Allicin derivative as well as preparation method and application thereof

Abstract

The preparation method specifically comprises the following steps: carrying out condensation reflux reaction on p-bromobenzyl bromide and 4-methyl benzene methanesulfonic acid sodium salt to obtain a white solid A1; 4-methoxybenzyl mercaptan and propargyl bromide are subjected to a reaction, and a compound A2 is obtained; the compound A2, AIBN and thioacetic acid are subjected to a reaction, and a compound A3 is obtained; reacting the compound A3, KOH and the white solid A1 to obtain a compound A4; a compound A4 and m-cpba are subjected to a reaction, and (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl) sulfinyl] propan-1-ene-1-yl] disulfide, namely the allicin derivative, is obtained. The allicin derivative has relatively high biological activity and stability; the obtained allicin derivative has remarkable anti-tumor activity, and new possibility is provided for application of allicin in the field of medicine.

Description

Technical Field

[0001] The invention belongs to the technical field of preparation of bioactive compounds, and in particular relates to a preparation method of an allicin derivative, and also relates to the allicin derivative and application thereof. Background Art

[0002] Allicin, a natural organic sulfur compound extracted from garlic, is widely used in medicine, agriculture and food industries due to its significant antibacterial, antifungal, antiviral and antiparasitic effects.

[0003] Alliin, the precursor of allicin, exists in an odorless state in unprocessed garlic. Once the garlic is chopped or mashed, alliin is converted into allicin by the action of alliinase. This process reveals the key steps in the formation of allicin in garlic. Traditional allicin extraction techniques include sulfur determination and mercuric nitrate precipitation, but these methods have limitations in extraction purity and detection sensitivity. The application of modern high performance liquid chromatography (HPLC) and gas chromatography (GC) has significantly improved the purity of allicin extraction and the sensitivity of detection, providing a powerful tool for in-depth research on its potential pharmacological effects and applications.

[0004] The biological activity of allicin is mainly attributed to its unique chemical structure, including active units such as thiosulfinate groups, allyl groups, sulfur atoms, disulfide bonds, and α,β-unsaturated thioethers. These groups work synergistically to give allicin significant biological functions. For example, the thiosulfinate group is crucial to the antibacterial and antifungal activity of allicin because it can react with sulfhydryl groups (-SH) in bacterial and fungal cells and interfere with their normal metabolic processes. The allyl group can interact with the enzyme system in the body to produce a series of biologically active substances. The presence of disulfide bonds makes the allicin molecule highly reactive and can react with proteins and other molecules in microbial cells.

[0005] Although allicin has a wide range of biological activities, its clinical application is limited by chemical instability, low bioavailability and potential toxicity. To overcome these limitations, researchers have tried to improve the performance of allicin through chemical modification, including the introduction of different functional groups to improve its stability and bioavailability, as well as to optimize its pharmacological effects. The synthesis of allicin derivatives has improved stability and bioavailability and reduced toxicity while retaining the biological activity of allicin. Summary of the invention

[0006] The purpose of the present invention is to provide a method for preparing an allicin derivative, which has high biological activity and stability.

[0007] The second object of the present invention is to provide allicin derivatives.

[0008] The third object of the present invention is to provide application of allicin derivatives in medicinal chemistry.

[0009] The technical scheme adopted by the present invention is a method for preparing an allicin derivative, which specifically comprises: using p-bromobenzyl bromide and sodium 4-methylbenzylmethanesulfonate to carry out a condensation reflux reaction to obtain a white solid A1; using p-methoxybenzyl mercaptan and propargyl bromide to carry out a reaction to obtain a compound A2; using compound A2, AIBN and thioacetic acid to carry out a reaction to obtain a compound A3; using compound A3, KOH and white solid A1 to carry out a reaction to obtain a compound A4; using compound A4 and m-cpba to carry out a reaction to obtain (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide, which is an allicin derivative.

[0010] The present invention is also characterized in that:

[0011] Follow the steps below to implement it:

[0012] Step 1, adding p-bromobenzyl bromide to acetonitrile, and then adding sodium 4-methylbenzene methanesulfonate, and performing condensation reflux reaction, and then reacting at room temperature, and detecting by thin layer chromatography. When p-bromobenzyl bromide is completely reacted, the reaction is stopped, the solvent is evaporated, washing, extraction, and the obtained crude product is purified by column chromatography to obtain a white solid A1;

[0013] Step 2, adding p-methoxybenzyl mercaptan to a methanol solution of KOH, stirring, adding a toluene solution containing propargyl bromide to the mixture, stirring, diluting with distilled water, and then extracting with ether three times, then adding a saturated sodium chloride aqueous solution to wash the combined organic phase, and then drying the organic extract with anhydrous magnesium sulfate, spin-drying the crude product and purifying it by column chromatography to obtain compound A2;

[0014] Step 3, dissolving compound A2 in degassed toluene, adding AIBN, heating the mixture, slowly dropping thioacetic acid dissolved in degassed toluene into the mixture over 1-2 hours, and continuing stirring until TLC shows a maximum conversion rate of the starting reactants. After the reaction is completed, evaporating the solvent to obtain a crude product, and then purifying by column chromatography to obtain compound A3;

[0015] Step 4, dissolving compound A3 in degassed methanol, placing in a low-temperature reactor and cooling to -35°C to -40°C, slowly injecting KOH dissolved in degassed methanol, stirring for 20-30 min, cooling the reaction mixture to -70°C to -80°C, injecting white solid A1 dissolved in degassed methanol into the reaction mixture, heating and stirring, adding saturated ammonium chloride aqueous solution after the reaction is completed, then extracting with dichloromethane, drying over anhydrous sodium sulfate, evaporating the organic solvent, and purifying the crude product by column chromatography to obtain compound A4;

[0016] Step 5, dissolving compound A4 in dichloromethane, cooling to -70°C to -80°C in a low-temperature reactor, adding m-cpba, reacting for 2-3 hours and then warming to room temperature, detecting by TLC until the raw material is consumed, adding saturated sodium bicarbonate solution, extracting with dichloromethane three times, drying with anhydrous sodium sulfate and evaporating the solvent, and purifying the crude product by column chromatography to obtain (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide, which is an allicin derivative.

[0017] In step 1, the reflux reaction temperature is 80-90° C., the reflux reaction time is 3-4 hours, the reaction time at room temperature is 10-12 hours, and the molar ratio of p-bromobenzyl bromide to sodium 4-methylbenzene methanesulfonate is 1-2:1.2-2.5.

[0018] In step 2, the first stirring time is 15-25 min; the second stirring time is 10-20 min; the molar ratio of p-methoxybenzyl mercaptan, KOH and propargyl bromide is 1-2:2.5-5:1.1-2.5.

[0019] In step 3, the heating temperature is 80-90° C.; the molar ratio of compound A2, AIBN, and thioacetic acid is 1-2: 0.05-0.1: 1.2-2.5.

[0020] In step 4, the heating and stirring temperature is 0-5°C, and the heating and stirring time is 2-3h; the molar ratio of compound A3, KOH, and white solid A1 is 1-2:1-2:1.1-2.5.

[0021] In step 5, the molar ratio of compound A4 and m-cpba is 1-2:1.2-2.5.

[0022] The second technical solution adopted by the present invention is the allicin derivative prepared by the preparation method of the allicin derivative.

[0023] The third technical solution adopted by the present invention is the application of allicin derivatives in pharmaceutical chemistry. The allicin derivatives have the function of inhibiting the proliferation of A549 tumor cells.

[0024] The beneficial effects of the present invention are as follows: the method of the present invention uses the structure of the allicin functional molecule as a guide, and through an organic chemical synthesis method, a natural product derivative functional molecule (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide can be obtained, and the allicin derivative has high biological activity and stability; in addition, the method is simple to operate, easy to implement, and cost-effective, and the product is easy to separate and purify; the obtained allicin derivative has significant anti-tumor activity, and provides new possibilities for the application of allicin in the medical field. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 The synthetic route of the (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide functional molecule of the present invention is as follows;

[0026] Figure 2a is a hydrogen nuclear magnetic resonance spectrum of the (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide functional molecule of the present invention;

[0027] Figure 2b is the carbon nuclear magnetic resonance spectrum of the (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide functional molecule of the present invention;

[0028] Figure 2c It is a high-resolution mass spectrum of the functional molecule of (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide of the present invention;

[0029] Figure 3 This is a CCK-8 cytotoxicity test diagram of the (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide functional molecule of the present invention;

[0030] Figure 4 This is a graph showing the test results of active oxygen confocal imaging of the (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide functional molecule of the present invention. DETAILED DESCRIPTION

[0031] The present invention is described in detail below with reference to the accompanying drawings and specific embodiments.

[0032] The preparation method of the allicin derivative of the present invention is as follows: Figure 1As shown, the synthesis route of the allicin derivative (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide;

[0033] Follow the steps below to implement it:

[0034] Step 1, adding p-bromobenzyl bromide to acetonitrile, and then adding sodium 4-methylbenzene methanesulfonate, and performing condensation reflux reaction, and then reacting at room temperature for 10-12 hours, and detecting by thin layer chromatography, when p-bromobenzyl bromide is completely reacted, the reaction is stopped, the solvent is evaporated, the product is washed with distilled water, and extracted with ethyl acetate for 2-3 times, and the obtained crude product is purified by column chromatography (PE:EA=10:1, v / v) to obtain a white solid A1;

[0035] The reflux reaction temperature is 80-90°C, and the reflux reaction time is 3-4h;

[0036] The molar ratio of p-bromobenzyl bromide to sodium 4-methylbenzene methanesulfonate is 1-2:1.2-2.5;

[0037] Step 2, add p-methoxybenzyl mercaptan to a methanol solution of KOH, stir for 15-25 minutes, add a toluene solution containing propargyl bromide (80wt% toluene solution) to the mixture, stir for 10-20 minutes, dilute with distilled water, and then extract with ether three times. Then add a saturated sodium chloride aqueous solution to wash the combined organic phase, and then dry the organic extract with anhydrous magnesium sulfate. After the ether is dried, the crude product is purified by column chromatography (PE: EA = 10: 1, v / v) to obtain compound A2;

[0038] The molar ratio of p-methoxybenzyl mercaptan, KOH and propargyl bromide is 1-2:2.5-5:1.1-2.5;

[0039] Step 3, dissolve compound A2 in degassed toluene, add AIBN, heat the mixture to 80-90°C, slowly drop thioacetic acid dissolved in degassed toluene into the mixture over 1-2 hours, and continue stirring until TLC shows the maximum conversion rate of the starting reactant. After the reaction is completed, evaporate the solvent to obtain a crude product, which is then purified by column chromatography (Hexane:EA=10:1, v / v) to obtain compound A3;

[0040] The molar ratio of compound A2, AIBN, and thioacetic acid is 1-2: 0.05-0.1: 1.2-2.5;

[0041] Step 4, dissolving compound A3 in degassed methanol, placing in a low temperature reactor and cooling to -35°C to -40°C, slowly injecting KOH dissolved in degassed methanol, stirring for 20-30 min, cooling the reaction mixture to -70°C to -80°C, injecting white solid A1 dissolved in degassed methanol into the reaction mixture, heating and stirring, the heating and stirring temperature is 0-5°C, the heating and stirring time is 2-3h, after the reaction is completed, adding saturated ammonium chloride aqueous solution, then extracting with dichloromethane, drying with anhydrous sodium sulfate, evaporating the organic solvent, and purifying the crude product by column chromatography (Hexane:EA=10:1, v / v) to obtain compound A4;

[0042] The molar ratio of compound A3, KOH, and white solid A1 is 1-2:1-2:1.1-2.5;

[0043] Step 5, dissolve compound A4 in dichloromethane, cool to -70°C to -80°C in a low temperature reactor, add m-cpba (meta-chloroperbenzoic acid), react for 2-3 hours and then warm to room temperature, detect by TLC until the raw material is consumed, add saturated sodium bicarbonate solution, extract three times with dichloromethane, dry with anhydrous sodium sulfate and evaporate the solvent. The crude product is purified by column chromatography (Hexane:EA=15:1, v / v) to obtain (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide (i.e. Figure 1 Compound A5) is the allicin derivative of the present invention;

[0044] The molar ratio of compound A4 and m-cpba is 1-2:1.2-2.5.

[0045] The product obtained by the method of the present invention is 1 H NMR, 13 C NMR, HRMS and other analyses verified the accuracy of its chemical structure, and the test results showed that the designed functional molecules were successfully prepared.

[0046] Depend on Figure 2a , Figure 2b and Figure 2c It can be seen that the hydrogen atom integrated area, the number of carbon atoms and the relative molecular mass of the synthesized product are consistent with the actual situation of (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide, and through Figure 2a The integrated area ratio of hydrogen atoms shows that E:Z is approximately 3:2. The above data verify the successful preparation of (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide bioactive molecules.

[0047] The (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide prepared by the method of the present invention has excellent performance in inhibiting the proliferation of A549 tumor cells and promoting the apoptosis of A549 tumor cells, and can be used for exploring the application fields of derivatives, including application research in medicine, agriculture and food additives.

[0048] Example 1

[0049] The preparation method of the allicin derivative of the present invention is specifically implemented according to the following steps:

[0050] Step 1, adding p-bromobenzyl bromide to acetonitrile, and then adding sodium 4-methylbenzene methanesulfonate, and performing condensation reflux reaction, and then reacting at room temperature for 10 hours, and detecting by thin layer chromatography. When p-bromobenzyl bromide is completely reacted, the reaction is stopped, the solvent is evaporated, the product is washed with distilled water, and extracted twice with ethyl acetate. The obtained crude product is purified by column chromatography to obtain a white solid A1;

[0051] The reflux reaction temperature was 80°C and the reflux reaction time was 3h;

[0052] The molar ratio of p-bromobenzyl bromide and sodium 4-methylbenzene methanesulfonate is 1:1.2;

[0053] Step 2, add p-methoxybenzyl mercaptan to a methanol solution of KOH, stir for 15 minutes, add a toluene solution containing propargyl bromide (80 wt% toluene solution) to the mixture, stir for 10 minutes, dilute with distilled water, and then extract with ether three times. Then add a saturated sodium chloride aqueous solution to wash the combined organic phase, and then dry the organic extract with anhydrous magnesium sulfate. After the ether is dried, the crude product is purified by column chromatography to obtain compound A2;

[0054] The molar ratio of p-methoxybenzyl mercaptan, KOH and propargyl bromide is 1:2.5:1.1;

[0055] Step 3, dissolve compound A2 in degassed toluene, add AIBN, heat the mixture to 80°C, slowly drop thioacetic acid dissolved in degassed toluene into the mixture over 1 hour, and continue stirring until TLC shows the maximum conversion rate of the starting reactant. After the reaction is completed, evaporate the solvent to obtain a crude product, which is then purified by column chromatography to obtain compound A3;

[0056] The molar ratio of compound A2, AIBN, and thioacetic acid is 1:0.05:1.2;

[0057] Step 4, dissolving compound A3 in degassed methanol, placing in a low-temperature reactor and cooling to -35°C, slowly injecting KOH dissolved in degassed methanol, stirring for 20 min, cooling the reaction mixture to -70°C, injecting white solid A1 dissolved in degassed methanol into the reaction mixture, heating and stirring, the heating and stirring temperature is 0°C, the heating and stirring time is 2 h, after the reaction is completed, adding saturated ammonium chloride aqueous solution, then extracting with dichloromethane, drying with anhydrous sodium sulfate, evaporating the organic solvent, and purifying the crude product by column chromatography to obtain compound A4;

[0058] The molar ratio of compound A3, KOH, and white solid A1 is 1:1:1.1;

[0059] Step 5, dissolve compound A4 in dichloromethane, cool to -70°C in a low-temperature reactor, add m-cpba, react for 2 hours and then warm to room temperature, detect by TLC until the raw material is consumed, add saturated sodium bicarbonate solution, extract three times with dichloromethane, dry with anhydrous sodium sulfate and evaporate the solvent. The crude product is purified by column chromatography to obtain (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide, which is the allicin derivative of the present invention;

[0060] The molar ratio of compound A4 and m-cpba is 1:1.2.

[0061] Example 2

[0062] The preparation method of the allicin derivative of the present invention is specifically implemented according to the following steps:

[0063] Step 1, adding p-bromobenzyl bromide to acetonitrile, and then adding sodium 4-methylbenzene methanesulfonate, and performing condensation reflux reaction, and then reacting at room temperature for 11 hours, and detecting by thin layer chromatography. When p-bromobenzyl bromide is completely reacted, the reaction is stopped, the solvent is evaporated, the product is washed with distilled water, and extracted with ethyl acetate for 3 times. The obtained crude product is purified by column chromatography to obtain a white solid A1;

[0064] The reflux reaction temperature was 85°C and the reflux reaction time was 3.5 h;

[0065] The molar ratio of p-bromobenzyl bromide and sodium 4-methylbenzene methanesulfonate is 2:1.8;

[0066] Step 2, add p-methoxybenzyl mercaptan to a methanol solution of KOH, stir for 20 minutes, add a toluene solution containing propargyl bromide (80 wt% toluene solution) to the mixture, stir for 10 minutes, dilute with distilled water, and then extract with ether three times. Then add a saturated sodium chloride aqueous solution to wash the combined organic phase, and then dry the organic extract with anhydrous magnesium sulfate. After the ether is dried, the crude product is purified by column chromatography to obtain compound A2;

[0067] The molar ratio of p-methoxybenzyl mercaptan, KOH, and propargyl bromide is 1.5:3:2;

[0068] Step 3, dissolve compound A2 in degassed toluene, add AIBN, heat the mixture to 80°C, slowly drop thioacetic acid dissolved in degassed toluene into the mixture over 1 hour, and continue stirring until TLC shows the maximum conversion rate of the starting reactant. After the reaction is completed, evaporate the solvent to obtain a crude product, which is then purified by column chromatography to obtain compound A3;

[0069] The molar ratio of compound A2, AIBN, and thioacetic acid is 1.5:0.08:2;

[0070] Step 4, dissolving compound A3 in degassed methanol, placing in a low-temperature reactor and cooling to -35°C, slowly injecting KOH dissolved in degassed methanol, stirring for 20 min, cooling the reaction mixture to -70°C, injecting white solid A1 dissolved in degassed methanol into the reaction mixture, heating and stirring, the heating and stirring temperature is 1°C, the heating and stirring time is 3 h, after the reaction is completed, adding saturated ammonium chloride aqueous solution, then extracting with dichloromethane, drying with anhydrous sodium sulfate, evaporating the organic solvent, and purifying the crude product by column chromatography to obtain compound A4;

[0071] The molar ratio of compound A3, KOH, and white solid A1 is 1.5:1.5:2;

[0072] Step 5, dissolve compound A4 in dichloromethane, cool to -70°C in a low-temperature reactor, add m-cpba, react for 2 hours and then warm to room temperature, detect by TLC until the raw material is consumed, add saturated sodium bicarbonate solution, extract three times with dichloromethane, dry with anhydrous sodium sulfate and evaporate the solvent. The crude product is purified by column chromatography to obtain (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide, which is the allicin derivative of the present invention;

[0073] The molar ratio of compound A4 and m-cpba is 2:1.2.

[0074] Example 3

[0075] The preparation method of the allicin derivative of the present invention is specifically implemented according to the following steps:

[0076] Step 1, adding p-bromobenzyl bromide to acetonitrile, and then adding sodium 4-methylbenzene methanesulfonate, and performing condensation reflux reaction, and then reacting at room temperature for 12 hours, and detecting by thin layer chromatography. When p-bromobenzyl bromide is completely reacted, the reaction is stopped, the solvent is evaporated, the product is washed with distilled water, and extracted with ethyl acetate for 3 times. The obtained crude product is purified by column chromatography to obtain a white solid A1;

[0077] The reflux reaction temperature was 85°C and the reflux reaction time was 3h;

[0078] The molar ratio of p-bromobenzyl bromide and sodium 4-methylbenzene methanesulfonate is 1:2.5;

[0079] Step 2, add p-methoxybenzyl mercaptan to a methanol solution of KOH, stir for 15 minutes, add a toluene solution containing propargyl bromide (80 wt% toluene solution) to the mixture, stir for 20 minutes, dilute with distilled water, and then extract with ether three times. Then add a saturated sodium chloride aqueous solution to wash the combined organic phase, and then dry the organic extract with anhydrous magnesium sulfate. After the ether is dried, the crude product is purified by column chromatography to obtain compound A2;

[0080] The molar ratio of p-methoxybenzyl mercaptan, KOH and propargyl bromide is 1.8:4:1.6;

[0081] Step 3, dissolve compound A2 in degassed toluene, add AIBN, heat the mixture to 88°C, slowly drop thioacetic acid dissolved in degassed toluene into the mixture over 2 hours, and continue stirring until TLC shows the maximum conversion rate of the starting reactant. After the reaction is completed, evaporate the solvent to obtain a crude product, which is then purified by column chromatography to obtain compound A3;

[0082] The molar ratio of compound A2, AIBN, and thioacetic acid is 2:0.09:2.2;

[0083] Step 4, dissolving compound A3 in degassed methanol, placing in a low-temperature reactor and cooling to -40°C, slowly injecting KOH dissolved in degassed methanol, stirring for 30 min, cooling the reaction mixture to -70°C, injecting white solid A1 dissolved in degassed methanol into the reaction mixture, heating and stirring, the heating and stirring temperature is 5°C, the heating and stirring time is 3 h, after the reaction is completed, adding saturated ammonium chloride aqueous solution, then extracting with dichloromethane, drying with anhydrous sodium sulfate, evaporating the organic solvent, and purifying the crude product by column chromatography to obtain compound A4;

[0084] The molar ratio of compound A3, KOH, and white solid A1 is 2:2:1.1;

[0085] Step 5, dissolve compound A4 in dichloromethane, cool to -70°C in a low temperature reactor, add m-cpba, react for 2 hours and then warm to room temperature, detect by TLC until the raw material is consumed, add saturated sodium bicarbonate solution, extract with dichloromethane three times, dry with anhydrous sodium sulfate and evaporate the solvent. The crude product is purified by column chromatography to obtain (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide (i.e. Figure 1 Compound A5) is the allicin derivative of the present invention;

[0086] The molar ratio of compound A4 and m-cpba is 1:2.5.

[0087] Example 4

[0088] The preparation method of the allicin derivative of the present invention is specifically implemented according to the following steps:

[0089] Step 1, adding p-bromobenzyl bromide to acetonitrile, and then adding sodium 4-methylbenzene methanesulfonate, and performing condensation reflux reaction, and then reacting at room temperature for 12 hours, and detecting by thin layer chromatography. When p-bromobenzyl bromide is completely reacted, the reaction is stopped, the solvent is evaporated, the product is washed with distilled water, and extracted with ethyl acetate for 3 times. The obtained crude product is purified by column chromatography to obtain a white solid A1;

[0090] The reflux reaction temperature was 90°C and the reflux reaction time was 4h;

[0091] The molar ratio of p-bromobenzyl bromide and sodium 4-methylbenzene methanesulfonate is 2:2.5;

[0092] Step 2, add p-methoxybenzyl mercaptan to a methanol solution of KOH, stir for 25 minutes, add a toluene solution containing propargyl bromide to the mixture, stir for 20 minutes, dilute with distilled water, and then extract with ether three times. Then add a saturated sodium chloride aqueous solution to wash the combined organic phase, and then dry the organic extract with anhydrous magnesium sulfate. After the ether is dried, the crude product is purified by column chromatography to obtain compound A2;

[0093] The molar ratio of p-methoxybenzyl mercaptan, KOH and propargyl bromide is 2:5:2.5;

[0094] Step 3, dissolve compound A2 in degassed toluene, add AIBN, heat the mixture to 90°C, slowly drop thioacetic acid dissolved in degassed toluene into the mixture over 2 hours, and continue stirring until TLC shows the maximum conversion rate of the starting reactant. After the reaction is completed, evaporate the solvent to obtain a crude product, which is then purified by column chromatography to obtain compound A3;

[0095] The molar ratio of compound A2, AIBN, and thioacetic acid is 2:0.1:2.5;

[0096] Step 4, dissolving compound A3 in degassed methanol, placing in a low-temperature reactor and cooling to -40°C, slowly injecting KOH dissolved in degassed methanol, stirring for 30 min, cooling the reaction mixture to -80°C, injecting white solid A1 dissolved in degassed methanol into the reaction mixture, heating and stirring, the heating and stirring temperature is 5°C, the heating and stirring time is 3 h, after the reaction is completed, adding saturated ammonium chloride aqueous solution, then extracting with dichloromethane, drying with anhydrous sodium sulfate, evaporating the organic solvent, and purifying the crude product by column chromatography to obtain compound A4;

[0097] The molar ratio of compound A3, KOH, and white solid A1 is 2:2:2.5;

[0098] Step 5, dissolve compound A4 in dichloromethane, cool to -80°C in a low temperature reactor, add m-cpba (meta-chloroperbenzoic acid), react for 2 hours and then warm to room temperature, detect by TLC until the raw material is consumed, add saturated sodium bicarbonate solution, extract three times with dichloromethane, dry with anhydrous sodium sulfate and evaporate the solvent. The crude product is purified by column chromatography (Hexane:EA=15:1, v / v) to obtain (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide (i.e. Figure 1 Compound A5) is the allicin derivative of the present invention;

[0099] The molar ratio of compound A4 and m-cpba is 2:2.5.

[0100] Example 5

[0101] The natural product derivative functional molecule (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide prepared by the method of the present invention was tested for biological activity in a tumor cell line, specifically:

[0102] Take a bottle of A549 cells in good growth state, prepare the cells into a cell suspension, and inoculate them into the corresponding wells of a 96-well plate, ensuring that an appropriate number of cells are added to each well. Subsequently, these wells are placed in a cell culture incubator for culture to promote cell attachment and growth. After 24-36h, add allicin and allicin derivatives (compound A5) of different concentrations (12.5μM, 25μM, 50μM, 100μM, 200μM, 300μM, 400μM, 1000μM, 2000μM) to the wells, and set up a control group. After 20-30 hours of drug treatment, add CCK-8 reagent to the wells and gently shake the culture plate to help the reagent mix with the culture medium; then put the 96-well plate back into the incubator and continue incubating for about 1-4 hours; after incubation, use an ELISA reader to measure the absorbance of each well at a wavelength of 450 nanometers. This absorbance value can be used as an indicator of the number of living cells; finally, by comparing the absorbance values ​​of the experimental group and the control group, calculate the survival rate of the cells under drug interference, and perform statistical analysis to evaluate the effects of allicin and allicin derivatives (compound A5) on the cells. During the entire experiment, attention should be paid to the consistency of cell seeding density, aseptic operation, the uniform amount of CCK-8 reagent added to each well, the consistency of the experimental conditions used for allicin and allicin derivatives (compound A5), and the setting of repeated wells to ensure the accuracy of the results;

[0103] A549 cells with good growth were selected, washed with phosphate buffered saline (PBS) buffer, digested with trypsin, and then added with culture medium to make a cell suspension, and then an appropriate amount of A549 cells were inoculated in a confocal dish. They were cultured in an incubator for 24-36 hours to adhere to the wall, the culture medium was removed, and then rinsed twice with PBS, and the cells were treated with culture medium containing 250μM allicin and allicin derivatives (compound A5) for 20-30 minutes, and then the commercial ROS probe DCFH-DA was added. Laser confocal microscopy was used for cell imaging, and cell fluorescence images of allicin and allicin derivatives (compound A5) were collected respectively.

[0104] Depend on Figure 3 It can be seen that compared with allicin, the allicin derivative (Compound A5) designed and synthesized by the present invention has better cytotoxicity against tumor cells. In cytotoxicity experiments at different concentrations, Compound A5 showed more excellent tumor cell cytotoxicity.

[0105] Example 6

[0106] Depend on Figure 4 It can be seen that compound A5 can induce tumor cells to produce more ROS than allicin. The mechanism by which the drug inhibits tumor cell proliferation by stimulating cells to produce reactive oxygen species (ROS) can be explained from the following aspects:

[0107] 1. Oxidative stress induces cell apoptosis: When drugs stimulate cells to produce excessive ROS, the balance between oxidation and anti-oxidation inside the cells is disrupted, damaging the cell's DNA, proteins and lipids, causing cell dysfunction. If the damage exceeds the cell's repair capacity, it will trigger the cell apoptosis program, thereby inhibiting the proliferation of tumor cells.

[0108] 2. Affecting the cell cycle: ROS can affect key regulatory proteins of the cell cycle, such as cyclins and cyclin-dependent kinases (CDKs). By oxidatively modifying these proteins, ROS can cause tumor cells to arrest in the DNA synthesis phase (S phase) or mitosis phase (M phase), thereby preventing the proliferation of tumor cells.

[0109] 3. Inhibit the metabolism of tumor cells: Tumor cells usually have high metabolic activity to support their rapid proliferation. ROS can interfere with the energy metabolism of tumor cells, such as mitochondrial function, reduce the production of ATP, and thus inhibit the metabolic activity and proliferation ability of tumor cells.

[0110] 4. Activate DNA repair mechanism: The right amount of reactive oxygen species (ROS) can trigger the DNA repair system inside the cell, such as nucleotide excision repair and base excision repair. However, when the ROS level is too high, DNA damage exceeds the load of the repair system, leading to the accumulation of gene mutations and cell dysfunction, thereby inhibiting the proliferation of tumor cells.

[0111] 5. Affecting signal transduction pathways: Reactive oxygen species (ROS) can act as second messengers and participate in the intracellular signal transduction process. They can oxidize and activate a variety of signal molecules, such as mitogen-activated protein kinases (MAPKs), phosphatidylinositol 3-kinase / protein kinase B (PI3K / Akt) and other signal pathways, and play a vital role in the regulation of cell proliferation, differentiation and apoptosis. By changing the activity of these signal pathways, ROS can inhibit the proliferation of tumor cells.

[0112] 6. Enhance immune response: ROS plays an important role in immune cells and can stimulate immune cells, such as macrophages and natural killer cells (NK cells), to improve their ability to identify and eliminate tumor cells. By enhancing the body's immune surveillance, ROS helps inhibit the escape and proliferation of tumor cells.

[0113] In summary, drugs can affect the biological behavior of tumor cells from multiple levels by stimulating cells to produce ROS, including inducing cell apoptosis, affecting cell cycle, inhibiting metabolism, activating DNA repair, regulating signal transduction and enhancing immune response, thereby achieving the purpose of inhibiting tumor cell proliferation. Figure 4) It can be seen that both compound A5 and allicin induce cells to produce ROS, indicating that both have biological properties of inhibiting tumor cell proliferation. In addition, according to the fluorescence intensity comparison of confocal imaging, it can be seen that at the same concentration (250μM), compound A5 has a stronger induction ability, which also shows that compound A5 has a higher ability to inhibit tumor cell proliferation by inducing cells to produce ROS than allicin.

Claims

1. A method for preparing a garlicin derivative, characterized in that: Specifically, p-bromobenzyl bromide and sodium 4-methylbenzylmethanesulfonate are used for condensation reflux reaction to obtain white solid A1; p-methoxybenzyl mercaptan and propargyl bromide are used for reaction to obtain compound A2; compound A2, AIBN and thioacetic acid are used for reaction to obtain compound A3; compound A3, KOH and white solid A1 are used for reaction to obtain compound A4; compound A4 and m-cpba are used for reaction to obtain (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide, which is an allicin derivative.

2. The method for preparing the allicin derivative according to claim 1, characterized in that: Follow the steps below to implement it: Step 1, adding p-bromobenzyl bromide to acetonitrile, and then adding sodium 4-methylbenzene methanesulfonate, and performing condensation reflux reaction, and then reacting at room temperature, and detecting by thin layer chromatography. When p-bromobenzyl bromide is completely reacted, the reaction is stopped, the solvent is evaporated, washing, extraction, and the obtained crude product is purified by column chromatography to obtain a white solid A1; Step 2, adding p-methoxybenzyl mercaptan to a methanol solution of KOH, stirring, adding a toluene solution containing propargyl bromide to the mixture, stirring, diluting with distilled water, and then extracting with ether three times, then adding a saturated sodium chloride aqueous solution to wash the combined organic phase, and then drying the organic extract with anhydrous magnesium sulfate, spin-drying the crude product and purifying it by column chromatography to obtain compound A2; Step 3, dissolving compound A2 in degassed toluene, adding AIBN, heating the mixture, slowly dropping thioacetic acid dissolved in degassed toluene into the mixture over 1-2 hours, and continuing stirring until TLC shows a maximum conversion rate of the starting reactants. After the reaction is completed, evaporating the solvent to obtain a crude product, and then purifying by column chromatography to obtain compound A3; Step 4, dissolving compound A3 in degassed methanol, placing in a low-temperature reactor and cooling to -35°C to -40°C, slowly injecting KOH dissolved in degassed methanol, stirring for 20-30 min, cooling the reaction mixture to -70°C to -80°C, injecting white solid A1 dissolved in degassed methanol into the reaction mixture, heating and stirring, adding saturated ammonium chloride aqueous solution after the reaction is completed, then extracting with dichloromethane, drying over anhydrous sodium sulfate, evaporating the organic solvent, and purifying the crude product by column chromatography to obtain compound A4; Step 5, dissolving compound A4 in dichloromethane, cooling to -70°C to -80°C in a low-temperature reactor, adding m-cpba, reacting for 2-3 hours and then warming to room temperature, detecting by TLC until the raw material is consumed, adding saturated sodium bicarbonate solution, extracting with dichloromethane three times, drying with anhydrous sodium sulfate and evaporating the solvent, and purifying the crude product by column chromatography to obtain (Z / E)-1-(4-bromophenyl)-2-[3-[(4-methoxyphenyl)sulfinyl]prop-1-en-1-yl] disulfide, which is an allicin derivative.

3. The method for preparing the allicin derivative according to claim 2, characterized in that: In the step 1, the reflux reaction temperature is 80-90° C., the reflux reaction time is 3-4 hours, the reaction time at room temperature is 10-12 hours, and the molar ratio of p-bromobenzyl bromide to sodium 4-methylbenzene methanesulfonate is 1-2:1.2-2.

5.

4. The method for preparing the allicin derivative according to claim 2, characterized in that: In the step 2, the first stirring time is 15-25 min; the second stirring time is 10-20 min; the molar ratio of p-methoxybenzyl mercaptan, KOH and propargyl bromide is 1-2: 2.5-5: 1.1-2.

5.

5. The method for preparing the allicin derivative according to claim 2, characterized in that: In the step 3, the heating temperature is 80-90° C.; the molar ratio of compound A2, AIBN and thioacetic acid is 1-2: 0.05-0.1：1.2-2.5。 6. The method for preparing the allicin derivative according to claim 2, characterized in that: In the step 4, the heating and stirring temperature is 0-5°C, and the heating and stirring time is 2-3h; the molar ratio of compound A3, KOH, and white solid A1 is 1-2:1-2:1.1-2.

5.

7. The method for preparing the allicin derivative according to claim 2, characterized in that: In the step 5, the molar ratio of compound A4 to m-cpba is 1-2:1.2-2.

5.

8. The allicin derivative prepared by the method for preparing the allicin derivative according to any one of claims 1 to 7.

9. Use of the allicin derivative according to any one of claims 1 to 7 in pharmaceutical chemistry.

10. The use according to claim 9, characterized in that Allicin derivatives have the function of inhibiting the proliferation of A549 tumor cells.

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