CN119707903A

CN119707903A - Isopentenyl flavonoid compound and preparation method and application thereof

Info

Publication number: CN119707903A
Application number: CN202410406511.7A
Authority: CN
Inventors: 胡文浩; 钱宇; 姜瑞; 赵婧煜; 叶静诗; 傅祥
Original assignee: Sun Yat Sen University
Current assignee: Sun Yat Sen University
Priority date: 2024-04-03
Filing date: 2024-04-07
Publication date: 2025-03-28

Abstract

本发明属于抗癌药物技术领域，具体公开了一种异戊烯基黄酮类化合物及其制备方法和应用，所述化合物具有式(1)所示的结构，所述方法首先使用氯甲基甲醚将2,4,6‑三羟基苯乙酮的2,4位的羟基进行保护、接着使用异戊烯基溴与上述产物进行取代反应、随后以N,N‑二乙基苯胺为溶剂进行微波反应发生克莱森重排、然后与相应的醛发生克莱森‑施密特反应生成查尔酮，最后再用碘关环并用盐酸脱去保护，制备得到所述化合物。本发明公开的所述化合物结构新颖、活性较佳、毒性小，该系列化合物作为抑制剂具有较好的抗肿瘤活性，对多种肿瘤细胞包括结直肠癌、乳腺癌、肺癌和肝癌等均具有较好的抑制活性，可应用于制备抗肿瘤药物，在抗肿瘤方面具有较好的应用前景。The present invention belongs to the technical field of anticancer drugs, and specifically discloses a prenyl flavonoid compound and a preparation method and application thereof, wherein the compound has a structure shown in formula (1), wherein the method first uses chloromethyl methyl ether to protect the hydroxyl groups at positions 2,4 of 2,4,6-trihydroxyacetophenone, then uses prenyl bromide to carry out substitution reaction with the above-mentioned product, then uses N,N-diethylaniline as a solvent to carry out microwave reaction to produce Claisen rearrangement, then reacts with the corresponding aldehyde to produce chalcone by Claisen-Schmidt reaction, and finally closes the ring with iodine and removes the protection with hydrochloric acid to prepare the compound. The compound disclosed by the present invention has novel structure, good activity and low toxicity, and the series of compounds have good antitumor activity as inhibitors, and have good inhibitory activity on a variety of tumor cells including colorectal cancer, breast cancer, lung cancer and liver cancer, etc., and can be applied to the preparation of antitumor drugs, and has good application prospects in antitumor.

Description

Isopentenyl flavonoid compound and preparation method and application thereof

Technical Field

The invention belongs to the technical field of anticancer drugs, and particularly relates to an isopentenyl flavonoid compound, a preparation method and application thereof.

Background

In nature, flavonoids are a wide variety of compounds that possess a variety of biological and effective medical uses, such as anti-cancer and anti-tumor activity, anti-inflammatory and anti-viral activity, antibacterial and antifungal activity, anti-cardiovascular disease, enzyme inhibition activity, anti-radical and antioxidant activity. In recent years, flavonoids have received attention from more and more researchers due to their broad spectrum of biological activity, broad natural distribution, diverse chemical structures and low toxicity. The isopentenyl flavone is a unique natural flavonoid compound and is characterized in that an isopentenyl side chain exists on a flavone skeleton. The C-prenylated flavone can enhance the affinity of the C-prenylated flavone to p-glycoprotein and the permeability of the C-prenylated flavone to cell membranes, and can obviously improve the biological activity of the C-prenylated flavone. And the flavonoid has no mental activity and does not lead patients to have dependence on drugs. Thus, isopentenyl flavonoids are becoming a research hotspot in the industry.

In 2019, rea et al reported that isopentenyl flavonoid compound B, which is a flavonoid compound of methallyl, was synthesized by catalyzing dimethylallyl diphosphate with isopentenyl transferase, patent WO2017091837 also discloses biosynthesis in the form of enzyme, but such biosynthesis method requires extraction of a large amount of enzyme from plants, is costly and is not easy for industrial scale-up.

Giovanni Appendino et al report the synthesis of isopentenyl flavonoids in A Regiodivergent Synthesis of Ring A C-Prenylflavones, but the catalysts used in the synthetic route are expensive, costly and not easily scaled up industrially.

At present, most of the synthesis of C-terminal isopentenyl flavone is to synthesize a flavone mother nucleus and then rearrange, so that the synthesis is difficult. The chemical synthesis method disclosed at present has the defects of long synthesis step, expensive catalyst and difficult purification. It is therefore an urgent need to develop a more efficient synthetic method.

Disclosure of Invention

In order to solve the defects and shortcomings in the prior art, the invention provides an isopentenyl flavonoid compound, and a preparation method and application thereof. The preparation method has the advantages of low cost and easy acquisition of raw materials, short reaction period, simple operation, higher reaction yield than the existing synthesis method and the like.

The invention firstly rearranges and synthesizes chalcone, finally closes the ring, is easier to purify, and can obtain a pair of isomers very stably in the closing process, so that the structure of the product is more diversified.

The invention provides an isopentenyl flavonoid compound, which has a structure shown in a formula (1):

Wherein R ¹ is isopentenyl or hydrogen, R ² is isopentenyl or hydrogen, and R ³ is hydroxy, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, halogen, trifluoromethyl, nitro, amino, morpholino, cyclopropane, cyclopentane, cyclobutane, hydroxymethyl, hydroxyethyl, or hydroxypropyl. Preferably, R ¹ is isopentenyl, R ² is isopentenyl, R ³ is hydroxy, methyl, isopropyl, methoxy, halogen, trifluoromethyl, morpholino ring.

The isopentenyl flavonoid compound disclosed by the invention comprises structures shown in the formulas (2) and (3):

Wherein R ³ is hydroxy, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, halogen, trifluoromethyl, nitro, amino, morpholino, cyclopropane, cyclopentane, cyclobutane, hydroxymethyl, hydroxyethyl, or hydroxypropyl. Preferably, R ³ is hydroxy, methyl, isopropyl, methoxy, halogen, trifluoromethyl, morpholino ring.

Further, the structure represented by the formula (2) is selected from any one of the following structures:

further, the structure represented by the formula (3) is selected from any one of the following structures:

The invention also provides a preparation method of the isopentenyl flavonoid compound, which comprises the steps of sequentially carrying out hydroxyl protection, substitution reaction, claisen rearrangement, claisen-schmitt reaction, ring closure and deprotection reaction to prepare the isopentenyl flavonoid compound, wherein the reaction process is shown in the following reaction formula (I):

the preparation method comprises the following steps:

The first step, protecting the hydroxyl of the 2, 4-position and the 4-position of 2,4, 6-trihydroxyacetophenone by using a hydroxyl protecting reagent in a first solvent under the action of organic alkali, and reacting to obtain a compound A;

secondly, in a second solvent, under the action of inorganic alkali, carrying out substitution reaction on a substitution reagent and the compound A to obtain a compound B;

thirdly, in a third solvent, carrying out microwave reaction on the compound B to generate claisen rearrangement to obtain a compound C;

Fourthly, in a fourth solvent, under the action of inorganic alkali, the compound C and aldehyde react to generate chalcone by claisen-Schmidt reaction;

and fifthly, in a fifth solvent, closing the ring of the chalcone by using an addition reagent, and deprotecting by using an acid in a sixth solvent, and reacting to obtain the final product of the isopentenyl flavonoid compound.

In particular, the method comprises the steps of,

In the first step, the organic base comprises one or more of N, N-diisopropylethylamine, triethylamine and the like, and preferably the organic base is N, N-Diisopropylethylamine (DIPEA).

In the first step, the hydroxyl protecting agent comprises chloromethyl methyl ether (MOMCl) and the like.

In the first step, the first solvent comprises one or more of dichloromethane, tetrahydrofuran, 1, 4-dioxane and the like, and preferably, the first solvent is Dichloromethane (DCM).

In the first step, when the 2,4, 6-trihydroxyacetophenone is 1 equivalent, the organic base is 2-3 equivalents, the hydroxyl protecting agent is 2-3 equivalents, the first solvent is 10 equivalents, preferably, the organic base is 2 equivalents, and the hydroxyl protecting agent is 2 equivalents.

In the first step, the temperature of the reaction is 0 ℃ to room temperature, and preferably the temperature of the reaction is 0 ℃.

In the first step, the reaction time is 6-8 hours, preferably 6 hours.

In the second step, the substitution reagent includes isopentenyl bromide and the like.

In the second step, the inorganic base includes one or more of potassium carbonate, cesium carbonate, calcium carbonate, sodium carbonate, and the like, and preferably, the inorganic base is potassium carbonate.

In the second step, the second solvent includes acetone or the like.

In the second step, the amount of the compound A is 1 equivalent, the amount of the substitution reagent is 1-2 equivalents, the amount of the inorganic base is 2-4 equivalents, and the amount of the second solvent is 10 equivalents, preferably, the amount of the substitution reagent is 1 equivalent, the amount of the inorganic base is 2 equivalents, and the amount of the second solvent is 10 equivalents.

In the second step, the temperature of the reaction is 60-70 ℃, and preferably, the temperature of the reaction is 65 ℃.

In the second step, the reaction time is 2-4 hours, preferably 2 hours.

In the third step, the third solvent is one or more of N, N-diethylaniline, N-dimethylaniline and the like, and preferably, the third solvent is N, N-diethylaniline.

In the third step, the amount of the compound B is 1 equivalent, and the amount of the third solvent is 10 equivalents.

In the third step, the temperature of the reaction is 200-220 ℃, and preferably, the temperature of the reaction is 200 ℃.

In the third step, the reaction time is 1-2 hours, preferably 1 hour.

In the fourth step, the aldehyde includes one or more of 2-chloro-4- (methoxymethoxy) benzaldehyde, 2-methoxy-4- (methoxymethoxy) benzaldehyde, etc., and preferably, the aldehyde is determined according to a specific substrate.

In the fourth step, the inorganic base comprises one or more of potassium hydroxide, sodium hydroxide and the like, and preferably, the base is potassium hydroxide.

In the fourth step, the fourth solvent comprises one or more of absolute ethyl alcohol, absolute methyl alcohol, ethylene glycol and the like, and preferably, the solvent is absolute ethyl alcohol.

In the fourth step, the amount of the compound C is 1 equivalent, the amount of the inorganic base is 4-6 equivalents, the amount of the aldehyde is 1 equivalent, and the amount of the fourth solvent is 10 equivalents, preferably, the amount of the base is 4 equivalents;

in the fourth step, the temperature of the reaction is room temperature.

In the fourth step, the reaction time is 3-6 hours, preferably 3 hours.

In the fifth step, the addition reagent includes iodine and the like.

In the fifth step, the acid comprises one or more of hydrochloric acid, sulfuric acid and the like, and preferably, the acid is hydrochloric acid.

In the fifth step, the fifth solvent includes dimethyl sulfoxide (DMSO), and the like.

In the fifth step, the sixth solvent comprises one or more of methanol, ethanol, ethylene glycol and the like, and preferably, the sixth solvent is methanol.

In the fifth step, the chalcone is 1 equivalent, the addition reagent is 0.1-0.2 equivalent, the fifth solvent is 10 equivalent, the sixth solvent is 10 equivalent, the acid used is 10% mass concentration, and preferably, the addition reagent is 0.1 equivalent.

In the fifth step, the temperature of the ring closing reaction is 110-120 ℃, and preferably, the temperature of the ring closing reaction is 110 ℃.

In the fifth step, the time of the ring closing reaction is 1.5-3 hours, and preferably, the time of the ring closing reaction is 1.5 hours.

In the fifth step, the reflux temperature of the deprotection reaction is 70-80 ℃, and preferably, the reflux temperature of the deprotection reaction is 80 ℃.

In the fifth step, the deprotection reaction time is 0.5-1 hour, and preferably, the deprotection reaction time is 0.5 hour.

The invention also provides a medicament/pharmaceutical composition comprising an isopentenyl flavonoid as described above.

In the present invention, the drug/pharmaceutical composition may be used alone or in combination with other drugs including PDL1 mab and the like.

In the present invention, the pharmaceutical composition further comprises an excipient, diluent, adjuvant, vehicle, or combination thereof.

In the present invention, the pharmaceutical composition is formulated as an injectable fluid, aerosol, cream, gel, pill, capsule, syrup, transdermal patch or excipient.

The invention also provides application of the isopentenyl flavonoid compound or the medicine/medicine composition in preparing antitumor medicines, medicines for inhibiting tumor cells and the like.

The isopentenyl flavonoid compound or the medicine/medicine composition is used for inhibiting the growth, migration, proliferation and the like of tumor cells and is used for promoting the apoptosis of the tumor cells.

Preferably, the tumor cells include, but are not limited to, human osteosarcoma cells (Sjsa-1), human colon cancer cells (HCT 116), human non-small cell lung cancer cells (a 549), human liver cancer cells (Hep G2).

In a specific embodiment, in the present invention, stable obtaining of the corresponding isomer can be achieved in one step of the iodinated ring, so that the structure of the compound is diversified.

The isopentenyl flavonoid compound can be used as a novel anticancer drug to promote apoptosis of cells by directly killing tumor cells and activating tumor immunity. Has good inhibitory activity on various cancer cell lines such as human osteosarcoma cell (Sjsa-1), human colon cancer cell (HCT 116), human non-small cell lung cancer cell (A549) and human liver cancer cell (Hep G2).

Compared with the prior art, the isopentenyl flavonoid compound has the beneficial effects that the isopentenyl flavonoid compound is novel in structure and good in activity, has good antitumor activity when being used as an inhibitor, has good inhibitory activity on various tumor cells (such as human osteosarcoma cells, human colon cancer cells, human non-small cell lung cancer cells, human liver cancer cells and the like), and can be applied to preparation of antitumor drugs. Has better application prospect in the aspects of antibiosis and anti-tumor. The chemical synthesis methods disclosed at present have the problems of long synthesis steps, expensive catalysts and difficult methods. The invention provides a total synthesis and derivative preparation method of isopentenyl flavonoid compounds. The preparation method has the advantages of low cost and easy acquisition of raw materials, short reaction period, simple operation and the like.

Detailed Description

The present invention will be described in further detail with reference to the following embodiments. The protection of the present invention is not limited to the following examples. Variations and advantages that would occur to one skilled in the art are included in the invention without departing from the spirit and scope of the inventive concept, and the scope of the invention is defined by the appended claims. The procedures, conditions, reagents, experimental methods, etc. for carrying out the present invention are common knowledge and common knowledge in the art, except for those specifically mentioned below, and the present invention is not limited in particular.

It will be apparent that the described embodiments are only some, but not all, embodiments of the invention. The following description of at least one exemplary embodiment is merely exemplary in nature and is in no way intended to limit the invention, its application, or uses. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

In all examples shown and discussed herein, any specific values should be construed as merely illustrative, and not a limitation. Thus, other examples of the exemplary embodiments may have different values.

The following describes the invention in more detail. The description of these embodiments is provided to assist understanding of the present invention, but is not intended to limit the present invention. In addition, the technical features of the embodiments of the present invention described below may be combined with each other as long as they do not collide with each other.

The invention belongs to the technical field of anticancer drugs, and in particular relates to an isopentenyl flavonoid compound, a preparation method and application thereof, wherein the compound has a structure shown in a formula (1), the method comprises the steps of firstly protecting hydroxyl groups at 2,4 positions of 2,4, 6-trihydroxyacetophenone by chloromethyl methyl ether, then carrying out substitution reaction on the product by using isopentenyl bromide, then carrying out microwave reaction on the product by using N, N-diethylaniline as a solvent to generate claisen rearrangement, then carrying out claisen-Schmidt reaction on the product and the corresponding aldehyde to generate chalcone, and finally removing the protection by using hydrochloric acid to obtain the compound by using iodine ring. The compound provided by the invention has novel structure, better activity and low toxicity, has better anti-tumor activity as an inhibitor, has better inhibition activity on various tumor cells including colorectal cancer, breast cancer, lung cancer, liver cancer and the like, and can be applied to preparation of anti-tumor drugs. Has better application prospect in the aspect of anti-tumor.

The experimental methods in the following examples, unless otherwise specified, were conventional, and the experimental materials used in the following examples, unless otherwise specified, were commercially available from conventional sources.

Wherein R ¹ is isopentenyl or hydrogen, R ² is isopentenyl or hydrogen, and R ³ is hydroxy, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, halogen, trifluoromethyl, nitro, amino, morpholino, cyclopropane, cyclopentane, cyclobutane, hydroxymethyl, hydroxyethyl, or hydroxypropyl.

The prenylflavonoids in this example include 36 compounds, and the structural formulae and molecular weights of these compounds are shown in Table 1.

TABLE 1 Structure and molecular weight of isopentenyl flavonoid Compounds

Example 1

Synthesis of Compounds 1 and 2:

the synthetic chemical reaction formula is shown below:

The specific synthesis method comprises the following steps:

Step 1,2, 4, 6-trihydroxyacetophenone (10.00 g,59.47 mmol) and N, N-diisopropylethylamine DIPEA (23.05 g,178.41 mmol) are weighed and dissolved in methylene chloride (100 mL), and stirred in an ice bath, after the solution is slightly clear, chloromethyl methyl ether MOMCl (14.36 g,178.41 mmol) is slowly added dropwise to the reaction system, and the temperature is slowly raised to room temperature and stirring is continued for 6 hours. After the reaction was completed, the reaction solution was quenched with saturated ammonium chloride, stirred at room temperature for 5 minutes, the organic phase was washed with water three times, finally the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin to give a pale yellow oily crude product, which was purified by column chromatography to give 13.60g of product A in 89% yield.

Step 2. Reaction product A (10.0 g,39.02 mmol) of the above reaction product was weighed and dissolved in acetone (100 mL), isopentenyl bromide (11.68 g,78.4 mmol) was added to the reaction system, and potassium carbonate (10.79 g,78.09 mmol) was reacted at a temperature elevated to reflux temperature for 2 hours. After the starting materials had reacted, the solid was removed by filtration and washed with ethyl acetate. After concentration in vacuo, purification by column chromatography gave 10.60g of product B in 84% yield.

Step 3 reaction product B (2.0 g,6.17 mmol) from the previous step was dissolved in N, N-diethylaniline (10 mL) and reacted for 1 hour at 200℃with microwaves. After the reaction of the raw materials was completed, 10% hydrochloric acid was added thereto to neutralize, pH was adjusted to neutrality, followed by extraction with ethyl acetate three times, and after washing with saturated sodium chloride solution water, the organic phase was dried over anhydrous sodium sulfate, filtered, and column-chromatographed to give 1.20g of pale yellow oily substance C in 60% yield.

Step 4, the above product C (2.0 g,6.17 mmol) was dissolved in absolute ethanol (20 mL), potassium hydroxide (1.38 g,24.66 mmol) was gradually added to the reaction system, stirred at room temperature, and after the potassium hydroxide was completely dissolved, 2-methoxy-4- (methoxymethoxy) benzaldehyde (1.21 g,6.17 mmol) was added and stirred at room temperature for 3 hours. After the reaction of the raw materials is completed, the reaction solution is concentrated, diluted by adding ethyl acetate, extracted by adding water, the water phase is back extracted by using ethyl acetate for three times, and the EA layers are combined and washed by saturated saline. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin-drying to give an oily crude product, which was purified by column chromatography to give 2.10g of a yellow oily product 1a in a yield of 68%.

Step 5. Reaction product 1a (1.00 g,1.99 mmol) from the previous step was placed in a reaction flask, dimethyl sulfoxide (20 mL) was added for dissolution, iodine (50.50 mg,0.17 mmol) was added, and the mixture was allowed to react at 110℃for 1.5 hours. After the reaction of the raw materials was completed, the reaction solution was diluted with ethyl acetate, extracted with water, washed with saturated sodium sulfate, combined with an organic phase, dried over anhydrous sodium sulfate, concentrated in vacuo to give a crude yellow oil, which was not further purified, added with methanol (20 mL) and 10% diluted hydrochloric acid solution (10 mL), refluxed at 80 ℃ for 0.5 hours, concentrated in vacuo, extracted with ethyl acetate, washed with saturated sodium sulfate, combined with an organic phase, dried over anhydrous sodium sulfate, dried over spin-dry solvent, separated and purified by column chromatography to give 110mg of compound 1 in 11% yield while 130mg of compound 2 was obtained in 13% yield. Compound 1¹H NMR(400MHz,DMSO-d₆)δ12.90(s,1H),10.54(s,2H),7.74(d,J＝8.6Hz,1H),6.76(s,1H),6.60(d,J＝2.2Hz,1H),6.55(dd,J＝8.6,2.2Hz,1H),6.27(s,1H),5.19–5.11(t,1H),3.89(s,3H),3.39(d,J＝7.0Hz,3H),1.71(s,3H),1.62(s,3H).HRMS(ESI):calculated for C₂₁H₂₀O₆[M+H]⁺＝369.1333;found 369.1335. Compound 2¹H NMR(400MHz,DMSO-d₆)δ13.21(s,1H),10.52(s,2H),7.78(d,J＝8.5Hz,1H),6.79(s,1H),6.60–6.55(m,2H),6.48(s,1H),5.18(t,J＝7.2Hz,1H),3.88(s,3H),3.22(d,J＝7.2Hz,2H),1.72(s,3H),1.62(s,3H).HRMS(ESI):calculated for C₂₁H₂₀O₆[M+H]⁺＝369.1333;found 369.1333.

Example 2

Synthesis of Compounds 3 and 4:

the synthetic chemical reaction formula is shown below:

The specific synthesis method comprises the following steps:

Step 1) the above-mentioned product C (2.0 g,6.17 mmol) was dissolved in absolute ethanol (20 mL), potassium hydroxide (1.38 g,24.66 mmol) was gradually added to the reaction system, stirred at room temperature, and after the potassium hydroxide was completely dissolved, 2-chloro-4- (methoxymethoxy) benzaldehyde (1.24 g,6.17 mmol) was added and stirred at room temperature for 3 hours. After the reaction of the raw materials is completed, the reaction solution is concentrated, diluted by adding ethyl acetate, extracted by adding water, the water phase is back extracted by using ethyl acetate for three times, and the EA layers are combined and washed by saturated saline. Finally, the organic phase is dried with anhydrous sodium sulfate, filtered, and the filtrate is dried by spin to obtain crude oil, and the crude oil is separated and purified by column chromatography to obtain 2.20g of yellow oily product 2a with the yield of 70%.

Step 2. The reaction product 2a (1.00 g,1.97 mmol) from the previous step was placed in a reaction flask, dimethyl sulfoxide (20 mL) was added for dissolution, iodine (50.06 mg,0.97 mmol) was added, and the mixture was allowed to react at 110℃for 1.5 hours. After the reaction of the raw materials was completed, the reaction solution was diluted with ethyl acetate, extracted with water, washed with saturated sodium sulfate, combined with an organic phase, dried over anhydrous sodium sulfate, concentrated in vacuo to give a crude yellow oil, which was not further purified, added with methanol (20 mL) and 10% diluted hydrochloric acid solution (10 mL), refluxed at 80 ℃ for 0.5 hours, concentrated in vacuo, extracted with ethyl acetate, washed with saturated sodium sulfate, combined with an organic phase, dried over anhydrous sodium sulfate, dried over spin-dry solvent, separated and purified by column chromatography to give 140mg of compound 3 in 14% yield while 126mg of compound 4 was obtained in 13% yield. Compound 3¹H NMR(400MHz,DMSO-d₆)δ12.70(s,1H),10.66(s,2H),7.61(d,J＝8.5Hz,1H),7.01(d,J＝2.3Hz,1H),6.90(dd,J＝8.6,2.4Hz,1H),6.47(s,1H),6.32(s,1H),5.11(t,J＝7.2Hz,1H),3.34(d,J＝7.2Hz,2H),1.58(d,J＝4.8Hz,6H).HRMS(ESI):calculated for C₂₀H₁₇ClO₅[M+H]⁺＝373.0837;found 373.0835. Compound 4¹H NMR(400MHz,DMSO-d₆)δ12.99(s,1H),7.61(d,J＝8.6Hz,1H),7.00(d,J＝2.4Hz,1H),6.90(dd,J＝8.6,2.4Hz,1H),6.47(s,1H),6.45(s,1H),5.17(t,1H),3.23(d,J＝7.2Hz,2H),1.72(s,3H),1.62(s,3H).HRMS(ESI):calculated for C₂₀H₁₇ClO₅[M+H]⁺＝373.0837;found 373.0840.

Example 3

Synthesis of compounds 5 and 6:

the synthetic chemical reaction formula is shown below:

The specific synthesis method comprises the following steps:

Step 1) the above-mentioned product C (2.0 g,6.17 mmol) was dissolved in absolute ethanol (20 mL), potassium hydroxide (1.38 g,24.66 mmol) was gradually added to the reaction system, stirred at room temperature, and 3-chloro-4- (methoxymethyloxy) benzaldehyde (1.24 g,6.17 mmol) was added after the potassium hydroxide was completely dissolved, and stirred at room temperature for 3 hours. After the reaction of the raw materials is completed, the reaction solution is concentrated, diluted by adding ethyl acetate, extracted by adding water, the water phase is back extracted by using ethyl acetate for three times, and the EA layers are combined and washed by saturated saline. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin-drying to give an oily crude product, which was purified by column chromatography to give 2.35g of a yellow oily product 3a in 75% yield.

Step 2. The reaction product 3a (1.00 g,1.97 mmol) from the previous step was placed in a reaction flask, dimethyl sulfoxide (20 mL) was added for dissolution, iodine (50.06 mg,0.97 mmol) was added, and the mixture was allowed to react at 110℃for 1.5 hours. After the reaction of the raw materials was completed, the reaction solution was diluted with ethyl acetate, extracted with water, washed with saturated sodium sulfate, combined with an organic phase, dried over anhydrous sodium sulfate, concentrated in vacuo to give a crude yellow oil, which was not further purified, added with methanol (20 mL) and 10% diluted hydrochloric acid solution (10 mL), refluxed at 80 ℃ for 0.5 hours, concentrated in vacuo, extracted with ethyl acetate, washed with saturated sodium sulfate, combined with an organic phase, dried over anhydrous sodium sulfate, dried over spin-dry solvent, separated and purified by column chromatography to give 112mg of compound 5 in 11% yield while 136mg of compound 6 was obtained in 14% yield. Compound 5¹H NMR(500MHz,DMSO-d₆)δ12.81(s,1H),10.87(s,2H),8.00(s,1H),7.86(d,J＝8.7Hz,1H),7.11(d,J＝8.6Hz,1H),6.85(s,1H),6.29(s,1H),5.18(t,J＝6.9Hz,1H),3.43(d,J＝7.0Hz,2H),1.78(s,3H),1.64(s,3H).HRMS(ESI):calculated for C₂₀H₁₇ClO₅[M+H]⁺＝373.0837;found 373.0838. Compound 6¹H NMR(400MHz,DMSO-d₆)δ13.14(s,1H),10.96(s,2H),8.06(d,J＝2.3Hz,1H),7.87(dd,J＝8.6,2.3Hz,1H),7.10(d,J＝8.7Hz,1H),6.84(s,1H),6.55(s,1H),5.18(t,1H),3.22(d,J＝7.2Hz,2H),1.72(s,3H),1.62(s,3H).HRMS(ESI):calculated for C₂₀H₁₇ClO₅[M+H]⁺＝373.0837;found 373.0840.

Example 4

Synthesis of Compounds 7 and 8:

the synthetic chemical reaction formula is shown below:

The specific synthesis method comprises the following steps:

Step 1, the above product C (2.0 g,6.17 mmol) was dissolved in absolute ethanol (20 mL), potassium hydroxide (1.38 g,24.66 mmol) was gradually added to the reaction system, stirred at room temperature, and after the potassium hydroxide was completely dissolved, 2, 5-dimethoxybenzaldehyde (1.02 g,6.17 mmol) was added thereto, and stirred at room temperature for 3 hours. After the reaction of the raw materials is completed, the reaction solution is concentrated, diluted by adding ethyl acetate, extracted by adding water, the water phase is back extracted by using ethyl acetate for three times, and the EA layers are combined and washed by saturated saline. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin-drying to give crude oil, which was purified by column chromatography to give 2.50g of yellow oily product 4a in 86% yield.

Step 2. The reaction product 4a (1.00 g,2.12 mmol) from the previous step was placed in a reaction flask, dimethyl sulfoxide (20 mL) was added for dissolution, iodine (53.71 mg,0.21 mmol) was added, and the mixture was allowed to react at 110℃for 1.5 hours. After the reaction of the raw materials was completed, the reaction solution was diluted with ethyl acetate, extracted with water, washed with saturated sodium sulfate, combined with an organic phase, dried over anhydrous sodium sulfate, concentrated in vacuo to give a crude yellow oil, which was not further purified, added with methanol (20 mL) and 10% diluted hydrochloric acid solution (10 mL), refluxed at 80 ℃ for 0.5 hours, concentrated in vacuo, extracted with ethyl acetate, washed with saturated sodium sulfate, combined with an organic phase, dried over anhydrous sodium sulfate, dried over spin-dry solvent, separated and purified by column chromatography to give 120mg of compound 7 in 12% yield while 145mg of compound 8 was obtained in 15% yield. Compound 7¹H NMR(400MHz,DMSO-d₆)δ13.05(s,1H),10.81(s,2H),7.49(t,J＝8.4Hz,1H),6.80(d,J＝8.5Hz,2H),6.39(s,1H),6.21(s,1H),5.17(t,1H),3.77(s,7H),3.23(d,J＝7.2Hz,2H),1.73(s,3H),1.63(s,3H).HRMS(ESI):calculated for C₂₂H₂₂O₆[M+H]⁺＝383.1489;found 383.1496. Compound 8¹H NMR(400MHz,DMSO-d₆)δ12.73(s,1H),7.49(t,J＝8.4Hz,1H),6.81(d,J＝8.5Hz,2H),6.30(s,1H),6.22(s,1H),5.10(t,J＝7.4Hz,1H),3.78(s,6H),3.22(d,J＝7.4Hz,2H),1.56(s,3H),1.49(s,3H).HRMS(ESI):calculated for C₂₂H₂₂O₆[M+H]⁺＝383.1489;found 383.1483.

Example 5

Synthesis of compounds 9 and 10:

the synthetic chemical reaction formula is shown below:

The specific synthesis method comprises the following steps:

Step 1, the above product C (2.0 g,6.17 mmol) was dissolved in absolute ethanol (20 mL), potassium hydroxide (1.38 g,24.66 mmol) was gradually added to the reaction system, stirred at room temperature, and 3, 4-dimethoxybenzaldehyde (1.02 g,6.17 mmol) was added after the potassium hydroxide was completely dissolved, and stirred at room temperature for 3 hours. After the reaction of the raw materials is completed, the reaction solution is concentrated, diluted by adding ethyl acetate, extracted by adding water, the water phase is back extracted by using ethyl acetate for three times, and the EA layers are combined and washed by saturated saline. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin-drying to give an oily crude product, which was purified by column chromatography to give 2.37g of a yellow oily product 5a in 81% yield.

Step 2. The reaction product 5a (1.00 g,2.12 mmol) from the previous step was placed in a reaction flask, dimethyl sulfoxide (20 mL) was added for dissolution, iodine (53.71 mg,0.21 mmol) was added, and the mixture was allowed to react at 110℃for 1.5 hours. After the reaction of the raw materials was completed, the reaction solution was diluted with ethyl acetate, extracted with water, washed with saturated sodium sulfate, combined with an organic phase, dried over anhydrous sodium sulfate, concentrated in vacuo to give a crude yellow oil, which was not further purified, added with methanol (20 mL) and 10% diluted hydrochloric acid solution (10 mL), refluxed at 80 ℃ for 0.5 hours, concentrated in vacuo, extracted with ethyl acetate, washed with saturated sodium sulfate, combined with an organic phase, dried over anhydrous sodium sulfate, dried over spin-dry solvent, separated and purified by column chromatography to give 160mg of compound 9 in 16% yield, while 155mg of compound 10 was obtained in 16% yield. Compound 9¹H NMR(500MHz,DMSO-d₆)δ12.85(s,1H),7.63(dd,J＝8.5,2.2Hz,1H),7.54(d,J＝2.2Hz,1H),7.14(d,J＝8.5Hz,1H),6.93(s,1H),6.29(s,1H),5.23(t,J＝6.9Hz,1H),3.85(d,J＝1.9Hz,6H),3.44(d,J＝6.9Hz,2H),1.75(s,3H),1.63(s,3H).HRMS(ESI):calculated for C₂₂H₂₂O₆[M+H]⁺＝383.1489;found 383.1492. Compound 10¹H NMR(400MHz,DMSO-d₆)δ13.18(s,1H),10.83(s,2H),7.67(dd,J＝8.5,2.2Hz,1H),7.55(d,J＝2.2Hz,1H),7.13(d,J＝8.6Hz,1H),6.95(s,1H),6.57(s,1H),5.18(m,1H),3.89(s,3H),3.85(s,3H),3.23(d,J＝7.2Hz,2H),1.73(s,3H),1.63(s,3H).HRMS(ESI):calculated for C₂₂H₂₂O₆[M+H]⁺＝383.1489;found383.1489.

Example 6

Synthesis of compounds 11 and 12:

the synthetic chemical reaction formula is shown below:

The specific synthesis method comprises the following steps:

Step 1, the above product C (2.0 g,6.17 mmol) was dissolved in absolute ethanol (20 mL), potassium hydroxide (1.38 g,24.66 mmol) was gradually added to the reaction system, stirred at room temperature, and after the potassium hydroxide was completely dissolved, 4-methoxy-3, 5-bis (methoxymethoxy) benzaldehyde (1.58 g,6.17 mmol) was added and stirred at room temperature for 3 hours. After the reaction of the raw materials is completed, the reaction solution is concentrated, diluted by adding ethyl acetate, extracted by adding water, the water phase is back extracted by using ethyl acetate for three times, and the EA layers are combined and washed by saturated saline. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin-drying to give crude oil, which was purified by column chromatography to give 2.43g of yellow oily product 6a in a yield of 70%.

Step 2. The reaction product 6a (1.00 g,1.78 mmol) from the previous step was placed in a reaction flask, dimethyl sulfoxide (20 mL) was added for dissolution, iodine (45.11 mg,0.18 mmol) was added, and the mixture was allowed to react at 110℃for 1.5 hours. After the reaction of the raw materials was completed, the reaction solution was diluted with ethyl acetate, extracted with water, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated in vacuo to give a crude yellow oil, which was not further purified, added with methanol (20 mL) and 10% diluted hydrochloric acid solution (10 mL), refluxed at 80 ℃ for 0.5 hours, concentrated in vacuo, extracted with ethyl acetate, washed with saturated brine, combined with the organic phase, dried over anhydrous sodium sulfate, dried over spin-dry solvent, separated and purified by column chromatography to give 142mg of compound 11 in 14% yield while 139mg of compound 12 was obtained in 14% yield. Compound 11¹H NMR(500MHz,DMSO-d₆)δ12.81(s,1H),10.64(s,1H),9.61(s,2H),6.99(s,2H),6.58(s,1H),6.28(s,1H),5.22(t,1H),3.77(s,3H),3.44(d,J＝7.2Hz,2H),1.76(s,3H),1.64(s,3H).HRMS(ESI):calculated for C₂₁H₂₀O₇[M+H]⁺＝385.1282;found385.1283. Compound 12¹H NMR(500MHz,DMSO-d₆)δ13.11(s,1H),9.59(s,3H),6.96(s,2H),6.58(s,1H),6.48(s,1H),5.18(t,1H),3.76(s,3H),3.22(d,J＝7.2Hz,2H),1.73(s,3H),1.63(s,3H).HRMS(ESI):calculated for C₂₁H₂₀O₇[M+H]⁺＝385.1282;found 385.1283.

Example 7

Synthesis of compounds 13 and 14:

the synthetic chemical reaction formula is shown below:

The specific synthesis method comprises the following steps:

Step 1, the above product C (2.0 g,6.17 mmol) was dissolved in absolute ethanol (20 mL), potassium hydroxide (1.38 g,24.66 mmol) was gradually added to the reaction system, stirred at room temperature, and after the potassium hydroxide was completely dissolved, 4-isopropylbenzaldehyde (0.91 g,6.17 mmol) was added, and stirred at room temperature for 3 hours. After the reaction of the raw materials is completed, the reaction solution is concentrated, diluted by adding ethyl acetate, extracted by adding water, the water phase is back extracted by using ethyl acetate for three times, and the EA layers are combined and washed by saturated saline. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin-drying to give crude oil, which was purified by column chromatography to give 2.17g of 7a as a yellow oily product in 77% yield.

Step 2. The reaction product 7a (1.00 g,2.20 mmol) from the previous step was placed in a reaction flask, dimethyl sulfoxide (20 mL) was added for dissolution, iodine (55.84 mg,0.22 mmol) was added, and the mixture was allowed to react at 110℃for 1.5 hours. After the reaction of the raw materials was completed, the reaction solution was diluted with ethyl acetate, extracted with water, washed with saturated sodium sulfate, combined with an organic phase, dried over anhydrous sodium sulfate, concentrated in vacuo to give a crude yellow oil, which was not further purified, added with methanol (20 mL) and 10% diluted hydrochloric acid solution (10 mL), refluxed at 80 ℃ for 0.5 hours, concentrated in vacuo, extracted with ethyl acetate, washed with saturated sodium sulfate, combined with an organic phase, dried over anhydrous sodium sulfate, dried over spin-dry solvent, separated and purified by column chromatography to give 128mg of compound 13 in 13% yield, while 145mg of compound 14 was obtained in 15% yield. Compound 13¹H NMR(500MHz,DMSO-d₆)δ13.12(s,1H),10.94(s,1H),7.96(d,J＝8.1Hz,2H),7.42(d,J＝8.1Hz,2H),6.89(s,1H),6.55(s,1H),5.18(t,1H),3.22(d,J＝7.2Hz,2H),2.97(hept,J＝7.0Hz,1H),1.72(s,3H),1.62(s,3H),1.22(d,J＝6.9Hz,6H).HRMS(ESI):calculated for C₂₃H₂₄O₄[M+H]⁺＝365.1747;found 365.1745. Compound 14¹H NMR(500MHz,DMSO-d₆)δ12.81(s,1H),10.87(s,1H),7.97(d,J＝8.2Hz,2H),7.46(d,J＝8.2Hz,2H),6.91(s,1H),6.31(s,1H),5.21(t,J＝6.9Hz,1H),3.45(d,J＝6.9Hz,2H),2.99(hept,J＝6.9Hz,1H),1.78(s,3H),1.64(s,3H),1.24(d,J＝6.9Hz,6H).HRMS(ESI):calculated for C₂₃H₂₄O₄[M+H]⁺＝365.1747;found365.1746.

Example 8

Synthesis of Compounds 15 and 16:

the synthetic chemical reaction formula is shown below:

The specific synthesis method comprises the following steps:

Step 1. The above-mentioned product C (2.0 g,6.17 mmol) was dissolved in absolute ethanol (20 mL), potassium hydroxide (1.38 g,24.66 mmol) was gradually added to the reaction system, stirred at room temperature, and after the potassium hydroxide was completely dissolved, 4- (4-morpholino) benzaldehyde (1.18 g,6.17 mmol) was added and stirred at room temperature for 3 hours. After the reaction of the raw materials is completed, the reaction solution is concentrated, diluted by adding ethyl acetate, extracted by adding water, the water phase is back extracted by using ethyl acetate for three times, and the EA layers are combined and washed by saturated saline. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin-drying to give crude oil, which was purified by column chromatography to give 2.11g of a yellow oily product 8a in 69% yield.

Step 2. The reaction product 8a (1.00 g,2.01 mmol) from the previous step was placed in a reaction flask, dimethyl sulfoxide (20 mL) was added for dissolution, iodine (51.01 mg,0.20 mmol) was added, and the mixture was allowed to react at 110℃for 1.5 hours. After the reaction of the raw materials was completed, the reaction solution was diluted with ethyl acetate, extracted with water, washed with saturated sodium sulfate, combined with an organic phase, dried over anhydrous sodium sulfate, concentrated in vacuo to give a crude yellow oil, which was not further purified, added with methanol (20 mL) and 10% diluted hydrochloric acid solution (10 mL), refluxed at 80 ℃ for 0.5 hours, concentrated in vacuo, extracted with ethyl acetate, washed with saturated sodium sulfate, combined with an organic phase, dried over anhydrous sodium sulfate, dried over spin-dry solvent, separated and purified by column chromatography to give 134mg of compound 15 in 13% yield, while 145mg of compound 16 was obtained in 15% yield. Compound 15¹H NMR(500MHz,DMSO-d₆)δ12.97(s,1H),10.77(s,1H),7.88(d,J＝8.7Hz,2H),7.06(d,J＝8.7Hz,2H),6.75(s,1H),6.28(s,1H),5.20(t,J＝7.0Hz,1H),3.75(t,J＝4.8Hz,4H),3.44(d,J＝7.0Hz,2H),3.30(t,J＝4.9Hz,4H),1.78(s,3H),1.64(s,3H).HRMS(ESI):calculated for C₂₄H₂₅NO₅[M+H]⁺＝408.1805;found 408.1806. Compound 16¹H NMR(500MHz,DMSO-d₆)δ13.28(s,1H),10.83(s,1H),7.91(d,J＝8.7Hz,2H),7.05(d,J＝8.8Hz,2H),6.77(s,1H),6.53(s,1H),5.19(t,J＝7.3Hz,1H),3.75(t,J＝4.8Hz,4H),3.29(t,J＝4.9Hz,4H),3.22(d,J＝7.2Hz,2H),1.73(s,3H),1.63(s,3H).HRMS(ESI):calculated for C₂₄H₂₅NO₅[M+H]⁺＝408.1805;found 408.1805.

Example 9

Synthesis of compounds 17 and 18:

the synthetic chemical reaction formula is shown below:

The specific synthesis method comprises the following steps:

Step 1, the above product C (2.0 g,6.17 mmol) was dissolved in absolute ethanol (20 mL), potassium hydroxide (1.38 g,24.66 mmol) was gradually added to the reaction system, stirred at room temperature, and after the potassium hydroxide was completely dissolved, 4-trifluoromethylbenzaldehyde (1.07 g,6.17 mmol) was added, and stirred at room temperature for 3 hours. After the reaction of the raw materials is completed, the reaction solution is concentrated, diluted by adding ethyl acetate, extracted by adding water, the water phase is back extracted by using ethyl acetate for three times, and the EA layers are combined and washed by saturated saline. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin-drying to give crude oil, which was purified by column chromatography to give 2.35g of a yellow oily product 9a in 79% yield.

Step 2. The reaction product 9a (1.00 g,2.08 mmol) from the previous step was placed in a reaction flask, dimethyl sulfoxide (20 mL) was added for dissolution, iodine (52.82 mg,0.21 mmol) was added, and the mixture was allowed to react at 110℃for 1.5 hours. After the reaction of the raw materials was completed, the reaction solution was diluted with ethyl acetate, extracted with water, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated in vacuo to give a crude yellow oil, which was not further purified, added with methanol (20 mL) and 10% diluted hydrochloric acid solution (10 mL), refluxed at 80 ℃ for 0.5 hours, concentrated in vacuo, extracted with ethyl acetate, washed with saturated brine, combined with the organic phase, dried over anhydrous sodium sulfate, dried over spin-dry solvent, separated and purified by column chromatography to give 142mg of compound 17 in 14% yield while 127mg of compound 18 was obtained in 13% yield. Compound 17¹H NMR(400MHz,DMSO-d₆)δ12.96(s,1H),10.98(s,1H),8.26(d,J＝8.2Hz,2H),7.91(d,J＝8.3Hz,2H),7.08(s,1H),6.58(s,1H),5.18(t,J＝7.2Hz,1H),3.22(d,J＝7.2Hz,2H),1.73(s,3H),1.63(s,3H).HRMS(ESI):calculated for C₂₁H₁₇F₃O₄[M+H]⁺＝391.1152;found 391.1151. Compound 18¹H NMR(500MHz,DMSO-d₆)δ12.66(s,1H),10.94(s,1H),8.24(d,J＝8.2Hz,2H),7.96(d,J＝8.2Hz,2H),7.09(s,1H),6.33(s,1H),5.20(t,J＝7.0Hz,1H),3.45(d,J＝7.0Hz,2H),1.76(s,3H),1.64(s,3H).HRMS(ESI):calculated for C₂₁H₁₇F₃O₄[M+H]⁺＝391.1152;found391.1154.

Example 10

Synthesis of Compounds 19 and 20:

the synthetic chemical reaction formula is shown below:

The specific synthesis method comprises the following steps:

Step 1, the above product C (2.0 g,6.17 mmol) was dissolved in absolute ethanol (20 mL), potassium hydroxide (1.38 g,24.66 mmol) was gradually added to the reaction system, stirred at room temperature, and after the potassium hydroxide was completely dissolved, 3-trifluoromethylbenzaldehyde (1.07 g,6.17 mmol) was added, and stirred at room temperature for 3 hours. After the reaction of the raw materials is completed, the reaction solution is concentrated, diluted by adding ethyl acetate, extracted by adding water, the water phase is back extracted by using ethyl acetate for three times, and the EA layers are combined and washed by saturated saline. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin-drying to give an oily crude product, which was purified by column chromatography to give 2.40g of a yellow oily product 10a in 81% yield.

Step 2. The reaction product 10a (1.00 g,2.08 mmol) from the previous step was placed in a reaction flask, dimethyl sulfoxide (20 mL) was added for dissolution, iodine (52.82 mg,0.21 mmol) was added, and the mixture was allowed to react at 110℃for 1.5 hours. After the reaction of the raw materials was completed, the reaction solution was diluted with ethyl acetate, extracted with water, washed with saturated sodium sulfate, combined with an organic phase, dried over anhydrous sodium sulfate, concentrated in vacuo to give a crude yellow oil, which was not further purified, added with methanol (20 mL) and 10% diluted hydrochloric acid solution (10 mL), refluxed at 80 ℃ for 0.5 hours, concentrated in vacuo, extracted with ethyl acetate, washed with saturated sodium sulfate, combined with an organic phase, dried over anhydrous sodium sulfate, dried over spin-dry solvent, separated and purified by column chromatography to give 150mg of compound 19 in 18% yield while obtaining 149mg of compound 20 in 15% yield. Compound 19¹H NMR(400MHz,DMSO-d₆)δ13.00(s,1H),10.96(s,1H),8.39–8.35(m,2H),7.96(d,J＝7.8Hz,1H),7.81(t,J＝8.1Hz,1H),7.15(s,1H),6.61(s,1H),5.19(t,J＝7.2Hz,1H),3.23(d,J＝7.2Hz,2H),1.73(s,3H),1.64(s,3H).HRMS(ESI):calculated for C₂₁H₁₇F₃O₄[M+H]⁺＝391.1152;found 391.1153. Compound 20¹H NMR(500MHz,DMSO-d₆)δ12.68(s,1H),10.94(s,1H),8.39–8.31(m,2H),7.98(d,J＝7.8Hz,1H),7.83(t,J＝7.9Hz,1H),7.15(s,1H),6.33(s,1H),5.20(t,J＝6.8Hz,1H),3.45(d,J＝6.8Hz,2H),1.74(s,3H),1.63(s,3H).HRMS(ESI):calculated for C₂₁H₁₇F₃O₄[M+H]⁺＝391.1152;found 391.1152.

Example 11

Synthesis of Compounds 21 and 22:

the synthetic chemical reaction formula is shown below:

The specific synthesis method comprises the following steps:

Step 1, the above product C (2.0 g,6.17 mmol) was dissolved in absolute ethanol (20 mL), potassium hydroxide (1.38 g,24.66 mmol) was gradually added to the reaction system, stirred at room temperature, and after the potassium hydroxide was completely dissolved, 2-trifluoromethylbenzaldehyde (1.07 g,6.17 mmol) was added, and stirred at room temperature for 3 hours. After the reaction of the raw materials is completed, the reaction solution is concentrated, diluted by adding ethyl acetate, extracted by adding water, the water phase is back extracted by using ethyl acetate for three times, and the EA layers are combined and washed by saturated saline. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin-drying to give an oily crude product, which was purified by column chromatography to give 2.27g of a yellow oily product 11a in 77% yield.

Step 2. The reaction product 11a (1.00 g,2.08 mmol) from the previous step was placed in a reaction flask, dimethyl sulfoxide (20 mL) was added for dissolution, iodine (52.82 mg,0.21 mmol) was added, and the mixture was allowed to react at 110℃for 1.5 hours. After the reaction of the raw materials was completed, the reaction solution was diluted with ethyl acetate, extracted with water, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated in vacuo to give a crude yellow oil, which was not further purified, added with methanol (20 mL) and 10% diluted hydrochloric acid solution (10 mL), refluxed at 80 ℃ for 0.5 hours, concentrated in vacuo, extracted with ethyl acetate, washed with saturated brine, combined with the organic phase, dried over anhydrous sodium sulfate, dried over spin-dry solvent, separated and purified by column chromatography to give 143mg of compound 21 in 14% yield while obtaining 133mg of compound 22 in 13% yield. Compound 21¹H NMR(500MHz,DMSO-d₆)δ12.90(s,1H),7.98(d,J＝7.6Hz,1H),7.87(d,J＝4.3Hz,2H),7.83(dd,J＝8.1,4.7Hz,1H),6.57(s,1H),6.43(s,1H),5.18(t,J＝7.1Hz,1H),3.23(d,J＝7.2Hz,2H),1.73(s,3H),1.63(s,3H).HRMS(ESI):calculated for C₂₁H₁₇F₃O₄[M+H]⁺＝391.1152;found 391.1152. Compound 22¹H NMR(400MHz,DMSO-d₆)δ12.89(s,1H),10.94(s,1H),7.97(d,J＝7.4Hz,1H),7.87(d,J＝4.2Hz,2H),7.85–7.79(m,1H),6.56(s,1H),6.43(s,1H),5.18(t,1H),3.24(d,J＝7.2Hz,2H),1.73(s,3H),1.63(s,3H).HRMS(ESI):calculated for C₂₁H₁₇F₃O₄[M+H]⁺＝391.1152;found 391.1151.

Example 12

Synthesis of Compounds 23 and 24:

the synthetic chemical reaction formula is shown below:

The specific synthesis method comprises the following steps:

Step 1, the above product C (2.0 g,6.17 mmol) was dissolved in absolute ethanol (20 mL), potassium hydroxide (1.38 g,24.66 mmol) was gradually added to the reaction system, stirred at room temperature, and after the potassium hydroxide was completely dissolved, 2-methylbenzaldehyde (1.07 g,6.17 mmol) was added, and stirred at room temperature for 3 hours. After the reaction of the raw materials is completed, the reaction solution is concentrated, diluted by adding ethyl acetate, extracted by adding water, the water phase is back extracted by using ethyl acetate for three times, and the EA layers are combined and washed by saturated saline. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin-drying to give an oily crude product, which was purified by column chromatography to give 2.31g of a yellow oily product 12a in 88% yield.

Step 2. The reaction product 12a (1.00 g,2.34 mmol) from the previous step was placed in a reaction flask, dimethyl sulfoxide (20 mL) was added for dissolution, iodine (59.51 mg,0.23 mmol) was added, and the mixture was allowed to react at 110℃for 1.5 hours. After the reaction of the raw materials was completed, the reaction solution was diluted with ethyl acetate, extracted with water, washed with saturated sodium sulfate, combined with an organic phase, dried over anhydrous sodium sulfate, concentrated in vacuo to give a crude yellow oil, which was not further purified, added with methanol (20 mL) and 10% diluted hydrochloric acid solution (10 mL), refluxed at 80 ℃ for 0.5 hours, concentrated in vacuo, extracted with ethyl acetate, washed with saturated sodium sulfate, combined with an organic phase, dried over anhydrous sodium sulfate, dried over spin-dry solvent, separated and purified by column chromatography to give 175mg of compound 23 in 18% yield while 158mg of compound 24 was obtained in 16% yield. Compound 23¹H NMR(500MHz,DMSO-d₆)δ13.05(s,1H),10.85(s,1H),7.59(dd,J＝7.6,1.4Hz,1H),7.48(td,J＝7.5,1.4Hz,1H),7.42–7.34(m,2H),6.47(s,1H),6.45(s,1H),5.18(tt,J＝7.2,1.6Hz,1H),3.24(d,J＝7.2Hz,2H),2.42(s,3H),1.73(s,3H),1.63(s,3H).HRMS(ESI):calculated for C₂₁H₂₀O₄[M+H]⁺＝337.1434;found 337.1431. Compound 24¹H NMR(500MHz,DMSO-d₆)δ12.77(s,1H),10.88(s,1H),7.58(d,J＝7.7Hz,1H),7.48(td,J＝7.5,1.4Hz,1H),7.42–7.33(m,2H),6.42(s,1H),6.33(s,1H),5.08(t,J＝7.0Hz,1H),3.31(d,J＝7.1Hz,2H),2.43(s,3H),1.58(s,3H),1.55(s,3H).HRMS(ESI):calculated for C₂₁H₂₀O₄[M+H]⁺＝337.1434;found337.1434.

Example 13

Synthesis of compounds 25 and 26:

the synthetic chemical reaction formula is shown below:

The specific synthesis method comprises the following steps:

Step 1, the above product C (2.0 g,6.17 mmol) was dissolved in absolute ethanol (20 mL), potassium hydroxide (1.38 g,24.66 mmol) was gradually added to the reaction system, stirred at room temperature, and 3-methylbenzaldehyde (1.07 g,6.17 mmol) was added after the potassium hydroxide was completely dissolved, and stirred at room temperature for 3 hours. After the reaction of the raw materials is completed, the reaction solution is concentrated, diluted by adding ethyl acetate, extracted by adding water, the water phase is back extracted by using ethyl acetate for three times, and the EA layers are combined and washed by saturated saline. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin-drying to give an oily crude product, which was purified by column chromatography to give 2.33g of a yellow oily product 13a in 89% yield.

Step 2. The reaction product 13a (1.00 g,2.34 mmol) from the previous step was placed in a reaction flask, dimethyl sulfoxide (20 mL) was added for dissolution, iodine (59.51 mg,0.23 mmol) was added, and the mixture was allowed to react at 110℃for 1.5 hours. After the reaction of the raw materials was completed, the reaction solution was diluted with ethyl acetate, extracted with water, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated in vacuo to give a crude yellow oil, which was not further purified, added with methanol (20 mL) and 10% diluted hydrochloric acid solution (10 mL), refluxed at 80 ℃ for 0.5 hours, concentrated in vacuo, extracted with ethyl acetate, washed with saturated brine, combined with the organic phase, dried over anhydrous sodium sulfate, dried over spin-dry solvent, separated and purified by column chromatography to give 150mg of compound 25 in 15% yield, and 145mg of compound 26 was obtained simultaneously in 15% yield. Compound 25¹H NMR(500MHz,DMSO-d₆)δ13.10(s,1H),10.93(s,1H),7.88(s,1H),7.84(d,J＝7.7Hz,1H),7.43(dt,J＝15.5,7.6Hz,2H),6.92(s,1H),6.57(s,1H),5.18(t,J＝7.3Hz,1H),3.22(d,J＝7.2Hz,2H),2.41(s,3H),1.73(s,3H),1.62(s,3H).HRMS(ESI):calculated for C₂₁H₂₀O₄[M+H]⁺＝337.1434;found337.1434. Compound 26¹H NMR(400MHz,DMSO-d₆)δ12.76(s,1H),7.84(s,1H),7.81(d,J＝7.6Hz,1H),7.43(dt,J＝14.0,7.5Hz,2H),6.89(s,1H),6.31(s,1H),5.19(t,J＝6.8Hz,1H),3.44(d,J＝6.9Hz,2H),2.39(s,3H),1.76(s,3H),1.63(s,3H).HRMS(ESI):calculated for C₂₁H₂₀O₄[M+H]⁺＝337.1434;found 337.1442.

Example 14

Synthesis of Compounds 27 and 28:

the synthetic chemical reaction formula is shown below:

The specific synthesis method comprises the following steps:

Step 1, the above product C (2.0 g,6.17 mmol) was dissolved in absolute ethanol (20 mL), potassium hydroxide (1.38 g,24.66 mmol) was gradually added to the reaction system, stirred at room temperature, and after the potassium hydroxide was completely dissolved, 2-chloro-benzaldehyde (0.87 g,6.17 mmol) was added, and stirred at room temperature for 3 hours. After the reaction of the raw materials is completed, the reaction solution is concentrated, diluted by adding ethyl acetate, extracted by adding water, the water phase is back extracted by using ethyl acetate for three times, and the EA layers are combined and washed by saturated saline. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin-drying to give crude oil, which was purified by column chromatography to give 2.14g of the yellow oily product 14a in 78% yield.

Step 2. The reaction product 14a (1.00 g,2.24 mmol) from the previous step was placed in a reaction flask, dimethyl sulfoxide (20 mL) was added for dissolution, iodine (56.79 mg,0.22 mmol) was added, and the mixture was allowed to react at 110℃for 1.5 hours. After the reaction of the raw materials was completed, the reaction solution was diluted with ethyl acetate, extracted with water, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated in vacuo to give a crude yellow oil, which was not further purified, added with methanol (20 mL) and 10% diluted hydrochloric acid solution (10 mL), refluxed at 80 ℃ for 0.5 hours, concentrated in vacuo, extracted with ethyl acetate, washed with saturated brine, combined with the organic phase, dried over anhydrous sodium sulfate, dried over spin-dry solvent, separated and purified by column chromatography to give 177mg of compound 27 in 18% yield while 161mg of compound 28 was obtained in 16% yield. Compound 27¹H NMR(500MHz,DMSO-d₆)δ12.63(s,1H),10.96(s,1H),7.77(dd,J＝7.7,1.7Hz,1H),7.68(d,J＝8.4Hz,1H),7.61(td,J＝7.7,1.7Hz,1H),7.54(td,J＝7.5,1.2Hz,1H),6.56(s,1H),6.34(s,1H),5.10(t,1H),3.32(d,J＝7.3Hz,2H),1.56(s,3H),1.53(s,3H).HRMS(ESI):calculated for C₂₀H₁₇ClO₄[M+H]⁺＝357.0888;found 357.0886. Compound 28¹H NMR(500MHz,DMSO-d₆)δ12.93(s,1H),10.99(s,1H),7.77(dd,J＝7.6,1.7Hz,1H),7.67(d,J＝7.9Hz,1H),7.61(td,J＝7.7,1.7Hz,1H),7.53(dd,J＝8.0,6.7Hz,1H),6.56(s,1H),6.47(s,1H),5.17(t,J＝7.0Hz,1H),3.23(d,J＝7.2Hz,2H),1.72(s,3H),1.62(s,3H).HRMS(ESI):calculated for C₂₀H₁₇ClO₄[M+H]⁺＝357.0888;found357.0888.

Example 15

Synthesis of compounds 29 and 30:

the synthetic chemical reaction formula is shown below:

The specific synthesis method comprises the following steps:

Step 1, the above product C (2.0 g,6.17 mmol) was dissolved in absolute ethanol (20 mL), potassium hydroxide (1.38 g,24.66 mmol) was gradually added to the reaction system, stirred at room temperature, and 3-chloro-benzaldehyde (0.87 g,6.17 mmol) was added after the potassium hydroxide was completely dissolved, and stirred at room temperature for 3 hours. After the reaction of the raw materials is completed, the reaction solution is concentrated, diluted by adding ethyl acetate, extracted by adding water, the water phase is back extracted by using ethyl acetate for three times, and the EA layers are combined and washed by saturated saline. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin-drying to give an oily crude product, which was purified by column chromatography to give 2.43g of a yellow oily product 15a in 88% yield.

Step 2. The reaction product 15a (1.00 g,2.24 mmol) from the previous step was placed in a reaction flask, dimethyl sulfoxide (20 mL) was added for dissolution, iodine (56.79 mg,0.22 mmol) was added, and the mixture was allowed to react at 110℃for 1.5 hours. After the reaction of the raw materials was completed, the reaction solution was diluted with ethyl acetate, extracted with water, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated in vacuo to give a crude yellow oil, which was not further purified, added with methanol (20 mL) and 10% diluted hydrochloric acid solution (10 mL), refluxed at 80 ℃ for 0.5 hours, concentrated in vacuo, extracted with ethyl acetate, washed with saturated brine, combined with the organic phase, dried over anhydrous sodium sulfate, dried over spin-dry solvent, separated and purified by column chromatography to give 162mg of compound 29 in 16% yield while obtaining 155mg of compound 30 in 16% yield. Compound 29¹H NMR(500MHz,DMSO-d₆)δ13.01(s,1H),10.98(s,1H),8.12(t,J＝1.9Hz,1H),8.02(d,J＝7.9Hz,1H),7.66(dd,J＝8.0,2.0Hz,1H),7.58(t,J＝7.9Hz,1H),7.04(s,1H),6.58(s,1H),5.17(t,J＝7.2Hz,1H),3.22(d,J＝7.2Hz,2H),1.72(s,3H),1.62(s,3H).HRMS(ESI):calculated for C₂₀H₁₇ClO₄[M+H]⁺＝357.0888;found 357.0886. Compound 30¹H NMR(500MHz,DMSO-d₆)δ12.67(s,1H),10.91(s,1H),8.02(t,J＝1.9Hz,1H),7.99(d,J＝8.0Hz,1H),7.66(dd,J＝8.2,1.4Hz,1H),7.59(t,J＝7.9Hz,1H),7.02(s,1H),6.31(s,1H),5.16(t,1H),3.42(d,J＝6.8Hz,2H),1.77(s,3H),1.64(s,3H).HRMS(ESI):calculated for C₂₀H₁₇ClO₄[M+H]⁺＝357.0888;found357.0889.

Example 16

Synthesis of compounds 31 and 32:

the synthetic chemical reaction formula is shown below:

The specific synthesis method comprises the following steps:

step 1, the above product C (2.0 g,6.17 mmol) was dissolved in absolute ethanol (20 mL), potassium hydroxide (1.38 g,24.66 mmol) was gradually added to the reaction system, stirred at room temperature, and after the potassium hydroxide was completely dissolved, 4-chloro-3-methylbenzaldehyde (0.95 g,6.17 mmol) was added, and stirred at room temperature for 3 hours. After the reaction of the raw materials is completed, the reaction solution is concentrated, diluted by adding ethyl acetate, extracted by adding water, the water phase is back extracted by using ethyl acetate for three times, and the EA layers are combined and washed by saturated saline. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin-drying to give crude oil, which was purified by column chromatography to give 2.22g of the yellow oily product 16a in 78% yield.

Step 2. The reaction product 16a (1.00 g,2.17 mmol) from the previous step was placed in a reaction flask, dimethyl sulfoxide (20 mL) was added for dissolution, iodine (55.06 mg,0.22 mmol) was added, and the mixture was allowed to react at 110℃for 1.5 hours. After the reaction of the raw materials was completed, the reaction solution was diluted with ethyl acetate, extracted with water, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated in vacuo to give a crude yellow oil, which was not further purified, added with methanol (20 mL) and 10% diluted hydrochloric acid solution (10 mL), refluxed at 80 ℃ for 0.5 hours, concentrated in vacuo, extracted with ethyl acetate, washed with saturated brine, combined with the organic phase, dried over anhydrous sodium sulfate, dried over spin-dry solvent, separated and purified by column chromatography to give 188mg of compound 31 in 19% yield while 173mg of compound 32 was obtained in 17% yield. Compound 31¹H NMR(500MHz,DMSO-d₆)δ13.04(s,1H),10.96(s,1H),8.06(d,J＝2.3Hz,1H),7.87(dd,J＝8.4,2.4Hz,1H),7.57(d,J＝8.4Hz,1H),6.95(s,1H),6.56(s,1H),5.17(t,J＝7.1Hz,1H),3.21(d,J＝7.2Hz,2H),2.41(s,3H),1.72(s,3H),1.62(s,3H).HRMS(ESI):calculated for C₂₁H₁₉ClO₄[M+H]⁺＝371.1045;found 371.1043. Compound 32¹H NMR(500MHz,DMSO-d₆)δ12.71(s,1H),10.88(s,1H),8.01(d,J＝2.3Hz,1H),7.84(dd,J＝8.4,2.3Hz,1H),7.61(d,J＝8.4Hz,1H),6.95(s,1H),6.30(s,1H),5.18(t,J＝6.9Hz,1H),3.43(d,J＝6.8Hz,2H),2.40(s,3H),1.76(s,3H),1.63(s,3H).HRMS(ESI):calculated for C₂₁H₁₉ClO₄[M+H]⁺＝371.1045;found 371.1049.

Example 17

Synthesis of Compounds 33 and 34:

the synthetic chemical reaction formula is shown below:

The specific synthesis method comprises the following steps:

Step 1) the above-mentioned product C (2.0 g,6.17 mmol) was dissolved in absolute ethanol (20 mL), potassium hydroxide (1.38 g,24.66 mmol) was gradually added to the reaction system, stirred at room temperature, and after the potassium hydroxide was completely dissolved, 4-chloro-3- (methoxymethoxy) benzaldehyde (1.24 g,6.17 mmol) was added and stirred at room temperature for 3 hours. After the reaction of the raw materials is completed, the reaction solution is concentrated, diluted by adding ethyl acetate, extracted by adding water, the water phase is back extracted by using ethyl acetate for three times, and the EA layers are combined and washed by saturated saline. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin-drying to give an oily crude product, which was purified by column chromatography to give 2.30g of a yellow oily product 17a in 74% yield.

Step 2. The reaction product 17a (1.00 g,1.97 mmol) from the previous step was placed in a reaction flask, dimethyl sulfoxide (20 mL) was added for dissolution, iodine (50.06 mg,0.20 mmol) was added, and the mixture was allowed to react at 110℃for 1.5 hours. After the reaction of the raw materials was completed, the reaction solution was diluted with ethyl acetate, extracted with water, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated in vacuo to give a crude yellow oil, which was not further purified, added with methanol (20 mL) and 10% diluted hydrochloric acid solution (10 mL), refluxed at 80 ℃ for 0.5 hours, concentrated in vacuo, extracted with ethyl acetate, washed with saturated brine, combined with the organic phase, dried over anhydrous sodium sulfate, dried over spin-dry solvent, separated and purified by column chromatography to give 175mg of compound 33 in 18% yield while 151mg of compound 34 was obtained in 15% yield. Compound 33¹H NMR(400MHz,DMSO-d₆)δ13.03(s,1H),10.80(s,1H),7.56(s,1H),7.50(s,2H),6.82(s,1H),6.51(s,1H),5.18(t,J＝7.2Hz,1H),3.22(d,J＝7.2Hz,2H),1.73(s,3H),1.62(s,3H).HRMS(ESI):calculated for C₂₀H₁₇ClO₅[M+H]⁺＝373.0837;found 373.0838. Compound 34¹H NMR(500MHz,DMSO-d₆)δ12.72(s,1H),10.80(s,2H),7.61(d,J＝2.1Hz,1H),7.52(d,J＝8.4Hz,1H),7.48(dd,J＝8.4,2.1Hz,1H),6.81(s,1H),6.30(s,1H),5.22(t,J＝7.2Hz,1H),3.44(d,J＝7.2Hz,2H),1.74(s,3H),1.62(s,3H).HRMS(ESI):calculated for C₂₀H₁₇ClO₅[M+H]⁺＝373.0837;found 373.0840.

Example 18

Synthesis of compounds 35 and 36:

the synthetic chemical reaction formula is shown below:

The specific synthesis method comprises the following steps:

Step 1) the above-mentioned product C (2.0 g,6.17 mmol) was dissolved in absolute ethanol (20 mL), potassium hydroxide (1.38 g,24.66 mmol) was gradually added to the reaction system, stirred at room temperature, and 3-methoxy-5- (methoxymethoxy) benzaldehyde (1.21 g,6.17 mmol) was added after the potassium hydroxide was completely dissolved, and stirred at room temperature for 3 hours. After the reaction of the raw materials is completed, the reaction solution is concentrated, diluted by adding ethyl acetate, extracted by adding water, the water phase is back extracted by using ethyl acetate for three times, and the EA layers are combined and washed by saturated saline. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin-drying to give crude oil, which was purified by column chromatography to give 2.03g of the yellow oily product 18a in 66% yield.

Step 2. The reaction product 18a (1.00 g,1.99 mmol) from the previous step was placed in a reaction flask, dimethyl sulfoxide (20 mL) was added for dissolution, iodine (50.50 mg,0.20 mmol) was added, and the mixture was allowed to react at 110℃for 1.5 hours. After the reaction of the raw materials was completed, the reaction solution was diluted with ethyl acetate, extracted with water, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated in vacuo to give a crude yellow oil, which was not further purified, added with methanol (20 mL) and 10% diluted hydrochloric acid solution (10 mL), refluxed at 80 ℃ for 0.5 hours, concentrated in vacuo, extracted with ethyl acetate, washed with saturated brine, combined with the organic phase, dried over anhydrous sodium sulfate, dried over spin-dry solvent, separated and purified by column chromatography to give 157mg of compound 35 in 16% yield while obtaining 133mg of compound 36 in 13% yield. Compound 35¹H NMR(400MHz,DMSO-d₆)δ12.75(s,1H),9.97(s,1H),7.02(t,J＝2.1Hz,2H),6.85(s,1H),6.56(d,J＝2.2Hz,1H),6.29(s,1H),3.79(s,3H),3.43(d,J＝7.0Hz,2H),1.75(s,3H),1.63(s,3H).HRMS(ESI):calculated for C₂₁H₂₀O₆[M+H]⁺＝369.1333;found 369.1329. Compound 36¹H NMR(400MHz,DMSO-d₆)δ13.07(s,1H),9.94(s,1H),7.01(d,J＝9.7Hz,1H),6.87(s,1H),6.57–6.50(m,1H),5.18(t,J＝14.1Hz,1H),3.80(s,3H),3.23(d,J＝7.1Hz,2H),1.73(s,3H),1.63(s,3H).HRMS(ESI):calculated for C₂₁H₂₀O₆[M+H]⁺＝369.1333;found369.1335.

Example 19 biological Activity test section:

determination of tumor cell inhibitory Activity of 36 prenylflavonoids in this example:

wherein the tumor refers to human colorectal cancer cells, human breast cancer cells, human non-small cell lung cancer cells and human osteosarcoma cells.

(1) The tumor cells used in the test were human osteosarcoma cell (Sjsa-1) (purchased from Guangzhou Sakuku Biotechnology Co., ltd.), human colon cancer cell (HCT 116) (purchased from Guangzhou Saku Biotechnology Co., ltd.), human non-small cell lung cancer cell (A549) (purchased from Guangzhou Saku Biotechnology Co., ltd.), human liver cancer cell (Hep G2) (Wuhan Punuo Seisaku Life technologies Co., ltd.)

(2) The test method comprises the following steps of determining proliferation inhibition effects of isopentenyl flavonoid compounds on human osteosarcoma cells, human colon cancer cells, human non-small cell lung cancer cells and human liver cancer cells by adopting a CCK-8 method:

1. The tumor cell strains are respectively prepared into single cell suspensions by using corresponding complete culture media, the concentration of the single cell suspensions is 50000 cells/mL, 100 mu L of the cell suspensions are inoculated into 96-well culture plates and placed into a CO ₂ incubator (37 ℃ C., 5% CO ₂, 95% air) for culturing for 24 hours, wherein human osteosarcoma cells (Sjsa-1), human colon cancer cells (HCT 116) and human non-small cell lung cancer cells (A549) are cultured by adopting 1640 culture media (containing 10% neonatal bovine serum and 1% double antibody), and human liver cancer cells (Hep G2) are cultured by adopting a DMEM culture media (containing 10% neonatal bovine serum, 1% double antibody and 0.01mg/mL insulin).

2. Compounds 1 to 36 were dissolved in DMSO and prepared as a 10mM stock solution, followed by dilution to a concentration of 5mM, 2.5mM, 1.25mM, 0.625mM, 0.3125mM, 0.16mM, 0.078mM, 0.039mM, respectively. 1.0 mu L of each concentration is added into each hole cell so that the final concentration is 1.0% of the concentration before adding, two parallel holes are arranged for each concentration of the compound, 1.0 mu L of DMSO is added into a negative control group and a blank control group, and the cells are placed in a CO ₂ incubator for culturing for 48 hours (the negative control group is 50000/mL cells and treated with 0.5 mu LDMSO for deducting the interference of DMSO contained in the original medicine, and the blank control group is free of cells and contains 100 mu L of complete culture medium for deducting the background interference).

3. After 48h of culture, the medium was removed and replaced with fresh medium, then 10. Mu.L of CCK-8 (Cell Counting Kits-8) reagent was added to each well of cells, and after incubation at 37℃for 2 hours, absorbance A at 450nm was measured using a Biotek multifunctional microplate reader, and the inhibition ratios of compounds 1 to 40 to tumor cell growth were calculated by [1- (A drug treatment group-A blank)/(A negative control group-A blank) ]. Times.100%, A being absorbance.

4. The results of the test are shown in Table 2 as a summary of the tumor inhibitory activity of some of the compounds. As shown in Table 2, the isopentenyl flavonoid compounds of the present invention have potential application value in the development of anticancer drugs against such 4 kinds of tumors in general.

TABLE 2 inhibition of several cancer cell lines by partial isopentenyl flavonoids

The results in Table 2 show that for A549, IC ₅₀ (μM) for compound 18 was 9.21, IC ₅₀ (μM) for compound 20 was 9.31, IC ₅₀ (μM) for compound 29 was 8.54, IC ₅₀ (μM) for compound 33 was 7.63, IC ₅₀ (μM) for compound 5 was 4.48 for HCT116, IC ₅₀ (μM) for compound 33 was 3.72, IC ₅₀ (μM) for compound 13 was 11.44, and IC ₅₀ (μM) for compound 33 was 8.96 for HepG 2. Therefore, the compound of the invention has good tumor inhibition activity and good application prospect in preparing anti-tumor drugs and drugs for inhibiting tumor cells.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

As used herein, the terms "comprising," "including," and "containing" are open-ended terms that include the teachings to which the present invention pertains, but do not exclude other aspects.

As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

The embodiments of the present invention have been described in detail above, but the present invention is not limited to the described embodiments. It will be apparent to those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, and yet fall within the scope of the invention.

Claims

1. An isopentenyl flavonoid compound, characterized in that the compound has a structure shown in formula (1):

In the formula, ^R1 is isopentenyl or hydrogen; ^R2 is isopentenyl or hydrogen; ^R3 is hydroxyl, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, halogen, trifluoromethyl, nitro, amino, morpholine ring, cyclopropane, cyclopentane, cyclobutane, hydroxymethyl, hydroxyethyl or hydroxypropyl.

2. The compound according to claim 1, characterized in that the compound comprises the structures shown in formula (2) and formula (3):

In the formula, R ³ is hydroxy, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, halogen, trifluoromethyl, nitro, amino, morpholine ring, cyclopropane, cyclopentane, cyclobutane, hydroxymethyl, hydroxyethyl, or hydroxypropyl.

3. The compound according to claim 2, characterized in that the structure represented by formula (2) is selected from any one of the following structures:

4. The compound according to claim 2, characterized in that the structure represented by formula (3) is selected from any one of the following structures:

5. A method for preparing the compound according to claim 1, characterized in that the preparation method sequentially carries out hydroxyl protection, substitution reaction, Claisen rearrangement, Claisen-Schmidt reaction, ring closure and deprotection reaction to prepare the isopentenyl flavonoid compound; the reaction process is shown in the following reaction formula (I):

6. The preparation method according to claim 5, characterized in that the preparation method comprises the following steps:

The first step: in a first solvent, under the action of an organic base, a hydroxyl protecting agent is used to protect the 2,4-hydroxyl groups of 2,4,6-trihydroxyacetophenone to obtain compound A;

Step 2: In a second solvent, under the action of an inorganic base, a substitution reagent is reacted with the compound A to obtain a compound B;

Step 3: In a third solvent, the compound B is then subjected to a microwave reaction to undergo Claisen rearrangement to obtain a compound C;

Step 4: In a fourth solvent, under the action of an inorganic base, the compound C reacts with an aldehyde to generate a chalcone by a Claisen-Schmidt reaction;

Step 5: In the fifth solvent, the chalcone is ring-closed with an addition reagent and then deprotected with an acid in the sixth solvent to obtain the final product, isopentenyl flavonoid compound.

7. The preparation method according to claim 6, characterized in that:

In the first step, the organic base includes one or two of N,N-diisopropylethylamine and triethylamine; and/or, the hydroxyl protecting agent includes chloromethyl methyl ether; and/or, the first solvent includes one or more of dichloromethane, tetrahydrofuran, and 1,4-dioxane; and/or, the 2,4,6-trihydroxyacetophenone is 1 equivalent, the organic base is 2 to 3 equivalents, the hydroxyl protecting agent is 2 to 3 equivalents, and the first solvent is 10 equivalents; and/or, the reaction temperature is 0°C to room temperature; and/or, the reaction time is 6 to 8 hours; and/or,

In the second step, the substitution reagent includes isopentenyl bromide; the inorganic base includes one or more of potassium carbonate, cesium carbonate, calcium carbonate, and sodium carbonate; the second solvent is acetone; and/or, the compound A is 1 equivalent, the inorganic base is 2 to 4 equivalents, the substitution reagent is 1 to 2 equivalents, and the second solvent is 10 equivalents; and/or, the reaction temperature is 60 to 70° C., and the reaction time is 2 to 4 hours; and/or,

In the third step, the third solvent includes one or two of N,N-diethylaniline and N,N-dimethylaniline; and/or, the compound B is 1 equivalent, and the third solvent is 10 equivalents; and/or, the reaction temperature is 200-220° C., and the reaction time is 1-2 hours; and/or,

In the fourth step, the aldehyde includes one or both of 2-chloro-4-(methoxymethoxy)benzaldehyde and 2-methoxy-4-(methoxymethoxy)benzaldehyde; the inorganic base includes one or both of potassium hydroxide and sodium hydroxide; the fourth solvent includes one or more of anhydrous ethanol, anhydrous methanol, and ethylene glycol; and/or, the compound C is 1 equivalent, the inorganic base is 4 to 6 equivalents, the aldehyde is 1 equivalent, and the fourth solvent is 10 equivalents; and/or, the reaction temperature is room temperature, and the reaction time is 3 to 6 hours; and/or,

In the fifth step, the addition reagent is iodine; the acid includes one or two of hydrochloric acid and sulfuric acid; the fifth solvent is dimethyl sulfoxide; the sixth solvent includes one or more of methanol, ethanol, and ethylene glycol; and/or, the chalcone is 1 equivalent, the addition reagent is 0.1-0.2 equivalents, the fifth solvent is 10 equivalents, the sixth solvent is 10 equivalents, and the acid used is 10% by mass concentration; and/or, the reaction temperature of the ring closure is 110-120° C., and the reaction time of the ring closure is 1.5-3 hours; and/or, the reflux temperature of the deprotection is 70-80° C., and the reaction time of the deprotection is 0.5-1 hour.

8. A medicine/pharmaceutical composition, characterized in that the medicine/pharmaceutical composition comprises the prenylated flavonoid compound according to any one of claims 1 to 4, or the prenylated flavonoid compound prepared by the preparation method according to any one of claims 5 to 7; and/or,

The drug/drug composition is used alone, and/or in combination with other drugs including PDL1 monoclonal antibody; and/or,

The pharmaceutical composition further comprises an excipient, a diluent, an adjuvant, a vehicle or a combination thereof; and/or,

The pharmaceutical composition is formulated as an injectable fluid, aerosol, cream, gel, pill, capsule, syrup, transdermal patch, or excipient.

9. Use of the isopentenyl flavonoid compound according to any one of claims 1 to 4, or the drug/drug composition according to claim 8 in the preparation of anti-tumor drugs or drugs for inhibiting tumor cells.

10. The use according to claim 9, characterized in that the tumor cells include human osteosarcoma cells Sjsa-1, human colon cancer cells HCT116, human non-small cell lung cancer cells A549, and human liver cancer cells Hep G2.