Two-sided needle base tetracyclic analogs-sulfonamide hybrids and their preparation and application

Abstract

This invention discloses a tetracyclic analogue-sulfonamide hybrid of *Zanthoxylum bungeanum* alkaloids, possessing the following general formula. The invention also provides a method for preparing the tetracyclic analogue-sulfonamide hybrid of *Zanthoxylum bungeanum* alkaloids, comprising dissolving compound 10 or compound 11 and triethylamine in dichloromethane, adding benzenesulfonyl chloride derivative 12a-12e at low temperature, stirring, and then stirring at room temperature until the starting material spot disappears. The reaction is monitored by TLC. After the reaction is complete, the product 13a-13e or 14a-14e is obtained by column chromatography separation and purification. The tetracyclic analogue-sulfonamide hybrid of *Zanthoxylum bungeanum* alkaloids provided by this invention has been shown in in vitro antitumor experiments to exhibit strong inhibitory effects on human cervical cancer cells in leukemia cells, human liver cancer cells, human lung cancer cells, human cervical cancer cells, and normal human liver cells.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical chemistry. More specifically, this invention relates to a tetracyclic analogue of ginseng, a sulfonamide hybrid, and its preparation and application. Background Technology

[0002] In recent years, with the increasing emphasis on "natural medicine therapy," the role of traditional Chinese medicine in prolonging the survival time and improving the quality of life of cancer patients has received more and more attention and recognition. Therefore, finding active ingredients from natural medicines that are significantly effective for humans, and modifying and transforming the structure of these active ingredients, has become one of the research hotspots in the field of cancer treatment in recent years.

[0003] The Chinese herb *Zanthoxylum nitidum* (Roxb.) DC., a plant in the Rutaceae family, is the dried root of the plant. It is mainly produced in Guangdong, Guangxi, and Fujian provinces. First recorded in the *Shennong Bencao Jing* (Shennong's Classic of Materia Medica), it is also known as *Manjiao*, *Liangbeizhen*, and *Rudi Jinniu*. *Zanthoxylum nitidum* has a bitter and pungent taste, is neutral in nature, and slightly toxic. It has the effects of regulating qi and relieving pain, promoting blood circulation and removing blood stasis, and dispelling wind and activating collaterals. *Zanthoxylum nitidum* contains various chemical components, mainly benzophenanthrene alkaloids, quinoline alkaloids, coumarins, lignans, organic acids, and others. Among these, alkaloids have advantages such as abundant sources, diverse structures, significant effects, and relatively few toxic side effects, making them a research hotspot in the field of natural products. Nitidine chloride (NC) is one of the important components of *Zanthoxylum nitidum* alkaloids. Compared to other chemical components of *Zanthoxylum nitidum*, NC has been studied more extensively, and its anticancer, anti-inflammatory, and antioxidant effects have been widely reported. Studies show that NC has a strong inhibitory effect on various tumor cells. Inhibition of Topo I activity may be one of its mechanisms of action. NC can insert into the DNA-Topo I complex via non-covalent insertion, causing irreversible DNA damage by blocking Topo I-mediated single-strand break repair, thereby affecting DNA replication in tumor cells and ultimately leading to apoptosis or necrosis, thus inhibiting tumor cell proliferation.

[0004] Sulfonamides contain certain nitrogen and sulfur atoms. These drugs were initially used for antibacterial therapy. With further research, it has been discovered that sulfonamides containing the classic structure of p-aminobenzenesulfonamide, after certain structural modifications, exhibit a variety of pharmacological activities, including antibacterial, anticancer, anti-HIV, anti-inflammatory, antiepileptic, and anticonvulsant effects, demonstrating their enormous pharmaceutical potential and development value. In recent years, increasing research has shown that introducing a sulfonamide group can effectively inhibit the proliferation of tumor cells. Therefore, introducing sulfonamide fragments into the tetracyclic analogue skeleton of thorn beetle alkaloid to screen for highly effective and low-toxicity antitumor compounds is of great significance for tumor treatment. Summary of the Invention

[0005] One object of the present invention is to solve at least the above-mentioned problems and to provide at least the advantages that will be described later.

[0006] One objective of this invention is to provide a tetracyclic analogue of succinate-sulfonamide hybrid, which has the advantages of novel structure and high efficacy and low toxicity in the treatment of tumors.

[0007] To achieve these objectives and other advantages of the present invention, a tetracyclic analogue-sulfonamide hybrid of thorn alkaloid is provided, having a structure of general formula I or general formula II.

[0008] The structural formula of general formula I is shown below:

[0009]

[0010] The structural formula of general formula II is shown below:

[0011]

[0012] Formula I includes compounds 13a-13e, and Formula II includes compounds 14a-14e;

[0013] In compound 13a, R1, R2, R3, and R4 are H atoms; in compound 14a, R1, R2, R3, and R4 are H atoms; in compound 13b, R1, R2, and R3 are H atoms, and R4 is Me; in compound 14b, R1, R2, and R3 are H atoms, and R4 is Me; in compound 13c, R1, R2, and R4 are Me, and R3 is H atom; in compound 14c, R1, R2, and R4 are Me, and R3 is H atom; in compound 13d, R1 and R2 are H atoms, and R3 and R4 are Cl atoms; in compound 14d, R1 and R2 are H atoms, and R3 and R4 are Cl atoms; in compound 13e, R1, R2, and R4 are H atoms, and R3 is F atom; in compound 14e, R1, R2, and R4 are H atoms, and R3 is F atom.

[0014] A method for preparing a tetracyclic analogue of thorn base-sulfonamide hybrid: Compound 10 or 11 and triethylamine are dissolved in dichloromethane, benzenesulfonyl chloride derivative 12a-12e is added at low temperature, the mixture is stirred and then stirred at room temperature until the starting material spot disappears, the reaction is monitored by TLC, and after the reaction is completed, the target products 13a-13e and / or 14a-14e are obtained by column chromatography separation and purification.

[0015] The structural formula of compound 10 is shown below:

[0016]

[0017] The structural formula of compound 11 is shown below:

[0018]

[0019] The general structural formulas of benzenesulfonyl chloride derivatives 12a-12e are shown below:

[0020]

[0021] In compound 12a, R1, R2, R3, and R4 are H atoms; in compound 12b, R1, R2, and R3 are H atoms, and R4 is Me; in compound 12c, R1, R2, and R4 are Me atoms, and R3 is a H atom; in compound 12d, R1 and R2 are H atoms, and R3 and R4 are Cl atoms; in compound 12e, R1, R2, and R4 are H atoms, and R3 is an F atom.

[0022] Preferably, under electromagnetic stirring, compound 8 or compound 9 is dissolved in an aqueous methylamine solution, stirred at room temperature, and the reaction is monitored by TLC. After the reaction is complete, dichloromethane is added for extraction, the organic phases are combined and washed with water, dried over anhydrous sodium sulfate overnight, and the filtrate is concentrated to obtain compound 10 or compound 11.

[0023] The structural formula of compound 8 is shown below:

[0024]

[0025] The structural formula of compound 9 is shown below:

[0026]

[0027] Preferably, compound 6 or compound 7 is added to a round-bottom flask, dissolved in acetonitrile, and phthalimide, potassium iodide and potassium carbonate are added to obtain a mixture. The mixture is refluxed and stirred under heating conditions, and the reaction is detected by TLC. After the reaction is completed, the mixture is separated and purified by column chromatography to obtain compound 8 or compound 9.

[0028] The structural formula of compound 6 is shown below:

[0029]

[0030] The structural formula of compound 7 is shown below:

[0031]

[0032] Preferably, under an inert gas protection, compound 4 or compound 5, palladium acetate, tri-o-methylphenylphosphine, and silver carbonate are added to a round-bottom flask, dissolved in ethyl acetate, and stirred under reflux under heating conditions. The reaction is monitored by TLC. After the reaction is completed, the brown insoluble matter is removed by hot filtration, the filtrate is collected, and purified by column chromatography to obtain compound 6 or compound 7.

[0033] The structural formula of compound 4 is as follows:

[0034]

[0035] The structural formula of compound 5 is shown below:

[0036]

[0037] Preferably, compound 3 is added to a round-bottom flask, dissolved in anhydrous DMF, sodium hydride is added, the mixture is stirred at low temperature, 1,4-dibromobutane or 1,6-dibromobutane is added, the mixture is stirred at room temperature, the reaction is monitored by TLC, after the reaction is completed, ice water is added to quench the reaction, ethyl acetate is extracted, the organic layers are combined, dried overnight with anhydrous sodium sulfate, the filtrate is concentrated after filtration, and the residue is separated and purified by column chromatography to obtain compound 4 or compound 5.

[0038] The structural formula of compound 3 is as follows:

[0039]

[0040] Preferably, naphthylamine is added to a round-bottom flask, dissolved in dichloromethane, followed by the addition of triethylamine and stirring. Then, o-iodobenzoyl is added, and the mixture is stirred at room temperature. The reaction is monitored by TLC. Once the reaction is complete, the mixture is filtered and dried to obtain compound 3.

[0041] An antitumor drug comprising the aforementioned tetracyclic analogue of ginseng alkaloid-sulfonamide hybrid.

[0042] Preferably, the sulfonamide heterozygote alkaloid tetracyclic analogue is formulated into an injection, tablet, pill, capsule, suspension or emulsion with pharmaceutically acceptable excipients.

[0043] Preferably, the excipients are one or more of the following: ethanol, propylene glycol, polyethylene glycol, diethylene glycol, triacetin, glycerol, dextrin, povidone, stearyl alcohol, stearic acid, microcrystalline cellulose, starch, lactose, mannitol, sodium bicarbonate, calcium carbonate, low-substituted hydroxypropyl methylcellulose, magnesium stearate, and talc.

[0044] An anticancer drug comprising the aforementioned tetracyclic analogue of succinate-sulfonamide hybrid.

[0045] Preferably, the cancer is cervical cancer or leukemia.

[0046] Application of sulfonamide heterozygote tetracyclic analogue in the preparation of drugs against human cervical cancer and leukemia.

[0047] The present invention has at least the following beneficial effects:

[0048] First, the present invention provides a tetracyclic analogue of thorn alkali-sulfonamide hybrid and its preparation method. The tetracyclic analogue of thorn alkali-sulfonamide hybrid has a novel structure and features high efficiency and low toxicity.

[0049] Secondly, the tetracyclic analogue of succinate-sulfonamide provided by this invention has excellent antitumor activity. Among them, compound 14d inhibited HL-60 leukemia cells by 69.64% at a concentration of 40 μmol / L; compounds 13a, 13e, and 14a-14e all inhibited human cervical cancer cells HeLa by more than 50%, indicating that compound 14d has a good inhibitory rate on leukemia cells and can be used to develop and prepare anti-leukemia drugs.

[0050] Third, in vitro cytotoxicity studies of the present invention show that compounds 13a, 13e, 14a-14e have a strong inhibitory effect on human cervical cancer cells and can be used to prepare anti-human cervical cancer drugs.

[0051] Other advantages, objectives and features of the present invention will become apparent in part from the following description, and in part from those skilled in the art through study and practice of the invention. Detailed Implementation

[0052] The present invention will now be described in further detail so that those skilled in the art can implement it based on the description.

[0053] It should be understood that terms such as “having,” “comprising,” and “including” as used herein do not exclude the presence or addition of one or more other elements or combinations thereof.

[0054] It should be noted that, unless otherwise specified, the experimental methods described in the following implementation plan are all conventional methods, and the reagents and materials described are all commercially available unless otherwise specified.

[0055] The synthetic route for the tetracyclic analogue-sulfonamide hybrid of the neem base of the present invention is shown below:

[0056]

[0057]

[0058] This invention provides a tetracyclic analogue of thorn-sulfonamide hybrid, having the structure of general formula I or general formula II.

[0059] The structural formula of general formula I is shown below:

[0060]

[0061] The structural formula of general formula II is shown below:

[0062]

[0063] Formula I includes compounds 13a-13e, and Formula II includes compounds 14a-14e;

[0064] In compound 13a, R1, R2, R3, and R4 are H atoms; in compound 14a, R1, R2, R3, and R4 are H atoms.

[0065] In compound 13b, R1, R2, and R3 are H atoms, and R4 is Me; in compound 14b, R1, R2, and R3 are H atoms, and R4 is Me.

[0066] In compound 13c, R1, R2, and R4 are Me atoms, and R3 is a H atom; in compound 14c, R1, R2, and R4 are Me atoms, and R3 is a H atom.

[0067] In compound 13d, R1 and R2 are H atoms, and R3 and R4 are Cl atoms; in compound 14d, R1 and R2 are H atoms, and R3 and R4 are Cl atoms.

[0068] In compound 13e, R1, R2, and R4 are H atoms, and R3 is an F atom; in compound 14e, R1, R2, and R4 are H atoms, and R3 is an F atom.

[0069] A method for preparing a tetracyclic analogue of thorn-based sulfonamide hybrid involves dissolving compound 10 or compound 11 and triethylamine in dichloromethane, adding benzenesulfonyl chloride derivatives 12a-12e at low temperature, stirring, and then stirring at room temperature until the starting material spot disappears. The reaction is monitored by TLC. After the reaction is complete, the product is purified by column chromatography to obtain the target product 13a-13e or 14a-14e. Preferably, the molar ratio of compound 10 or compound 11, triethylamine, and benzenesulfonyl chloride derivative is 0.1-1:1-5:0.1-1.

[0070] The structural formula of compound 10 is shown below:

[0071]

[0072] The structural formula of compound 11 is shown below:

[0073]

[0074] The general structural formulas of benzenesulfonyl chloride derivatives 12a-12e are shown below:

[0075]

[0076] In compound 12a, R1, R2, R3, and R4 are H atoms;

[0077] In compound 12b, R1, R2, and R3 are H atoms, and R4 is Me.

[0078] In compound 12c, R1, R2, and R4 are Me atoms, and R3 is a H atom;

[0079] In compound 12d, R1 and R2 are H atoms, and R3 and R4 are Cl atoms;

[0080] In compound 12e, R1, R2, and R4 are H atoms, and R3 is an F atom.

[0081] In another technical solution, under electromagnetic stirring, compound 8 and / or compound 9 are dissolved in an aqueous methylamine solution, stirred at room temperature, and the reaction is monitored by TLC. After the reaction is completed, dichloromethane is added for extraction, the organic phases are combined and washed with water, dried overnight with anhydrous sodium sulfate, and the filtrate is concentrated to obtain compound 10 or compound 11.

[0082] The structural formula of compound 8 is shown below:

[0083]

[0084] The structural formula of compound 9 is shown below:

[0085]

[0086] In another technical solution, compound 6 or compound 7 is added to a round-bottom flask, dissolved in acetonitrile, and then phthalimide, potassium iodide, and potassium carbonate are added to obtain a mixture. The mixture is refluxed and stirred under heating conditions, and the reaction is detected by TLC. After the reaction is completed, the mixture is separated and purified by column chromatography to obtain compound 8 and / or compound 9. Preferably, the molar ratio of compound 6 or compound 7, phthalimide, potassium iodide, and potassium carbonate is 0.1-5:0.1-5:0.05-0.5:1-10, and the heating temperature is 60-90℃.

[0087] The structural formula of compound 6 is shown below:

[0088]

[0089] The structural formula of compound 7 is shown below:

[0090]

[0091] In another technical solution, under inert gas protection, compound 4 or compound 5, palladium acetate, tri-o-methylphenylphosphine (POT), and silver carbonate are added to a round-bottom flask, dissolved in ethyl acetate, and stirred under reflux under heating. The reaction is monitored by TLC. After the reaction is complete, the brown insoluble matter is removed by hot filtration, and the filtrate is collected and purified by column chromatography to obtain compound 6 or compound 7. Preferably, the molar ratio of compound 4 or compound 5, palladium acetate, tri-o-methylphenylphosphine (POT), and silver carbonate is 1-5:1-5:0.1-0.5:5-10; the heating temperature is 50-80℃.

[0092] The structural formula of compound 4 is as follows:

[0093]

[0094] The structural formula of compound 5 is shown below:

[0095]

[0096] In another technical solution, compound 3 is added to a round-bottom flask, dissolved in anhydrous DMF (N,N-dimethylformamide), sodium hydride is added, and the mixture is stirred at low temperature. Then, 1,4-dibromobutane or 1,6-dibromobutane is added, and the mixture is stirred at room temperature. The reaction is monitored by TLC. After the reaction is complete, ice water is added to quench the reaction. The mixture is extracted with ethyl acetate, and the organic layers are combined. The mixture is dried overnight with anhydrous sodium sulfate, filtered, and the filtrate is concentrated. The residue is separated and purified by column chromatography to obtain compound 4 or compound 5. Preferably, the molar ratio of compound 3, sodium hydride, and 1,4-dibromobutane or 1,6-dibromobutane is 0.5-2:1-5:1-5.

[0097] The structural formula of compound 3 is as follows:

[0098]

[0099] In another technical solution, naphthylamine is added to a round-bottom flask, dissolved in dichloromethane, followed by the addition of triethylamine and stirring. Then, o-iodobenzoyl chloride is added and stirred at room temperature. The reaction is monitored by TLC. Once the reaction is complete, the mixture is filtered and dried to obtain compound 3. Preferably, the molar ratio of naphthylamine, triethylamine, and o-iodobenzoyl chloride is 0.5-5:1-5:0.5-5.

[0100] An antitumor drug comprising the aforementioned tetracyclic analogue of ginseng alkaloid-sulfonamide hybrid.

[0101] In another technical solution, the sulfonamide heterozygote tetracyclic analogue is formulated with pharmaceutically acceptable excipients into injections, tablets, pills, capsules, suspensions, or emulsions.

[0102] In another technical solution, the excipients are one or more of the following: ethanol, propylene glycol, polyethylene glycol, diethylene glycol, triacetin, glycerol, dextrin, povidone, stearyl alcohol, stearic acid, microcrystalline cellulose, starch, lactose, mannitol, sodium bicarbonate, calcium carbonate, low-substituted hydroxypropyl methylcellulose, magnesium stearate, and talc.

[0103] The application of the aforementioned succinate tetracyclic analog-sulfonamide hybrid in the preparation of antitumor drugs.

[0104] The application of the aforementioned succinate tetracyclic analog-sulfonamide hybrid in the preparation of drugs that inhibit topoisomerase I.

[0105] <Example 1>

[0106] A method for preparing a tetracyclic analogue of thorn-sulfonamide 13a includes the following steps:

[0107] Step 1: Take a 150 mL round-bottom flask, weigh out o-iodobenzoic acid (5.633 mmol), dissolve it in 80 mL of dichloromethane (DCM), and stir in an ice bath at 0 °C under nitrogen protection. Slowly add oxaloyl chloride (11.3 mmol) dropwise over 5 min, and finally add 1-3 drops of N,N-dimethylformamide (DMF). Stir for 5 min, then transfer to room temperature and stir for 2 h. Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, directly distill under reduced pressure to obtain compound 2.

[0108] The structural formula of compound 2 is shown below:

[0109]

[0110] Step 2: Take a 50 mL round-bottom flask, weigh and add naphthylamine (0.5 mmol), dissolve it in 20 mL dichloromethane, add triethylamine (1 mmol), stir for 5 min, then add o-iodobenzoyl chloride (0.5 mmol), stir at room temperature for 2 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), after the reaction is complete, filter and dry to obtain compound 3;

[0111] Step 3: Take a 50 mL round-bottom flask, weigh out compound 3 (0.5 mmol), dissolve it in 5 mL of anhydrous DMF, add sodium hydride (60%, 1 mmol), stir in an ice well at 0 °C for 1 h, then add 1,4-dibromobutane (1 mmol), stir at room temperature for 8 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), after the reaction is complete, add 50 mL of ice water to quench the reaction, extract three times with ethyl acetate (30 mL × 3), combine the organic layers, dry with anhydrous sodium sulfate overnight, filter and concentrate the filtrate, and separate and purify the residue by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 4;

[0112] Step 4: Under N2 protection, take a 50 mL round-bottom flask, add compound 4 (0.5 mmol), palladium acetate (0.05 mmol), tris(o-methylphenyl)phosphine (POT, 0.1 mmol), and silver carbonate (0.5 mmol), dissolve in 30 mL ethyl acetate, stir and reflux at 50 °C for 12 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 4:1), after the reaction is complete, filter while hot to remove brown insoluble matter, collect the filtrate, and separate and purify by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 6;

[0113] Step 5: Take a 50 mL round-bottom flask, weigh out compound 6 (2 mmol), dissolve it in 20 mL acetonitrile, and add phthalimide (0.294 g, 2 mmol), potassium iodide (0.0332 g, 0.2 mmol), and potassium carbonate (0.829 g, 6 mmol). Reflux the mixture at 60 °C for 36 h, and monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, purify the mixture by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound 8.

[0114] Step 6: Under electromagnetic stirring, compound 8 (3 mmol) was dissolved in 30 mL of 40% methylamine aqueous solution and stirred at room temperature for 12 h. The reaction was monitored by TLC (dichloromethane:methanol = 90:1). After the reaction was completed, dichloromethane was added for extraction (3 × 10 mL). The organic phases were combined and washed with water. The mixture was dried over anhydrous sodium sulfate overnight. After filtration, the filtrate was concentrated to obtain compound 10.

[0115] Step 7: Dissolve compound 10 (0.1 mmol) and triethylamine (1 mmol) in 15 mL of dichloromethane. Add benzenesulfonyl chloride derivative 12a (0.1 mmol) at 0 °C. Stir for 5 min and then stir at room temperature until the starting material spot disappears (about 3 h). Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, purify by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain the target product compound 13a.

[0116] Compound 13a, namely N-(4-(6-oxobenzo[c]phenanthridin-5(6H)-yl)butyl)benzenesulfonamide, is a yellow solid with a yield of 63% and an mp of 102.7-103.6 °C. HRMS (ESI) m / z: calcd for C27H24N2O3S[M+H]+: 457.1586, FOUND: 457.1588. 1H NMR (500MHz, CDCl3) δ8.52(d,J=7.3Hz,1H,ArH),8.27(d,J=8.1Hz,1H,ArH),8.21(d,J=8.7Hz,1H ,ArH),8.15(d,J=8.2Hz,1H,ArH),7.90(d,J=7.0Hz,1H,ArH),7.81(d,J=7.1Hz,3H,ArH),7.74(d, J=8.7Hz,1H,ArH),7.60(t,J=7.5Hz,1H,ArH),7.58–7.47(m,3H,ArH),7.44(t,J=7.5Hz,2H,ArH) ,5.07(s,1H,-NH),4.50(s,2H,-CH2-),2.92(t,J=6.7Hz,2H,-CH2-),2.07–1.67(m,4H,-CH2-×2). 13 C NMR (126MHz, CDCl3) δ164.8,139.9,135.7,134.7,133.8,132.9,132.5,129.0,128.7,128.6,128. 1,127.0,126.5,125.6,125.4,124.9,124.5,124.3,122.1,119.9,117.9,51.1,42.4,26.7,25.7.

[0117] <Example 2>

[0118] A method for preparing a tetracyclic analogue of thorn-sulfonamide 13b includes the following steps:

[0119] Step 1: Take a 150 mL round-bottom flask, weigh out o-iodobenzoic acid (5.633 mmol), dissolve it in 80 mL of dichloromethane (DCM), and stir in an ice bath at 0 °C under nitrogen protection. Slowly add oxaloyl chloride (11.3 mmol) dropwise over 5 min, and finally add 1-3 drops of N,N-dimethylformamide (DMF). Stir for 5 min, then transfer to room temperature and stir for 2 h. Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, directly distill under reduced pressure to obtain compound 2.

[0120] Step 2: Take a 50 mL round-bottom flask, weigh and add naphthylamine (1 mmol), dissolve it in 20 mL dichloromethane, add triethylamine (1.9 mmol), stir for 5 min, then add o-iodobenzoyl chloride (1.2 mmol), stir at room temperature for 2 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), after the reaction is complete, filter and dry to obtain compound 3;

[0121] Step 3: Take a 50 mL round-bottom flask, weigh out compound 3 (0.86 mmol), dissolve it in 5 mL of anhydrous DMF, add sodium hydride (60%, 2.7 mmol), stir in an ice well at 0 °C for 1 h, then add 1,4-dibromobutane (2.58 mmol), stir at room temperature for 8 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), after the reaction is complete, add 50 mL of ice water to quench the reaction, extract three times with ethyl acetate (30 mL × 3), combine the organic layers, dry with anhydrous sodium sulfate overnight, filter and concentrate the filtrate, and separate and purify the residue by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 4;

[0122] Step 4: Under N2 protection, take a 50 mL round-bottom flask, add compound 4 (1 mmol), palladium acetate (0.1 mmol), tris(o-methylphenyl)phosphine (0.4 mmol), and silver carbonate (2 mmol), dissolve in 30 mL ethyl acetate, stir and reflux at 60 °C for 12 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 4:1), after the reaction is complete, filter while hot to remove brown insoluble matter, collect the filtrate, and separate and purify by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 6;

[0123] Step 5: Take a 50 mL round-bottom flask, weigh out compound 6 (2 mmol), dissolve it in 20 mL acetonitrile, and add phthalimide (2 mmol), potassium iodide (0.2 mmol), and potassium carbonate (6 mmol). Reflux the mixture at 80 °C for 36 h, and monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, separate and purify the compound by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound 8.

[0124] Step 6: Under electromagnetic stirring, compound 8 (3 mmol) was dissolved in 30 mL of 40% methylamine aqueous solution and stirred at room temperature for 12 h. The reaction was monitored by TLC (dichloromethane:methanol = 90:1). After the reaction was completed, dichloromethane was added for extraction (3 × 10 mL). The organic phases were combined and washed with water. The mixture was dried over anhydrous sodium sulfate overnight. After filtration, the filtrate was concentrated to obtain compound 10.

[0125] Step 7: Dissolve compound 10 (0.5 mmol) and triethylamine (3.6 mmol) in 15 mL of dichloromethane. Add benzenesulfonyl chloride derivative 12b (0.5 mmol) at 0 °C. Stir for 5 min and then stir at room temperature until the starting material spot disappears (about 3 h). Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, purify by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain the target product compound 13b.

[0126] Compound 13b, namely

[0127] 4-methyl-N-(4-(6-oxobenzo[c]phenanthridin-5(6H)-yl)butyl)benzenesulfonamide: white solid, yield 60%, mp 164.8-165.2℃; HRMS(ESI) m / z: calcd for C28H26N2O3S[M+Na]+: 493.1562, FOUND: 493.1559. 1 H NMR (500MHz, CDCl3) δ8.51(d,J=7.3Hz,1H,ArH),8.26(d,J=8.1Hz,1H,ArH),8.21(d,J=8.7Hz,1H,ArH),8.15(d,J=8.5H z,1H,ArH),7.90(d,J=9.0Hz,1H,ArH),7.79(t,J=7.6Hz,1H,ArH),7.73(d,J=8.6Hz,1H,ArH),7.69(d,J=8.2Hz,2H,ArH) ,7.60(t,J=7.5Hz,1H,ArH),7.54(q,J=5.3,3.5Hz,2H,ArH),7.23(d,J=7.9Hz,2H,ArH),4.93(s,1H,-NH),4.55–4.45(m, 2H,-CH2-),2.89(t,J=6.8Hz,2H,-CH2-),2.38(s,3H,-CH3),1.91(p,J=7.6Hz,2H,-CH2-),1.41(p,J=6.8Hz,2H,-CH2-). 13 C NMR (126MHz, CDCl3) δ164.8,143.2,136.9,135.8,134.7,133.8,132.8,129.6,128.7,128.6,128.1, 127.1,126.5,125.6,125.4,124.9,124.5,124.3,122.1,119.9,117.8,51.2,42.5,26.7,25.7,21.5.

[0128] <Example 3>

[0129] A method for preparing a tetracyclic analogue of thorn-sulfonamide 13c includes the following steps:

[0130] Step 1: Take a 150 mL round-bottom flask, weigh out 5.633 mmol of o-iodobenzoic acid, dissolve it in 80 mL of dichloromethane (DCM), and stir in an ice bath at 0 °C under nitrogen protection. Slowly add 11.3 mmol of oxaloyl chloride over 5 min, and finally add 1-3 drops of N,N-dimethylformamide (DMF). Stir for 5 min, then transfer to room temperature and stir for 2 h. Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, directly distill under reduced pressure to obtain compound 2.

[0131] Step 2: Take a 150 mL round-bottom flask, weigh and add naphthylamine (5 mmol), dissolve it in 80 mL dichloromethane, add triethylamine (5 mmol), stir for 5 min, add o-iodobenzoyl chloride (5 mmol), stir at room temperature for 2 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), when the reaction is complete, filter and dry to obtain compound 3;

[0132] Step 3: Take a 150 mL round-bottom flask, weigh out compound 3 (2 mmol), dissolve it in 50 mL of anhydrous DMF, add sodium hydride (60%, 5 mmol), stir in an ice well at 0 °C for 1 h, then add 1,4-dibromobutane (5 mmol), stir at room temperature for 8 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), after the reaction is complete, add 50 mL of ice water to quench the reaction, extract three times with ethyl acetate (30 mL × 3), combine the organic layers, dry with anhydrous sodium sulfate overnight, filter and concentrate the filtrate, and separate and purify the residue by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 4;

[0133] Step 4: Under N2 protection, take a 50 mL round-bottom flask, add compound 4 (1 mmol), palladium acetate (0.044 g, 0.1 mmol), tris(o-methylphenyl)phosphine (0.122 g, 0.4 mmol), and silver carbonate (0.548 g, 2 mmol), dissolve in 30 mL ethyl acetate, stir and reflux at 80 °C for 12 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 4:1), after the reaction is complete, filter while hot to remove brown insoluble matter, collect the filtrate, and separate and purify by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 6;

[0134] Step 5: Take a 150 mL round-bottom flask, weigh out compound 6 (5 mmol), dissolve it in 20 mL acetonitrile, and add phthalimide (5 mmol), potassium iodide (0.5 mmol), and potassium carbonate (10 mmol). Reflux the mixture at 90 °C for 36 h, and monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, purify the mixture by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound 8.

[0135] Step 6: Under electromagnetic stirring, compound 8 (3 mmol) was dissolved in 30 mL of 40% methylamine aqueous solution and stirred at room temperature for 12 h. The reaction was monitored by TLC (dichloromethane:methanol = 90:1). After the reaction was completed, dichloromethane was added for extraction (3 × 10 mL). The organic phases were combined and washed with water. The mixture was dried over anhydrous sodium sulfate overnight. After filtration, the filtrate was concentrated to obtain compound 10.

[0136] Step 7: Dissolve compound 10 (0.1 mmol) and triethylamine (1 mmol) in 15 mL of dichloromethane. Add benzenesulfonyl chloride derivative 12c (0.1 mmol) at 0 °C. Stir for 5 min and then stir at room temperature until the starting material spot disappears (about 3 h). Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, purify by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain the target product compound 13c.

[0137] Compound 13c, namely 2,4,6-trimethyl-N-(4-(6-oxobenzo[c]phenanthridin-5(6H)-yl)butyl)benzenesulfonamide: white solid, yield 61%, mp 182.5-182.9℃; HRMS(ESI) m / z: calcd for C30H30N2O3S[M+Na]+: 521.1828, FOUND: 521.1829. 1 H NMR (500MHz, CDCl3) δ8.51(d,J=7.9Hz,1H,ArH),8.27(d,J=8.1Hz,1H,ArH),8.22(d,J=8.7Hz,1H,ArH),8.14(d,J= 7.9Hz,1H,ArH),7.91(d,J=7.4Hz,1H,ArH),7.80(t,J=7.5Hz,1H,ArH),7.74(d,J=8.6Hz,1H,ArH),7.60(t,J=7.5Hz ,1H,ArH),7.53(p,J=6.6Hz,2H,ArH),6.88(s,2H,ArH),4.87(s,1H,-NH),4.56–4.46(m,2H,-CH2-),2.85(t,J=6.6H z,2H,-CH2-),2.58(s,6H,-CH3×2),2.25(s,3H,-CH3),1.98–1.84(m,2H,-CH2-),1.38(p,J=8.3,7.0Hz,2H,-CH2-). 13CNMR (126MHz, CDCl3) δ164.8,142.0,138.9,135.8,134.7,133.8,133.6,132.8,131.9,128.7,128.6,12 8.1,126.5,125.7,125.4,124.9,124.5,124.3,122.1,119.9,117.8,51.2,41.8,26.8,25.8,22.9,20.9.

[0138] <Example 4>

[0139] A method for preparing a tetracyclic analogue of neem root alkaloid - sulfonamide hybrid 13d includes the following steps:

[0140] Step 1: Take a 150 mL round-bottom flask, weigh out o-iodobenzoic acid (1 mmol), dissolve it in 80 mL of dichloromethane (DCM), and stir in an ice bath at 0 °C under nitrogen protection. Slowly add oxaloyl chloride (4 mmol) dropwise over 5 min, and finally add 1-3 drops of N,N-dimethylformamide (DMF). Stir for 5 min, then transfer to room temperature and stir for 2 h. Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, directly distill under reduced pressure to obtain compound 2.

[0141] Step 2: Take a 50 mL round-bottom flask, weigh and add naphthylamine (2 mmol), dissolve it in 30 mL dichloromethane, add triethylamine (3 mmol), stir for 5 min, add o-iodobenzoyl chloride (3 mmol), stir at room temperature for 2 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), after the reaction is complete, filter and dry to obtain compound 3;

[0142] Step 3: Take a 50 mL round-bottom flask, weigh out compound 3 (1.5 mmol), dissolve it in 10 mL of anhydrous DMF, add sodium hydride (60%, 4 mmol), stir in an ice well at 0 °C for 1 h, then add 1,4-dibromobutane (4 mmol), stir at room temperature for 8 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), after the reaction is complete, add 50 mL of ice water to quench the reaction, extract three times with ethyl acetate (30 mL × 3), combine the organic layers, dry with anhydrous sodium sulfate overnight, filter and concentrate the filtrate, and separate and purify the residue by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 4;

[0143] Step 4: Under N2 protection, take a 50 mL round-bottom flask, add compound 4 (1 mmol), palladium acetate (0.2 mmol), tris(o-methylphenyl)phosphine (0.5 mmol), and silver carbonate (4 mmol), dissolve in 30 mL ethyl acetate, stir and reflux at 60 °C for 12 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 4:1), after the reaction is complete, filter while hot to remove brown insoluble matter, collect the filtrate, and separate and purify by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 6;

[0144] Step 5: Take a 50 mL round-bottom flask, weigh out compound 6 (4 mmol), dissolve it in 30 mL acetonitrile, and add phthalimide (4 mmol), potassium iodide (0.3 mmol), and potassium carbonate (8 mmol). Reflux the mixture at 80 °C for 36 h, and monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, purify the mixture by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound 8.

[0145] Step 6: Under electromagnetic stirring, compound 8 (3 mmol) was dissolved in 30 mL of 40% methylamine aqueous solution and stirred at room temperature for 12 h. The reaction was monitored by TLC (dichloromethane:methanol = 90:1). After the reaction was completed, dichloromethane was added for extraction (3 × 10 mL). The organic phases were combined and washed with water. The mixture was dried over anhydrous sodium sulfate overnight. After filtration, the filtrate was concentrated to obtain compound 10.

[0146] Step 7: Dissolve compound 10 (0.8 mmol) and triethylamine (4 mmol) in 15 mL of dichloromethane. Add benzenesulfonyl chloride derivative 12d (0.8 mmol) at 0 °C. Stir for 5 min and then stir at room temperature until the starting material spot disappears (about 3 h). Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, purify by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain the target product compound 13d.

[0147] Compound 13d, namely 3,4-dichloro-N-(4-(6-oxobenzo[c]phenanthridin-5(6H)-yl)butyl)benzenesulfonamide: white solid, yield 85%, mp 165.2-166.3℃; HRMS(ESI) m / z: calcd for C27H22Cl2N2O3S[M+Na]+: 547.0626, FOUND: 547.0628. 1H NMR (500MHz, CDCl3) δ8.52(d,J=7.9Hz,1H,ArH),8.27(d,J=8.1Hz,1H,ArH),8.22(d,J=8.8Hz,1H,ArH),8.16(d,J=8 .3Hz,1H,ArH),7.96(d,J=1.9Hz,1H,ArH),7.91(d,J=7.1Hz,1H,ArH),7.80(t,J=7.0Hz,1H,ArH),7.74(d,J=8.6Hz, 1H,ArH),7.66(d,J=8.4Hz,1H,ArH),7.60(t,J=7.5Hz,1H,ArH),7.54(dt,J=14.1,7.7Hz,3H,ArH),5.60(s,1H,-NH) ,4.57–4.44(m,2H,-CH2-),2.97(t,J=6.3Hz,2H,-CH2-),1.96(p,J=7.3Hz,2H,-CH2-),1.47(p,J=6.8Hz,2H,-CH2-). 13 C NMR (126MHz, CDCl3) δ164.9,140.0,137.2,135.6,134.7,133.84,133.6,133.0,131.0,129.1,128.7,128 .6,128.1,126.6,126.1,125.5,125.4,124.8,124.5,124.4,122.1,119.9,117.9,50.9,42.4,26.4,25.6.

[0148] <Example 5>

[0149] A method for preparing a tetracyclic analogue of neem root alkaloid—sulfonamide hybrid 13e, comprising the following steps:

[0150] Step 1: Take a 150 mL round-bottom flask, weigh out o-iodobenzoic acid (5.633 mmol), dissolve it in 80 mL of dichloromethane (DCM), and stir in an ice bath at 0 °C under nitrogen protection. Slowly add oxaloyl chloride (11.3 mmol) dropwise over 5 min, and finally add 1-3 drops of N,N-dimethylformamide (DMF). Stir for 5 min, then transfer to room temperature and stir for 2 h. Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, directly distill under reduced pressure to obtain compound 2.

[0151] Step 2: Take a 50 mL round-bottom flask, weigh and add naphthylamine (1 mmol), dissolve it in 20 mL dichloromethane, add triethylamine (3 mmol), stir for 5 min, add o-iodobenzoyl chloride (2 mmol), stir at room temperature for 2 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), when the reaction is complete, filter and dry to obtain compound 3;

[0152] Step 3: Take a 50 mL round-bottom flask, weigh out compound 3 (0.5 mmol), dissolve it in 5 mL of anhydrous DMF, add sodium hydride (60%, 3 mmol), stir in an ice well at 0 °C for 1 h, then add 1,4-dibromobutane (3 mmol), stir at room temperature for 8 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), after the reaction is complete, add 50 mL of ice water to quench the reaction, extract three times with ethyl acetate (30 mL × 3), combine the organic layers, dry with anhydrous sodium sulfate overnight, filter and concentrate the filtrate, and separate and purify the residue by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 4;

[0153] Step 4: Under N2 protection, take a 50 mL round-bottom flask, add compound 4 (1 mmol), palladium acetate (0.3 mmol), tris(o-methylphenyl)phosphine (0.3 mmol), and silver carbonate (1 mmol), dissolve in 30 mL ethyl acetate, stir and reflux at 60 °C for 12 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 4:1), after the reaction is complete, filter while hot to remove brown insoluble matter, collect the filtrate, and purify by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 6;

[0154] Step 5: Take a 50 mL round-bottom flask, weigh out compound 6 (4 mmol), dissolve it in 20 mL acetonitrile, and add phthalimide (3 mmol), potassium iodide (0.4 mmol) and potassium carbonate (8 mmol). Reflux the mixture at 80 °C for 36 h. Detect the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, separate and purify by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound 8.

[0155] Step 6: Under electromagnetic stirring, compound 8 (3 mmol) was dissolved in 30 mL of 40% methylamine aqueous solution and stirred at room temperature for 12 h. The reaction was monitored by TLC (dichloromethane:methanol = 90:1). After the reaction was completed, dichloromethane was added for extraction (3 × 10 mL). The organic phases were combined and washed with water. The mixture was dried over anhydrous sodium sulfate overnight. After filtration, the filtrate was concentrated to obtain compound 10.

[0156] Step 7: Dissolve compound 10 (0.8 mmol) and triethylamine (4 mmol) in 15 mL of dichloromethane. Add benzenesulfonyl chloride derivative 13e (1 mmol) at 0 °C. Stir for 5 min and then stir at room temperature until the starting material spot disappears (about 3 h). Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, purify by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain the target product compound 13e.

[0157] Compound 13e, i.e.

[0158] 3-fluoro-N-(4-(6-oxobenzo[c]phenanthridin5(6H)-yl)butyl)benzenesulfonamide: Orange-red solid, 70% yield, mp 95.2-96.1℃; HRMS(ESI) m / z: calcd for C27H23FN2O3S[M+Na]+: 497.1311, FOUND: 497.1310. 1 H NMR (500MHz, CDCl3) δ8.51(d,J=7.9Hz,1H,ArH),8.26(d,J=8.1Hz,1H,ArH),8.21(d,J=8.7Hz,1H,ArH),8.15(d,J =7.7Hz,1H,ArH),7.91(d,J=7.8Hz,1H),7.79(t,J=7.6Hz,1H,ArH),7.73(d,J=8.6Hz,1H,ArH),7.61(q,J=7.5Hz, 2H, ArH), 7.54 (s, 3H, ArH), 7.43 (q, J=7.8, 7.0Hz, 1H, ArH), 7.21 (t, J=8.2Hz, 1H, ArH), 5.44 (s, 1H, -NH), 4.53–4. 47(m,2H,-CH2-), 2.95(q,J=6.1Hz,2H,-CH2-), 1.92(p,J=7.3Hz,2H,-CH2-), 1.44(dt,J=13.9,6.8Hz,2H,-CH2-). 13 C NMR (126MHz, CDCl3) δ164.8,161.3,142.1,142.1,135.7,134.7,133.8,132.9,130.8,128.7,128.6,128. 1,126.6,125.5,125.4,124.8,124.5,124.4,122.1,119.9,117.9,114.5,114.3,51.0,42.4,26.6,25.7.

[0159] <Example 6>

[0160] A method for preparing a tetracyclic analogue of thorn-sulfonamide 14a includes the following steps:

[0161] Step 1: Take a 150 mL round-bottom flask, weigh out 5 mmol of o-iodobenzoic acid, dissolve it in 80 mL of dichloromethane (DCM), and stir in an ice bath at 0 °C under nitrogen protection. Slowly add 11 mmol of oxaloyl chloride over 5 min, and finally add 1-3 drops of N,N-dimethylformamide (DMF). Stir for 5 min, then turn to room temperature and stir for 2 h. Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, directly distill under reduced pressure to obtain compound 2.

[0162] Step 2: Take a 50 mL round-bottom flask, weigh and add naphthylamine (0.5 mmol), dissolve it in 20 mL dichloromethane, add triethylamine (1 mmol), stir for 5 min, then add o-iodobenzoyl chloride (0.5 mmol), stir at room temperature for 2 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), after the reaction is complete, filter and dry to obtain compound 3;

[0163] Step 3: Take a 50 mL round-bottom flask, weigh out compound 3 (0.5 mmol), dissolve it in 5 mL of anhydrous DMF, add sodium hydride (60%, 1 mmol), stir in an ice well at 0 °C for 1 h, then add 1,6-dibromobutane (1 mmol), stir at room temperature for 8 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), after the reaction is complete, add 50 mL of ice water to quench the reaction, extract three times with ethyl acetate (30 mL × 3), combine the organic layers, dry with anhydrous sodium sulfate overnight, filter and concentrate the filtrate, and separate and purify the residue by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 5;

[0164] Step 4: Under N2 protection, take a 50 mL round-bottom flask, add compound 5 (0.5 mmol), palladium acetate (0.05 mmol), tris(o-methylphenyl)phosphine (0.1 mmol), and silver carbonate (0.5 mmol), dissolve in 30 mL ethyl acetate, stir and reflux at 50 °C for 12 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 4:1), after the reaction is complete, filter while hot to remove brown insoluble matter, collect the filtrate, and separate and purify by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 7;

[0165] Step 5: Take a 50 mL round-bottom flask, weigh out compound 7 (1 mmol), dissolve it in 20 mL acetonitrile, and add phthalimide (1 mmol), potassium iodide (0.1 mmol), and potassium carbonate (5 mmol); reflux the mixture at 60 °C for 36 h, and detect the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, separate and purify by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound 9;

[0166] Step 6: Under electromagnetic stirring, compound 9 (3 mmol) was dissolved in 30 mL of 40% methylamine aqueous solution and stirred at room temperature for 12 h. The reaction was monitored by TLC (dichloromethane:methanol = 90:1). After the reaction was completed, dichloromethane was added for extraction (3 × 10 mL). The organic phases were combined and washed with water. The mixture was dried over anhydrous sodium sulfate overnight. After filtration, the filtrate was concentrated to obtain compound 11.

[0167] Step 7: Dissolve compound 11 (0.1 mmol) and triethylamine (1 mmol) in 15 mL of dichloromethane. Add benzenesulfonyl chloride derivative 12a (0.1 mmol) at 0 °C. Stir for 5 min and then stir at room temperature until the starting material spot disappears (about 3 h). Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, purify by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain the target product compound 14a.

[0168] Compound 14a, namely N-(6-(6-oxobenzo[c]phenanthridin-5(6H)-yl)hexyl)benzenesulfonamide: yellow oil, yield 73%; HRMS(ESI) m / z: calcd for C29H28N2O3S[M+H]+: 485.1189, FOUND: 485.1902. 1H NMR (500MHz, CDCl3) δ8.53(d,J=7.9Hz,1H,ArH),8.27(d,J=8.1Hz,1H,ArH),8.21(dd,J=16.2,8.3Hz,2H,ArH),7 .91(d,J=7.9Hz,1H,ArH),7.87–7.77(m,3H,ArH),7.73(d,J=8.6Hz,1H,ArH),7.60(t,J=7.5Hz,1H,ArH),7.58–7 .50(m,3H,ArH),7.47(t,J=7.4Hz,2H,ArH),5.30(s,1H,-NH),4.56–4.52(m,2H,-CH2-),2.83(q,J=6.7Hz,2H,-C H2-),1.82(p,J=7.1Hz,2H,-CH2-),1.32(dt,J=13.0,6.1Hz,2H,-CH2-),1.14(dq,J=19.7,7.2Hz,4H,-CH2-×2). 13 C NMR (126MHz, CDCl3) δ164.8,139.9,136.0,134.7,133.8,132.8,132.5,129.0,128.6,128.0,127.0, 126.5,125.8,125.2,125.1,124.7,124.2,122.0,120.0,117.8,51.7,42.9,29.2,28.4,26.0,25.7.

[0169] <Example 7>

[0170] A method for preparing a tetracyclic analogue of thorn-sulfonamide 14b includes the following steps:

[0171] Step 1: Take a 150 mL round-bottom flask, weigh out 5 mmol of o-iodobenzoic acid, dissolve it in 80 mL of dichloromethane (DCM), and stir in an ice bath at 0 °C under nitrogen protection. Slowly add 11 mmol of oxaloyl chloride over 5 min, and finally add 1-3 drops of N,N-dimethylformamide (DMF). Stir for 5 min, then turn to room temperature and stir for 2 h. Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, directly distill under reduced pressure to obtain compound 2.

[0172] Step 2: Take a 50 mL round-bottom flask, weigh and add naphthylamine (1 mmol), dissolve it in 20 mL dichloromethane, add triethylamine (1.9 mmol), stir for 5 min, then add o-iodobenzoyl chloride (1.2 mmol), stir at room temperature for 2 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), after the reaction is complete, filter and dry to obtain compound 3;

[0173] Step 3: Take a 50 mL round-bottom flask, weigh out compound 3 (0.86 mmol), dissolve it in 5 mL of anhydrous DMF, add sodium hydride (60%, 2.7 mmol), stir in an ice well at 0 °C for 1 h, then add 1,6-dibromobutane (2.58 mmol), stir at room temperature for 8 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), after the reaction is complete, add 50 mL of ice water to quench the reaction, extract three times with ethyl acetate (30 mL × 3), combine the organic layers, dry with anhydrous sodium sulfate overnight, filter and concentrate the filtrate, and separate and purify the residue by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 5;

[0174] Step 4: Under N2 protection, take a 50 mL round-bottom flask, add compound 5 (1 mmol), palladium acetate (0.1 mmol), tris(o-methylphenyl)phosphine (0.4 mmol), and silver carbonate (2 mmol), dissolve in 30 mL ethyl acetate, stir and reflux at 60 °C for 12 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 4:1), after the reaction is complete, filter while hot to remove brown insoluble matter, collect the filtrate, and separate and purify by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 7;

[0175] Step 5: Take a 50 mL round-bottom flask, weigh out compound 7 (2 mmol), dissolve it in 20 mL acetonitrile, and add phthalimide (2 mmol), potassium iodide (0.2 mmol), and potassium carbonate (6 mmol). Reflux the mixture at 80 °C for 36 h, and monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, separate and purify the compound by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound 9.

[0176] Step 6: Under electromagnetic stirring, compound 9 (3 mmol) was dissolved in 30 mL of 40% methylamine aqueous solution and stirred at room temperature for 12 h. The reaction was monitored by TLC (dichloromethane:methanol = 90:1). After the reaction was completed, dichloromethane was added for extraction (3 × 10 mL). The organic phases were combined and washed with water. The mixture was dried over anhydrous sodium sulfate overnight. After filtration, the filtrate was concentrated to obtain compound 11.

[0177] Step 7: Dissolve compound 11 (0.5 mmol) and triethylamine (3.6 mmol) in 15 mL of dichloromethane. Add benzenesulfonyl chloride derivative 12b (0.5 mmol) at 0 °C. Stir for 5 min and then stir at room temperature until the starting material spot disappears (about 3 h). Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, purify by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain the target product compound 14b.

[0178] Compound 14b, namely

[0179] 4-methyl-N-(6-(6-oxobenzo[c]phenanthridin-5(6H)-yl)hexyl)benzenesulfonamide: Yellow oil, yield 80%; HRMS(ESI) m / z: calcd for C30H30N2O3S[M+Na]+: 521.1875, FOUND: 521.1870. 1 H NMR (500MHz, CDCl3) δ8.53(d,J=7.9Hz,1H,ArH),8.27(d,J=8.1Hz,1H,ArH),8.21(dd,J=14.9,8.4Hz,2H,ArH),7.90 (d,J=8.3Hz,1H,ArH),7.79(t,J=7.6Hz,1H,ArH),7.72(dd,J=14.1,8.4Hz,3H,ArH),7.60(t,J=7.5Hz,1H,ArH),7.53 (p,J=6.7Hz,2H,ArH),7.27(s,2H,ArH),5.30(s,1H,-NH),4.58–4.51(m,2H,-CH2-),2.81(q,J=6.7Hz,2H,-CH2-),2. 39(s,3H,-CH3),1.83(p,J=6.9,6.5Hz,2H,-CH2-),1.32(dt,J=13.8,6.7Hz,2H,-CH2-),1.18–1.07(m,4H,-CH2-×2). 13 C NMR (126MHz, CDCl3) δ165.3,165.0,135.9,135.6,134.7,133.85,132.9,132.8,130.5,129.2,128.6,128.5,128.1 ,126.5,126.1,125.7,125.3,125.0,124.7,124.3,122.1,120.0,117.9,51.5,45.6,39.9,28.9,28.4,25.9,25.8.

[0180] <Example 8>

[0181] A method for preparing a tetracyclic analogue of thorn-sulfonamide 14c includes the following steps:

[0182] Step 1: Take a 150 mL round-bottom flask, weigh out 5 mmol of o-iodobenzoic acid, dissolve it in 80 mL of dichloromethane (DCM), and stir in an ice bath at 0 °C under nitrogen protection. Slowly add 11 mmol of oxaloyl chloride over 5 min, and finally add 1-3 drops of N,N-dimethylformamide (DMF). Stir for 5 min, then turn to room temperature and stir for 2 h. Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, directly distill under reduced pressure to obtain compound 2.

[0183] Step 2: Take a 150 mL round-bottom flask, weigh and add naphthylamine (5 mmol), dissolve it in 80 mL dichloromethane, add triethylamine (5 mmol), stir for 5 min, add o-iodobenzoyl chloride (5 mmol), stir at room temperature for 2 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), when the reaction is complete, filter and dry to obtain compound 3;

[0184] Step 3: Take a 150 mL round-bottom flask, weigh out compound 3 (2 mmol), dissolve it in 10 mL of anhydrous DMF, add sodium hydride (60%, 5 mmol), stir in an ice well at 0 °C for 1 h, then add 1,6-dibromobutane (5 mmol), stir at room temperature for 8 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), after the reaction is complete, add 80 mL of ice water to quench the reaction, extract three times with ethyl acetate (30 mL × 3), combine the organic layers, dry with anhydrous sodium sulfate overnight, filter and concentrate the filtrate, and separate and purify the residue by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 5;

[0185] Step 4: Under N2 protection, take a 150 mL round-bottom flask, add compound 5 (2 mmol), palladium acetate (0.5 mmol), tris(o-methylphenyl)phosphine (0.5 mmol), and silver carbonate (5 mmol), dissolve in 60 mL ethyl acetate, stir and reflux at 80 °C for 12 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 4:1), after the reaction is complete, filter while hot to remove brown insoluble matter, collect the filtrate, and separate and purify by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 7;

[0186] Step 5: Take a 150 mL round-bottom flask, weigh out compound 7 (5 mmol), dissolve it in 50 mL acetonitrile, and add phthalimide (5 mmol), potassium iodide (0.5 mmol), and potassium carbonate (10 mmol). Reflux the mixture at 90 °C for 36 h. Detect the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, separate and purify by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound 9.

[0187] Step 6: Under electromagnetic stirring, compound 9 (3 mmol) was dissolved in 30 mL of 40% methylamine aqueous solution and stirred at room temperature for 12 h. The reaction was monitored by TLC (dichloromethane:methanol = 90:1). After the reaction was completed, dichloromethane was added for extraction (3 × 10 mL). The organic phases were combined and washed with water. The mixture was dried over anhydrous sodium sulfate overnight. After filtration, the filtrate was concentrated to obtain compound 11.

[0188] Step 7: Dissolve compound 11 (1 mmol) and triethylamine (5 mmol) in 20 mL of dichloromethane. Add benzenesulfonyl chloride derivative 12c (1 mmol) at 0 °C. Stir for 5 min and then stir at room temperature until the starting material spot disappears (about 3 h). Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, separate and purify by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain the target product compound 14c.

[0189] Compound 14c, namely 2,4,6-trimethyl-N-(6-(6-oxobenzo[c]phenanthridin-5(6H)-yl)hexyl)benzenesulfonamide: yellow oil, yield 69%; HRMS(ESI) m / z: calcd for C32H34N2O3S[M+Na]+: 549.2188, FOUND: 549.2189. 1H NMR (500MHz, CDCl3) δ8.53(d,J=7.9Hz,1H,ArH),8.27(d,J=8.1Hz,1H,ArH),8.20(dd,J=16.2,8.5Hz,2H,ArH),7.90(d ,J=7.3Hz,1H,ArH),7.79(t,J=7.6Hz,1H,ArH),7.73(d,J=8.6Hz,1H,ArH),7.60(t,J=7.5Hz,1H,ArH),7.53(p,J=6.7Hz ,2H,ArH),6.91(s,2H,ArH),5.30(s,1H,-NH),4.58–4.49(m,2H,-CH2-),2.76(t,J=6.9Hz,2H,-CH2-),2.59(s,6H,-CH 3×2),2.25(s,3H,-CH3),1.86–1.78(m,2H,-CH2-),1.30(q,J=8.3,7.5Hz,2H,-CH2-),1.12(t,J=16.8Hz,4H,-CH2-×2). 13 C NMR (126MHz, CDCl3) δ164.8,142.0,139.0,136.0,134.7,133.8,133.6,132.7,131.9,128.6,128.0,126.5 ,125.8,125.2,125.1,124.7,124.1,122.0,120.0,117.8,51.7,42.3,29.3,28.4,26.1,25.9,22.9,20.9.

[0190] <Example 9>

[0191] A method for preparing a tetracyclic analogue of neem root alkaloid - sulfonamide hybrid 14d includes the following steps:

[0192] Step 1: Take a 150 mL round-bottom flask, weigh out 5 mmol of o-iodobenzoic acid, dissolve it in 80 mL of dichloromethane (DCM), and stir in an ice bath at 0 °C under nitrogen protection. Slowly add 11 mmol of oxaloyl chloride over 5 min, and finally add 1-3 drops of N,N-dimethylformamide (DMF). Stir for 5 min, then turn to room temperature and stir for 2 h. Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, directly distill under reduced pressure to obtain compound 2.

[0193] Step 2: Take a 50 mL round-bottom flask, weigh and add naphthylamine (3 mmol), dissolve it in 30 mL dichloromethane, add triethylamine (4 mmol), stir for 5 min, add o-iodobenzoyl chloride (4 mmol), stir at room temperature for 2 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), after the reaction is complete, filter and dry to obtain compound 3;

[0194] Step 3: Take a 50 mL round-bottom flask, weigh out compound 3 (1 mmol), dissolve it in 5 mL of anhydrous DMF, add sodium hydride (60%, 4 mmol), stir in an ice well at 0 °C for 1 h, then add 1,6-dibromobutane (3 mmol), stir at room temperature for 8 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), after the reaction is complete, add 50 mL of ice water to quench the reaction, extract three times with ethyl acetate (30 mL × 3), combine the organic layers, dry with anhydrous sodium sulfate overnight, filter and concentrate the filtrate, and separate and purify the residue by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 5;

[0195] Step 4: Under N2 protection, take a 50 mL round-bottom flask, add compound 5 (1.5 mmol), palladium acetate (0.3 mmol), tris(o-methylphenyl)phosphine (0.3 mmol), and silver carbonate (4 mmol), dissolve in 30 mL ethyl acetate, stir and reflux at 60 °C for 12 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 4:1), after the reaction is complete, filter while hot to remove brown insoluble matter, collect the filtrate, and separate and purify by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 7;

[0196] Step 5: Take a 50 mL round-bottom flask, weigh out compound 7 (3 mmol), dissolve it in 30 mL acetonitrile, and add phthalimide (4 mmol), potassium iodide (0.4 mmol), and potassium carbonate (8 mmol). Reflux the mixture at 80 °C for 36 h, and monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, purify the compound by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound 9.

[0197] Step 6: Under electromagnetic stirring, compound 9 (3 mmol) was dissolved in 30 mL of 40% methylamine aqueous solution and stirred at room temperature for 12 h. The reaction was monitored by TLC (dichloromethane:methanol = 90:1). After the reaction was completed, dichloromethane was added for extraction (3 × 10 mL). The organic phases were combined and washed with water. The mixture was dried over anhydrous sodium sulfate overnight. After filtration, the filtrate was concentrated to obtain compound 11.

[0198] Step 7: Dissolve compound 11 (0.8 mmol) and triethylamine (2 mmol) in 15 mL of dichloromethane. Add benzenesulfonyl chloride derivative 12d (0.8 mmol) at 0 °C. Stir for 5 min and then stir at room temperature until the starting material spot disappears (about 3 h). Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, purify by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain the target product compound 14d.

[0199] Compound 14d, namely 3,4-dichloro-N-(6-(6-oxobenzo[c]phenanthridin-5(6H)-yl)hexyl)benzenesulfonamide: white solid, yield 79%, mp 174.5-174.9℃; HRMS(ESI) m / z: calcd for C29H26Cl2N2O3S[M+Na]+: 575.0939, FOUND: 575.0936. 1 H NMR (500MHz, CDCl3) δ8.50(d,J=7.8Hz,1H,ArH),8.28(d,J=8.1Hz,1H,ArH),8.23(d,J=8.4Hz,2H,ArH),7.91(s,2H,ArH ),7.79(t,J=7.4Hz,1H,ArH),7.74(d,J=8.6Hz,1H,ArH),7.64(d,J=8.3Hz,1H,ArH),7.59(t,J=7.5Hz,1H,ArH),7.57–7. 50(m,2H,ArH),7.47(d,J=8.3Hz,1H,ArH),6.51(s,1H,-NH),4.61(t,J=7.2Hz,2H,-CH2-),3.32(q,J=6.6Hz,2H,-CH2-) ,1.88(p,J=7.2Hz,2H,-CH2-), 1.52(p,J=7.0Hz,2H,-CH2-), 1.33(p,J=7.6Hz,2H,-CH2-), 1.21(p,J=7.3Hz,2H,-CH2-). 13 C NMR (126MHz, CDCl3) δ165.3,165.0,135.9,135.6,134.7,134.5,133.8,132.9,132.8,130.5,128.6,128.5,128 .1,126.5,126.1,125.7,125.3,125.0,124.7,124.3,122.1,120.0,117.9,51.5,39.9,28.9,28.4,25.9,25.8.

[0200] <Example 10>

[0201] A method for preparing a tetracyclic analogue of thorn alkaloid - sulfonamide hybrid 14e, comprising the following steps:

[0202] Step 1: Take a 150 mL round-bottom flask, weigh out 5 mmol of o-iodobenzoic acid, dissolve it in 80 mL of dichloromethane (DCM), and stir in an ice bath at 0 °C under nitrogen protection. Slowly add 11 mmol of oxaloyl chloride over 5 min, and finally add 1-3 drops of N,N-dimethylformamide (DMF). Stir for 5 min, then turn to room temperature and stir for 2 h. Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, directly distill under reduced pressure to obtain compound 2.

[0203] Step 2: Take a 50 mL round-bottom flask, weigh and add naphthylamine (1 mmol), dissolve it in 20 mL dichloromethane, add triethylamine (1.58 mL, 1.9 mmol), stir for 5 min, then add o-iodobenzoyl chloride (1.2 mmol), stir at room temperature for 2 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), after the reaction is complete, filter and dry to obtain compound 3;

[0204] Step 3: Take a 50 mL round-bottom flask, weigh out compound 3 (1 mmol), dissolve it in 5 mL of anhydrous DMF, add sodium hydride (60%, 3 mmol), stir in an ice well at 0 °C for 1 h, then add 1,6-dibromobutane (3 mmol), stir at room temperature for 8 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1), after the reaction is complete, add 50 mL of ice water to quench the reaction, extract three times with ethyl acetate (30 mL × 3), combine the organic layers, dry with anhydrous sodium sulfate overnight, filter and concentrate the filtrate, and separate and purify the residue by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 5;

[0205] Step 4: Under N2 protection, take a 50 mL round-bottom flask, add compound 4 or 5 (1 mmol), palladium acetate (0.1 mmol), tris(o-methylphenyl)phosphine (0.4 mmol), and silver carbonate (2 mmol), dissolve in 30 mL ethyl acetate, stir and reflux at 60 °C for 12 h, monitor the reaction by TLC (petroleum ether: ethyl acetate = 4:1), after the reaction is complete, filter while hot to remove brown insoluble matter, collect the filtrate, and separate and purify by column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain compound 7;

[0206] Step 5: Take a 50 mL round-bottom flask, weigh out compound 7 (2 mmol), dissolve it in 20 mL acetonitrile, and add phthalimide (2 mmol), potassium iodide (0.2 mmol), and potassium carbonate (6 mmol). Reflux the mixture at 85 °C for 36 h. Detect the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, separate and purify the compound by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound 9.

[0207] Step 6: Under electromagnetic stirring, compound 9 (3 mmol) was dissolved in 30 mL of 40% methylamine aqueous solution and stirred at room temperature for 12 h. The reaction was monitored by TLC (dichloromethane:methanol = 90:1). After the reaction was completed, dichloromethane was added for extraction (3 × 10 mL). The organic phases were combined and washed with water. The mixture was dried over anhydrous sodium sulfate overnight. After filtration, the filtrate was concentrated to obtain compound 11.

[0208] Step 7: Dissolve compound 11 (0.5 mmol) and triethylamine (3.6 mmol) in 15 mL of dichloromethane. Add benzenesulfonyl chloride derivative 12e (0.5 mmol) at 0 °C. Stir for 5 min and then stir at room temperature until the starting material spot disappears (about 3 h). Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction is complete, purify by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain the target product compound 14e.

[0209] Compound 14e, namely 3-fluoro-N-(6-(6-oxobenzo[c]phenanthridin-5(6H)-yl)hexyl)benzenesulfonamide: yellow oil, yield 71%; HRMS(ESI) m / z: calcd for C29H27FN2O3S[M+H]+: 503.1805, FOUND: 503.1802. 1H NMR (500MHz, CDCl3) δ8.52(d,J=7.9Hz,1H,ArH),8.26(d,J=8.1Hz,1H,ArH),8.20(dd,J=13.2,8.4Hz,2H,ArH),7.90(d,J=7. 9Hz,1H,ArH),7.78(t,J=7.6Hz,1H,ArH),7.73(d,J=8.6Hz,1H,ArH),7.60(dd,J=19.3,7.6Hz,2H,ArH),7.53(dt,J=12.9,7.2 Hz,3H,ArH),7.46(q,J=7.9Hz,1H,ArH),7.23(t,J=8.1Hz,1H,ArH),4.85(t,J=5.8Hz,1H,-NH),4.61–4.50(m,2H,-CH2-),2.8 5(q,J=6.6Hz,2H,-CH2-),1.82(p,J=7.2Hz,2H,-CH2-),1.34(p,J=6.9Hz,2H,-CH2-),1.15(dp,J=23.4,7.0Hz,4H,-CH2-×2). 13 C NMR (126MHz, CDCl3) δ164.8,163.4,142.2,142.1,134.7,133.8,132.8,130.9,130.8,128.6,128.6,128.0,125 .7,125.2,125.0,122.7,122.0,120.0,119.8,119.6,117.8,114.5,114.3,51.6,43.0,29.2,28.4,26.0,25.6.

[0210] <In vitro antitumor activity experiment>

[0211] In vitro antitumor screening using the MTS method

[0212] 1) Cell seeding: Prepare a single-cell suspension using culture medium (DMEM or RMPI1640) containing 10% fetal bovine serum, and seed 3,000 to 15,000 cells per well into a 96-well plate, with a volume of 100 μl per well. The cells should be seeded and cultured 12 to 24 hours in advance.

[0213] 2) Add the test compound solution: Dissolve the compound separately with DMSO, set the initial concentration to 40 μM, and perform preliminary screening on five types of cells: leukemia cells (HL-60), human liver cancer cells (HepG2), human lung cancer cells (H460), human cervical cancer cells (HeLa), and human normal liver cells (LO2). The final volume of each well is 200 μl, and each treatment has 3 replicates.

[0214] 3) Color development: After culturing at 37°C for 48 hours, discard the culture medium in the wells of adherent cells, and add 20 μl of MTS solution and 100 μl of culture medium to each well; discard 100 μl of culture supernatant in the wells of HL-60 suspension cells, and add 20 μl of MTS solution to each well; add 20 μl of MTS solution directly to each well of MT-4 suspension cells; set up 3 blank replicates (a mixture of 20 μl of MTS solution and 100 μl of culture medium), and continue incubation for 2-4 hours to allow the reaction to proceed fully before measuring the absorbance.

[0215] 4) Colorimetric analysis: Select a wavelength of 492nm, use a multi-functional microplate reader (MULTISKAN FC) to read the absorbance values ​​of each well, record the results, and after data processing, obtain the inhibition rate results as shown in Table 1.

[0216] 5) IC 50 IC50 calculation: Compounds with inhibition rates greater than 50% from Table 1 were dissolved in DMSO and screened at concentrations of 40, 8, 1.6, 0.32, and 0.064 μM, with a final volume of 200 μl per well and three replicates for each treatment. Two positive compounds, cisplatin (DDP) and nifedipine chloride (NC), were included in each experiment. Cell growth curves were plotted with concentration on the x-axis and cell viability on the y-axis. The IC50 of the compounds was calculated using the Reed-Muench method. 50 The values ​​are shown in Table 2.

[0217] Table 1. Inhibition rate of the tetracyclic analogue of purslane-sulfonamide hybrid on selected cells at a concentration of 40 μmol / L.

[0218]

[0219]

[0220] Table 2 IC50 of tetracyclic analogues of pamoate alkaloids-sulfonamide hybrids 50 value

[0221]

[0222] As shown in Table 1, the tetracyclic analogues of *Zanthoxylum nitidum* and sulfonamide hybrids 13a-14e of this invention exhibit selective inhibitory effects on human cervical cancer cells (HeLa). Compounds 13a, 13e, and 14a-14e showed inhibition rates greater than 50% against HeLa cells, indicating that they possess good inhibitory effects. Except for compound 14d (82.78%), the tetracyclic analogues of *Zanthoxylum nitidum* and sulfonamide hybrids showed low toxicity to normal human hepatocytes (LO2), with inhibition rates ranging from 20.39% to 47.25%.

[0223] As shown in Table 2, most sulfonamide derivatives (13a, 13e, 14a-14e) exhibited selective inhibition against human cervical cancer cells (HeLa), and their IC50 values ​​were [missing data]. 50 The values ​​were 19.47±0.19 μM, 11.75±1.16 μM, 9.27±1.45 μM, 7.39±0.24 μM, 1.83±0.24 μM, 17.48±2.80 μM, and 4.20±0.116 μM, respectively, among which the IC50 of compound 14c was... 50 (1.83±0.24μM) lowest, compared to NC IC 50 The value (1.48±0.11μM) was similar to that of the positive control drug cisplatin (DDP), and the IC50 value was similar. 50 The concentration of the anti-tumor activity was low (2.01±0.14 μM), and it is worth mentioning that, except for compound 14d, this series of compounds showed almost no toxicity to normal LO2 cells, indicating great potential for development into anti-human cervical cancer drugs. Furthermore, only compound 14d in this series exhibited excellent anti-tumor activity against leukemia cells (HL-60), with an IC50 value of 2.01±0.14 μM. 50 The value (0.96±0.01μM) is much lower than that of NC (13.47±0.38μM) and cisplatin (2.44±0.08μM), indicating that it has great potential to be developed into an anti-leukemia drug.

[0224] Although embodiments of the present invention have been disclosed above, they are not limited to the applications listed in the specification and embodiments. They can be applied to various fields suitable for the present invention. For those skilled in the art, other modifications can be easily made. Therefore, without departing from the general concept defined by the claims and their equivalents, the present invention is not limited to the specific details and illustrations shown and described herein.

Claims

1. A quaternary analogue of strictosidine - sulphonamide hybrid characterised in that, having the structure of general formula I or general formula II, The structural formula of general formula I is as follows: The structural formula of general formula II is as follows: General formula I is compounds 13a-13e, and general formula II is compounds 14a-14e; In compound 13a, R1, R2, R3, and R4 are H atoms; in compound 14a, R1, R2, R3, and R4 are H atoms; In compound 13b, R1, R2, and R3 are H atoms, and R4 is Me; in compound 14b, R1, R2, and R3 are H atoms, and R4 is Me; In compound 13c, R1, R2, and R4 are Me, and R3 is H atom; in compound 14c, R1, R2, and R4 are Me, and R3 is H atom; In compound 13d, R1 and R2 are H atoms, and R3 and R4 are Cl atoms; in compound 14d, R1 and R2 are H atoms, and R3 and R4 are Cl atoms; In compound 13e, R1 and R2, and R4 are H atoms, and R3 is F atom; in compound 14e, R1 and R2, and R4 are H atoms, and R3 is F atom.

2. The process for the preparation of a diterbina base tetralone analog-sulfonamide hybrid as claimed in claim 1 wherein, Compound 10 or compound 11 and triethylamine are dissolved in dichloromethane, and benzene sulfonyl chloride derivatives 12a-12e are added at low temperature, and after stirring, the mixture is stirred at room temperature until the raw material point disappears, and the reaction is monitored by TLC; after the reaction is completed, the target product 13a-13e or 14a-14e is obtained by column chromatography separation and purification; The structural formula of compound 10 is as follows: The structural formula of compound 11 is as follows: The structural formula of benzene sulfonyl chloride derivatives 12a-12e is as follows: In compound 12a, R1, R2, R3, R4are H atoms; in compound 12b, R1, R2, R3are H atoms, and R4is Me; in compound 12c, R1, R2, R4are Me, and R3is H atom; in compound 12d, R1, R2are H atoms, and R3, R4are Cl atoms; in compound 12e, R1, R2, R4are H atoms, and R3is F atom.

3. The method for preparing the tetracyclic analogue-sulfonamide hybrid of thorn alkaloid as described in claim 2, characterized in that, Compound 8 or compound 9 is dissolved in aqueous methylamine under electromagnetic stirring, and stirred at room temperature, and the reaction is monitored by TLC; after the reaction is completed, dichloromethane is added for extraction, the organic phases are combined and washed with water, dried with anhydrous sodium sulfate overnight, filtered, and the filtrate is concentrated to obtain compound 10 or compound 11; The structural formula of compound 8 is as follows: The structural formula of compound 9 is as follows:

4. The process for the preparation of a diterbina base tetralone analog-sulfonamide hybrid as claimed in claim 3 wherein, Compound 6 or compound 7 is added to a round-bottom flask, dissolved in acetonitrile, and o-phthalic imide, potassium iodide, and potassium carbonate are added to obtain a mixture, which is stirred under heating and reflux, and the reaction is monitored by TLC; after the reaction is completed, the mixture is separated and purified by column chromatography to obtain compound 8 or compound 9; The structural formula of compound 6 is as follows: The structural formula of compound 7 is as follows:

5. The process for the preparation of a diterbina base tetralone analog-sulfonamide hybrid as claimed in claim 4 wherein, Compound 4 or compound 5, palladium acetate, tri-o-methylphenyl phosphine, and silver carbonate are added to a round-bottom flask under inert gas protection, dissolved in ethyl acetate, and stirred under heating and reflux, and the reaction is monitored by TLC; after the reaction is completed, brown insoluble substances are removed by hot filtration, the filtrate is collected, and compound 6 or compound 7 is obtained by column chromatography separation and purification; The structural formula of compound 4 is as follows: The structural formula of compound 5 is as follows:

6. The process for the preparation of a diterbina base tetralone analog-sulfonamide hybrid as claimed in claim 5 wherein, Into a round bottom flask, compound 3 was dissolved in anhydrous DMF, stirred at low temperature, 1,4-dibromobutane or 1,6-dibromobutane was added, stirred at room temperature, the reaction was monitored by TLC, after the reaction was completed, the reaction was quenched by adding ice water, extracted by ethyl acetate, the organic layers were combined, dried over anhydrous sodium sulfate overnight, the filtrate was concentrated after filtration, and the residue was separated and purified by a chromatography column to obtain compound 4 or compound 5; The structural formula of compound 3 is as follows:

7. The process for the preparation of a diterbina base tetralone analog-sulfonamide hybrid as claimed in claim 6 wherein, Into a round bottom flask, compound 3 was dissolved in anhydrous DMF, stirred at low temperature, 1,4-dibromobutane or 1,6-dibromobutane was added, stirred at room temperature, the reaction was monitored by TLC, after the reaction was completed, the reaction was quenched by adding ice water, extracted by ethyl acetate, the organic layers were combined, dried over anhydrous sodium sulfate overnight, the filtrate was concentrated after filtration, and the residue was separated and purified by a chromatography column to obtain compound 4 or compound 5; 8. An anticancer drug, characterized by, The quaternary alkaloid-sulfonamide hybrid of claim 1.

9. The anticancer drug according to claim 8, characterized in that, The cancer is human cervical cancer, leukemia.

10. Use of compound 13a, compound 13e, 14a-14e in the quaternary alkaloid-sulfonamide hybrid of claim 1 in the preparation of a drug for resisting human cervical cancer.

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