Isochromanone compound, pharmaceutical composition comprising same, and use of compound

Abstract

Disclosed in the present invention are an isochromanone compound, a pharmaceutical composition comprising same, and a use of the compound. The structure of the compound is as shown in formula I. The compound can inhibit the activity of PCSK9. In vivo and in vitro experiments show that the compound has lipid-lowering activity, and has pharmacokinetic properties conducive to being manufactured as a drug.

Description

Isochrysantheminate compounds, their pharmaceutical compositions and applications Technical Field

[0001] This invention relates to an isochorone compound, pharmaceutical compositions thereof, and applications, and more particularly to an isochorone compound having PCSK9 inhibitory activity, pharmaceutical compositions thereof, and applications. Background Technology

[0002] Dyslipidemia mainly refers to abnormal levels of triglycerides and cholesterol in blood plasma, which play an important role in the occurrence and development of atherosclerosis. Classical familial hypercholesterolemia (FH) is characterized by elevated levels of total plasma cholesterol, especially low-density lipoprotein cholesterol (LDL-C). These cholesterol metabolism disorders increase the risk of diseases such as atherosclerosis and coronary heart disease, and this condition has long been considered an independent risk factor for cardiovascular disease.

[0003] Statins are currently the first-line lipid-lowering drugs in clinical practice, and have a good lipid-lowering effect. However, they also have certain limitations. Some patients will experience "statin intolerance". Common clinical side effects include myositis, myalgia, and liver damage. Some patients cannot achieve satisfactory lipid-lowering effects even when using the maximum dose.

[0004] PCSK9 is a FH-related protease. It exists in the endoplasmic reticulum of hepatocytes and other cells as a soluble prozyme precursor. After autocatalytic hydrolysis, it forms mature PCSK9, which is secreted into the plasma. Its regulation is primarily influenced by a membrane-bound transcription factor that regulates cellular cholesterol homeostasis. The low-density lipoprotein receptor (LDLR) expressed on the surface of liver cells is the primary determinant of LDL-C levels. LDLR on the cell surface can bind to LDL-C in plasma, mediating the uptake of LDL-C by cells and its transport to lysosomes for degradation. LDLR then recirculates to the cell surface, continuing its LDL-C-lowering effect. PCSK9 in the blood can specifically bind to LDLR on the cell surface, forming a complex and being transported to lysosomes, leading to accelerated degradation of lysosomes and increased LDL-C levels. Therefore, PCSK9 plays a crucial regulatory role in maintaining cholesterol homeostasis in the body.

[0005] PCSK9 inhibitors include siRNA, mAbs, small molecule antagonists, ASOs, and antibody-mimicking proteins. Among these, monoclonal antibody drugs are expensive and have a single administration method, significantly reducing long-term patient adherence. Currently, there are no PCSK9 small molecule inhibitors on the market, failing to meet clinical needs. Summary of the Invention

[0006] Objectives of the invention: The first objective of this invention is to provide an isochorone compound with PCSK9 inhibitory activity; the second objective is to provide a pharmaceutical composition containing the compound; and the third objective is to provide a pharmaceutical application of the compound and the pharmaceutical composition thereof.

[0007] Technical solution: The isochromatic ketone compound or its cis-trans isomer, or a pharmaceutically acceptable salt, as described in this invention, wherein the compound has the structure of Formula I.

[0008] In the aforementioned structure:

[0009] R1 and R2 are each independently selected from hydrogen, halogen, CN, NO2, NH2, OH, CF3, CH2OH, COOH, and -O[(CH2)] m O] r R5, substituted or unsubstituted, includes C1-C6 straight-chain or branched alkyl, C1-C6 straight-chain or branched alkoxy, C2-C6 straight-chain or branched alkenyl, C2-C6 straight-chain or branched alkynyl, 6-10 aryl, benzyl, 3-12 heterocyclic, 3-7 cycloalkyl, C2-C6 straight-chain or branched alkylphenyl, C2-C6 straight-chain or branched alkyl 5-7 heteroaryl, -OR5, -NR5R6, -SO2NR5R6, -CONR5R6, -COOR5, -OOCR5, -NHCOR5, -SO2R5, -OSO2R5, m selected from 1, 2, 3, 4, r selected from 1, 2, 3, 4;

[0010] R3 is selected from one or more hydrogens, halogens, hydroxyl groups, substituted or unsubstituted straight-chain or branched C1-C6 alkyl groups, substituted or unsubstituted straight-chain or branched C1-C6 alkoxy groups, substituted or unsubstituted 3-12 membered cycloalkyl groups, substituted or unsubstituted 6-10 membered aryl groups, substituted or unsubstituted 3-12 membered heterocyclic groups, substituted or unsubstituted 3-7 membered cycloalkyl groups, cyano, nitro, carboxyl, mercapto, NR5R6, -CONR5R6, -SO2R5, -SO2NR5R6, -OSO2R5, -OCOR5;

[0011] R4 is selected from hydrogen, amino group, hydroxyl group, substituted or unsubstituted straight-chain or branched C1-C6 alkyl group, substituted or unsubstituted C1-C6 straight-chain or branched alkoxy group, substituted or unsubstituted 6-10 aryl group, substituted or unsubstituted 5-7 heterocyclic group, substituted or unsubstituted C1-C6 straight-chain or branched alkylphenyl group, substituted or unsubstituted 3-12 cycloalkyl group, substituted or unsubstituted C2-C 10 Straight-chain or branched acyl groups, substituted or unsubstituted C2-C 10Straight-chain or branched ester group, amino group, C1-C6 straight-chain or branched alkylamine group, substituted or unsubstituted C1-C6 straight-chain or branched amide group, -OSO2R5, -CH2OSO2R5, -OCOR5;

[0012] R5 and R6 are each independently selected from hydrogen, substituted or unsubstituted 6-10 aryl groups, C1-C4 straight-chain or branched alkyl groups, C1-C4 straight-chain or branched alkenyl groups, C1-C4 straight-chain or branched alkynyl groups, substituted or unsubstituted 3-12 heterocyclic groups, substituted or unsubstituted 3-7 cycloalkyl groups, and C1-C6 straight-chain or branched hydroxyalkyl groups.

[0013] The substituents are selected from one or more halogens, unsubstituted or halogenated or 3-6 membered cycloalkyl-substituted C1-C6 straight-chain or branched alkyl groups, C1-C4 straight-chain or branched alkoxy groups, 3-6 membered cycloalkyl groups, C1-C4 straight-chain or branched alkyl-substituted amino groups, hydroxyl groups, cyano groups, nitro groups, =O groups, hydroxyl-C1-C6 straight-chain or branched alkyl groups, carboxyl groups, mercapto groups, unsubstituted or 1-3 halogenated or hydroxyl-substituted 5-10 membered aryl groups, unsubstituted or halogenated 5-7 membered heterocyclic groups, unsubstituted or halogenated C2-C6 straight-chain or branched acyl groups, C1-C6 hydroxyalkyl groups, -NR5R6, -NHCOR5, -SO2R5, -OSO2R5, -SO2NR5R6, -COOR5, -OCOR5;

[0014] The heterocyclic group contains 1-3 cyclic heteroatoms selected from oxygen, sulfur, and nitrogen;

[0015] The heteroaryl group contains 1-4 cyclic heteroatoms selected from oxygen, sulfur, and nitrogen.

[0016] The term "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), and iodine (I) atoms.

[0017] The term "alkyl" refers to a straight-chain or branched saturated hydrocarbon group having the stated number of carbon atoms. The term "C1-C6 aliphatic chain" refers to a straight-chain or branched saturated hydrocarbon group having 1-6 carbon atoms. C1-C6 alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, 2,2-dimethylbutyl, and 2,3-dimethylbutyl.

[0018] Preferably, in the structure described above:

[0019] R1 and R2 are each independently selected from hydrogen, halogen, CN, NH2, OH, CH2OH, COOH, and -O[(CH2)] m O] rR5, substituted or unsubstituted C1-C4 straight-chain or branched alkyl, substituted or unsubstituted C1-C4 straight-chain or branched alkoxy, substituted or unsubstituted benzyl, substituted or unsubstituted benzyloxy, -OR5, -NR5R6, -SO2NR5R6, -CONR5R6, -COOR5, -OOCR5, -NHCOR5, -SO2R5, -OSO2R5, m is selected from 1, 2, 3, r is selected from 1, 2;

[0020] R5 and R6 are each independently selected from hydrogen, substituted or unsubstituted 6-10 aryl groups, C1-C4 straight-chain or branched alkyl groups, C1-C4 straight-chain or branched alkenyl groups, C1-C4 straight-chain or branched alkynyl groups, substituted or unsubstituted 3-12 heterocyclic groups, substituted or unsubstituted 3-7 cycloalkyl groups, and C1-C6 straight-chain or branched hydroxyalkyl groups.

[0021] The substituents are selected from one or more halogens, unsubstituted or halogenated or 3-6 membered cycloalkyl-substituted C1-C6 straight-chain or branched alkyl groups, C1-C4 straight-chain or branched alkoxy groups, 3-6 membered cycloalkyl groups, C1-C4 straight-chain or branched alkyl-substituted amino groups, hydroxyl groups, cyano groups, nitro groups, =O groups, hydroxyl-C1-C6 straight-chain or branched alkyl groups, carboxyl groups, mercapto groups, unsubstituted or 1-3 halogenated or hydroxyl-substituted 5-10 membered aryl groups, unsubstituted or halogenated 5-7 membered heterocyclic groups, unsubstituted or halogenated C2-C6 straight-chain or branched acyl groups, C1-C6 hydroxyalkyl groups, -NR5R6, -NHCOR5, -SO2R5, -OSO2R5, -SO2NR5R6, -COOR5, -OCOR5;

[0022] The heterocyclic group contains 1-3 oxygen, sulfur, and nitrogen ring heteroatoms.

[0023] Preferably, in the structure described above:

[0024] R3 is selected from one or two hydrogens, hydroxyl groups, halogens, amino groups, substituted or unsubstituted C1-C4 straight-chain or branched alkyl groups, substituted or unsubstituted C1-C4 straight-chain or branched alkoxy groups, substituted or unsubstituted 3-6 membered cycloalkyl groups, substituted or unsubstituted 6-10 membered aryl groups, substituted or unsubstituted 3-12 membered heterocyclic groups, substituted or unsubstituted 3-7 membered cycloalkyl groups, and -NR5R6.

[0025] R5 and R6 are each independently selected from hydrogen, substituted or unsubstituted 6-10 aryl groups, C1-C4 straight-chain or branched alkyl groups, C1-C4 straight-chain or branched alkenyl groups, C1-C4 straight-chain or branched alkynyl groups, substituted or unsubstituted 3-12 heterocyclic groups, substituted or unsubstituted 3-7 cycloalkyl groups, and C1-C6 straight-chain or branched hydroxyalkyl groups.

[0026] R4 is selected from C1-C3 straight-chain or branched alkyl, hydroxymethyl, acetyl, and allyl groups;

[0027] The substituents are selected from one or more halogens, unsubstituted or halogenated or 3-6 membered cycloalkyl-substituted C1-C6 straight-chain or branched alkyl groups, C1-C4 straight-chain or branched alkoxy groups, 3-6 membered cycloalkyl groups, C1-C4 straight-chain or branched alkyl-substituted amino groups, hydroxyl groups, cyano groups, nitro groups, =O groups, hydroxyl-C1-C6 straight-chain or branched alkyl groups, carboxyl groups, mercapto groups, unsubstituted or 1-3 halogenated or hydroxyl-substituted 5-10 membered aryl groups, unsubstituted or halogenated 5-7 membered heterocyclic groups, unsubstituted or halogenated C2-C6 straight-chain or branched acyl groups, C1-C6 hydroxyalkyl groups, -NR5R6, -NHCOR5, -SO2R5, -OSO2R5, -SO2NR5R6, -COOR5, -OCOR5.

[0028] Preferably, in the structure described above:

[0029] R1 and R2 are each independently selected from hydrogen, F, Cl, Br, I, CN, NH2, OH, CH2OH, COOH, C1-C3 straight-chain or branched alkoxy, halogen-substituted C1-C3 straight-chain or branched alkoxy, substituted or unsubstituted benzyloxy, -CONH2, -SO2NH2, -COOCH2CH3, -NHCOCH2CH3;

[0030] R3 is selected from one hydrogen atom, F, Cl, Br, I, CH3, CN, OCH3, CF3, -COCH2CH3, COOH, NH2, -NHCOCH2CH3;

[0031] R4 is selected from hydrogen, CH3, -CH2OH, -COCH3, -SO2R5, -CH2OSO2R5, and -CH2OCOR5;

[0032] R5 is selected from substituted or unsubstituted 6-10 aryl groups, C1-C3 straight-chain or branched alkyl groups;

[0033] The substituents are selected from one or more halogens, unsubstituted or halogenated or 3-6 membered cycloalkyl-substituted C1-C6 straight-chain or branched alkyl groups, C1-C4 straight-chain or branched alkoxy groups, 3-6 membered cycloalkyl groups, C1-C4 straight-chain or branched alkyl-substituted amino groups, hydroxyl groups, cyano groups, nitro groups, =O groups, hydroxyl-C1-C6 straight-chain or branched alkyl groups, carboxyl groups, mercapto groups, unsubstituted or 1-3 halogenated or hydroxyl-substituted 5-10 membered aryl groups, unsubstituted or halogenated 5-7 membered heterocyclic groups, unsubstituted or halogenated C2-C6 straight-chain or branched acyl groups, C1-C6 hydroxyalkyl groups, -NR5R6, -NHCOR5, -SO2R5, -OSO2R5, -SO2NR5R6, -COOR5, -OCOR5;

[0034] R5 and R6 are each independently selected from hydrogen, substituted or unsubstituted 6-10 aryl groups, C1-C4 straight-chain or branched alkyl groups, C1-C4 straight-chain or branched alkenyl groups, C1-C4 straight-chain or branched alkynyl groups, substituted or unsubstituted 3-12 heterocyclic groups, substituted or unsubstituted 3-7 cycloalkyl groups, and C1-C6 straight-chain or branched hydroxyalkyl groups.

[0035] Preferably, in the structure described above:

[0036] Selected from

[0037] Preferably, in the structure described above:

[0038] Selected from

[0039] Further preferred, in the aforementioned structure:

[0040] and Forming Z or E type isomers, of which, except and combination, and combination, and Aside from combinations that form Z or E isomers, all other combinations form Z isomers.

[0041] Further optimization, and It forms a Z-type isomer.

[0042] Preferably, the isochromic ketone compound or its cis-trans isomers and pharmaceutically acceptable salts are selected from the Z-isomers.

[0043] Preferably, the isochoric ketone compound or its cis-trans isomer, or a pharmaceutically acceptable salt thereof, is selected from any one of the following compounds:

[0044] The pharmaceutically acceptable salts described in this invention are salts formed by a compound of Formula I or its cis-trans isomers with any of the following acids: hydrochloric acid, carbonic acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, acetic acid, propionic acid, formic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzylsulfonic acid, ferulic acid, benzylsulfonic acid, aspartic acid, glutamic acid; or salts formed with any of the following bases: hydroxides, carbonates, alcoholic bases, alkyl bases, or organic amines containing lithium, sodium, potassium, calcium, magnesium, or ammonium ions.

[0045] Generally, such salts are prepared by reacting these compounds in their free acid or base form with a stoichiometric amount of a suitable base or acid in water, an organic solvent, or a mixture of both. Non-aqueous media such as ethers, ethyl acetate, ethanol, isopropanol, or acetonitrile are generally preferred.

[0046] The method for preparing the heterochromatic compounds or their cis-trans isomers described in this invention is selected from any of the following methods:

[0047] Method 1:

[0048] The specific reaction process is as follows:

[0049] Step a: Dissolve raw material 1-1 in acetone, then add potassium carbonate to the mixture, slowly add benzyl bromide while stirring, and heat the reaction for 2-8 hours to obtain intermediate 1-2;

[0050] Step b: Add intermediate 1-2 to methanol, slowly add pyridine tribromide, and react at room temperature to obtain intermediate 1-3;

[0051] Step c: Add intermediates 1-3 to methanol, add sodium borohydride, and react to obtain intermediates 1-4;

[0052] Step d: Place intermediates 1-4 in a reaction flask, add toluene to dissolve them, add 50% KOH solution, tert-butyl bromoacetate, and a catalytic amount of tetra-n-butylammonium bromide, and react to obtain intermediates 1-5;

[0053] Step e: Dissolve intermediates 1-5 in ethyl acetate, then add a methanol solution of sodium methoxide, concentrate the solution, and then add acetonitrile and water to continue the reaction to obtain intermediates 1-6.

[0054] Step f: Dissolve intermediates 1-6 in dichloromethane, add a catalytic amount of N,N-dimethylformamide, and add oxaloyl chloride dropwise at 0°C. After the addition is complete, slowly restore the reaction solution to room temperature and react for 0.5 h, then concentrate. Add N,O-dimethylhydroxylamine hydrochloride and potassium carbonate to dry acetonitrile to form a turbid solution. Dissolve the concentrated product in an appropriate amount of acetonitrile, and slowly add it dropwise to the turbid solution using a constant pressure dropping funnel. React at room temperature for 2 h to obtain intermediates 1-7;

[0055] Step g: Dissolve intermediates 1-7 in tetrahydrofuran under nitrogen protection, slowly add tert-butyllithium at -78°C, react at this temperature for 20 min, quench the reaction with saturated ammonium chloride solution, and separate and purify to obtain pale yellow solids 1-8.

[0056] Step h: The obtained intermediates 1-8 are subjected to aldol condensation reaction with substituted indolecarboxaldehyde, and filtered to obtain intermediates 1-9 or products A14, C1-C4.

[0057] Step i: Dissolve the obtained intermediates 1-9 in dry dichloromethane under nitrogen protection, slowly add boron trichloride at -78°C, and react at this temperature for 4 hours. Quench the reaction with methanol, and obtain products A1-A12, D1-D6, E1-E10, A13, D7, and E11 by separation and purification.

[0058] Method 2:

[0059] The specific method is as follows:

[0060] Step j: Dissolve the obtained intermediates 1-8 in methanol and add palladium on carbon. React under H2 atmosphere for 12 h. After filtration and column chromatography, obtain intermediates 1-12.

[0061] Step k: Dissolve the obtained intermediate 1-12 in methanol, add potassium hydroxide, then add the substituted product, react at 70°C overnight, and obtain intermediate 1-13 by filtration and column chromatography;

[0062] Step 1: The obtained intermediates 1-13 are subjected to aldol condensation reaction with substituted indolecarboxaldehyde, and the intermediates 1-14d, 1-14e or products B1, B3, B4 are obtained by filtration.

[0063] Step m: Dissolve the obtained intermediate 1-14 in methanol and add palladium on carbon. React under H2 atmosphere for 12 h. After filtration and column chromatography, obtain products B2 and B5.

[0064] The definitions of R1, R2, R3, and R4 are as described above.

[0065] Compound I obtained by the above method is combined with a pharmaceutically acceptable acid to form a pharmaceutically acceptable salt.

[0066] The pharmaceutical composition of the present invention comprises the isochorone compound or its cis-trans isomer as described in the present invention, a pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier.

[0067] Preferably, the pharmaceutically acceptable carrier is an excipient widely used in the pharmaceutical manufacturing industry. Excipients primarily serve to provide a safe, stable, and functional pharmaceutical composition, and may also provide methods for dissolving the active ingredient at a desired rate after administration to a subject, or for promoting effective absorption of the active ingredient after administration to a subject. The pharmaceutical excipient may be an inert filler, or may provide a function such as stabilizing the overall pH of the composition or preventing degradation of the active ingredient. The pharmaceutical excipient may include one or more of the following: binders, suspending agents, emulsifiers, diluents, fillers, granulators, adhesives, disintegrants, lubricants, anti-adhesion agents, flow aids, wetting agents, gelling agents, absorption delay agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavoring agents, and sweeteners.

[0068] The pharmaceutical compositions of the present invention can be prepared using any method known to those skilled in the art, based on the disclosure. For example, conventional mixing, dissolving, granulation, emulsification, grinding, encapsulation, embedding, or lyophilization processes.

[0069] The pharmaceutical compositions of this invention can be administered in any form, including by injection (intravenous), mucosal, oral (solid and liquid formulations), inhalation, ocular, rectal, topical, or parenteral (infusion, injection, implantation, subcutaneous, intravenous, intra-arterial, intramuscular) administration. The pharmaceutical compositions of this invention can also be controlled-release or sustained-release dosage forms (e.g., liposomes or microspheres). Examples of solid oral formulations include, but are not limited to, powders, capsules, tablets, soft capsules, and tablets. Examples of liquid formulations for oral or mucosal administration include, but are not limited to, suspensions, emulsions, elixirs, and solutions. Examples of topical formulations include, but are not limited to, emulsions, gels, ointments, creams, patches, pastes, foams, lotions, drops, or serum preparations. Examples of parenteral formulations include, but are not limited to, solutions for injection, dry powder formulations that can be dissolved or suspended in a pharmaceutically acceptable carrier, suspensions for injection, and emulsions for injection. Examples of other suitable formulations of the pharmaceutical composition include, but are not limited to, eye drops and other ophthalmic preparations; aerosols, such as nasal sprays or inhalers; liquid dosage forms suitable for parenteral administration; suppositories; and tablets.

[0070] The isochromic ketone compounds or their cis-trans isomers, pharmaceutically acceptable salts, or pharmaceutical compositions thereof described in this invention are used in the preparation of drugs for the production of preprotein convertase subtilisin 9 inhibitors.

[0071] Preferably, the drug is a drug for the prevention and / or treatment of hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, fatty liver disease, atherosclerosis, and obesity.

[0072] Further preferably, the drug is a drug that lowers total cholesterol, low-density lipoprotein cholesterol, and triglycerides.

[0073] Further preferably, the drug is one that increases liver LDL receptor expression, inhibits PCSK9 expression, and activates AMPK protein kinase.

[0074] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages:

[0075] The compound designed in this invention can effectively inhibit the activity of PCSK9, with an optimal inhibition rate of over 90%. It also exhibits significant lipid-lowering effects both in vivo and in vitro, including downregulating the levels of total cholesterol, triglycerides, and low-density lipoprotein. Furthermore, it possesses pharmacokinetic properties that are beneficial for drug development and has promising clinical application prospects in the treatment of metabolic diseases. Detailed Implementation

[0076] The technical solution of the present invention will be further described below with reference to the embodiments.

[0077] Unless otherwise specified, all starting materials used in this invention are commercially available.

[0078] Example 1: (Z)-3-((1H-indol-2-yl)methylene)-6-hydroxy-7-methoxyisocyano-4-one (A1)

[0079] (1) Synthesis of intermediate 4-benzyloxy-5-methoxybenzaldehyde (a2)

[0080] 4-Hydroxy-5-methoxybenzaldehyde (a1, 20.00 g, 86.56 mmol) was dissolved in acetone, and potassium carbonate (23.93 g, 173.13 mmol) and benzyl bromide (17.77 g, 103.88 mmol) were slowly added. After the addition was complete, the mixture was stirred at 65 °C for 3 h. After the reaction was completed by TLC monitoring, the reaction solution was filtered, and the filtrate was collected and concentrated under reduced pressure to give compound a2 (24.05 g, 74.73 mmol), with a yield of 86.33%. 1 H NMR (400MHz, CDCl3) δ7.48–7.40(m,2H),7.36(dq,J=6.8,1.1Hz,1H),7.35–7.28(m,2H),5.14(t,J=1.0Hz,1H),3.88(s,2H). 13C NMR (100MHz, CDCl3) δ191.84,152.18,148.94,136.68,130.13,128.76,128. 48,128.15,120.96,119.08,111.82,71.21,56.55.LRMS(ESI,m / z):243[M+H] + .

[0081] (2) Synthesis of intermediate 4-benzyloxy-2-bromo-5-methoxybenzaldehyde (a3)

[0082] 4-Benzyloxy-5-methoxybenzaldehyde (a2, 24.05 g, 74.73 mmol) was dissolved in methanol, and solid pyridine tribromide (63.90 g, 198.54 mmol) was slowly added. The mixture was stirred at room temperature for 3 h. After the reaction was completed by TLC monitoring, the reaction solution was evaporated to dryness to a muddy state, solidified, and the product was washed with toluene. The filtrate was collected and concentrated under reduced pressure to give compound a3 (18.00 g, 56.05 mmol), with a yield of 56.46%.

[0083] 1 H NMR (400MHz, CDCl3) δ7.48–7.40(m,1H),7.39–7.32(m,1H),7.35–7.28(m,1H),5.14(t,J=1.0Hz,1H),3.88(s,1H). 13 C NMR (100MHz, CDCl3) δ191.84,152.18,148.94,136.68,130.13,128.76,128. 48,128.15,120.96,119.08,111.82,71.21,56.55.LRMS(ESI,m / z):322[M+H] + .

[0084] (3) Synthesis of intermediate [4-(benzyloxy)-2-bromo-5-methoxyphenyl]methanol (a4)

[0085] 4-Benzyloxy-2-bromo-5-methoxybenzaldehyde (a3, 18.00 g, 56.05 mmol) was dissolved in methanol, and NaBH4 (3.18 g, 84.07 mmol) was slowly added under ice bath conditions, with stirring for 1 h. After the reaction was complete as monitored by TLC, the reaction solution was evaporated to dryness, the reaction was quenched with ammonium chloride, and the mixture was extracted with ethyl acetate and water. The solution was concentrated under reduced pressure to give compound a4 (17.6 g, 54.46 mmol), in 97.17% yield.

[0086] 1H NMR(400MHz, CDCl3)δ7.43(ddq,J=6.9,1.8,0.9Hz,2H),7.39–7.32(m,2H),7.35–7.28(m,1H),7.13(s,1H),6 .96(t,J=1.1Hz,1H),5.14(t,J=0.9Hz,2H),4.70(dd,J=6.3,1.0Hz,2H),4.05(t,J=6.3Hz,1H),3.86(s,2H). 13 C NMR (100MHz, CDCl3) δ148.22,148.05,136.70,134.51,128.54,128.48,128. 15,118.40,116.68,112.48,71.21,62.85,56.14.LRMS(ESI,m / z):324[M+H] + .

[0087] (4) Synthesis of intermediate ({[4-(benzyloxy)-2-bromo-5-methoxyphenyl]methyl}oxy)acetic acid-2-methylpropyl-2-yl ester (a5)

[0088] [4-(benzyloxy)-2-bromo-5-methoxyphenyl]methanol (a4, 17.60 g, 54.46 mmol) was dissolved in toluene, and tert-butyl bromoacetate (15.93 g, 81.69 mmol), tetrabutylammonium bromide (526.68 g, 1.63 mmol), and 50% KOH (15.28 g, 272.29 mmol) were added. The reaction was carried out in an oil bath at 60 °C for 4 h. After the reaction was completed, the mixture was extracted with ethyl acetate and water by TLC. The compound a5 (18.00 g, 41.16 mmol) was obtained by concentration under reduced pressure, with a yield of 75.58%.

[0089] 1 H NMR(400MHz, CDCl3)δ7.42(ddt,J=7.8,1.9,1.0Hz,2H),7.39–7.32(m,2H),7.34–7.27(m,1H),7.11(s,1H), 6.97(t,J=1.0Hz,1H),5.13(t,J=1.0Hz,2H),4.65(d,J=0.9Hz,2H),4.14(s,1H),3.84(s,2H),1.42(s,6H). 13C NMR (100MHz, CDCl3) δ170.04,148.65,148.01,136.69,133.10,128.50,128.24,127.53,11 8.48,116.85,111.78,82.09,71.30,70.74,67.06,56.16,27.79.LRMS(ESI,m / z):438[M+H] + .

[0090] (5) Synthesis of intermediate ({[4-(benzyloxy)-2-bromo-5-methoxyphenyl]methyl}oxy)acetic acid (a6)

[0091] ({[4-(benzyloxy)-2-bromo-5-methoxyphenyl]methyl}oxy)acetic acid-2-methylpropyl-2-yl ester (a5, 18.00 g, 41.16 mmol) was dissolved in methanol, and a methanol solution of sodium methoxide (4.45 g, 82.32 mmol) was slowly added. A suitable amount of water was then added, and the reaction was carried out at room temperature for 5 min. After the reaction was complete as monitored by TLC, the methanol was evaporated to dryness, and a small amount of EA and a larger amount of water were added for extraction once. While stirring, concentrated hydrochloric acid was slowly added to the aqueous layer obtained from the extraction to adjust the pH to 2. EA was added again for extraction 2–3 times. After 3 extractions, the mixture was washed with saturated NaCl, dried over anhydrous sodium sulfate, and concentrated to obtain compound a6 (14.68 g, 38.51 mmol), with a yield of 93.56%.

[0092] 1 H NMR(400MHz, CDCl3)δ7.43(ddt,J=7.2,1.8,1.0Hz,2H),7.39–7.32(m,2H),7.34–7.27(m,1H),7.12( s,1H),6.98(t,J=1.0Hz,1H),5.13(t,J=1.0Hz,2H),4.65(d,J=0.9Hz,2H),4.11(s,1H),3.85(s,2H). 13 C NMR (100MHz, CDCl3) δ173.65,148.45,147.99,136.67,132.51,128.48,128.14,127 .53,118.61,116.42,112.43,71.21,70.81,66.95,56.17.LRMS(ESI,m / z):382[M+H] + .

[0093] (6) Synthesis of intermediate 2-({[4-(benzyloxy)-2-bromo-5-methoxyphenyl]methyl}oxy)-N-methoxy-N-methylacetamide (a7)

[0094] {[4-(benzyloxy)-2-bromo-5-methoxyphenyl]methyl}oxy)acetic acid (a6, 14.68 g, 38.51 mmol) was dissolved in dichloromethane, and O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethylurea hexafluorophosphate (19.03 g, 50.06 mmol) and N,N-diisopropylethylamine (7.47 g, 57.7 mmol) were slowly added. The reaction was allowed to proceed for 12 h. After the reaction was completed as monitored by TLC, dichloromethane and a large amount of water were added, and the mixture was extracted three times. The extract was washed with saturated NaCl, dried over anhydrous sodium sulfate, and concentrated to give compound a7 (12 g, 28.28 mmol), with a yield of 73.45%.

[0095] 1 H NMR (400MHz, CDCl3) δ7.42(ddt,J=7.1,2.0,1.0Hz,2H),7.35(ddt,J=7.6,6.6,0.9Hz,2H),7.34–7.26(m,1H),7.12(s,1H) ,6.97(t,J=1.0Hz,1H),5.13(t,J=1.0Hz,2H),4.65(d,J=0.9Hz,2H),4.15(s,1H),3.84(s,2H),3.55(s,2H),3.03(s,2H). 13 C NMR (100MHz, CDCl3) δ169.53,148.65,148.01,136.69,132.63,128.50,128.27,127.53,11 8.48,116.71,112.45,71.24,70.92,65.26,61.23,56.16,32.45.LRMS(ESI,m / z):425[M+H] + .

[0096] (7) Synthesis of intermediate 6-(benzyloxy)-7-methoxy-3,4-dihydro-1H-isochroman-4-one (a8)

[0097] 2-({[4-(benzyloxy)-2-bromo-5-methoxyphenyl]methyl}oxy)-N-methoxy-N-methylacetamide (a7, 6 g, 17.37 mmol) was placed in a three-necked flask, anhydrous tetrahydrofuran was added, and under nitrogen protection, tert-butyllithium (2.23 g, 34.74 mmol) was slowly added at -78 °C. The reaction was carried out at this temperature for 20 min, and the reaction was quenched by adding saturated ammonium chloride solution. The mixture was separated and purified to obtain a white solid a8, with a yield of 26.32%.

[0098] 1H NMR(400MHz, CDCl3) δ7.42(ddtt,J=5.7,3.6,2.2,1.0Hz,3H),7.39–7.32(m,4H),7.29(s,1H),7.05( t,J=1.1Hz,1H),5.13(t,J=1.0Hz,3H),4.98(d,J=0.9Hz,2H),4.75(s,1H),4.68(s,1H),3.83(s,3H). 13 CNMR (100MHz, CDCl3) δ195.78,152.23,147.99,136.62,135.27,128.66,128.35,128 .14,127.38,111.65,109.83,72.24,71.24,70.10,56.17.LRMS(ESI,m / z):285[M+H] + .

[0099] (8) Synthesis of intermediate (Z)-3-((1H-indol-2-yl)methylene)-6-(benzyloxy)-7-methoxyisocyano-4-one (a9)

[0100] 6-(benzyloxy)-7-methoxy-3,4-dihydro-1H-isochroman-4-one (a8, 0.15 g, 527.59 μmol) was dissolved in tetrahydrofuran, 10% KOH was added, followed by 1H-indole-2-carboxaldehyde, and the reaction was continued until a solid precipitated. After filtration, a yellow solid a9 (0.12 g, 291.65 μmol) was obtained, with a yield of 55.28%.

[0101] 1 H NMR(400MHz, CDCl3)δ9.80(s,1H),7.56(dt,J=7.5,1.6Hz,1H),7.45–7.39(m,3H),7.42–7.32(m,3H),7.34–7.27(m,1H),7.25(td,J=7.5,1.6Hz,1 H),7.16(td,J=7.5,1.5Hz,1H),7.10(t,J=1.0Hz,1H),6.84(s,1H),6.58 –6.55(m,1H),5.29(d,J=1.0Hz,2H),5.12(d,J=1.1Hz,2H),3.83(s,2H). 13C NMR (100MHz, CDCl3) δ179.69,151.28,148.46,142.18,137.56,137.03,136.83,131.36,128.51,128.39,128.17,127.66, 125.57,122.61,120.55,120.52,112.73,111.85,109.46,109.09,104.44,71.91,71.32,56.16.LRMS(ESI,m / z):412[M+H] + .

[0102] (9) Synthesis of (Z)-3-((1H-indol-2-yl)methylene)-6-hydroxy-7-methoxyisocyano-4-one (A1)

[0103] (Z)-3-((1H-indol-2-yl)methylene)-6-(benzyloxy)-7-methoxyisocyano-4-one (a9, 0.12 g, 291.65 μmol) was placed in a three-necked flask and dissolved in anhydrous dichloromethane. Under nitrogen protection, boron trichloride was slowly added at -78 °C. After the reaction was completed by TLC monitoring, water was added to quench the reaction. The mixture was extracted three times with dichloromethane-water, washed with saturated NaCl, dried over anhydrous sodium sulfate, concentrated, and then subjected to column chromatography to give compound A1 (0.054 g, 168.05 mmol), yield 57.62%, and A13 (0.01 g, mmol), yield 10.67%.

[0104] 1 H NMR (400MHz, CDCl3) δ7.67(s,1H),7.55(dt,J=7.4,1.7Hz,1H),7.39(dd,J=7.4,1.6Hz,1H),7.31(s,1H),7.22(td,J=7.5,1.6H z,1H),7.15(td,J=7.5,1.5Hz,1H),7.03(t,J=1.0Hz,1H),6.80(s,1H),6.58–6.55(m,1H),5.29(d,J=1.0Hz,2H),3.83(s,2H). 13 C NMR (100MHz, CDCl3) δ179.93,150.63,147.01,142.31,137.59,136.79,128.91,126.48,125.56,12 2.43,120.74,120.38,112.40,111.69,109.49,108.54,103.87,71.83,56.03.HRMS(ESI)m / z:calcd for C 19 H15 NO4[M+H] + 322.1001, found 322.1018.

[0105] Example 2: (Z)-6-hydroxy-7-methoxy-3-((5-methoxy-1H-indol-2-yl)methylene)isocyanine-4-one (A2)

[0106] Intermediate a8 was dissolved in tetrahydrofuran, and 10% KOH was added, followed by the addition of 5-methoxy-1H-indole-2-carboxaldehyde. The reaction was continued until a solid precipitated. The mixture was filtered to obtain a yellow solid intermediate. Compound A2 was synthesized from the intermediate using the same method as compound A1.

[0107] 1 H NMR (400MHz, CDCl3) δ9.81 (s, 1H), 7.65 (s, 1H), 7.31 (s, 1H), 7.25 (d, J = 7.5Hz, 1H), 7.19 (t, J = 1.5Hz, 1H), 7.03 (t, J = 1. 0Hz, 1H), 6.81 (dd, J = 7.5, 1.5Hz, 1H), 6.66 (s, 1H), 6.48 (d, J = 1.5Hz, 1H), 5.29 (d, J = 0.9Hz, 2H), 3.84 (d, J = 10.3Hz, 7H). 13 C NMR (100MHz, CDCl3) δ179.75,156.75,150.34,146.91,142.31,137.82,133.77,128.95,127.90,127.0 9,113.21,112.56,112.40,109.51,108.71,106.80,103.74,71.82,56.01,55.27.HRMS(ESI)m / z:calcd for C 20 H 17 NO5[M+H] + 352.1107, found 352.1101.

[0108] Example 3: (Z)-6-hydroxy-7-methoxy-3-((6-methoxy-1H-indol-2-yl)methylene)isocyanine-4-one (A3)

[0109] Intermediate a8 was dissolved in tetrahydrofuran, and 10% KOH was added, followed by the addition of 6-methoxy-1H-indole-2-carboxaldehyde. The reaction was continued until a solid precipitated. The mixture was filtered to obtain a yellow solid intermediate. Compound A3 was synthesized from the intermediate using the same method as compound A1.

[0110] 1 H NMR(400MHz, CDCl3)δ9.82(s,1H),7.83(dd,J=7.4,1.6Hz,1H),7.65(s,1H),7.31(s,1H),7.03(t,J=1.0Hz ,1H),6.91–6.83(m,2H),6.66(s,1H),6.54(d,J=1.4Hz,1H),5.29(d,J=0.9Hz,2H),3.84(d,J=3.5Hz,7H). 13 C NMR (100MHz, CDCl3) δ179.75,157.67,150.34,146.91,142.31,138.22,137.24,128.95,127.09,123.3 6,121.36,112.40,110.21,109.51,108.68,103.74,95.60,71.82,56.01,55.37.HRMS(ESI)m / z:calcd for C 20 H 17 NO5[M+H] + 352.1107, found 352.1009.

[0111] Example 4: (Z)-6-hydroxy-7-methoxy-3-((6-methoxy-1H-indol-2-yl)methylene)isocyanine-4-one (A4)

[0112] Intermediate a8 was dissolved in tetrahydrofuran, and 10% KOH was added, followed by 5,6,7-trimethoxy-1H-indole-2-carboxaldehyde. The reaction was continued until a solid precipitated. After filtration, a yellow solid intermediate was obtained. Compound A5 was synthesized from the intermediate using the same method as compound A1.

[0113] 1 H NMR (400MHz, CDCl3) δ9.61 (s, 1H), 7.76 (s, 1H), 7.32 (s, 1H), 7.07 (d, J = 1.4Hz, 1H), 7.03 (t, J = 1.0Hz, 1H) ,6.90(s,1H),6.46(d,J=1.6Hz,1H),5.29(d,J=1.1Hz,2H),3.89(d,J=12.5Hz,5H),3.82(d,J=6.6Hz,6H). 13C NMR (100MHz, CDCl3) δ179.81,150.35,148.48,147.05,142.17,139.34,138.62,137.05,129.15,127.01,126. 55,125.87,112.69,109.28,109.12,104.13,103.82,71.86,60.93,60.86,56.25,56.06.HRMS(ESI)m / z:calcd for C 22 H 21 NO7[M+H] + 412.1309, found 412.1325.

[0114] Example 5: (Z)-3-((5-fluoro-1H-indol-2-yl)methylene)-6-hydroxy-7-methoxyisocyano-4-one (A5)

[0115] Intermediate a8 was dissolved in tetrahydrofuran, and 10% KOH was added, followed by 5-fluoro-1H-indole-2-carboxaldehyde. The reaction was continued until a solid precipitated. The mixture was filtered to obtain a yellow solid intermediate. Compound A5 was synthesized from the intermediate using the same method as compound A1.

[0116] 1 H NMR(400MHz, CDCl3)δ9.78(s,1H),7.62(s,1H),7.46–7.37(m,2H),7.33(s,1H),7.02(t,J=0.9Hz,1 H),6.98(td,J=7.8,1.5Hz,1H),6.66(s,1H),6.54–6.50(m,1H),5.29(d,J=0.9Hz,2H),3.84(s,2H). 13 CNMR (100MHz, CDCl3) δ179.61,159.08,157.06,150.34,147.01,142.31,138.05,135.26,135.23,128.91,127.39,127.32,127.1 5,113.37,113.30,113.24,113.08,112.83,109.68,109.63,109.61,108.59,108.43,103.74,71.61,56.01.HRMS(ESI)m / z:calcd for C 19 H 14 FNO4[M+H] + 340.0907, found 340.0900.

[0117] Example 6: (Z)-3-((6-fluoro-1H-indol-2-yl)methylene)-6-hydroxy-7-methoxyisocyano-4-one (A6)

[0118] Intermediate a8 was dissolved in tetrahydrofuran, and 10% KOH was added, followed by the addition of 6-fluoro-1H-indole-2-carboxaldehyde. The reaction was continued until a solid precipitated. After filtration, a yellow solid intermediate was obtained. Compound A6 was synthesized from the intermediate using the same method as compound A1.

[0119] 1 H NMR(400MHz, CDCl3)δ7.89(ddd,J=7.8,5.0,1.6Hz,1H),7.62(s,1H),7.33(s,1H),7.27–7.21(m,1H),7.18(td ,J=7.7,1.5Hz,1H),7.02(t,J=0.9Hz,1H),6.66(s,1H),6.58–6.54(m,1H),5.29(d,J=0.9Hz,2H),3.84(s,2H). 13 C NMR (100MHz, CDCl3) δ179.61,161.02,159.00,150.34,147.01,142.31,138.99,138.93,138.45,128.91,127.15,123.95,1 23.93,123.12,123.06,112.83,112.29,112.13,109.68,109.24,103.74,97.47,97.31,71.61,56.01.HRMS(ESI)m / z:calcd for C 19 H 14 FNO4[M+H] + 340.0907, found 340.0919.

[0120] Example 7: (Z)-3-((6-fluoro-1H-indol-2-yl)methylene)-6-hydroxy-7-methoxyisocyano-4-one (A7)

[0121] Intermediate a8 was dissolved in tetrahydrofuran, and 10% KOH was added, followed by the addition of 7-fluoro-1H-indole-2-carboxaldehyde. The reaction was continued until a solid precipitated. The intermediate was then filtered to obtain a yellow solid. Compound A7 was synthesized from the intermediate using the same method as compound A1.

[0122] 1H NMR(400MHz, CDCl3)δ7.65–7.59(m,2H),7.44(td,J=7.5,5.0Hz,1H),7.33(s,1H),7.0 4–6.96(m,2H),6.89(s,1H),6.64(d,J=1.4Hz,1H),5.29(d,J=0.9Hz,2H),3.84(s,2H). 13 C NMR (100MHz, CDCl3) δ179.61,152.68,150.66,150.34,147.01,142.25,136.95,136.86,128.91,128.34,128.18,127.15,127.00,12 6.94,122.43,122.36,117.70,117.67,112.83,111.57,111.41,109.84,109.81,109.68,103.87,71.61,56.01.HRMS(ESI)m / z:calcd for C 19 H 14 FNO4[M+H] + 340.0907, found 340.0900.

[0123] Example 8: (Z)-3-((5-bromo-1H-indol-2-yl)methylethylenedimethyl)-6-hydroxy-7-methoxyisocyano-4-one (A8)

[0124] Intermediate a8 was dissolved in tetrahydrofuran, and 10% KOH was added, followed by the addition of 5-bromo-1H-indole-2-carboxaldehyde. The reaction was continued until a solid precipitated. After filtration, a pale yellow solid intermediate was obtained. Compound A8 was synthesized from the intermediate using the same method as compound A1.

[0125] 1 H NMR(400MHz, CDCl3)δ9.71(s,1H),7.88(t,J=1.7Hz,1H),7.63(s,1H),7.33–7.25(m,2H),7.22(dd,J=7.5, 1.5Hz,1H),7.03(t,J=1.0Hz,1H),6.66(s,1H),6.60(d,J=1.8Hz,1H),5.29(d,J=0.9Hz,2H),3.83(s,3H). 13C NMR (100MHz, CDCl3) δ179.89,150.41,147.14,142.31,137.82,136.87,128.98,128.41,127.17,12 7.14,123.25,115.78,113.97,112.78,109.34,108.91,103.52,71.83,56.04.HRMS(ESI)m / z:calcd for C 19 H 14 BrNO4[M+H] + 400.0106, found 400.0118.

[0126] Example 9: (Z)-3-((6-bromo-1H-indol-2-yl)methylethylene)-6-hydroxy-7-methoxyisocyano-4-one (A9)

[0127] Intermediate a8 was dissolved in tetrahydrofuran, and 10% KOH was added, followed by the addition of 6-bromo-1H-indole-2-carboxaldehyde. The reaction was continued until a solid precipitated. After filtration, a pale yellow solid intermediate was obtained. Compound A9 was synthesized from the intermediate using the same method as compound A1.

[0128] 1 H NMR(400MHz, CDCl3)δ7.88(dd,J=7.5,1.6Hz,1H),7.72–7.68(m,1H),7.63(s,1H),7.56(dd,J=7.5,1.5Hz,1 H),7.31(s,1H),7.03(d,J=1.0Hz,1H),6.66(s,1H),6.62–6.58(m,1H),5.29(d,J=0.9Hz,2H),3.83(s,2H). 13 C NMR (100MHz, CDCl3) δ179.89,150.41,147.14,142.31,138.22,137.37,128.98,127.17,124.83,12 4.44,123.38,115.84,114.74,112.78,109.38,109.34,103.52,71.83,56.04.HRMS(ESI)m / z:calcd for C 19 H 14 BrNO4[M+H] + 400.0106, found 400.0122.

[0129] Example 10: (Z)-3-((7-bromo-1H-indol-2-yl)methylethylene)-6-hydroxy-7-methoxy-isochroman-4-one (A10)

[0130] Intermediate a8 was dissolved in tetrahydrofuran, and 10% KOH was added, followed by the addition of 7-bromo-1H-indole-2-carboxaldehyde. The reaction was continued until a solid precipitated. After filtration, a pale yellow solid intermediate was obtained. Compound A10 was synthesized from the intermediate using the same method as compound A1.

[0131] 1 H NMR (400MHz, CDCl3) δ7.69(dt,J=7.5,1.6Hz,1H),7.63(s,1H),7.35(t,J=7.5Hz,1H),7.32–7.25(m ,2H),7.03(t,J=1.0Hz,1H),6.85(s,1H),6.72(d,J=1.6Hz,1H),5.29(d,J=1.0Hz,2H),3.83(s,2H). 13 CNMR (100MHz, CDCl3) δ179.89,150.41,147.14,142.26,137.84,135.13,128.98,128.24,127.50,1 27.17,122.05,120.48,112.78,110.08,109.34,103.65,97.07,71.83,56.04.HRMS(ESI)m / z:calcd for C 19 H 14 BrNO4[M+H] + 400.0106, found 400.0122.

[0132] Example 11: (Z)-6-hydroxy-3-((1-(hydroxymethyl)-1H-indol-2-yl)methylethylene)-7-methoxyisocyano-4-one (A11)

[0133] Intermediate a8 was dissolved in tetrahydrofuran, and 10% KOH was added, followed by the addition of 1-hydroxymethyl-1H-indole-2-carboxaldehyde. The reaction was continued until a solid precipitated. After filtration, a pale yellow solid intermediate was obtained. Compound A11 was synthesized from the intermediate using the same method as compound A1.

[0134] 1H NMR(400MHz, CDCl3) δ7.64(d,J=7.0Hz,2H),7.57–7.52(m,1H),7.30–7.19(m,3H),7.03(t,J=1.0Hz,1H),6.95( s,1H),6.86(d,J=1.7Hz,1H),6.12(t,J=5.3Hz,1H),5.56(d,J=5.2Hz,2H),5.29(d,J=0.9Hz,2H),3.83(s,2H). 13 C NMR (100MHz, CDCl3) δ179.90,150.34,148.04,146.91,138.50,136.86,128.95,128.51,127.02,124.7 4,122.15,121.31,112.40,110.61,110.42,109.51,105.44,72.20,69.46,56.01.HRMS(ESI)m / z:calcd for C 20 H 17 NO5[M+H] + 352.1107, found 352.1101.

[0135] Example 12: Synthesis of (Z)-6-hydroxy-7-methoxy-3-((7-methyl-1H-indol-2-yl)methylethylene)-isochroman-4-one (A12)

[0136] Intermediate a8 was dissolved in tetrahydrofuran, and 10% KOH was added, followed by the addition of 7-methyl-1H-indole-2-carboxaldehyde. The reaction was continued until a solid precipitated. After filtration, a pale yellow solid intermediate was obtained. Compound A12 was synthesized from the intermediate using the same method as compound A1.

[0137] 1 H NMR (400MHz, CDCl3) δ9.58(s,1H),7.62(s,1H),7.57(dt,J=7.5,1.6Hz,1H),7.33(s,1H),7.18(t,J=7.5Hz,1H) ,7.06–7.00(m,2H),6.85(s,1H),6.80–6.76(m,1H),5.29(d,J=0.9Hz,2H),3.84(s,2H),2.59(d,J=0.7Hz,3H).. 13C NMR (100MHz, CDCl3) δ179.61,150.39,147.01,142.25,137.78,136.98,128.91,128.28,127.15,124.6 4,122.96,120.93,118.00,112.83,109.68,109.35,103.89,71.62,56.02,17.23.HRMS(ESI)m / z:calcd for C 20 H 17 NO4[M+H] + 336.1158, found 336.1142.

[0138] Example 13: Synthesis of (E)-3-((1H-indol-2-yl)methylene)-6-hydroxy-7-methoxyisocyano-4-one (A13)

[0139] Compound A13 was synthesized according to the same method as compound A1.

[0140] 1 H NMR (400MHz, CDCl3) δ7.67(s,1H),7.55(dt,J=7.1,1.6Hz,1H),7.39(dd,J=7.4,1.6Hz,1H),7.31(s,1H),7.22(td,J=7.5,1.6H z,1H),7.15(td,J=7.5,1.5Hz,1H),7.03(t,J=1.0Hz,1H),6.58–6.55(m,1H),6.17(s,1H),5.29(d,J=1.0Hz,2H),3.83(s,2H). 13 C NMR (100MHz, CDCl3) δ180.40,150.63,147.01,142.31,136.83,136.79,128.91,126.48,125.56,12 2.43,120.74,120.38,112.40,111.69,109.49,108.54,104.47,71.83,56.03.HRMS(ESI)m / z:calcd for C 19 H 15 NO4[M+H] + 322.1001, found 322.1018.

[0141] Example 14: Synthesis of (Z)-3-((1H-indol-2-yl)methylene)-6-(benzyloxy)-7-methoxyisocyano-4-one (A14)

[0142] 6-(benzyloxy)-7-methoxy-3,4-dihydro-1H-isochroman-4-one (0.15 g, 527.59 μmol) was dissolved in tetrahydrofuran, 10% KOH was added, followed by 1H-indole-2-carboxaldehyde, and the reaction was continued until a solid precipitated. After filtration, a yellow solid A14 (0.15 g, 364.56 μmol) was obtained, with a yield of 69.10%.

[0143] 1 H NMR(400MHz, CDCl3)δ9.80(s,1H),7.56(dt,J=7.5,1.6Hz,1H),7.45–7.39(m,3H),7.42–7.32(m,3H),7.34–7.27(m,1H),7.25(td,J=7.5,1.6Hz,1 H),7.16(td,J=7.5,1.5Hz,1H),7.10(t,J=1.0Hz,1H),6.84(s,1H),6.58 –6.55(m,1H),5.29(d,J=1.0Hz,2H),5.12(d,J=1.1Hz,2H),3.83(s,2H). 13 C NMR (100MHz, CDCl3) δ179.69,151.28,148.46,142.18,137.56,137.03,136.83,131.36,128.51,128.39,128.17,127.6 6,125.57,122.61,120.55,120.52,112.73,111.85,109.46,109.09,104.44,71.91,71.32,56.16.HRMS(ESI)m / z:calcd for C 26 H 21 NO4[M+H] + 412.1471, found 412.1462.

[0144] Example 15: Synthesis of (Z)-3-((1H-indol-2-yl)methylene)-7-methoxy-6-(3-methoxypropoxy)isochroman-4-one (B1)

[0145] (1) Synthesis of intermediate 6-hydroxy-7-methoxy-3,4-dihydro-1H-isochroman-4-one (b1)

[0146] Intermediate a8 (0.80 g, 2.81 mmol) was dissolved in methanol in a 100 mL single-necked flask, and ammonium formate (0.53 g, 8.44 mmol) and palladium on carbon were added. The mixture was reacted at 80 °C for 3 h. After the reaction was complete as monitored by TLC, the mixture was filtered. The filtrate was concentrated to give a brownish-yellow solid, yielding intermediate b1 (0.52 g, 2.69 mmol), with a yield of 95.53%.

[0147] 1 H NMR (400MHz, CDCl3) δ7.58 (s, 4H), 7.24 (s, 3H), 6.99 (t, J = 0.9Hz, 4H), 4.98 (d, J = 1.1Hz, 8H), 4.79 (s, 6H), 3.87 (s, 9H). 13 C NMR(100MHz, CDCl3)δ195.34,150.58,146.24,132.93,128.95,112.57,110.28,72.17,70.26,56.11.LRMS(ESI,m / z):195[M+H] + .

[0148] (2) Synthesis of intermediate 7-methoxy-6-[(3-methoxypropyl)oxy]-3,4-dihydro-1H-isochroman-4-one (b2)

[0149] Intermediate b1 (0.20 g, 1.03 mmol) was dissolved in acetonitrile in a 100 mL single-necked flask, and 1-bromo-3-methoxypropane (0.19 g, 1.24 mmol) and potassium carbonate (0.28 g, 2.06 mmol) were added. The mixture was reacted at 80 °C for 6 h. After the reaction was completed by TLC monitoring, the mixture was filtered. The filtrate was concentrated and subjected to column chromatography to give intermediate b2 (0.18 g, 675.95 μmol), with a yield of 65.63%.

[0150] 1 H NMR (400MHz, CDCl3) δ7.27(s,1H),7.05(t,J=1.0Hz,1H),4.98(d,J=0.9Hz,2H),4.72(s,1H), 4.13(t,J=7.1Hz,2H),3.83(s,2H),3.52(t,J=7.1Hz,2H),3.19(s,2H),2.01(p,J=7.1Hz,2H). 13C NMR (100MHz, CDCl3) δ195.17,151.09,148.18,136.14,127.77,111.56,109.8 4,72.22,70.20,69.98,67.70,58.37,56.15,30.20.LRMS(ESI,m / z):267[M+H] + .

[0151] (3) Synthesis of (Z)-3-((1H-indol-2-yl)methylene)-7-methoxy-6-(3-methoxypropoxy)isocyano-4-one (B1)

[0152] Intermediate b2, 10% KOH, and 1H-indole-2-carboxaldehyde were reacted according to the method in Example 1 to obtain the target compound B1.

[0153] 1 H NMR (400MHz, CDCl3) δ9.78 (s, 1H), 7.56 (dt, J = 7.5, 1.7Hz, 1H), 7.43–7.35 (m, 2H) ,7.24(td,J=7.4,1.6Hz,1H),7.16(td,J=7.4,1.5Hz,1H),7.08(t,J=1.0Hz,1H), 6.84(s,1H),6.58–6.55(m,1H),5.29(d,J=1.1Hz,2H),4.19(t,J=7.1Hz,2H),3.8 3(s,2H),3.52(t,J=7.1Hz,2H),3.18(s,2H),2.05(ddq,J=22.4,14.4,7.1Hz,2H). 13 C NMR (100MHz, CDCl3) δ179.85,150.27,149.41,142.23,137.56,136.77,131.56,127.98,125.58,122.70,120. 53,112.64,111.69,109.46,108.85,103.87,71.89,69.86,67.56,58.37,56.16,30.17.HRMS(ESI)m / z:calcd for C 23 H 23 NO5[M+H] + 393.1576, found 393.1588.

[0154] Example 16: Synthesis of (Z)-3-((1H-indol-2-yl)methylene)-6-(3-hydroxypropoxy)-7-methoxyisocyanate-4-one (B2)

[0155] Intermediate b1 and 3-bromoprop-1-ol were used to obtain intermediate 6-[(3-hydroxypropyl)oxy]-7-methoxy-3,4-dihydro-1H-isochroman-4-one according to the method of Example 15.

[0156] 1 H NMR (400MHz, CDCl3) δ7.27(s,1H),7.05(t,J=1.0Hz,1H),4.98(d,J=0.9Hz,2H),4.72(s,2H),4.15 (t,J=7.1Hz,2H),4.10(t,J=6.9Hz,1H),3.83(s,2H),3.76(q,J=7.0Hz,2H),2.01(p,J=7.1Hz,2H). 13 C NMR (100MHz, CDCl3) δ7.27(s,1H),7.05(t,J=1.0Hz,1H),4.98(d,J=0.9Hz,2H),4.72(s,2H),4.15(t,J=7.1Hz, 2H),4.10(t,J=6.9Hz,1H),3.83(s,2H),3.76(q,J=7.0Hz,2H),2.01(p,J=7.1Hz,2H).LRMS(ESI,m / z):253[M+H] + .

[0157] The intermediate 6-{[3-(benzyloxy)propyl]-7-methoxy-3,4-dihydro-1H-isochroman-4-one was obtained by means of intermediate 6-[(3-hydroxypropyl)oxy]-7-methoxy-3,4-dihydro-1H-isochroman-4-one, benzyl bromide, and potassium carbonate according to the method of Example 1.

[0158] 1 H NMR (400MHz, CDCl3) δ7.37–7.31(m,1H),7.31–7.23(m,2H),7.07(t,J=1.0Hz,0H),4.98(d,J=1.1Hz,1H) ,4.42(t,J=1.0Hz,1H),4.20(t,J=7.1Hz,1H),3.83(s,1H),3.58(t,J=7.1Hz,1H),2.03(p,J=7.2Hz,1H). 13C NMR (100MHz, CDCl3) δ196.18,152.19,148.73,138.02,136.14,128.34,128.01,127.92,127.5 2,111.56,109.78,72.96,71.60,70.20,67.73,67.68,56.11,29.69.LRMS(ESI,m / z):343[M+H] + .

[0159] The intermediate 6-{[3-(benzyloxy)propyl]oxy}-7-methoxy-3,4-dihydro-1H-isochroman-4-one was prepared by mixing intermediate 6-{[3-(benzyloxy)propyl]oxy}-3-[(Z)-1H-indol-2-methylmethyl alkyl]-7-methoxy-3,4-dihydro-1H-isochroman-4-one with 10% KOH and 1H-indol-2-carboxaldehyde according to the method of Example 1.

[0160] 1 H NMR (400MHz, CDCl3) δ7.40–7.25(m,10H),7.29–7.20(m,1H),7.11(d,J=1.0Hz,1H),6.89(d,J=1.3Hz,1H),5.31(d,J=1. 0Hz,2H),4.42(t,J=1.0Hz,2H),4.16(t,J=7.1Hz,2H),3.85–3.80(m,4H),3.57(t,J=7.1Hz,2H),2.04(p,J=7.1Hz,2H). 13 C NMR (100MHz, CDCl3) δ7.40–7.25(m,10H),7.29–7.20(m,1H),7.11(d,J=1.0Hz,1H),6.89(d,J=1.3Hz,1H),5.31(d,J=1.0Hz,2H),4.4 2(t,J=1.0Hz,2H),4.16(t,J=7.1Hz,2H),3.85–3.80(m,4H),3.57(t,J=7.1Hz,2H),2.04(p,J=7.1Hz,2H).LRMS(ESI,m / z):469[M+H] + .

[0161] Compound B2 was synthesized from the intermediate 6-{[3-(benzyloxy)propyl]oxy}-3-[(Z)-1H-indol-2-ylmethylylidene]-7-methoxy-3,4-dihydro-1H-isochroman-4-one according to the synthetic scheme of compound A1.

[0162] 1H NMR (400MHz, CDCl3) δ9.78 (s, 1H), 7.56 (dt, J = 7.3, 1.6Hz, 1H), 7.43–7.35 ( m,2H),7.24(td,J=7.5,1.6Hz,1H),7.16(td,J=7.5,1.5Hz,1H),7.08(t,J=1 .0Hz,1H),6.84(s,1H),6.58–6.55(m,1H),5.29(d,J=1.1Hz,2H),4.17(dt,J =9.2,7.0Hz,3H),3.83(s,2H),3.76(q,J=7.1Hz,2H),2.03(p,J=7.1Hz,2H). 13 CNMR (100MHz, CDCl3) δ179.87,151.27,149.11,142.28,137.56,136.77,131.56,128.14,125.56,122.34,120. 77,120.61,112.64,111.69,109.29,108.85,103.87,71.89,67.44,59.71,56.17,32.24.HRMS(ESI)m / z:calcd for C 22 H 21 NO5[M+H] + 380.1420, found 380.1408.

[0163] Example 17: Synthesis of (Z)-3-((1H-indol-2-yl)methylene)-6-((4-fluorobenzyl)oxo)-7-methoxyisocyano-4-one (B3)

[0164] Intermediate b1 and 4-(bromomethyl)-1-fluorobenzene were synthesized using the same method as compound B1 to obtain target compound B3.

[0165] 1 H NMR(400MHz, CDCl3)δ9.81(s,1H),7.56(dt,J=7.5,1.6Hz,1H),7.51(ddt,J=7.3,5.0,1.0Hz,2H),7.42–7.36(m,2H),7.25(td,J=7.5,1.6Hz,1H ),7.16(td,J=7.6,1.3Hz,3H),7.10(t,J=1.0Hz,1H),6.84(s,1H),6.58 –6.55(m,1H),5.28(d,J=1.0Hz,2H),5.11(t,J=1.0Hz,2H),3.83(s,2H). 13C NMR (100MHz, CDCl3) δ179.81,150.82,148.46,142.18,137.56,137.02,131.09,129.92,129.86,128.39,125.58,122.7 0,121.02,120.61,115.59,115.43,112.73,111.88,109.53,108.49,104.44,71.91,71.39,56.16.HRMS(ESI)m / z:calcd for C 26 H 20 FNO4[M+H] + 430.1376, found 430.1379.

[0166] Example 18: Synthesis of (Z)-3-((1H-indol-2-yl)methylene)-7-methoxy-6-(3-morpholinopropoxy)isochroman-4-one (B4)

[0167] Intermediate B1 and 4-(3-chloropropyl)-1,4-oxazacyclohexane were synthesized using the same method as compound B1 to obtain the target compound B4.

[0168] 1 H NMR(400MHz, CDCl3)δ9.80(s,1H),7.56(dt,J=7.7,1.6Hz,1H),7.43–7.35(m,2H),7.25(td,J=7.5,1.6Hz,1H),7.16(td,J=7.5,1.5Hz,1H),7.09 (t,J=1.0Hz,1H),6.84(s,1H),6.58–6.55(m,1H),5.28(d,J=0.9Hz,2H) ,4.71(s,2H),3.84(s,2H),3.71(t,J=7.1Hz,4H),2.83(t,J=7.1Hz,4H). 13 C NMR (100MHz, CDCl3) δ179.72,149.70,149.20,142.18,137.56,137.03,132.59,128.89,125.58,122.65,120. 57,120.52,115.12,111.85,109.52,109.09,104.44,88.21,71.91,66.36,56.16,50.85.HRMS(ESI)m / z:calcd for C 24 H 24 N₂O₅[M+H] +421.1685, found 421.1675.

[0169] Example 19: Synthesis of (Z)-3-((1H-indol-2-yl)methylene)-6-hydroxyisochroman-4-one (B5)

[0170] 4-hydroxybenzaldehyde was synthesized according to the method of Example 1 to obtain intermediate c9, and c9 was synthesized according to the method of compound A1 to obtain compound B5.

[0171] 1 H NMR (400MHz, CDCl3) δ9.67(s,1H),9.28(s,1H),7.56(dt,J=7.4,1.6Hz,1H),7.38(dd,J=7.5,1.5Hz,1H),7.28(dt,J=7.4,1.0Hz,1H) ,7.25–7.18(m,2H),7.15(td,J=7.5,1.6Hz,1H),6.87(s,1H),6.82(dd,J=7.4,1.6Hz,1H),6.58–6.55(m,1H),5.28(d,J=1.1Hz,2H). 13 C NMR (100MHz, CDCl3) δ180.26,157.73,142.30,137.59,136.76,131.57,128.65,128.26,125 .62,122.61,120.71,119.38,112.92,111.93,109.11,104.33,71.61.HRMS(ESI)m / z:calcd for C 18 H 13 NO3[M+H]+292.0895, found 292.0889.

[0172] Example 20: Synthesis of (Z)-3-((1H-indol-2-yl)methylylidene)-6,7-dimethoxy-isochroman-4-one (C1)

[0173] The target compound C1 was synthesized from 2-bromo-4,5-dimethoxybenzene-1-carboxaldehyde according to the method in Example 1.

[0174] 1H NMR (400MHz, CDCl3) δ7.56 (dt, J=7.5, 1.6Hz, 1H), 7.44–7.36 (m, 2H), 7.23 (td, J=7.5, 1.5Hz, 1H), 7.15 (td, J=7.5, 1.5Hz,1H),7.08(t,J=1.0Hz,1H),6.84(s,1H),6.58–6.55(m,1H),5.29(d,J=0.9Hz,2H),3.91(s,2H),3.83(s,2H). 13 C NMR (100MHz, CDCl3) δ179.54,153.04,150.26,142.27,137.55,136.86,129.85,127.88,125.56,122.3 9,120.54,120.47,111.69,111.19,109.20,108.60,103.87,71.66,56.38,55.96.HRMS(ESI)m / z:calcd for C 20 H 17 NO4[M+H] + 336.1158, found 336.1147.

[0175] Example 21: Synthesis of (Z)-3-((1H-indol-2-yl)methylylidene)-6,7-dimethoxy-isochroman-4-one (C2)

[0176] The target compound C2 was synthesized from 2-bromo-4,5-dimethoxybenzene-1-carboxaldehyde and 5-methoxy-1H-indole-2-carboxaldehyde according to the method in Example 1.

[0177] 1 H NMR (400MHz, CDCl3) δ9.81(s,1H),7.41(s,1H),7.25(d,J=7.5Hz,1H),7.20(t,J=1.6Hz,1H),7.08(t,J=1.0Hz,1H),6.8 1(dd,J=7.5,1.6Hz,1H),6.66(s,1H),6.48(d,J=1.5Hz,1H),5.29(d,J=0.9Hz,2H),3.91(s,2H),3.85(d,J=16.1Hz,5H). 13C NMR (100MHz, CDCl3) δ179.63,155.48,153.00,150.26,142.28,137.86,134.90,129.90,127.90,127.82,1 13.25,112.52,111.16,109.82,108.95,104.64,103.74,71.89,56.38,56.01,55.27.HRMS(ESI)m / z:calcd for C 21 H 19 NO5[M+H] + 366.1263, found 366.1254.

[0178] Example 22: Synthesis of (Z)-6,7-dimethoxy-3-((6-methoxy-1H-indol-2-yl)methyl methyl alkyl)-isochroman-4-one (C3)

[0179] The target compound C3 was synthesized from 2-bromo-4,5-dimethoxybenzene-1-carboxaldehyde and 6-methoxy-1H-indole-2-carboxaldehyde according to the method in Example 1.

[0180] 1 H NMR (400MHz, CDCl3) δ9.80 (s, 1H), 7.83 (dd, J = 7.4, 1.6Hz, 1H), 7.41 (s, 1H), 7.08 (t, J = 1.0Hz, 1H), 6. 91–6.83(m,2H),6.66(s,1H),6.54(d,J=1.4Hz,1H),5.29(d,J=1.0Hz,2H),3.91(s,3H),3.83(s,5H). 13 C NMR (100MHz, CDCl3) δ179.63,157.67,153.00,150.26,142.28,138.27,137.24,129.90,127.82,123.21,1 21.36,111.16,110.12,109.82,108.78,103.74,95.60,71.89,56.38,56.01,55.37.HRMS(ESI)m / z:calcd for C 21 H 19 NO5[M+H] + 366.1263, found 366.1269.

[0181] Example 23: Synthesis of (Z)-6,7-dimethoxy-3-(5,6,7-trimethoxy-1H-indol-2-yl)methyl methyl alkylene]-isochroman-4-one (C4)

[0182] The target compound C4 was obtained by synthesizing 2-bromo-4,5-dimethoxybenzene-1-carboxaldehyde and 5,6,7-trimethoxy-1H-indole-2-carboxaldehyde according to Example 1.

[0183] 1 H NMR(400MHz, CDCl3)δ9.49(s,1H),7.39(s,1H),7.11–7.05(m,2H),6.90(s,1H),6.46( d,J=1.6Hz,1H),5.28(d,J=0.9Hz,2H),3.89(d,J=14.6Hz,8H),3.83(d,J=11.7Hz,6H). 13 C NMR (100MHz, CDCl3) δ179.81,152.88,149.10,148.91,142.14,139.14,138.62,137.12,129.76,127.82,126.39, 125.87,111.16,109.50,109.12,104.13,103.82,71.91,60.93,60.86,56.38,56.25,56.01.HRMS(ESI)m / z:calcd for C 23 H 23 NO7[M+H] + 426.1475, found 426.1461.

[0184] Example 24: Synthesis of (Z)-3-((1H-indol-2-yl)methylylidene)-7-hydroxy-6-methoxyisocyano-4-one (D1)

[0185] The target compound D1 was obtained by synthesizing 2-bromo-5-hydroxy-4-methoxybenzene-1-carboxaldehyde using the same method as compound A1.

[0186] 1H NMR (400MHz, CDCl3) δ8.26(s,1H),7.55(dt,J=7.3,1.6Hz,1H),7.39(dd,J=7.4,1.5Hz,1H),7.29(s,1H),7.22(td,J=7.5,1.6H z,1H),7.15(td,J=7.5,1.5Hz,1H),6.99(t,J=1.0Hz,1H),6.80(s,1H),6.58–6.55(m,1H),5.24(d,J=0.9Hz,2H),3.88(s,2H). 13 C NMR (100MHz, CDCl3) δ179.89,148.81,147.15,142.27,137.59,136.79,129.25,126.12,125.56,12 2.43,120.74,120.38,112.89,111.69,111.21,108.54,103.87,71.70,56.66.HRMS(ESI)m / z:calcd for C 19 H 15 NO4[M+H] + 322.1001, found 322.1014.

[0187] Example 25: Synthesis of (Z)-7-hydroxy-6-methoxy-3-((5-methoxy-1H-indol-2-yl)methylethylene)-isochroman-4-one (D2)

[0188] 2-Bromo-5-hydroxy-4-methoxybenzyl-1-carboxaldehyde and 5-methoxy-1H-indole-2-carboxaldehyde were synthesized into target compound D2 according to the synthetic scheme of compound A1.

[0189] 1 H NMR (400MHz, CDCl3) δ9.81 (s, 1H), 8.21 (s, 1H), 7.29 (s, 1H), 7.25 (d, J = 7.5Hz, 1H), 7.19 (t, J = 1.6Hz, 1H), 6.99 (t, J = 1. 0Hz, 1H), 6.81 (dd, J = 7.5, 1.5Hz, 1H), 6.66 (s, 1H), 6.48 (d, J = 1.5Hz, 1H), 5.24 (d, J = 0.9Hz, 2H), 3.87 (d, J = 17.4Hz, 5H). 13C NMR (100MHz, CDCl3) δ179.70,156.75,149.94,147.13,142.28,137.82,133.77,129.28,127.90,127.3 1,113.21,112.76,112.56,111.23,108.71,106.80,103.74,71.69,56.65,55.27.HRMS(ESI)m / z:calcd for C 20 H 17 NO5[M+H] + 352.1107, found 352.1118.

[0190] Example 26: Synthesis of (Z)-7-hydroxy-6-methoxy-3-((6-methoxy-1H-indol-2-yl)methylethylene)-isochroman-4-one (D3)

[0191] The target compound D3 was obtained by synthesizing 2-bromo-5-hydroxy-4-methoxybenzyl-1-carboxaldehyde and 6-methoxy-1H-indole-2-carboxaldehyde using the same method as compound A1.

[0192] 1 H NMR (400MHz, CDCl3) δ9.82(s,1H),8.21(s,1H),7.83(dd,J=7.4,1.6Hz,1H),7.29(s,1H),6.99(t,J=1.0Hz, 1H),6.91–6.83(m,2H),6.66(s,1H),6.54(d,J=1.4Hz,1H),5.24(d,J=0.9Hz,2H),3.89(s,3H),3.84(s,3H). 13 C NMR (100MHz, CDCl3) δ179.70,157.67,149.94,147.13,142.28,138.22,137.24,129.28,127.31,123.3 6,121.36,112.76,111.23,110.21,108.68,103.74,95.60,71.69,56.65,55.37.HRMS(ESI)m / z:calcd for C 20 H 17 NO5[M+H] + 352.1107, found 352.1009.

[0193] Example 27: Synthesis of (Z)-7-hydroxy-6-methoxy-3-((5,6,7-trimethoxy-1H-indol-2-yl)methylethylene)-isochroman-4-one (D4)

[0194] The target compound D4 was obtained by synthesizing 2-bromo-5-hydroxy-4-methoxybenzyl-1-carboxaldehyde and 5,6,7-trimethoxy-1H-indole-2-carboxaldehyde using the same method as compound A1.

[0195] 1 H NMR (400MHz, CDCl3) δ9.61(s,1H),8.52(s,1H),7.23(s,1H),7.07(d,J=1.4Hz,1H),7.01(t,J=1.1H z,1H),6.90(s,1H),6.46(d,J=1.6Hz,1H),5.23(d,J=0.9Hz,2H),3.92–3.86(m,10H),3.81(s,3H). 13 CNMR (100MHz, CDCl3) δ179.76,148.48,148.17,147.27,142.14,139.34,138.62,137.05,129.29,127.02,126. 55,125.87,112.52,111.24,109.12,104.13,103.82,71.74,60.93,60.86,56.66,56.25.HRMS(ESI)m / z:calcd for C 22 H 21 NO7[M+H] + 412.1318, found 412.1309.

[0196] Example 28: Synthesis of (Z)-7-hydroxy-3-((1-(hydroxymethyl)-1H-indol-2-yl)methylethylene)-6-methoxyisocyano-4-one (D5)

[0197] The target compound D5 was obtained by synthesizing 2-bromo-5-hydroxy-4-methoxybenzyl-1-carboxaldehyde and 1-(hydroxymethyl)indole-2-carboxaldehyde according to the same method used for compound A1.

[0198] 1H NMR(400MHz, CDCl3)δ8.21(s,1H),7.64(dt,J=8.0,1.8Hz,1H),7.57–7.52(m,1H),7.29–7.19(m,3H),6.99(t,J=1.0Hz,1 H), 6.95 (s, 1H), 6.86 (d, J = 1.7Hz, 1H), 6.12 (t, J = 5.3Hz, 1H), 5.56 (d, J = 5.2Hz, 2H), 5.24 (d, J = 0.9Hz, 2H), 3.89 (s, 2H). 13 C NMR (100MHz, CDCl3) δ179.85,149.94,148.01,147.13,138.50,136.86,129.28,128.51,127.24,124.7 4,122.15,121.31,112.76,111.23,110.61,110.42,105.44,72.07,69.46,56.65.HRMS(ESI)m / z:calcd for C 20 H 17 NO5[M+H] + 352.1107, found 352.1114.

[0199] Example 29: Synthesis of (Z)-7-hydroxy-6-methoxy-3-((1-methyl-1H-indol-2-yl)methylethylene)isochroman-4-one (D6)

[0200] The target compound D6 was obtained by synthesizing 2-bromo-5-hydroxy-4-methoxybenzyl-1-carboxaldehyde and 1-methylindole-2-carboxaldehyde using the same method as compound A1.

[0201] 1 H NMR (400MHz, CDCl3) δ8.07(s,1H),7.67(dt,J=7.6,1.7Hz,1H),7.39(dd,J=7.3,1.7Hz,1H),7.29–7. 19(m,3H),6.99(t,J=1.0Hz,1H),6.81–6.74(m,2H),5.24(d,J=0.9Hz,2H),3.89(s,3H),3.79(s,3H). 13C NMR (100MHz, CDCl3) δ179.77,148.68,147.99,147.11,139.52,137.52,129.25,127.69,127.28,123.2 4,121.19,121.19,112.80,111.21,109.88,107.91,104.86,71.84,56.65,31.70.HRMS(ESI)m / z:calcd for C 20 H 17 NO4[M+H] + 336.1158, found 336.1142.

[0202] Example 30: Synthesis of (E)-3-((1H-indol-2-yl)methylene)-7-hydroxy-6-methoxyisocyano-4-one (D7)

[0203] The target compound D7 can be obtained by following the synthetic scheme of compound D1.

[0204] 1 H NMR (400MHz, CDCl3) δ8.26(s,1H),7.55(dt,J=7.1,1.6Hz,1H),7.39(dd,J=7.4,1.5Hz,1H),7.29(s,1H),7.22(td,J=7.5,1.6H z,1H),7.15(td,J=7.5,1.5Hz,1H),6.99(t,J=1.0Hz,1H),6.58–6.55(m,1H),6.17(s,1H),5.24(d,J=0.9Hz,2H),3.88(s,2H). 13 C NMR (100MHz, CDCl3) δ180.36,148.81,147.15,142.27,136.83,136.79,129.25,126.12,125.56,12 2.43,120.74,120.38,112.89,111.69,111.21,108.54,104.47,71.70,56.66.HRMS(ESI)m / z:calcd for C 20 H 17 NO4[M+H] + 336.1158, found 336.1151.

[0205] Example 31: Synthesis of (Z)-3-((1H-indol-2-yl)methylene)-6,7-dihydroxyisochroman-4-one (E1)

[0206] (1) Synthesis of intermediate 4,5-bis(benzyloxy)benzaldehyde (e2)

[0207] 4,5-Dihydroxybenzyl-1-carboxaldehyde (20.00 g, 144.80 mmol) was dissolved in acetone, and potassium carbonate (40.02 g, 289.60 mmol) and benzyl bromide (54.49 g, 318.56 mmol) were slowly added. After the addition was complete, the mixture was stirred at 60 °C for 3 h. After the reaction was completed as monitored by TLC, the reaction solution was filtered, and the filtrate was collected and concentrated under reduced pressure to give compound e2 (35.42 g, 111.25 mmol), with a yield of 69.54%.

[0208] 1 H NMR (400MHz, CDCl3) δ9.85 (s, 1H), 7.48–7.39 (m, 6H), 7.39–7.27 (m, 6H), 7.08–7.03 (m, 1H), 5.13 (dt, J = 11.5, 1.0Hz, 4H). 13 C NMR (100MHz, CDCl3) δ190.94,150.70,148.52,136.87,136.46,130.85,128.50,128.48,128.46,128. 13,128.11,127.75,127.67,127.58,126.27,114.44,112.27,71.06,71.03.LRMS(ESI,m / z):319[M+H] + .

[0209] (2) Synthesis of intermediate 4,5-bis(benzyloxy)-2-bromobenzene-1-carboxaldehyde (e3)

[0210] 4,5-bis(benzyloxy)-2-bromobenzene-1-carboxaldehyde (35.42 g, 111.25 mmol) was dissolved in methanol, and solid pyridine tribromide (71.16 g, 222.51 mmol) was slowly added. The mixture was stirred at room temperature for 3 h. After the reaction was completed as monitored by TLC, the reaction solution was evaporated to a slurry state, solidified, and the product was washed with toluene. The filtrate was collected and concentrated under reduced pressure to give compound e3 (28.62 g, 72.04 mmol), with a yield of 64.75%.

[0211] 1 H NMR (400MHz, CDCl3) δ7.47–7.39(m,4H),7.39–7.26(m,6H),5.13(dt,J=5.1,1.0Hz,3H). 13C NMR (100MHz, CDCl3) δ190.83,151.43,147.91,136.68,136.67,130.46,128.51,128.50,128.48,128. 13,128.11,127.70,127.62,127.53,121.14,119.03,113.12,71.21,71.21.LRMS(ESI,m / z):397[M+H] + .

[0212] (3) Synthesis of intermediate [4-(benzyloxy)-2-bromo-5-methoxyphenyl]methanol (e4)

[0213] 4,5-bis(benzyloxy)-2-bromobenzene-1-carboxaldehyde (28.62 g, 72.04 mmol) was dissolved in methanol, and NaBH4 (5.45 g, 144.08 mmol) was slowly added under ice bath conditions, with stirring for 1 h. After the reaction was complete as monitored by TLC, the reaction solution was evaporated to dryness, the reaction was quenched with ammonium chloride, and the mixture was extracted with ethyl acetate and water. The solution was concentrated under reduced pressure to give compound e4 (26.90 g, 54.46 mmol), in 93.52% yield.

[0214] 1 H NMR(400MHz, CDCl3)δ7.42(dtd,J=8.1,1.9,1.0Hz,4H),7.39–7.32(m,4H),7.34–7.26(m,2H),7.14(s,1H ), 6.95 (t, J = 1.0Hz, 1H), 5.13 (dt, J = 3.5, 1.0Hz, 4H), 4.70 (dd, J = 6.3, 1.0Hz, 2H), 4.05 (t, J = 6.2Hz, 1H). 13 C NMR (100MHz, CDCl3) δ148.16,148.09,136.70,136.69,134.82,128.51,128.50,128.48,128.13,128. 11,127.70,127.62,127.53,118.50,116.56,113.68,71.21,71.17,62.94.LRMS(ESI,m / z):399[M+H] + .

[0215] (4) Synthesis of intermediate ({[4,5-bis(benzyloxy)-2-bromophenyl]methyl}oxy)acetic acid-2-methylpropyl-2-yl ester (e5)

[0216] [4-(benzyloxy)-2-bromo-5-methoxyphenyl]methanol (26.90 g, 54.46 mmol) was dissolved in toluene, and tert-butyl bromoacetate (19.71 g, 101.06 mmol), tetrabutylammonium bromide (0.65 g, 2.02 mmol), and 50% KOH (37.80 g, 336.85 mmol) were added. The reaction was carried out in an oil bath at 60 °C for 4 h. After the reaction was completed, the mixture was extracted with ethyl acetate and water by TLC. The compound e5 (32.84 g, 63.96 mmol) was obtained by concentration under reduced pressure, with a yield of 94.94%.

[0217] 1 H NMR(400MHz, CDCl3) δ7.42(ddt,J=7.1,1.8,1.0Hz,2H),7.42–7.32(m,2H),7.34–7.27(m,1H),6. 94(t,J=1.0Hz,0H),5.13(dt,J=4.9,1.0Hz,2H),4.64(d,J=1.1Hz,1H),4.15(s,1H),1.41(s,3H). 13 C NMR (100MHz, CDCl3) δ170.10,148.22,147.97,136.87,136.86,133.40,128.49,128.47,128.45,128.23,128.20,12 7.75,127.68,127.63,118.46,116.63,114.05,82.12,71.31,71.23,70.68,67.06,27.80.LRMS(ESI,m / z):513[M+H] + .

[0218] (5) Synthesis of intermediate ({[4,5-bis(benzyloxy)-2-bromophenyl]methyl}oxy)acetic acid (a6)

[0219] ({[4,5-bis(benzyloxy)-2-bromophenyl]methyl}oxy)acetic acid-2-methylpropyl-2-yl ester (32.84 g, 63.96 mmol) was dissolved in methanol, and a methanol solution of sodium methoxide (6.91 g, 127.92 mmol) was slowly added. A suitable amount of water was then added, and the reaction was carried out at room temperature for 5 min. After the reaction was complete as monitored by TLC, the methanol was evaporated, and a small amount of EA and a larger amount of water were added for extraction once. While stirring, concentrated hydrochloric acid was slowly added to the aqueous layer obtained from the extraction to adjust the pH to 2. EA was added for extraction three times, followed by washing with saturated NaCl, drying over anhydrous sodium sulfate, and concentration to obtain compound e6 (27.64 g, 60.44 mmol), with a yield of 94.49%.

[0220] 1H NMR(400MHz, CDCl3)δ7.42(ddq,J=7.9,1.8,0.9Hz,4H),7.39–7.32(m,4H),7.34–7.27(m,2H),7. 12(s,1H),6.95(t,J=1.0Hz,1H),5.12(dt,J=6.0,0.9Hz,4H),4.68(d,J=1.0Hz,2H),4.08(s,2H). 13 C NMR (100MHz, CDCl3) δ173.66,148.13,148.07,136.72,136.70,133.35,128.53,128.51,128.49,128.32,128 .29,127.72,127.64,127.56,118.58,116.87,114.04,71.30,71.21,70.91,66.95.LRMS(ESI,m / z):457[M+H] + .

[0221] (6) Synthesis of intermediate 2-({[4,5-bis(benzyloxy)-2-bromophenyl]methyl}oxy)-N-methoxy-N-methylacetamide (e7)

[0222] ({[4,5-bis(benzyloxy)-2-bromophenyl]methyl}oxy)acetic acid (27.64 g, 60.44 mmol) was dissolved in dichloromethane, and O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethylurea hexafluorophosphate (29.88 g, 78.57 mmol) and N,N-diisopropylethylamine (11.72 g, 90.66 mmol) were slowly added. The reaction was allowed to proceed for 12 h. After the reaction was completed as monitored by TLC, dichloromethane and a large amount of water were added, and the mixture was extracted three times. The extract was washed with saturated NaCl, dried over anhydrous sodium sulfate, and concentrated to give compound e7 (18.13 g, 45.98 mmol), with a yield of 76.08%.

[0223] 1 H NMR(400MHz, CDCl3)δ7.42(ddt,J=7.7,1.9,0.9Hz,5H),7.39–7.27(m,8H),7.11(s,1H),6.94(t,J=1 .1Hz,1H),5.12(dt,J=4.6,1.0Hz,5H),4.64(d,J=1.0Hz,2H),4.14(s,2H),3.54(s,3H),3.03(s,3H). 13C NMR (100MHz, CDCl3) δ169.76,147.97,147.92,136.87,136.86,133.40,128.51,128.49,128.47,128.20,128.17,12 7.74,127.67,127.62,118.46,116.63,114.06,71.31,71.23,71.00,65.33,61.23,32.44.LRMS(ESI,m / z):500[M+H] + .

[0224] (7) Synthesis of intermediate 6,7-bis(benzyloxy)-3,4-dihydro-1H-isochroman-4-one (e8)

[0225] 2-({[4,5-bis(benzyloxy)-2-bromophenyl]methyl}oxy)-N-methoxy-N-methylacetamide (5 g, 11.86 mmol) was placed in a three-necked flask, anhydrous tetrahydrofuran was added, and under nitrogen protection, tert-butyllithium (1.52 g, 23.73 mmol) was slowly added at -78 °C. The reaction was carried out at this temperature for 20 min, and the reaction was quenched with saturated ammonium chloride solution. The mixture was extracted three times with ethyl acetate-water, washed with saturated NaCl, dried over anhydrous sodium sulfate, concentrated, and then subjected to column chromatography to give a white solid e8 (1.14 g, 3.16 mmol), in 26.66% yield.

[0226] 1 H NMR(400MHz, CDCl3)δ7.43(ddt,J=7.5,1.8,1.0Hz,4H),7.39–7.32(m,4H),7.34–7.27(m,3 H),7.02(t,J=1.0Hz,1H),5.13(dt,J=5.3,1.1Hz,4H),4.98(d,J=1.1Hz,2H),4.74(s,2H). 13 C NMR (100MHz, CDCl3) δ196.18,149.33,147.54,136.68,136.67,136.59,128.54,128.52,128.50,128.12,128 .10,127.75,127.67,127.59,127.57,111.75,110.68,71.83,71.26,71.21,69.78.LRMS(ESI,m / z):361[M+H] + .

[0227] (8) Synthesis of intermediate 6,7-bis(benzyloxy)-3-[(Z)-1H-indol-2-ylmethylylidene]-3,4-dihydro-1H-isochroman-4-one (e9)

[0228] 6,7-bis(benzyloxy)-3,4-dihydro-1H-isochroman-4-one (0.15 g, 416.19 μmol) was dissolved in tetrahydrofuran, 10% KOH was added, followed by 1H-indole-2-carboxaldehyde, and the reaction was continued until a solid precipitated. After filtration, a yellow solid e9 (0.09 g, 184.59 μmol) was obtained, with a yield of 44.35%.

[0229] 1 H NMR (400MHz, CDCl3) δ9.80 (s, 1H), 7.55 (dt, J = 7.2, 1.7Hz, 1H), 7.42 (ddd, J =4.8,2.6,1.0Hz,2H),7.41(s,1H),7.43–7.30(m,9H),7.33–7.28(m,1H),7. 30–7.21(m,1H),7.16(td,J=7.5,1.6Hz,1H),7.05(t,J=1.0Hz,1H),6.84(s, 1H), 6.58–6.55 (m, 1H), 5.29 (d, J=1.1Hz, 2H), 5.12 (dt, J=16.3, 1.0Hz, 4H). 13 C NMR (100MHz, CDCl3) δ180.23,149.22,148.10,142.19,137.64,136.86,136.75,131.44,128.90,128.46,128.44,128.42,128. 16,128.14,127.78,127.70,127.63,125.70,122.53,121.30,120.87,112.82,111.59,110.20,109.94,105.01,71.91,71.29.

[0230] (9) Synthesis of (Z)-3-((1H-indol-2-yl)methylene)-6,7-dihydroxyisochroman-4-one (E1)

[0231] 0.09 g (184.59 μmol) of 6,7-bis(benzyloxy)-3-[(Z)-1H-indol-2-ylmethylylidene]-3,4-dihydro-1H-isochroman-4-one was placed in a three-necked flask and dissolved in anhydrous dichloromethane. Under nitrogen protection, boron trichloride was slowly added at -78 °C. After the reaction was completed by TLC monitoring, water was added to quench the reaction. The mixture was extracted three times with dichloromethane-water, washed with saturated NaCl, dried over anhydrous sodium sulfate, concentrated, and then subjected to column chromatography to give compound E1 (0.032 g, 104.13 mmol), in a yield of 56.41%.

[0232] 1 H NMR (400MHz, CDCl3) δ8.64(s,2H),7.56(dt,J=7.3,1.6Hz,2H),7.39(dd,J=7.2,1.6Hz,2H),7.27(s,2H),7.22(td,J=7. 5,1.7Hz,2H),7.15(td,J=7.4,1.5Hz,2H),6.94(t,J=1.0Hz,2H),6.80(s,2H),6.58–6.55(m,2H),5.24(d,J=1.1Hz,4H). 13 C NMR (100MHz, CDCl3) δ180.17,147.61,145.27,142.25,137.59,136.76,128.74,126.54,125.56 ,122.41,121.28,120.38,113.37,112.91,111.93,109.11,103.87,71.44.HRMS(ESI)m / z:calcd for C 18 H 13 NO4[M+H] + 308.0845, found 308.0836.

[0233] Example 32: Synthesis of (Z)-3-((5-fluoro-1H-indol-2-yl)methylene)-6,7-dihydroxyisochroman-4-one (E2)

[0234] Compound E2 was obtained by synthesizing intermediate e8,5-fluoroindole-2-carboxaldehyde using the same method as compound E1.

[0235] 1H NMR (400MHz, CDCl3) δ8.64(s,2H),7.56(dt,J=7.3,1.6Hz,2H),7.39(dd,J=7.2,1.6Hz,2H),7.27(s,2H),7.22(td,J=7. 5,1.7Hz,2H),7.15(td,J=7.4,1.5Hz,2H),6.94(t,J=1.0Hz,2H),6.80(s,2H),6.58–6.55(m,2H),5.24(d,J=1.1Hz,4H). 13 C NMR (100MHz, CDCl3) δ180.17,147.61,145.27,142.25,137.59,136.76,128.74,126.54,125.56 ,122.41,121.28,120.38,113.37,112.91,111.93,109.11,103.87,71.44.HRMS(ESI)m / z:calcd for C 18 H 12 FNO4[M+H] + 326.0750, found 326.0741.

[0236] Example 33: Synthesis of (Z)-3-((6-fluoro-1H-indol-2-yl)methylene)-6,7-dihydroxyisochroman-4-one (E3)

[0237] The intermediate e8,6-fluoroindole-2-carboxaldehyde was synthesized using the same method as compound E1 to obtain compound E3.

[0238] 1 H NMR(400MHz, CDCl3)δ8.67(s,2H),7.90–7.84(m,2H),7.27(d,J=1.4Hz,1H),7.25(s,3H),7.17(td ,J=7.7,1.5Hz,2H),6.92(t,J=1.0Hz,2H),6.66(s,2H),6.57–6.54(m,2H),5.24(d,J=1.1Hz,4H). 13C NMR (100MHz, CDCl3) δ179.94,161.02,159.00,147.61,145.28,142.31,139.32,139.26,138.50,128.46,125.81,123.9 2,123.90,123.00,122.93,113.10,112.91,112.65,112.49,108.91,103.74,97.37,97.21,71.65.HRMS(ESI)m / z:calcd for C 18 H 12 FNO4[M+H] + 326.0750, found 326.0762.

[0239] Example 34: Synthesis of (Z)-3-((7-fluoro-1H-indol-2-yl)methylene)-6,7-dihydroxyisochroman-4-one (E4)

[0240] The intermediate e8,7-fluoroindole-2-carboxaldehyde was synthesized using the same method as compound E1 to obtain compound E4.

[0241] 1 H NMR (400MHz, CDCl3) δ8.67(s,1H),7.62(dt,J=7.4,1.5Hz,1H),7.41(td,J=7.5,5.0Hz,1H),7.25(s,1H),7. 01(td,J=7.7,1.6Hz,1H),6.92(t,J=1.0Hz,1H),6.85(s,1H),6.64(d,J=1.4Hz,1H),5.24(d,J=1.1Hz,2H). 13 C NMR (100MHz, CDCl3) δ179.94,152.68,150.66,147.61,145.28,142.25,137.06,136.97,128.46,128.34,128.18,127.37,127.30 ,125.81,123.06,123.00,117.74,117.72,113.10,112.91,111.56,111.40,109.05,109.03,103.87,71.65.HRMS(ESI)m / z:calcd for C 18 H 12 FNO4[M+H] + 326.0750, found 326.0756.

[0242] Example 35: Synthesis of (Z)-3-((5-bromo-1H-indol-2-yl)methylene)-6,7-dihydroxyisochroman-4-one (E5)

[0243] The intermediate e8, 5-bromoindole-2-carboxaldehyde was synthesized using the same method as compound E1 to obtain compound E5.

[0244] 1 H NMR(400MHz, CDCl3)δ9.71(s,1H),8.71(s,1H),7.93–7.89(m,1H),7.29(d,J=7.5Hz,1H),7.25(s,1H),7.2 1(dd,J=7.5,1.5Hz,1H),6.94(t,J=1.0Hz,1H),6.66(s,1H),6.60(d,J=1.8Hz,1H),5.24(d,J=1.1Hz,2H). 13 C NMR (100MHz, CDCl3) δ180.25,147.69,145.28,142.31,137.82,136.87,128.51,128.41,127.14 ,126.97,123.25,115.63,113.97,113.03,113.02,108.91,103.52,71.66.HRMS(ESI)m / z:calcd for C 18 H 12 BrNO4[M+H] + 385.9950, found 385.9943.

[0245] Example 36: Synthesis of (Z)-3-((6-bromo-1H-indol-2-yl)methylene)-6,7-dihydroxyisochroman-4-one (E6)

[0246] The intermediate e8, 6-bromoindole-2-carboxaldehyde was synthesized using the same method as compound E1 to obtain compound E6.

[0247] 1 H NMR(400MHz, CDCl3)δ8.71(s,1H),7.88(dd,J=7.5,1.6Hz,1H),7.72–7.68(m,1H),7.55(dd,J=7.5,1 .5Hz,1H),7.25(s,1H),6.94(d,J=1.1Hz,1H),6.66(s,1H),6.62–6.58(m,1H),5.24(d,J=1.1Hz,2H). 13C NMR (100MHz, CDCl3) δ180.25,147.69,145.28,142.31,138.22,137.37,128.51,126. 97,124.83,123.91,123.39,115.88,114.68,113.03,113.02,109.15,103.52,71.66.

[0248] HRMS(ESI)m / z:calcd for C 18 H 12 BrNO4[M+H] + 385.9950, found 385.9941.

[0249] Example 37: Synthesis of (Z)-3-((7-bromo-1H-indol-2-yl)methylene)-6,7-dihydroxyisochroman-4-one (E7)

[0250] Intermediate e8, 7-bromoindole-2-carboxaldehyde was synthesized using the same method as compound E1 to obtain compound E6.

[0251] 1 H NMR(400MHz, CDCl3)δ8.71(s,1H),7.72(dt,J=7.3,1.5Hz,1H),7.31(t,J=7.4Hz,1H),7.28–7 .23(m,2H),6.94(t,J=1.0Hz,1H),6.85(s,1H),6.72(d,J=1.4Hz,1H),5.24(d,J=1.1Hz,2H). 13 C NMR (100MHz, CDCl3) δ180.25,147.69,145.28,142.26,137.84,135.08,128.51,128.24,127.42 ,126.97,121.74,120.74,113.03,113.02,110.08,103.65,97.07,71.66.HRMS(ESI)m / z:calcd for C 18 H 12 BrNO4[M+H] + 385.9950, found 385.9961.

[0252] Example 38: Synthesis of (Z)-3-((5-methoxy-1H-indol-2-yl)methylene)-6,7-dihydroxyisochroman-4-one (E8)

[0253] Compound E8 was obtained by synthesizing intermediate e8, 5-methoxy-1H-indole-2-carboxaldehyde, using the same method as compound E1.

[0254] 1 H NMR (400MHz, CDCl3) δ9.82(s,1H),8.63(s,1H),7.27–7.22(m,2H),7.17(t,J=1.6Hz,1H),6.92(t,J=1.0Hz ,1H),6.81(dd,J=7.5,1.5Hz,1H),6.66(s,1H),6.48(d,J=1.5Hz,1H),5.24(d,J=1.1Hz,2H),3.85(s,3H). 13 C NMR (100MHz, CDCl3) δ179.62,156.75,147.50,145.26,142.31,137.85,134.10,128.46,127.88,12 6.57,113.11,113.10,113.06,112.57,108.22,104.92,103.74,71.45,55.28.HRMS(ESI)m / z:calcd for C 19 H 15 NO5[M+H] + 338.0950, found 338.0942.

[0255] Example 39: Synthesis of (Z)-3-((6-methoxy-1H-indol-2-yl)methylene)-6,7-dihydroxyisochroman-4-one (E9)

[0256] Intermediate e8, 6-methoxy-1H-indole-2-carboxaldehyde was synthesized using the same method as compound E1 to obtain compound E9.

[0257] 1 H NMR (400MHz, CDCl3) δ9.82(s,1H),8.63(s,1H),7.80(dd,J=7.6,1.5Hz,1H),7.25(s,1H),6.92(t,J=1 .0Hz,1H),6.90–6.83(m,2H),6.66(s,1H),6.54(d,J=1.4Hz,1H),5.24(d,J=1.1Hz,2H),3.84(s,3H). 13C NMR (100MHz, CDCl3) δ179.62,157.76,147.50,145.26,142.31,138.25,137.17,128.46,126.57,12 3.37,121.37,113.10,113.06,110.66,108.72,103.74,95.53,71.45,55.37.HRMS(ESI)m / z:calcd for C 19 H 15 NO5[M+H] + 338.0950, found 338.0946.

[0258] Example 40: Synthesis of (Z)-3-((5,6,7-trimethoxy-1H-indol-2-yl)methylene)-6,7-dihydroxyisochroman-4-one (E10)

[0259] Compound E10 was obtained by synthesizing the intermediate e8,5,6,7-trimethoxy-1H-indole-2-carboxaldehyde using the same method as compound E1.

[0260] 1 H NMR (400MHz, CDCl3) δ9.61 (s, 1H), 8.80 (s, 1H), 7.25 (s, 1H), 7.09 (d, J = 1.6Hz, 1H), 6.94–6. 88(m,2H),6.46(d,J=1.6Hz,1H),5.23(d,J=0.9Hz,2H),3.89(d,J=13.9Hz,6H),3.82(s,3H). 13 C NMR (100MHz, CDCl3) δ179.92,148.47,147.45,145.34,142.21,139.34,138.62,138.40,128.78,126.38,1 25.91,125.00,113.08,112.80,110.40,103.82,103.56,71.66,60.93,60.86,56.28.HRMS(ESI)m / z:calcd for C 21 H 19 NO7[M+H] + 397.1162, found 339.1169.

[0261] Example 41: Synthesis of (E)-3-((1H-indol-2-yl)methylene)-6,7-dihydroxyisochroman-4-one (E11)

[0262] Compound E11 was obtained using the same synthetic method as compound E1.

[0263] 1 H NMR (400MHz, CDCl3) δ8.64(s,1H),7.56(dt,J=7.3,1.6Hz,1H),7.39(dd,J=7.2,1.6Hz,1H),7.27(s,1H),7.22(td,J=7. 5,1.7Hz,1H),7.15(td,J=7.4,1.5Hz,1H),6.94(t,J=1.0Hz,1H),6.58–6.55(m,1H),6.17(s,1H),5.24(d,J=1.1Hz,2H). 13 C NMR (100MHz, CDCl3) δ180.54,147.61,145.27,142.25,136.83,136.76,128.74,126.54,125.56 ,122.41,121.28,120.38,113.37,112.91,111.93,109.11,104.47,71.44.HRMS(ESI)m / z:calcd for C 18 H 13 NO4[M+H] + 308.0845, found 308.0856.

[0264] Example 42: Evaluation of the inhibition rate of the compound against PCSK9

[0265] 1. Experimental Methods

[0266] HepG2 cells were seeded in 12-well plates and cultured in DMEM containing 10% FBS for 24 h. The old culture medium was then discarded, and the medium was replaced with fresh DMEM containing 10% FBS. The test compound (working concentration 10 μM) was added, and the cells were incubated for 48 h. The PCSK9 level in the supernatant was detected using a human PCSK9 ELISA kit according to the manufacturer's instructions. In short, a sample / standard and antibody cocktail was added to each well and incubated for 2 h. After washing three times with 1× wash buffer, the cells were incubated with TMB developing solution for 15 min. Stop solution was then added, and the OD value at 450 nm for each well was measured using a microplate reader. PCSK9 inhibition rate = (1 - PCSK9 content in the test sample group / PCSK9 content in the control group) × 100%.

[0267] 2. Experimental Results

[0268] Table 1. Inhibitory activity of compounds against PCSK9 (10 μM)

[0269] As shown in Table 1, compounds A1, A3, A5, A10, A12, B5, and E1 exhibited PCSK9 inhibition rates greater than 55%, demonstrating superior PCSK9 regulatory activity.

[0270] Example 43: Evaluation of the in vitro lipid-lowering activity of the compound

[0271] 1. Experimental Methods

[0272] HepG2 cells were seeded in 24-well plates and cultured in DMEM containing 10% FBS for 24 h. The old culture medium was then discarded, and the cells were replaced with fresh DMEM containing 2% defatted serum. The test compound (working concentration 10 μM, 3 parallel wells per sample) was added, and the cells were incubated for 24 h. The old culture medium was then discarded, and DMEM containing DiI-LDL (20 μg / ml) (400 μl / well) was added, and the cells were incubated for 3 h. The DMEM containing DiI-LDL was discarded, and the cells were carefully washed twice with PBS containing 0.4% BSA, followed by three washes with PBS to remove excess DiI-LDL. Trypsin was added for digestion, and the cells were resuspended. After centrifugation, the cells were resuspended in PBS. The mean fluorescence intensity was measured using a flow cytometer via the PE channel. LDL uptake rate = (test sample group - blank group) / (control group - blank group) × 100%.

[0273] 2. Experimental Results

[0274] Table 2. In vitro lipid-lowering activity of the compounds

[0275] As shown in Table 2, compounds A1, A12, B5, and E1 had an uptake rate of Dil-LDL greater than 190%, exhibiting good in vitro lipid-lowering activity and promoting the uptake of Dil-LDL by HepG2 cells.

[0276] Example 44: Pharmacokinetic Evaluation of the Compound

[0277] 1. Experimental Methods

[0278] Rats were randomly divided into two groups: one group received a single intravenous administration of 1.5 mg / kg, and the other group received a gavage administration of 30 mg / kg. Blood samples of approximately 0.2 mL were collected from the orbital cavity at 30 min, 1 h, 2 h, 3 h, 4 h, 6 h, 8 h, 10 h, and 24 h post-administration. Blood samples were collected using heparin sodium anticoagulant tubes, centrifuged at 3000 rpm for 10 min, and plasma was separated into labeled 1.5 mL EP tubes and stored at -20°C for analysis. After all samples were completely thawed, 90 μL was added to a 1.5 mL EP tube, followed by 10 μL of tolbutamide internal standard solution (5 ng / mL), and then 300 μL of acetonitrile solution. The tube was vortexed for 5 min, centrifuged at 13500 rpm for 5 min, and 200 μL was transferred to a vial for analysis using LC-MS / MS.

[0279] 2. Experimental Results

[0280] Table 3. Pharmacokinetic parameters of representative compounds (n=4)

[0281] As shown in Table 3, compounds A1, A12, B5, and E1 have good pharmacokinetic properties and good drug-likeness.

[0282] Example 45: Evaluation of the in vivo lipid-lowering activity of the compound

[0283] 1. Experimental Methods

[0284] ApoE - / - Mice (6-8 weeks old, male, 20g) were acclimatized for one week with a high-fat diet. After 8 weeks of high-fat feeding, blood was collected from the tail to measure serum levels of total cholesterol (CHOL), triglycerides (TG), and low-density lipoprotein (LDL-C). Mice were then divided into four groups based on their lipid levels: a model control group, an A1 gavage group, an A12 gavage group, and a B5 gavage group. All groups continued to be fed a high-fat diet. A compound was prepared using 5% DMSO + 20% Tween 80 + 65% saline and administered by gavage once daily (50 mg / kg) for four weeks. After four weeks, body weight was measured, blood was collected, and the animals were euthanized. Organs were collected and stored at -80°C.

[0285] 2. Experimental Results

[0286] Table 4. In vivo lipid-lowering activity studies of the compounds.

[0287] As shown in Table 4, compounds A1, A12, and B5 all exhibited good in vivo lipid-lowering activity in the hyperlipidemia model, significantly reducing the levels of total cholesterol (CHOL), triglycerides (TG), and low-density lipoprotein (LDL-C) in the serum of hyperlipidemic mice.

Claims

1. A ketone compound or its cis-trans isomer, or a pharmaceutically acceptable salt thereof, characterized in that it has the structure of Formula I. in: R1 and R2 are each independently selected from hydrogen, halogen, CN, NO2, NH2, OH, CF3, CH2OH, COOH, and -O[(CH2)] m O] r R5, substituted or unsubstituted, includes C1-C6 straight-chain or branched alkyl, C1-C6 straight-chain or branched alkoxy, C2-C6 straight-chain or branched alkenyl, C2-C6 straight-chain or branched alkynyl, 6-10 aryl, benzyl, 3-12 heterocyclic, 3-7 cycloalkyl, C2-C6 straight-chain or branched alkylphenyl, C2-C6 straight-chain or branched alkyl 5-7 heteroaryl, -OR5, -NR5R6, -SO2NR5R6, -CONR5R6, -COOR5, -OOCR5, -NHCOR5, -SO2R5, -OSO2R5, m selected from 1, 2, 3, 4, r selected from 1, 2, 3, 4; R3 is selected from one or more hydrogens, halogens, hydroxyl groups, substituted or unsubstituted straight-chain or branched C1-C6 alkyl groups, substituted or unsubstituted straight-chain or branched C1-C6 alkoxy groups, substituted or unsubstituted 3-12 membered cycloalkyl groups, substituted or unsubstituted 6-10 membered aryl groups, substituted or unsubstituted 3-12 membered heterocyclic groups, substituted or unsubstituted 3-7 membered cycloalkyl groups, cyano, nitro, carboxyl, mercapto, NR5R6, -CONR5R6, -SO2R5, -SO2NR5R6, -OSO2R5, -OCOR5; R4 is selected from hydrogen, amino group, hydroxyl group, substituted or unsubstituted straight-chain or branched C1-C6 alkyl group, substituted or unsubstituted C1-C6 straight-chain or branched alkoxy group, substituted or unsubstituted 6-10 aryl group, substituted or unsubstituted 5-7 heterocyclic group, substituted or unsubstituted C1-C6 straight-chain or branched alkylphenyl group, substituted or unsubstituted 3-12 cycloalkyl group, substituted or unsubstituted C2-C 10 Straight-chain or branched acyl groups, substituted or unsubstituted C2-C 10 Straight-chain or branched ester group, amino group, C1-C6 straight-chain or branched alkylamine group, substituted or unsubstituted C1-C6 straight-chain or branched amide group, -OSO2R5, -CH2OSO2R5, -OCOR5; R5 and R6 are each independently selected from hydrogen, substituted or unsubstituted 6-10 aryl groups, C1-C4 straight-chain or branched alkyl groups, C1-C4 straight-chain or branched alkenyl groups, C1-C4 straight-chain or branched alkynyl groups, substituted or unsubstituted 3-12 heterocyclic groups, substituted or unsubstituted 3-7 cycloalkyl groups, and C1-C6 straight-chain or branched hydroxyalkyl groups. The substituents are selected from one or more halogens, unsubstituted or halogenated or 3-6 membered cycloalkyl-substituted C1-C6 straight-chain or branched alkyl groups, C1-C4 straight-chain or branched alkoxy groups, 3-6 membered cycloalkyl groups, C1-C4 straight-chain or branched alkyl-substituted amino groups, hydroxyl groups, cyano groups, nitro groups, =O groups, hydroxyl-C1-C6 straight-chain or branched alkyl groups, carboxyl groups, mercapto groups, unsubstituted or 1-3 halogenated or hydroxyl-substituted 5-10 membered aryl groups, unsubstituted or halogenated 5-7 membered heterocyclic groups, unsubstituted or halogenated C2-C6 straight-chain or branched acyl groups, C1-C6 hydroxyalkyl groups, -NR5R6, -NHCOR5, -SO2R5, -OSO2R5, -SO2NR5R6, -COOR5, -OCOR5; The heterocyclic group contains 1-3 cyclic heteroatoms selected from oxygen, sulfur, and nitrogen; The heteroaryl group contains 1-4 cyclic heteroatoms selected from oxygen, sulfur, and nitrogen.

2. The isochromatic ketone compound or its cis-trans isomer, or a pharmaceutically acceptable salt, according to claim 1, characterized in that, In the aforementioned structure: R1 and R2 are each independently selected from hydrogen, halogen, CN, NH2, OH, CH2OH, COOH, and -O[(CH2)] m O] r R5, substituted or unsubstituted C1-C4 straight-chain or branched alkyl, substituted or unsubstituted C1-C4 straight-chain or branched alkoxy, substituted or unsubstituted benzyl, substituted or unsubstituted benzyloxy, -OR5, -NR5R6, -SO2NR5R6, -CONR5R6, -COOR5, -OOCR5, -NHCOR5, -SO2R5, -OSO2R5, m is selected from 1, 2, 3, r is selected from 1, 2; R5 and R6 are each independently selected from hydrogen, substituted or unsubstituted 6-10 aryl groups, C1-C4 straight-chain or branched alkyl groups, C1-C4 straight-chain or branched alkenyl groups, C1-C4 straight-chain or branched alkynyl groups, substituted or unsubstituted 3-12 heterocyclic groups, substituted or unsubstituted 3-7 cycloalkyl groups, and C1-C6 straight-chain or branched hydroxyalkyl groups. The substituents are selected from one or more halogens, unsubstituted or halogenated or 3-6 membered cycloalkyl-substituted C1-C6 straight-chain or branched alkyl groups, C1-C4 straight-chain or branched alkoxy groups, 3-6 membered cycloalkyl groups, C1-C4 straight-chain or branched alkyl-substituted amino groups, hydroxyl groups, cyano groups, nitro groups, =O groups, hydroxyl-C1-C6 straight-chain or branched alkyl groups, carboxyl groups, mercapto groups, unsubstituted or 1-3 halogenated or hydroxyl-substituted 5-10 membered aryl groups, unsubstituted or halogenated 5-7 membered heterocyclic groups, unsubstituted or halogenated C2-C6 straight-chain or branched acyl groups, C1-C6 hydroxyalkyl groups, -NR5R6, -NHCOR5, -SO2R5, -OSO2R5, -SO2NR5R6, -COOR5, -OCOR5; The heterocyclic group contains 1-3 oxygen, sulfur, and nitrogen ring heteroatoms.

3. The isochromatic ketone compound or its cis-trans isomer, or a pharmaceutically acceptable salt, according to claim 1, characterized in that... In the aforementioned structure: R3 is selected from one or two hydrogens, hydroxyl groups, halogens, amino groups, substituted or unsubstituted C1-C4 straight-chain or branched alkyl groups, substituted or unsubstituted C1-C4 straight-chain or branched alkoxy groups, substituted or unsubstituted 3-6 membered cycloalkyl groups, substituted or unsubstituted 6-10 membered aryl groups, substituted or unsubstituted 3-12 membered heterocyclic groups, substituted or unsubstituted 3-7 membered cycloalkyl groups, and -NR5R6. R5 and R6 are each independently selected from hydrogen, substituted or unsubstituted 6-10 aryl groups, C1-C4 straight-chain or branched alkyl groups, C1-C4 straight-chain or branched alkenyl groups, C1-C4 straight-chain or branched alkynyl groups, substituted or unsubstituted 3-12 heterocyclic groups, substituted or unsubstituted 3-7 cycloalkyl groups, and C1-C6 straight-chain or branched hydroxyalkyl groups. R4 is selected from C1-C3 straight-chain or branched alkyl, hydroxymethyl, acetyl, and allyl groups; The substituents are selected from one or more halogens, unsubstituted or halogenated or 3-6 membered cycloalkyl-substituted C1-C6 straight-chain or branched alkyl groups, C1-C4 straight-chain or branched alkoxy groups, 3-6 membered cycloalkyl groups, C1-C4 straight-chain or branched alkyl-substituted amino groups, hydroxyl groups, cyano groups, nitro groups, =O groups, hydroxyl-C1-C6 straight-chain or branched alkyl groups, carboxyl groups, mercapto groups, unsubstituted or 1-3 halogenated or hydroxyl-substituted 5-10 membered aryl groups, unsubstituted or halogenated 5-7 membered heterocyclic groups, unsubstituted or halogenated C2-C6 straight-chain or branched acyl groups, C1-C6 hydroxyalkyl groups, -NR5R6, -NHCOR5, -SO2R5, -OSO2R5, -SO2NR5R6, -COOR5, -OCOR5.

4. The isochromatic ketone compound or its cis-trans isomer, or a pharmaceutically acceptable salt, according to claim 1, characterized in that, In the aforementioned structure: R1 and R2 are each independently selected from hydrogen, F, Cl, Br, I, CN, NH2, OH, CH2OH, COOH, C1-C3 straight-chain or branched alkoxy, halogen-substituted C1-C3 straight-chain or branched alkoxy, substituted or unsubstituted benzyloxy, -CONH2, -SO2NH2, -COOCH2CH3, -NHCOCH2CH3; R3 is selected from one hydrogen atom, F, Cl, Br, I, CH3, CN, OCH3, CF3, -COCH2CH3, COOH, NH2, -NHCOCH2CH3; R4 is selected from hydrogen, CH3, -CH2OH, -COCH3, -SO2R5, -CH2OSO2R5, and -CH2OCOR5; R5 is selected from substituted or unsubstituted 6-10 aryl groups, C1-C3 straight-chain or branched alkyl groups; The substituents are selected from one or more halogens, unsubstituted or halogenated or 3-6 membered cycloalkyl-substituted C1-C6 straight-chain or branched alkyl groups, C1-C4 straight-chain or branched alkoxy groups, 3-6 membered cycloalkyl groups, C1-C4 straight-chain or branched alkyl-substituted amino groups, hydroxyl groups, cyano groups, nitro groups, =O groups, hydroxyl-C1-C6 straight-chain or branched alkyl groups, carboxyl groups, mercapto groups, unsubstituted or 1-3 halogenated or hydroxyl-substituted 5-10 membered aryl groups, unsubstituted or halogenated 5-7 membered heterocyclic groups, unsubstituted or halogenated C2-C6 straight-chain or branched acyl groups, C1-C6 hydroxyalkyl groups, -NR5R6, -NHCOR5, -SO2R5, -OSO2R5, -SO2NR5R6, -COOR5, -OCOR5; R5 and R6 are each independently selected from hydrogen, substituted or unsubstituted 6-10 aryl groups, C1-C4 straight-chain or branched alkyl groups, C1-C4 straight-chain or branched alkenyl groups, C1-C4 straight-chain or branched alkynyl groups, substituted or unsubstituted 3-12 heterocyclic groups, substituted or unsubstituted 3-7 cycloalkyl groups, and C1-C6 straight-chain or branched hydroxyalkyl groups.

5. The isochromatic ketone compound or its cis-trans isomer, or a pharmaceutically acceptable salt, according to claim 1, characterized in that, The compound is selected from any one of the following compounds:

6. A pharmaceutical composition, characterized in that, It comprises the isochromic ketone compound of claim 1 or its cis-trans isomer, a pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier.

7. The use of the isochorone compound of claim 1 or its cis-trans isomer, a pharmaceutically acceptable salt, or the pharmaceutical composition of claim 6 in the preparation of a medicament for the production of a preprotein convertase subtilisin 9 inhibitor.

8. The application according to claim 7, characterized in that, The aforementioned medication is for the prevention and / or treatment of hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, fatty liver disease, atherosclerosis, and obesity.

9. The application according to claim 8, characterized in that, The aforementioned drug is used to lower total cholesterol, low-density lipoprotein cholesterol, and triglycerides.

10. The application according to claim 8, characterized in that, The drug described is one that increases the expression of hepatic LDL receptors, inhibits PCSK9 expression, and activates AMPK protein kinase.

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