazole-containing myricetin derivative are shown in Table 1, and the nuclear magnetic resonance hydrogen spectrum (C: (C))1H NMR), carbon spectrum (13C NMR) and fluorine Spectroscopy (19F NMR) data are shown in table 2.
TABLE 1 physicochemical Properties of the example Compounds I1-I22
TABLE 2 NMR data for target compounds I1-I22
Example 23 test for inhibiting cancer cell activity of object compounds I1-I22 (taking a549 lung cancer cells as an example):
1 test method
(1) Cell culture
The A549 cells used in the experiment contain 5% CO2Culturing and subculturing in a constant temperature incubator at 37 ℃, and culturing in RPMI1640 medium containing 10% fetal calf serum, 100U/mL penicillin and 100 mu g/mL streptomycin; the passages were digested with 0.25% pancreatin-EDTA and passaged twice a week. DMSO was used as a negative control group, gemcitabine was used as a positive control, and the effect was observed on cells in the logarithmic growth phase.
(2) MTT colorimetric method
A549 cells in logarithmic phase are digested by 0.25% pancreatin-EDTA to prepare single cell suspension with a certain concentration, the single cell suspension is inoculated into a 96-well plate according to 4000 cells/well, and 200 mu L of cell suspension is added into each well. 24h later, fresh medium containing different concentrations of compound and corresponding solvent control was added at 200. mu.L per well (DMSO final concentration)<0.1%), setting 5 dose groups for each tested compound, continuously culturing at 37 ℃ for 72h, adding 20 mu L of 5mg/mL MTT solution into each hole, continuously culturing for 4h, discarding supernatant, adding 200 mu L DMSO into each hole to dissolve MTT Formazan (Formazan) precipitate, uniformly oscillating by a micro oscillator, measuring 570nm optical density value (OD) by an enzyme labeling instrument, taking tumor cells treated by solvent control as a control group, calculating the inhibition rate of the compound on the tumor cells by the following formula, and calculating IC by SPSS software50：
(3) Statistical method
The experimental results were analyzed by SPSS11.5 using One-Way ANOVA method, and P <0.05 indicates significant differences between the data.
2 inhibition of A549 lung cancer cell activity test result
TABLE 3 inhibition ratio (%) -of Compounds I1-I22 against A549 Lung cancer cells at the set concentrations, respectively
Compound (I)
n
R
IC50(μM)
Compound (I)
n
R
IC50(μM)
Ⅰ1
3
4-CH3-Ph
1.56
Ⅰ12
4
4-CH3-Ph
1.90
Ⅰ2
3
4-Cl-Ph
3.38
Ⅰ13
4
4-Cl-Ph
5.30
Ⅰ3
3
3-Cl-Ph
17.59
Ⅰ14
4
3-Cl-Ph
28.23
Ⅰ4
3
Furan-2-yl
11.35
Ⅰ15
4
Furan-2-yl
2.16
Ⅰ5
3
Thiophene-2-yl
2.52
Ⅰ16
4
Thiophene-2-yl
2.22
Ⅰ6
3
3-F-Ph
4.47
Ⅰ17
4
3-F-Ph
1.60
Ⅰ7
3
3-Br-Ph
23.64
Ⅰ18
4
3-Br-Ph
37.97
Ⅰ8
3
Pyridine-2-yl
27.36
Ⅰ19
4
Pyridine-2-yl
19.24
Ⅰ9
3
Ph
2.16
Ⅰ20
4
Ph
4.92
Ⅰ10
3
3-NO2-Ph
13.29
Ⅰ21
4
3-NO2-Ph
25.48
Ⅰ11
3
4-NO2-Ph
16.33
Ⅰ22
4
4-NO2-Ph
10.45
Gefitinib
-
-
10.38
Gefitinib
-
-
10.38
The MTT method is adopted to carry out A549 lung cancer cell activity test on the synthesized compound, and the test result of Table 3 shows that: some compounds show good inhibitory activity, and the inhibition rate (IC) of the compounds I1, I2, I5, I6, I9, I12, I15, I16, I17 and I20 on A549 lung cancer cells50) The values were 1.56,3.38,2.52, 4.47,2.16,1.90,2.16,2.22,1.60, and 4.92. mu.M, respectively, which are superior to the control gefitinib (10.38. mu.M).
The experimental activity data show that the myricetin derivative containing benzimidazole has a certain inhibition effect on A549 lung cancer cells, wherein a part of target compounds show excellent activity on the A549 lung cancer cells, can be used as a potential lung cancer cell inhibition drug, and has a good application prospect.
In summary, the present invention is only a preferred embodiment, and is not limited to any form, and any simple modification, equivalent change and modification made to the above embodiment according to the technical essence of the present invention are within the scope of the technical solution of the present invention without departing from the technical solution of the present invention.
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