Isopentenyl flavonoid compound as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of anti-cancer drugs, and particularly discloses an isopentenyl flavonoid compound and a preparation method and application thereof.The isopentenyl flavonoid compound is of a structure shown in the formula (1), and according to the method, firstly, chloromethyl methyl ether is used for reacting 2, 4, 6-trihydroxyacetophenone with 2, 4, 6-trihydroxyacetophenone to obtain 2, 4, 6-trihydroxyacetophenone; the preparation method comprises the following steps: protecting hydroxyl at the position 4 by using a reaction solvent, then carrying out a substitution reaction on isopentenyl bromide and the product, then carrying out a microwave reaction by using N, N-diethyl aniline as a solvent to generate Claisen rearrangement, then carrying out a Claisen-Schmidt reaction with corresponding aldehyde to generate chalcone, and finally carrying out ring closing by using iodine and carrying out deprotection by using hydrochloric acid to obtain the chalcone. And preparing to obtain the compound. The compound disclosed by the invention is novel in structure, relatively good in activity and small in toxicity, has relatively good anti-tumor activity as an inhibitor, has relatively good inhibitory activity on various tumor cells including colorectal cancer, breast cancer, lung cancer, liver cancer and the like, can be applied to preparation of anti-tumor drugs, and has wide application prospects. Good application prospects are realized in the anti-tumor aspect.

Description

Technical Field

[0001] The invention belongs to the technical field of anticancer drugs, and specifically relates to an isopentenyl flavonoid compound and a preparation method and application thereof. Background Art

[0002] In nature, there are many types of flavonoids, which have a variety of biological activities and effective medical applications, such as anticancer and antitumor activities, anti-inflammatory and antiviral activities, antibacterial and antifungal activities, anti-cardiovascular diseases, enzyme inhibition activities, anti-free radical and antioxidant activities. In recent years, flavonoids have attracted more and more attention from researchers due to their broad-spectrum biological activities, wide natural distribution, diverse chemical structures and low toxicity. Isopentenyl flavonoids are a unique class of natural flavonoid compounds, characterized by the presence of isopentenyl side chains on the flavonoid skeleton. C-isopentenylated flavonoids can enhance their affinity for p-glycoprotein and permeability to cell membranes, and can significantly improve their biological activity. In addition, this type of flavonoid has no psychoactive activity and will not make patients dependent on drugs. Therefore, isopentenyl flavonoids have gradually become a research hotspot in the industry.

[0003] In 2019, Rea et al. reported the use of isopentenyl transferase to catalyze dimethylallyl diphosphate to synthesize the isopentenyl flavonoid compound cannaflavonoid B. Patent WO2017091837 also discloses biosynthesis in the form of enzymes, but this type of biosynthesis method requires the extraction of a large amount of enzymes from plants, which is costly and not easy to scale up industrially.

[0004] Giovanni Appendino et al. reported the synthesis of isopentenyl flavonoids in A Regiodivergent Synthesis of Ring A C-Prenylflavones, but the catalyst used in their synthesis route was expensive, the cost was high and it was not easy to scale up industrially.

[0005] At present, the synthesis of C-terminal prenylated flavonoids is mostly done by first synthesizing the flavonoid nucleus and then rearranging it, which makes the synthesis difficult. The problems of the currently disclosed chemical synthesis methods are that the synthesis steps are long, the catalyst is expensive, and it is difficult to purify. Therefore, it is urgent to develop a more efficient synthesis method. Summary of the invention

[0006] In order to solve the defects and shortcomings of the prior art, the present invention provides an isopentenyl flavonoid compound and a preparation method and application thereof. The preparation method has the advantages of cheap and readily available raw materials, short reaction cycle, simple operation, and a higher reaction yield than the existing synthesis method.

[0007] The present invention first performs rearrangement and then synthesizes chalcone, and finally performs ring closure, which is easier to purify. In addition, the present invention can stably obtain a pair of isomers during the ring closure process, so that the structure of the product is more diverse.

[0008] The present invention provides an isopentenyl flavonoid compound, wherein the compound has a structure shown in formula (1):

[0009]

[0010] In the formula, R 1 isopentenyl, hydrogen; R 2 isopentenyl, hydrogen; R 3 is hydroxy, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, halogen, trifluoromethyl, nitro, amino, morpholine ring, cyclopropane, cyclopentane, cyclobutane, hydroxymethyl, hydroxyethyl, hydroxypropyl. Preferably, R 1 isopentenyl; R 2 isopentenyl; R 3 It is hydroxy, methyl, isopropyl, methoxy, halogen, trifluoromethyl, or morpholine ring.

[0011] The isopentenyl flavonoid compounds of the present invention include the structures shown in formula (2) and formula (3):

[0012]

[0013] In the formula, R 3 is hydroxy, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, halogen, trifluoromethyl, nitro, amino, morpholine ring, cyclopropane, cyclopentane, cyclobutane, hydroxymethyl, hydroxyethyl, hydroxypropyl. Preferably, R 3 It is hydroxy, methyl, isopropyl, methoxy, halogen, trifluoromethyl, or morpholine ring.

[0014] Furthermore, the structure represented by formula (2) is selected from any one of the following structures:

[0015]

[0016]

[0017] Furthermore, the structure represented by formula (3) is selected from any one of the following structures:

[0018]

[0019]

[0020] The present invention also provides a method for preparing the prenylated flavonoid compound as described above, wherein the method comprises sequentially performing hydroxyl protection, substitution reaction, Claisen rearrangement, Claisen-Schmidt reaction, ring closure and deprotection reaction to prepare the prenylated flavonoid compound, and the reaction process is shown in the following reaction formula (I):

[0021]

[0022] The preparation method comprises the following steps:

[0023] 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;

[0024] 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;

[0025] 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;

[0026] 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;

[0027] 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.

[0028] Specifically,

[0029] In the first step, the organic base includes one or more of N,N-diisopropylethylamine, triethylamine, etc.; preferably, the organic base is N,N-diisopropylethylamine (DIPEA).

[0030] In the first step, the hydroxyl protecting agent includes chloromethyl methyl ether (MOMCl) and the like.

[0031] In the first step, the first solvent includes one or more of dichloromethane, tetrahydrofuran, 1,4-dioxane, etc.; preferably, the first solvent is dichloromethane (DCM).

[0032] In the first step, when 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; preferably, the organic base is 2 equivalents, and the hydroxyl protecting agent is 2 equivalents.

[0033] In the first step, the reaction temperature is 0°C to room temperature; preferably, the reaction temperature is 0°C.

[0034] In the first step, the reaction time is 6 to 8 hours; preferably, the reaction time is 6 hours.

[0035] In the second step, the substitution reagent includes isopentenyl bromide and the like.

[0036] In the second step, the inorganic base includes one or more of potassium carbonate, cesium carbonate, calcium carbonate, sodium carbonate, etc. Preferably, the inorganic base is potassium carbonate.

[0037] In the second step, the second solvent includes acetone and the like.

[0038] In the second step, the compound A is 1 equivalent, the substitution reagent is 1-2 equivalents, the inorganic base is 2-4 equivalents, and the second solvent is 10 equivalents; preferably, the substitution reagent is 1 equivalent, the inorganic base is 2 equivalents, and the second solvent is 10 equivalents.

[0039] In the second step, the reaction temperature is 60-70°C; preferably, the reaction temperature is 65°C.

[0040] In the second step, the reaction time is 2 to 4 hours; preferably, the reaction time is 2 hours.

[0041] In the third step, the third solvent is one or more of N,N-diethylaniline, N,N-dimethylaniline, etc.; preferably, the third solvent is N,N-diethylaniline.

[0042] In the third step, the compound B is 1 equivalent, and the third solvent is 10 equivalents.

[0043] In the third step, the reaction temperature is 200-220°C; preferably, the reaction temperature is 200°C.

[0044] In the third step, the reaction time is 1 to 2 hours; preferably, the reaction time is 1 hour.

[0045] In the fourth step, the aldehyde includes one or more of 2-chloro-4-(methoxymethoxy)benzaldehyde, 2-methoxy-4-(methoxymethoxy)benzaldehyde, etc.; preferably, the aldehyde is determined according to the specific substrate.

[0046] In the fourth step, the inorganic base includes one or more of potassium hydroxide, sodium hydroxide, etc.; preferably, the base is potassium hydroxide.

[0047] In the fourth step, the fourth solvent includes one or more of anhydrous ethanol, anhydrous methanol, ethylene glycol, etc.; preferably, the solvent is anhydrous ethanol.

[0048] In the fourth step, 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; preferably, the base is 4 equivalents;

[0049] In the fourth step, the reaction temperature is room temperature.

[0050] In the fourth step, the reaction time is 3 to 6 hours; preferably, the reaction time is 3 hours.

[0051] In the fifth step, the addition reagent includes iodine and the like.

[0052] In the fifth step, the acid includes one or more of hydrochloric acid, sulfuric acid, etc.; preferably, the acid is hydrochloric acid.

[0053] In the fifth step, the fifth solvent includes dimethyl sulfoxide (DMSO) and the like.

[0054] In the fifth step, the sixth solvent includes one or more of methanol, ethanol, ethylene glycol, etc.; preferably, the sixth solvent is methanol.

[0055] In the fifth step, the chalcone is 1 equivalent, the addition reagent is 0.1-0.2 equivalent, the fifth solvent is 10 equivalents, the sixth solvent is 10 equivalents, and the acid used is 10% mass concentration; preferably, the addition reagent is 0.1 equivalent.

[0056] In the fifth step, the temperature of the ring-closing reaction is 110-120°C; preferably, the temperature of the ring-closing reaction is 110°C.

[0057] In the fifth step, the ring-closure reaction time is 1.5 to 3 hours; preferably, the ring-closure reaction time is 1.5 hours.

[0058] In the fifth step, the reflux temperature of the deprotection reaction is 70-80°C; preferably, the reflux temperature of the deprotection reaction is 80°C.

[0059] In the fifth step, the deprotection reaction time is 0.5 to 1 hour; preferably, the deprotection reaction time is 0.5 hour.

[0060] The present invention also provides a medicine / pharmaceutical composition, which comprises the prenylated flavonoid compound as described above.

[0061] In the present invention, the drug / drug composition can be used alone or in combination with other drugs including PDL1 monoclonal antibody.

[0062] In the present invention, the pharmaceutical composition further comprises an excipient, a diluent, an adjuvant, a vehicle or a combination thereof.

[0063] In the present invention, the pharmaceutical composition is formulated into an injectable fluid, an aerosol, a cream, a gel, a pill, a capsule, a syrup, a transdermal patch or an excipient.

[0064] The present invention also provides the use of the above-mentioned isopentenyl flavonoid compound or the above-mentioned drug / drug composition in the preparation of anti-tumor drugs, drugs for inhibiting tumor cells, etc.

[0065] The isopentenyl flavonoid compound, or the drug / drug composition is used to inhibit the growth, migration, proliferation, etc. of tumor cells, and to promote the apoptosis of tumor cells.

[0066] Preferably, the tumor cells include but are not limited to 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).

[0067] In a specific embodiment, in the present invention, the corresponding isomers can be stably obtained in the iodine ring closure step, so that the structure of the compound has diversity.

[0068] The isopentenyl flavonoid compound of the present invention can be used as a new type of anticancer drug, which can promote cell apoptosis by directly killing tumor cells and activating tumor immunity. It has good inhibitory activity against various cancer cell lines such as 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).

[0069] Compared with the prior art, the beneficial effects of the present invention are as follows: the present invention provides an isopentenyl flavonoid compound, the compound has a novel structure, good activity, and the compound has good anti-tumor activity as an inhibitor, and has good inhibitory activity against a variety of tumor cells (such as human osteosarcoma cells, human colon cancer cells, human non-small cell lung cancer cells, human liver cancer cells, etc.), and can be used to prepare anti-tumor drugs. It has good application prospects in antibacterial and anti-tumor aspects. The problems of the currently disclosed chemical synthesis methods are long synthesis steps, expensive catalysts, and difficult methods. The present invention provides a method for the total synthesis of an isopentenyl flavonoid compound and its derivative preparation. This preparation method has the advantages of cheap and readily available raw materials, short reaction cycle, and simple operation. DETAILED DESCRIPTION

[0070] The present invention is further described in detail in conjunction with the following embodiments. The protection content of the present invention is not limited to the following embodiments. Without departing from the spirit and scope of the inventive concept, changes and advantages that can be thought of by those skilled in the art are included in the present invention, and the attached claims are the scope of protection. The process, conditions, reagents, experimental methods, etc. for implementing the present invention, except for the contents specifically mentioned below, are all common knowledge and common common sense in the art, and the present invention has no special restrictions.

[0071] Obviously, the described embodiments are only some embodiments of the present invention, rather than all embodiments. The following description of at least one exemplary embodiment is actually only illustrative and is by no means intended to limit the present invention and its application or use. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0072] In all examples shown and discussed herein, any specific values ​​should be interpreted as merely exemplary and not as limiting. Therefore, other examples of the exemplary embodiments may have different values.

[0073] The specific embodiments of the present invention are further described below. It should be noted that the description of these embodiments is used to help understand the present invention, but does not constitute a limitation of the present invention. In addition, the technical features involved in the various embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0074] The present invention belongs to the technical field of anticancer drugs, and specifically relates to an isopentenyl flavonoid compound and a preparation method and application thereof. The compound has a structure shown in formula (1). The method first uses chloromethyl methyl ether to protect the hydroxyl groups at positions 2,4,6-trihydroxyacetophenone, then uses isopentenyl bromide to carry out a substitution reaction with the above product, then uses N,N-diethylaniline as a solvent to carry out a microwave reaction to produce a Claisen rearrangement, then reacts with the corresponding aldehyde to produce a Claisen-Schmidt reaction to generate a chalcone, and finally uses iodine to close the ring and remove the protection with hydrochloric acid to prepare the compound. The compound provided by the present invention has a novel structure, good activity and low toxicity. The series of compounds have good antitumor activity as inhibitors, and have good inhibitory activity against a variety of tumor cells including colorectal cancer, breast cancer, lung cancer and liver cancer, and can be used to prepare antitumor drugs. It has good application prospects in antitumor.

[0075] The experimental methods in the following examples are conventional methods unless otherwise specified, and the experimental materials used in the following examples are commercially available unless otherwise specified.

[0076] The present invention provides an isopentenyl flavonoid compound, wherein the compound has a structure shown in formula (1):

[0077]

[0078] In the formula, R 1 isopentenyl, hydrogen; R 2 isopentenyl, hydrogen; R 3 It is hydroxy, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, halogen, trifluoromethyl, nitro, amino, morpholine ring, cyclopropane, cyclopentane, cyclobutane, hydroxymethyl, hydroxyethyl, or hydroxypropyl.

[0079] The isopentenyl flavonoid compounds in this example include 36 compounds, and the structural formulas and molecular weights of these compounds are shown in Table 1.

[0080] Table 1 Structural formula and molecular weight of isopentenyl flavonoid compounds

[0081]

[0082]

[0083]

[0084]

[0085]

[0086]

[0087]

[0088] Example 1

[0089] Synthesis of compounds 1 and 2:

[0090] The chemical reaction formula of the synthesis is as follows:

[0091]

[0092] The specific synthesis method comprises the following steps:

[0093] Step 1: Weigh 2,4,6-trihydroxyacetophenone (10.00 g, 59.47 mmol) and N,N-diisopropylethylamine DIPEA (23.05 g, 178.41 mmol) and dissolve them in dichloromethane (100 mL). Stir in an ice bath. After the solution is slightly clarified, slowly add chloromethyl methyl ether MOMCl (14.36 g, 178.41 mmol) to the reaction system. The temperature slowly rises to room temperature and continues to stir for 6 hours. After the reaction is completed, the reaction solution is quenched with saturated ammonium chloride, stirred at room temperature for 5 minutes, and the organic phase is washed with water three times. Finally, the organic phase is dried with anhydrous sodium sulfate, filtered, and the filtrate is dried to obtain a pale yellow oily crude product, which is purified by column chromatography to obtain 13.60 g of product A with a yield of 89%.

[0094] Step 2: Weigh the product A (10.0 g, 39.02 mmol) of the previous step, dissolve it in acetone (100 mL), add isopentenyl bromide (11.68 g, 78.4 mmol) and potassium carbonate (10.79 g, 78.09 mmol) to the reaction system, and raise the temperature to reflux temperature for 2 hours. After the raw materials react, filter out the solid and wash with ethyl acetate. After vacuum concentration, purify by column chromatography to obtain 10.60 g of product B with a yield of 84%.

[0095] Step 3: The product B (2.0 g, 6.17 mmol) of the previous step was dissolved in N, N-diethylaniline (10 mL) and reacted at 200°C for 1 hour in a microwave oven. After the reaction of the raw materials was completed, 10% hydrochloric acid was added thereto for neutralization, the pH was adjusted to neutral, and then extracted with ethyl acetate three times, washed with saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate, filtered, and column chromatography was performed to obtain 1.20 g of light yellow oil C, with a yield of 60%.

[0096] Step 4: The above product C (2.0 g, 6.17 mmol) was dissolved in anhydrous ethanol (20 mL), potassium hydroxide (1.38 g, 24.66 mmol) was gradually added to the reaction system, and stirred at room temperature. 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 raw materials reacted, the reaction solution was concentrated and diluted with ethyl acetate, extracted with water, and then the aqueous phase was back-extracted with ethyl acetate three times, the EA layers were combined, and washed with saturated brine. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was spin-dried to obtain an oily crude product. The crude product was separated and purified by column chromatography to obtain 2.10 g of yellow oily product 1a with a yield of 68%.

[0097] Step 5: Place the product 1a (1.00 g, 1.99 mmol) of the previous reaction in a reaction bottle, add dimethyl sulfoxide (20 mL) to dissolve, then add iodine (50.50 mg, 0.17 mmol), and place at 110°C to react for 1.5 hours. After the raw materials have reacted, dilute the reaction solution with ethyl acetate, extract with water, wash with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, and concentrate in vacuo to obtain a crude yellow oil. No further purification is required. Methanol (20 mL) and 10% dilute hydrochloric acid solution (10 mL) are added thereto. After reflux at 80°C for 0.5 hours, vacuum concentrate, extract with ethyl acetate, wash with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, then spin dry the solvent, separate and purify by column chromatography to obtain 110 mg of compound 1 with a yield of 11%, and 130 mg of compound 2 with a yield of 13%. Compound 1 1 H NMR (400 MHz, DMSO-d 6 )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 C 21 H 20 O 6 [M+H] + =369.1333; found 369.1335. Compound 2 1 H NMR (400 MHz, DMSO-d 6 )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 C 21 H 20 O 6 [M+H] + =369.1333; found369.1333.

[0098] Example 2

[0099] Synthesis of compounds 3 and 4:

[0100] The chemical reaction formula of the synthesis is as follows:

[0101]

[0102] The specific synthesis method comprises the following steps:

[0103] Step 1: Dissolve the above product C (2.0 g, 6.17 mmol) in anhydrous ethanol (20 mL), gradually add potassium hydroxide (1.38 g, 24.66 mmol) to the reaction system, stir at room temperature, add 2-chloro-4-(methoxymethoxy)benzaldehyde (1.24 g, 6.17 mmol) after potassium hydroxide is completely dissolved, and stir at room temperature for 3 hours. After the raw materials react, the reaction solution is concentrated and diluted with ethyl acetate, extracted with water, and then the aqueous phase is back-extracted with ethyl acetate three times, the EA layers are combined, and washed with saturated brine. Finally, the organic phase is dried over anhydrous sodium sulfate, filtered, and the filtrate is spin-dried to obtain an oily crude product. The crude product is separated and purified by column chromatography to obtain 2.20 g of yellow oily product 2a, with a yield of 70%.

[0104] Step 2: Place the product 2a (1.00 g, 1.97 mmol) of the previous step in a reaction bottle, add dimethyl sulfoxide (20 mL) to dissolve, then add iodine (50.06 mg, 0.97 mmol), and place at 110°C for reaction for 1.5 hours. After the reaction of the raw materials is complete, dilute the reaction solution with ethyl acetate, extract with water, wash with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, and concentrate in vacuo to obtain a crude yellow oil. No further purification is required. Methanol (20 mL) and 10% dilute hydrochloric acid solution (10 mL) are added thereto. After reflux at 80°C for 0.5 hours, vacuum concentrate, extract with ethyl acetate, wash with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, then spin dry the solvent, separate and purify by column chromatography to obtain 140 mg of compound 3 with a yield of 14%. At the same time, 126 mg of compound 4 is obtained with a yield of 13%. Compound 3 1 H NMR (400 MHz, DMSO-d 6 )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 C 20 H 17 C1O 5 [M+H]+ =373.0837; found 373.0835. Compound 4 1 H NMR (400 MHz, DMSO-d 6 )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 C 20 H 17 C1O 5 [M+H] + =373.0837; found 373.0840.

[0105] Example 3

[0106] Synthesis of compounds 5 and 6:

[0107] The chemical reaction formula of the synthesis is as follows:

[0108]

[0109] The specific synthesis method comprises the following steps:

[0110] Step 1: Dissolve the above product C (2.0 g, 6.17 mmol) in anhydrous ethanol (20 mL), gradually add potassium hydroxide (1.38 g, 24.66 mmol) to the reaction system, stir at room temperature, add 3-chloro-4-(methoxymethoxy)benzaldehyde (1.24 g, 6.17 mmol) after potassium hydroxide is completely dissolved, and stir at room temperature for 3 hours. After the raw materials react, the reaction solution is concentrated and diluted with ethyl acetate, extracted with water, and then the aqueous phase is back-extracted with ethyl acetate three times, the EA layers are combined, and washed with saturated brine. Finally, the organic phase is dried over anhydrous sodium sulfate, filtered, and the filtrate is spin-dried to obtain an oily crude product. The crude product is separated and purified by column chromatography to obtain 2.35 g of yellow oily product 3a with a yield of 75%.

[0111] Step 2: Place the product 3a (1.00 g, 1.97 mmol) of the previous reaction in a reaction bottle, add dimethyl sulfoxide (20 mL) to dissolve, then add iodine (50.06 mg, 0.97 mmol), and place at 110°C for reaction for 1.5 hours. After the raw materials have reacted, dilute the reaction solution with ethyl acetate, extract with water, wash with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, and concentrate in vacuo to obtain a crude yellow oil. No further purification is required. Methanol (20 mL) and 10% dilute hydrochloric acid solution (10 mL) are added thereto. After reflux at 80°C for 0.5 hours, vacuum concentrate, extract with ethyl acetate, wash with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, then spin dry the solvent, separate and purify by column chromatography to obtain 112 mg of compound 5 with a yield of 11%. At the same time, 136 mg of compound 6 is obtained with a yield of 14%. Compound 5 1 H NMR (500 MHz, DMSO-d 6 )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 forC 20 H 17 C1O 5 [M+H] + =373.0837; found 373.0838. Compound 6 1 H NMR (400 MHz, DMSO-d 6 )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),6.8 4(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 C 20 H 17 C1O 5 [M+H] + =373.0837; found 373.0840.

[0112] Example 4

[0113] Synthesis of compounds 7 and 8:

[0114] The chemical reaction formula of the synthesis is as follows:

[0115]

[0116] The specific synthesis method comprises the following steps:

[0117] Step 1: Dissolve the above product C (2.0 g, 6.17 mmol) in anhydrous ethanol (20 mL), gradually add potassium hydroxide (1.38 g, 24.66 mmol) to the reaction system, stir at room temperature, add 2,5-dimethoxybenzaldehyde (1.02 g, 6.17 mmol) after potassium hydroxide is completely dissolved, and stir at room temperature for 3 hours. After the raw materials react, concentrate the reaction solution and dilute it with ethyl acetate, extract it with water, and then back-extract the aqueous phase with ethyl acetate three times, combine the EA layers, and wash with saturated brine. Finally, the organic phase is dried over anhydrous sodium sulfate, filtered, and the filtrate is spin-dried to obtain an oily crude product. The crude product is separated and purified by column chromatography to obtain 2.50 g of yellow oily product 4a with a yield of 86%.

[0118] Step 2: Place the product 4a (1.00 g, 2.12 mmol) of the previous reaction in a reaction bottle, add dimethyl sulfoxide (20 mL) to dissolve, then add iodine (53.71 mg, 0.21 mmol), and place at 110°C to react for 1.5 hours. After the raw materials have reacted, dilute the reaction solution with ethyl acetate, extract with water, wash with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, and concentrate in vacuo to obtain a crude yellow oil. No further purification is required. Methanol (20 mL) and 10% dilute hydrochloric acid solution (10 mL) are added thereto. After reflux at 80°C for 0.5 hours, vacuum concentrate, extract with ethyl acetate, wash with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, then spin dry the solvent, separate and purify by column chromatography to obtain 120 mg of compound 7 with a yield of 12%. At the same time, 145 mg of compound 8 is obtained with a yield of 15%. Compound 7 1 H NMR (400 MHz, DMSO-d 6 )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 C 22 H 22 O 6 [M+H] +=383.1489; found 383.1496. Compound 8 1 H NMR (400 MHz, DMSO-d 6 )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 forC 22 H 22 O 6 [M+H] + =383.1489; found 383.1483.

[0119] Example 5

[0120] Synthesis of compounds 9 and 10:

[0121] The chemical reaction formula of the synthesis is as follows:

[0122]

[0123] The specific synthesis method comprises the following steps:

[0124] Step 1: Dissolve the above product C (2.0 g, 6.17 mmol) in anhydrous ethanol (20 mL), gradually add potassium hydroxide (1.38 g, 24.66 mmol) to the reaction system, stir at room temperature, add 3,4-dimethoxybenzaldehyde (1.02 g, 6.17 mmol) after potassium hydroxide is completely dissolved, and stir at room temperature for 3 hours. After the raw materials react, concentrate the reaction solution and dilute it with ethyl acetate, extract it with water, and then back-extract the aqueous phase with ethyl acetate three times, combine the EA layers, and wash with saturated brine. Finally, the organic phase is dried over anhydrous sodium sulfate, filtered, and the filtrate is spin-dried to obtain an oily crude product. The crude product is separated and purified by column chromatography to obtain 2.37 g of yellow oily product 5a with a yield of 81%.

[0125] Step 2: Place the product 5a (1.00 g, 2.12 mmol) of the previous reaction in a reaction bottle, add dimethyl sulfoxide (20 mL) to dissolve, then add iodine (53.71 mg, 0.21 mmol), and place at 110°C to react for 1.5 hours. After the raw materials have reacted, dilute the reaction solution with ethyl acetate, extract with water, wash with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, and concentrate in vacuo to obtain a crude yellow oil. No further purification is required. Methanol (20 mL) and 10% dilute hydrochloric acid solution (10 mL) are added thereto. After reflux at 80°C for 0.5 hours, vacuum concentrate, extract with ethyl acetate, wash with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, then spin dry the solvent, separate and purify by column chromatography to obtain 160 mg of compound 9 with a yield of 16%. At the same time, 155 mg of compound 10 is obtained with a yield of 16%. Compound 9 1 H NMR (500 MHz, DMSO-d 6 )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 C 22 H 22 O 6 [M+H] + =383.1489; found 383.1492. Compound 10 1 HNMR (400 MHz, DMSO-d 6 )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),6.5 7(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 forC 22 H 22 O 6 [M+H] + =383.1489; found383.1489.

[0126] Example 6

[0127] Synthesis of compounds 11 and 12:

[0128] The chemical reaction formula of the synthesis is as follows:

[0129]

[0130] The specific synthesis method comprises the following steps:

[0131] Step 1: Dissolve the above product C (2.0 g, 6.17 mmol) in anhydrous ethanol (20 mL), gradually add potassium hydroxide (1.38 g, 24.66 mmol) to the reaction system, stir at room temperature, add 4-methoxy-3,5-bis(methoxymethoxy)benzaldehyde (1.58 g, 6.17 mmol) after the potassium hydroxide is completely dissolved, and stir at room temperature for 3 hours. After the raw materials react, the reaction solution is concentrated and diluted with ethyl acetate, extracted with water, and then the aqueous phase is back-extracted with ethyl acetate three times, the EA layers are combined, and washed with saturated brine. Finally, the organic phase is dried over anhydrous sodium sulfate, filtered, and the filtrate is spin-dried to obtain an oily crude product. The crude product is separated and purified by column chromatography to obtain 2.43 g of yellow oily product 6a with a yield of 70%.

[0132] Step 2: Place the product 6a (1.00 g, 1.78 mmol) of the previous step in a reaction bottle, add dimethyl sulfoxide (20 mL) to dissolve, then add iodine (45.11 mg, 0.18 mmol), and place at 110°C to react for 1.5 hours. After the raw materials react, dilute the reaction solution with ethyl acetate, extract with water, wash with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, and concentrate in vacuo to obtain a crude yellow oil. No further purification is required. Methanol (20 mL) and 10% dilute hydrochloric acid solution (10 mL) are added thereto. After reflux at 80°C for 0.5 hours, vacuum concentrate, extract with ethyl acetate, wash with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, then spin dry the solvent, separate and purify by column chromatography to obtain 142 mg of compound 11 with a yield of 14%. At the same time, 139 mg of compound 12 is obtained with a yield of 14%. Compound 11 1 H NMR (500 MHz, DMSO-d 6 )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 C 21 H 20 O 7[M+H] + =385.1282; found 385.1283. Compound 12 1 H NMR (500 MHz, DMSO-d 6 )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 C 21 H 20 O 7 [M+H] + =385.1282;found385.1283.

[0133] Example 7

[0134] Synthesis of compounds 13 and 14:

[0135] The chemical reaction formula of the synthesis is as follows:

[0136]

[0137] The specific synthesis method comprises the following steps:

[0138] Step 1: Dissolve the above product C (2.0 g, 6.17 mmol) in anhydrous ethanol (20 mL), gradually add potassium hydroxide (1.38 g, 24.66 mmol) to the reaction system, stir at room temperature, add 4-isopropylbenzaldehyde (0.91 g, 6.17 mmol) after the potassium hydroxide is completely dissolved, and stir at room temperature for 3 hours. After the raw materials react, concentrate the reaction solution and dilute it with ethyl acetate, extract it with water, and then back-extract the aqueous phase with ethyl acetate three times, combine the EA layers, and wash with saturated brine. Finally, the organic phase is dried over anhydrous sodium sulfate, filtered, and the filtrate is spin-dried to obtain an oily crude product. The crude product is separated and purified by column chromatography to obtain 2.17 g of yellow oily product 7a with a yield of 77%.

[0139] Step 2: Place the product 7a (1.00 g, 2.20 mmol) of the previous step in a reaction bottle, add dimethyl sulfoxide (20 mL) to dissolve, then add iodine (55.84 mg, 0.22 mmol), and place at 110°C for reaction for 1.5 hours. After the reaction of the raw materials is complete, dilute the reaction solution with ethyl acetate, extract with water, wash with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, and concentrate in vacuo to obtain a crude yellow oil. No further purification is required. Methanol (20 mL) and 10% dilute hydrochloric acid solution (10 mL) are added thereto. After reflux at 80°C for 0.5 hours, vacuum concentrate, extract with ethyl acetate, wash with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, then spin dry the solvent, separate and purify by column chromatography to obtain 128 mg of compound 13 with a yield of 13%. At the same time, 145 mg of compound 14 is obtained with a yield of 15%. Compound 13 1 H NMR (500 MHz, DMSO-d 6 )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 C 23 H 24 O 4 [M+H] + =365.1747; found 365.1745. Compound 14 1 HNMR (500 MHz, DMSO-d 6 )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 C 23 H 24 O 4 [M+H] + =365.1747;found365.1746.

[0140] Example 8

[0141] Synthesis of compounds 15 and 16:

[0142] The chemical reaction formula of the synthesis is as follows:

[0143]

[0144] The specific synthesis method comprises the following steps:

[0145] Step 1: Dissolve the above product C (2.0 g, 6.17 mmol) in anhydrous ethanol (20 mL), gradually add potassium hydroxide (1.38 g, 24.66 mmol) to the reaction system, stir at room temperature, add 4-(4-morpholine)benzaldehyde (1.18 g, 6.17 mmol) after the potassium hydroxide is completely dissolved, and stir at room temperature for 3 hours. After the raw materials react, the reaction solution is concentrated and diluted with ethyl acetate, extracted with water, and then the aqueous phase is back-extracted with ethyl acetate three times, the EA layers are combined, and washed with saturated brine. Finally, the organic phase is dried over anhydrous sodium sulfate, filtered, and the filtrate is spin-dried to obtain an oily crude product. The crude product is separated and purified by column chromatography to obtain 2.11 g of yellow oily product 8a with a yield of 69%.

[0146] Step 2: Place the product 8a (1.00 g, 2.01 mmol) of the previous step in a reaction bottle, add dimethyl sulfoxide (20 mL) to dissolve, then add iodine (51.01 mg, 0.20 mmol), and place at 110°C for reaction for 1.5 hours. After the reaction of the raw materials is complete, dilute the reaction solution with ethyl acetate, extract with water, wash with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, and concentrate in vacuo to obtain a crude yellow oil. No further purification is required. Methanol (20 mL) and 10% dilute hydrochloric acid solution (10 mL) are added thereto. After reflux at 80°C for 0.5 hours, vacuum concentrate, extract with ethyl acetate, wash with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, then spin dry the solvent, separate and purify by column chromatography to obtain 134 mg of compound 15 with a yield of 13%. At the same time, 145 mg of compound 16 is obtained with a yield of 15%. Compound 15 1 H NMR (500 MHz, DMSO-d 6)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,1 H),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 C 24 H 25 NO 5 [M+H] + =408.1805; found 408.1806. Compound 16 1 H NMR (500 MHz, DMSO-d 6 )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,1 H),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 C 24 H 25 NO 5 [M+H] + =408.1805; found 408.1805.

[0147] Example 9

[0148] Synthesis of compounds 17 and 18:

[0149] The chemical reaction formula of the synthesis is as follows:

[0150]

[0151] The specific synthesis method comprises the following steps:

[0152] Step 1: Dissolve the above product C (2.0 g, 6.17 mmol) in anhydrous ethanol (20 mL), gradually add potassium hydroxide (1.38 g, 24.66 mmol) to the reaction system, stir at room temperature, add 4-trifluoromethylbenzaldehyde (1.07 g, 6.17 mmol) after the potassium hydroxide is completely dissolved, and stir at room temperature for 3 hours. After the raw materials react, concentrate the reaction solution and dilute it with ethyl acetate, extract it with water, and then back-extract the aqueous phase with ethyl acetate three times, combine the EA layers, and wash with saturated brine. Finally, the organic phase is dried over anhydrous sodium sulfate, filtered, and the filtrate is spin-dried to obtain an oily crude product. The crude product is separated and purified by column chromatography to obtain 2.35 g of yellow oily product 9a with a yield of 79%.

[0153] Step 2: Place the product 9a (1.00 g, 2.08 mmol) of the previous step in a reaction bottle, add dimethyl sulfoxide (20 mL) to dissolve, then add iodine (52.82 mg, 0.21 mmol), and place at 110°C for reaction for 1.5 hours. After the reaction of the raw materials is complete, dilute the reaction solution with ethyl acetate, extract with water, wash with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, and concentrate in vacuo to obtain a crude yellow oil. No further purification is required. Methanol (20 mL) and 10% dilute hydrochloric acid solution (10 mL) are added thereto. After reflux at 80°C for 0.5 hours, vacuum concentrate, extract with ethyl acetate, wash with saturated brine, combine the organic phases, dry over anhydrous sodium sulfate, then spin dry the solvent, and separate and purify by column chromatography to obtain 142 mg of compound 17 with a yield of 14%. At the same time, 127 mg of compound 18 is obtained with a yield of 13%. Compound 17 1 H NMR (400 MHz, DMSO-d 6 )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,1 H),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 C 21 H 17 F 3 O 4 [M+H] + =391.1152; found 391.1151. Compound 18 1 H NMR (500 MHz, DMSO-d 6)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):calculatedfor C 21 H 17 F 3 O 4 [M+H] + =391.1152; found391.1154.

[0154] Example 10

[0155] Synthesis of compounds 19 and 20:

[0156] The chemical reaction formula of the synthesis is as follows:

[0157]

[0158] The specific synthesis method comprises the following steps:

[0159] Step 1: Dissolve the above product C (2.0 g, 6.17 mmol) in anhydrous ethanol (20 mL), gradually add potassium hydroxide (1.38 g, 24.66 mmol) to the reaction system, stir at room temperature, add 3-trifluoromethylbenzaldehyde (1.07 g, 6.17 mmol) after the potassium hydroxide is completely dissolved, and stir at room temperature for 3 hours. After the raw materials react, concentrate the reaction solution and dilute it with ethyl acetate, extract it with water, and then back-extract the aqueous phase with ethyl acetate three times, combine the EA layers, and wash with saturated brine. Finally, the organic phase is dried over anhydrous sodium sulfate, filtered, and the filtrate is spin-dried to obtain an oily crude product. The crude product is separated and purified by column chromatography to obtain 2.40 g of yellow oily product 10a with a yield of 81%.

[0160] Step 2: The product 10a (1.00 g, 2.08 mmol) of the previous step was placed in a reaction bottle, dimethyl sulfoxide (20 mL) was added to dissolve, and iodine (52.82 mg, 0.21 mmol) was added, and the mixture was placed at 110°C for reaction 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, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated in vacuo to obtain a crude yellow oil. No further purification was required. Methanol (20 mL) and 10% dilute hydrochloric acid solution (10 mL) were added thereto. After reflux at 80°C for 0.5 hours, the mixture was concentrated in vacuo, extracted with ethyl acetate, washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was then spin-dried. The mixture was separated and purified by column chromatography to obtain 150 mg of compound 19 with a yield of 18%. At the same time, 149 mg of compound 20 was obtained with a yield of 15%. Compound 19 1 H NMR (400 MHz, DMSO-d 6 )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 C 21 H 17 F 3 O 4 [M+H] + =391.1152; found 391.1153. Compound 20 1 H NMR (500 MHz, DMSO-d 6 )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 C 21 H 17 F 3 O 4 [M+H] + =391.1152; found391.1152.

[0161] Embodiment 11

[0162] Synthesis of compounds 21 and 22:

[0163] The chemical reaction formula of the synthesis is as follows:

[0164]

[0165] The specific synthesis method comprises the following steps:

[0166] Step 1: Dissolve the above product C (2.0 g, 6.17 mmol) in anhydrous ethanol (20 mL), gradually add potassium hydroxide (1.38 g, 24.66 mmol) to the reaction system, stir at room temperature, add 2-trifluoromethylbenzaldehyde (1.07 g, 6.17 mmol) after the potassium hydroxide is completely dissolved, and stir at room temperature for 3 hours. After the raw materials react, concentrate the reaction solution and dilute it with ethyl acetate, extract it with water, and then back-extract the aqueous phase with ethyl acetate three times, combine the EA layers, and wash with saturated brine. Finally, the organic phase is dried over anhydrous sodium sulfate, filtered, and the filtrate is spin-dried to obtain an oily crude product. The crude product is separated and purified by column chromatography to obtain 2.27 g of yellow oily product 11a with a yield of 77%.

[0167] Step 2: The product 11a (1.00 g, 2.08 mmol) of the previous step was placed in a reaction bottle, dimethyl sulfoxide (20 mL) was added to dissolve, and iodine (52.82 mg, 0.21 mmol) was added, and the mixture was placed at 110°C for reaction 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, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated in vacuo to obtain a crude yellow oil. No further purification was required. Methanol (20 mL) and 10% dilute hydrochloric acid solution (10 mL) were added thereto. After reflux at 80°C for 0.5 hours, the mixture was concentrated in vacuo, extracted with ethyl acetate, washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was then spin-dried. The mixture was separated and purified by column chromatography to obtain 143 mg of compound 21 with a yield of 14%. At the same time, 133 mg of compound 22 was obtained with a yield of 13%. Compound 21 1 H NMR (500 MHz, DMSO-d 6 )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 C 21 H 17F 3 O 4 [M+H] + =391.1152; found 391.1152. Compound 22 1 H NMR (400 MHz, DMSO-d 6 )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 C 21 H 17 F 3 O 4 [M+H] + =391.1152; found391.1151.

[0168] Example 12

[0169] Synthesis of compounds 23 and 24:

[0170] The chemical reaction formula of the synthesis is as follows:

[0171]

[0172] The specific synthesis method comprises the following steps:

[0173] Step 1: The above product C (2.0 g, 6.17 mmol) was dissolved in anhydrous ethanol (20 mL), potassium hydroxide (1.38 g, 24.66 mmol) was gradually added to the reaction system, and stirred at room temperature. 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 was completed, the reaction solution was concentrated and diluted with ethyl acetate, extracted with water, and then the aqueous phase was back-extracted with ethyl acetate three times, the EA layers were combined, and washed with saturated brine. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was spin-dried to obtain an oily crude product. The crude product was separated and purified by column chromatography to obtain 2.31 g of yellow oily product 12a with a yield of 88%.

[0174] Step 2: The product 12a (1.00 g, 2.34 mmol) of the previous step was placed in a reaction bottle, dimethyl sulfoxide (20 mL) was added to dissolve, and iodine (59.51 mg, 0.23 mmol) was added, and the mixture was placed at 110°C for reaction 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, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated in vacuo to obtain a crude yellow oil. No further purification was required. Methanol (20 mL) and 10% dilute hydrochloric acid solution (10 mL) were added thereto. After reflux at 80°C for 0.5 hours, the mixture was concentrated in vacuo, extracted with ethyl acetate, washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was then dried by spin drying. The mixture was separated and purified by column chromatography to obtain 175 mg of compound 23 with a yield of 18%. At the same time, 158 mg of compound 24 was obtained with a yield of 16%. Compound 23 1 H NMR (500 MHz, DMSO-d 6 )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),6.4 5(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 C 21 H 20 O 4 [M+H] + =337.1434; found 337.1431. Compound 24 1 H NMR (500 MHz, DMSO-d 6 )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):calculatedfor C 21 H 20 O 4 [M+H] + =337.1434; found337.1434.

[0175] Embodiment 13

[0176] Synthesis of compounds 25 and 26:

[0177] The chemical reaction formula of the synthesis is as follows:

[0178]

[0179] The specific synthesis method comprises the following steps:

[0180] Step 1: The above product C (2.0 g, 6.17 mmol) was dissolved in anhydrous ethanol (20 mL), potassium hydroxide (1.38 g, 24.66 mmol) was gradually added to the reaction system, and stirred at room temperature. After the potassium hydroxide was completely dissolved, 3-methylbenzaldehyde (1.07 g, 6.17 mmol) was added, and stirred at room temperature for 3 hours. After the reaction of the raw materials was completed, the reaction solution was concentrated and diluted with ethyl acetate, extracted with water, and then the aqueous phase was back-extracted with ethyl acetate three times, the EA layers were combined, and washed with saturated brine. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was spin-dried to obtain an oily crude product. The crude product was separated and purified by column chromatography to obtain 2.33 g of yellow oily product 13a with a yield of 89%.

[0181] Step 2: The product 13a (1.00 g, 2.34 mmol) of the previous step was placed in a reaction bottle, dimethyl sulfoxide (20 mL) was added to dissolve, and iodine (59.51 mg, 0.23 mmol) was added, and the mixture was placed at 110°C for reaction 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, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated in vacuo to obtain a crude yellow oil. No further purification was required. Methanol (20 mL) and 10% dilute hydrochloric acid solution (10 mL) were added thereto. After reflux at 80°C for 0.5 hours, the mixture was concentrated in vacuo, extracted with ethyl acetate, washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was then dried by spin drying. The mixture was separated and purified by column chromatography to obtain 150 mg of compound 25 with a yield of 15%. At the same time, 145 mg of compound 26 was obtained with a yield of 15%. Compound 25 1 H NMR (500 MHz, DMSO-d 6 )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 C21 H 20 O 4 [M+H] + =337.1434; found337.1434. Compound 26 1 H NMR (400 MHz, DMSO-d 6 )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 C 21 H 20 O 4 [M+H] + =337.1434; found 337.1442.

[0182] Embodiment 14

[0183] Synthesis of compounds 27 and 28:

[0184] The chemical reaction formula of the synthesis is as follows:

[0185]

[0186] The specific synthesis method comprises the following steps:

[0187] Step 1: The above product C (2.0 g, 6.17 mmol) was dissolved in anhydrous ethanol (20 mL), potassium hydroxide (1.38 g, 24.66 mmol) was gradually added to the reaction system, and stirred at room temperature. 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 was completed, the reaction solution was concentrated and diluted with ethyl acetate, extracted with water, and then the aqueous phase was back-extracted with ethyl acetate three times, the EA layers were combined, and washed with saturated brine. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was spin-dried to obtain an oily crude product. The crude product was separated and purified by column chromatography to obtain 2.14 g of yellow oily product 14a with a yield of 78%.

[0188] Step 2: The product 14a (1.00 g, 2.24 mmol) of the previous step was placed in a reaction bottle, dimethyl sulfoxide (20 mL) was added to dissolve, and iodine (56.79 mg, 0.22 mmol) was added, and the mixture was placed at 110°C for reaction 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, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated in vacuo to obtain a crude yellow oil. No further purification was required. Methanol (20 mL) and 10% dilute hydrochloric acid solution (10 mL) were added thereto. After reflux at 80°C for 0.5 hours, the mixture was concentrated in vacuo, extracted with ethyl acetate, washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was then dried by spin drying. The mixture was separated and purified by column chromatography to obtain 177 mg of compound 27 with a yield of 18%. At the same time, 161 mg of compound 28 was obtained with a yield of 16%. Compound 27 1 H NMR (500 MHz, DMSO-d 6 )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 C 20 H 17 C1O 4 [M+H] + =357.0888; found 357.0886. Compound 28 1 H NMR (500 MHz, DMSO-d 6 )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 C 20 H 17 C1O 4 [M+H] + =357.0888; found357.0888.

[0189] Embodiment 15

[0190] Synthesis of compounds 29 and 30:

[0191] The chemical reaction formula of the synthesis is as follows:

[0192]

[0193] The specific synthesis method comprises the following steps:

[0194] Step 1: Dissolve the above product C (2.0 g, 6.17 mmol) in anhydrous ethanol (20 mL), gradually add potassium hydroxide (1.38 g, 24.66 mmol) to the reaction system, stir at room temperature, add 3-chloro-benzaldehyde (0.87 g, 6.17 mmol) after the potassium hydroxide is completely dissolved, and stir at room temperature for 3 hours. After the raw materials react, the reaction solution is concentrated and diluted with ethyl acetate, extracted with water, and then the aqueous phase is back-extracted with ethyl acetate three times, the EA layers are combined, and washed with saturated brine. Finally, the organic phase is dried over anhydrous sodium sulfate, filtered, and the filtrate is spin-dried to obtain an oily crude product. The crude product is separated and purified by column chromatography to obtain 2.43 g of yellow oily product 15a with a yield of 88%.

[0195] Step 2: The product 15a (1.00 g, 2.24 mmol) of the previous step was placed in a reaction bottle, dimethyl sulfoxide (20 mL) was added to dissolve, and iodine (56.79 mg, 0.22 mmol) was added, and the mixture was placed at 110°C for reaction 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, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated in vacuo to obtain a crude yellow oil. No further purification was required. Methanol (20 mL) and 10% dilute hydrochloric acid solution (10 mL) were added thereto. After reflux at 80°C for 0.5 hours, the mixture was concentrated in vacuo, extracted with ethyl acetate, washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was then spin-dried. The mixture was separated and purified by column chromatography to obtain 162 mg of compound 29 with a yield of 16%. At the same time, 155 mg of compound 30 was obtained with a yield of 16%. Compound 29 1 H NMR (500 MHz, DMSO-d 6)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,1 H),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 C 20 H 17 C1O 4 [M+H] + =357.0888; found 357.0886. Compound 30 1 H NMR (500 MHz, DMSO-d 6 )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=7.9 Hz,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 C 20 H 17 C1O 4 [M+H] + =357.0888; found357.0889.

[0196] Example 16

[0197] Synthesis of compounds 31 and 32:

[0198] The chemical reaction formula of the synthesis is as follows:

[0199]

[0200] The specific synthesis method comprises the following steps:

[0201] Step 1: The above product C (2.0 g, 6.17 mmol) was dissolved in anhydrous ethanol (20 mL), potassium hydroxide (1.38 g, 24.66 mmol) was gradually added to the reaction system, and stirred at room temperature. 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 was completed, the reaction solution was concentrated and diluted with ethyl acetate, extracted with water, and then the aqueous phase was back-extracted with ethyl acetate three times, the EA layers were combined, and washed with saturated brine. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was spin-dried to obtain an oily crude product. The crude product was separated and purified by column chromatography to obtain 2.22 g of yellow oily product 16a with a yield of 78%.

[0202] Step 2: The product 16a (1.00 g, 2.17 mmol) of the previous step was placed in a reaction bottle, dimethyl sulfoxide (20 mL) was added to dissolve, and iodine (55.06 mg, 0.22 mmol) was added, and the mixture was placed at 110°C for reaction 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, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated in vacuo to obtain a crude yellow oil. No further purification was required. Methanol (20 mL) and 10% dilute hydrochloric acid solution (10 mL) were added thereto. After reflux at 80°C for 0.5 hours, the mixture was concentrated in vacuo, extracted with ethyl acetate, washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was then dried by spin drying. The mixture was separated and purified by column chromatography to obtain 188 mg of compound 31 with a yield of 19%. At the same time, 173 mg of compound 32 was obtained with a yield of 17%. Compound 31 1 H NMR (500 MHz, DMSO-d 6 )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 C 21 H 19 C1O 4 [M+H] + =371.1045; found 371.1043. Compound 32 1 HNMR (500 MHz, DMSO-d 6)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 forC 21 H 19 C1O 4 [M+H] + =371.1045; found 371.1049.

[0203] Embodiment 17

[0204] Synthesis of compounds 33 and 34:

[0205] The chemical reaction formula of the synthesis is as follows:

[0206]

[0207] The specific synthesis method comprises the following steps:

[0208] Step 1: The above product C (2.0 g, 6.17 mmol) was dissolved in anhydrous ethanol (20 mL), potassium hydroxide (1.38 g, 24.66 mmol) was gradually added to the reaction system, and stirred at room temperature. 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 was completed, the reaction solution was concentrated and diluted with ethyl acetate, extracted with water, and then the aqueous phase was back-extracted with ethyl acetate three times, the EA layers were combined, and washed with saturated brine. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was spin-dried to obtain an oily crude product. The crude product was separated and purified by column chromatography to obtain 2.30 g of yellow oily product 17a with a yield of 74%.

[0209] Step 2: The product 17a (1.00 g, 1.97 mmol) of the previous step was placed in a reaction bottle, dimethyl sulfoxide (20 mL) was added to dissolve, and iodine (50.06 mg, 0.20 mmol) was added, and the mixture was placed at 110°C for reaction 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, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated in vacuo to obtain a crude yellow oil. No further purification was required. Methanol (20 mL) and 10% dilute hydrochloric acid solution (10 mL) were added thereto. After reflux at 80°C for 0.5 hours, the mixture was concentrated in vacuo, extracted with ethyl acetate, washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was then dried by spin drying. The mixture was separated and purified by column chromatography to obtain 175 mg of compound 33 with a yield of 18%. At the same time, 151 mg of compound 34 was obtained with a yield of 15%. Compound 33 1 H NMR (400 MHz, DMSO-d 6 )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 C 20 H 17 C1O 5 [M+H] + =373.0837; found 373.0838. Compound 34 1 H NMR (500 MHz, DMSO-d 6 )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 C 20 H 17 C1O 5 [M+H] + =373.0837; found 373.0840.

[0210] Embodiment 18

[0211] Synthesis of compounds 35 and 36:

[0212] The chemical reaction formula of the synthesis is as follows:

[0213]

[0214] The specific synthesis method comprises the following steps:

[0215] Step 1: The above product C (2.0 g, 6.17 mmol) was dissolved in anhydrous ethanol (20 mL), potassium hydroxide (1.38 g, 24.66 mmol) was gradually added to the reaction system, and stirred at room temperature. After the potassium hydroxide was completely dissolved, 3-methoxy-5-(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 was completed, the reaction solution was concentrated and diluted with ethyl acetate, extracted with water, and then the aqueous phase was back-extracted with ethyl acetate three times, the EA layers were combined, and washed with saturated brine. Finally, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was spin-dried to obtain an oily crude product. The crude product was separated and purified by column chromatography to obtain 2.03 g of yellow oily product 18a with a yield of 66%.

[0216] Step 2: The product 18a (1.00 g, 1.99 mmol) of the previous step was placed in a reaction bottle, dimethyl sulfoxide (20 mL) was added to dissolve, and iodine (50.50 mg, 0.20 mmol) was added, and the mixture was placed at 110°C for reaction 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, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated in vacuo to obtain a crude yellow oil. No further purification was required. Methanol (20 mL) and 10% dilute hydrochloric acid solution (10 mL) were added thereto. After reflux at 80°C for 0.5 hours, the mixture was concentrated in vacuo, extracted with ethyl acetate, washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was then dried by spin drying. The mixture was separated and purified by column chromatography to obtain 157 mg of compound 35 with a yield of 16%. At the same time, 133 mg of compound 36 was obtained with a yield of 13%. Compound 35 1 H NMR (400 MHz, DMSO-d 6 )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 C 21 H 20 O 6 [M+H] + =369.1333; found 369.1329. Compound 361 H NMR (400 MHz, DMSO-d 6 )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=1 4.1Hz,1H),3.80(s,3H),3.23(d,J=7.1Hz,2H),1.73(s,3H),1.63(s,3H).HRMS(ESI):calculated for C 21 H 20 O 6 [M+H] + =369.1333; found369.1335.

[0217] Example 19 Biological Activity Test Part:

[0218] Determination of the inhibitory activity of 36 isopentenyl flavonoid compounds on tumor cells in this example:

[0219] Wherein, the tumor refers to human colorectal cancer cells, human breast cancer cells, human non-small cell lung cancer cells, and human osteosarcoma cells.

[0220] (1) Tumor cells used in the test: human osteosarcoma cells (Sjsa-1) (purchased from Guangzhou Saiku Biotechnology Co., Ltd.), human colon cancer cells (HCT116) (purchased from Guangzhou Saiku Biotechnology Co., Ltd.), human non-small cell lung cancer cells (A549) (purchased from Guangzhou Saiku Biotechnology Co., Ltd.), human hepatocellular carcinoma cells (Hep G2) (Wuhan Pronosai Life Science Technology Co., Ltd.)

[0221] (2) Test method: The CCK-8 method was used to determine the proliferation inhibitory effect of prenylated flavonoids on human osteosarcoma cells, human colon cancer cells, human non-small cell lung cancer cells and human liver cancer cells. The specific test process is as follows:

[0222] 1. Prepare the above tumor cell lines into single cell suspensions with corresponding complete culture medium. The concentration of single cell suspension is 50,000 cells / mL. Take 100 μL of cell suspension and inoculate it into 96-well culture plates. Place them in CO 2 Incubator (37°C, 5% CO 2, 95% air) for 24 h; human osteosarcoma cells (Sjsa-1), human colon cancer cells (HCT116), and human non-small cell lung cancer cells (A549) were cultured in 1640 medium (containing 10% newborn calf serum, 1% double antibody); human hepatoma cells (HepG2) were cultured in DMEM medium (containing 10% newborn calf serum, 1% double antibody, 0.01 mg / mL insulin).

[0223] 2. Dissolve compounds 1-36 in DMSO and prepare 10mM stock solution, then dilute to 5mM, 2.5mM, 1.25mM, 0.625mM, 0.3125mM, 0.16mM, 0.078mM, 0.039mM. Take 1.0μL of each concentration and add it to each well of cells, so that the final concentration is 1.0% of the concentration before addition. Set up two parallel wells for each concentration of compound. Add 1.0μL of DMSO to the negative control group and blank control group and place them in CO 2 The cells were cultured in an incubator for 48 hours (the negative control group contained 50,000 cells / mL and was treated with 0.5 μL DMSO to deduct the interference of DMSO contained in the original drug; the blank control group contained no cells and 100 μL complete culture medium to deduct background interference).

[0224] 3. After 48 h of culture, the culture medium was removed and replaced with fresh culture medium, and then 10 μL of CCK-8 (Cell Counting Kits-8) reagent was added to each well of cells. After incubation at 37°C for 2 hours, the absorbance A at 450 nm was measured using a Biotek multi-function microplate reader to calculate the inhibition rate of tumor cell growth by compounds 1 to 40; the inhibition rate was calculated as follows: [1-(A drug treatment group-A blank control) / (A negative control group-A blank control)]×100%, where A is the absorbance.

[0225] 4. Test results: The tumor inhibition activities of some compounds are summarized in Table 2. As shown in Table 2, in general, the prenylated flavonoid compounds of the present invention have potential application value in the research and development of anticancer drugs for these four types of tumors.

[0226] Table 2 Inhibitory effects of some prenylated flavonoids on various cancer cell lines

[0227]

[0228]

[0229] The results in Table 2 show that for A549, the IC 50 (μM) was 9.21, and the IC of compound 20 50(μM) was 9.31, and the IC of compound 29 50 (μM) was 8.54, and the IC of compound 33 50 (μM) was 7.63; for HCT116, the IC 50 (μM) was 4.48, and the IC of compound 33 50 (μM) was 3.72; for HepG2, the IC 50 (μM) was 11.44, and the IC of compound 33 50 (μM) is 8.96. It can be seen that the compound of the present invention exhibits good tumor inhibitory activity and has good application prospects in the preparation of anti-tumor drugs and drugs for inhibiting tumor cells.

[0230] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0231] As used in the present invention, the terms "include" and "comprising" are open expressions, that is, including the contents specified in the present invention but not excluding other contents.

[0232] As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items.

[0233] The embodiments of the present invention are described in detail above, but the present invention is not limited to the described embodiments. For those skilled in the art, various changes, modifications, substitutions and variations of these embodiments are made without departing from the principles and spirit of the present invention, and still fall within the protection scope of the present invention.

Claims

1. An isopentenyl flavonoid compound, characterized in that: The compound has a structure shown in formula (1): In the formula, R 1 isopentenyl, hydrogen; R 2 isopentenyl, hydrogen; R 3 It is hydroxy, 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 includes the structures shown in formula (2) and formula (3): In the formula, R 3 It 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 undergoes 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 drug / drug composition, characterized in that The drug / 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.

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