Diselenide compound and application thereof in preparation of pesticide and medicine

Abstract

The invention relates to a diselenide compound and application of the diselenide compound in preparation of pesticides and medicines, in particular to application of the diselenide compound in preparation of plant virus resisting agents and agricultural bactericides, and application of the diselenide compound in preparation of anti-cancer medicines, especially medicines for resisting lung adenocarcinoma, liver cancer, pancreatic cancer and breast cancer. The diselenide compound disclosed by the invention has a very good prevention effect on tobacco mosaic virus (TMV) under the concentration of 500mg / L, and the living body passivation prevention effect can reach 55.0%. The diselenide compound disclosed by the invention has a very good prevention and treatment effect on tomato early blight, wheat scab, rice blast, phytophthora capsici, sclerotinia rot of colza, cucumber gray mold, rice sheath blight disease, cucumber fusarium wilt, peanut brown spot, apple ring spot, wheat sharp eyespot, corn southern leaf blight, watermelon anthracnose and rice bakanae disease at the concentration of 50mg / L; and the bacteriostatic activity on the rice blast can reach 100%. The diselenide compound disclosed by the invention has a very strong inhibition effect on various cancer cells, and the IC50 of the diselenide compound on pancreatic cancer cells can reach 4.33 mu M.

Description

Technical Field

[0001] This invention relates to a diselenyl ether compound and its use in the preparation of pesticides and pharmaceuticals, specifically to the use of a diselenyl ether compound in the preparation of antiviral agents for plants and agricultural fungicides, and in the preparation of anticancer drugs, particularly those for lung adenocarcinoma, liver cancer, pancreatic cancer, and breast cancer. Technical Background

[0002] Selenium, belonging to group VIA, has an abundance of only 0.5 parts per hundred million in the Earth's crust, classifying it as a rare and dispersed nonmetallic element. It was first discovered in 1817 by the Swedish chemist Berzelius and formally identified and named in 1818. In 1957, the National Institutes of Health in the United States first discovered that selenium is an important protective factor against nutritional liver necrosis, thus initiating research into the relationship between selenium and health (Schwarz K, et al. Proc. Soc. Exp. Biol. Med. 1957, 95(4), 621-625). In 1973, Rotruck et al. proposed that selenium is an important component of glutathione peroxidase (GPx), an enzyme closely related to immunity, aging, antioxidation, and anticancer activity, thus establishing selenium as an essential trace element for the human body at the molecular level (Rotruck JT, et al. Science, 1973, 179(4073), 588-590). Selenium deficiency in the human body can lead to premature aging, Keshan disease, Kashin-Beck disease, lethargy, and increased susceptibility to colds. Severe selenium deficiency can also cause cardiomyopathy and heart failure. On the other hand, excessive selenium intake can cause symptoms such as dulled pain sensation in the skin, numbness in the limbs, dizziness, loss of appetite, hair loss, thickened nails, and indigestion.

[0003] China possesses more than one-third of the world's selenium reserves, with selenium mines mainly distributed in the central, southern, and northwestern regions of the country. The Shuanghe Yutangba selenium deposit in Enshi Prefecture, Hubei Province, is the only proven independent selenium deposit in the world. Although the abundance of selenium is comparable to that of precious metals such as gold and platinum, its industrial price is not high. In June 2025, industrial selenium was only 220 yuan per kilogram, which facilitates the research and development of selenium-containing bioactive compounds.

[0004] Overall, selenium is a noteworthy chemical element, and its introduction into organic compound molecules has significant research value. In recent years, the excellent biological activities of organoselenium compounds have been gradually discovered, and their research value has also received increasing attention. For example, ebuselenline was discovered early on to have antioxidant, anti-inflammatory, antiviral, and cardiovascular protective activities, and has entered the clinical trial stage (Zhou Jingxuan et al., Chinese Journal of Clinical Pharmacology and Therapeutics, 2020, 25(2), 233-240). Another example is diphenyldiselenoether, which has relatively low toxicity and anti-tumor and anti-diabetic effects, and continuous research has been conducted in this area (Wang Xing et al., Acta Pharmaceutica Sinica, 2022, 57(11), 3259-3267). Wang Jianguo et al. designed and synthesized a class of organic compounds containing selenium sulfide structures and found that some of these compounds (such as compound A) showed good anti-tobacco mosaic virus effects (Shang MH, et al. Pest Manag. Sci. 2023, 79, 1885-1896). Wang Jianguo and others also designed a class of asymmetric aromatic diselenyl ether compounds, which have good passivation, therapeutic and protective effects against TMV (Chinese Invention Patent Application No. 202510199516.1, not yet published).

[0005] Summary of the Invention

[0006] The purpose of this invention is to provide a diselenyl ether compound and its uses in the preparation of pesticides and pharmaceuticals, particularly its uses in the preparation of antiviral agents for plants and agricultural fungicides, and its uses in the preparation of anticancer drugs, especially those for lung adenocarcinoma, liver cancer, pancreatic cancer, and breast cancer. One diselenyl ether compound of this invention is...

[0007]

[0008]

[0009]

[0010] And the lithium, sodium, potassium and ammonium salts of the above compounds.

[0011] The diselenyl ether compounds SJR-1 to SJR-16 of the present invention are obtained by the following reaction formulas.

[0012]

[0013] This process uses ebuselenoline and its derivatives or analogs as raw material 1, and substituted selenophenol as raw material 2. Raw material 2 is dissolved in acetone, and then raw material 1 is added and stirred at room temperature. The reaction is completed in about 5 to 10 minutes, and the product is purified by column chromatography to obtain the target compound 3.

[0014] The diselenyl ether compounds SJR-17 to SJR-52 of the present invention are obtained by the following reaction formula.

[0015]

[0016] This scenario uses a selenocyanate compound as raw material 1 and a substituted selenophenol as raw material 2. Raw material 2 and a trace amount of aluminum oxide powder are dissolved in acetone, followed by the addition of raw material 1. The mixture is stirred at room temperature and the reaction is completed in about 5 to 10 minutes. The mixture is then purified by column chromatography to obtain the target compound 3.

[0017] The diselenyl ether compounds SJR-53 to SJR-68 of the present invention are obtained by the following reaction formulas.

[0018]

[0019] This scenario uses compound 1 and substituted selenophenol as raw material 2. Compound 1 is dissolved in acetone and dimethyl sulfoxide, followed by the addition of raw material 2 and stirring at room temperature. The reaction is completed in about 5 to 10 minutes, during which the system changes from turbid to clear and transparent. After concentration and filtration, hexane is added to finally precipitate the product, yielding the target compound 3.

[0020] Through greenhouse studies on the antiviral activity against plant viruses, the diselenyl ether compounds of this invention effectively inhibit tobacco mosaic virus, pepper virus, rice virus, tomato virus, sweet potato virus, potato virus, cucurbit viruses, and maize dwarf mosaic virus, etc., and can effectively control viral diseases in various crops such as tobacco, pepper, rice, tomato, melons, grains, vegetables, and legumes, especially suitable for controlling tobacco mosaic virus. The diselenyl ether compounds of this invention have excellent protective efficacy against tobacco mosaic virus at a concentration of 500 mg / L, with a live inactivation efficacy of up to 55.0%.

[0021] Through in vitro inhibitory activity studies on agricultural fungi, the diselenyl ether compounds of this invention exhibit excellent control effects against early blight of tomato, scab of wheat, rice blast, Phytophthora blight of pepper, sclerotinia rot of rapeseed, gray mold of cucumber, sheath blight of rice, wilt of cucumber, brown spot of peanut, ring rot of apple, sheath blight of wheat, small leaf spot of corn, anthracnose of watermelon, and bakanae disease of rice. At a concentration of 50 mg / L, the compounds of this invention achieve 100% fungicidal activity against rice blast fungus.

[0022] Bioactivity studies using cell models demonstrated that the diselenyl ether compounds of this invention exhibit excellent anti-cancer effects, showing significant inhibitory effects against lung adenocarcinoma cells, liver cancer cells, pancreatic cancer cells, and breast cancer cells. The compounds of this invention also inhibit the IC50 of pancreatic cancer cells. 50 The value can reach 4.33 μM.

[0023] This invention also provides an antiviral agent for plants. At an effective dosage, this antiviral agent can effectively inhibit tobacco mosaic virus, pepper virus, rice virus, tomato virus, sweet potato virus, potato virus, cucurbit viruses, and maize dwarf mosaic virus, etc. It can effectively prevent and control viral diseases in various crops such as tobacco, pepper, rice, wheat, corn, tomato, melons, grains, vegetables, and beans, and is especially suitable for the prevention and control of tobacco mosaic disease.

[0024] This antiviral agent for plants may contain the aforementioned diselenyl ether compounds and one or more agriculturally acceptable carriers. The carriers include diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorbents, lubricants, or synergists conventional in the pesticide field. Its formulation may be an emulsifiable concentrate, wettable powder, soluble powder, water-in-oil emulsion, microemulsion, aqueous solution, suspension concentrate, microcapsule, or water-dispersible granule.

[0025] This invention also provides an agricultural fungicide that, at an effective dosage, has excellent control efficacy against agricultural pathogens, particularly early blight of tomato, scab of wheat, rice blast, Phytophthora blight of pepper, sclerotinia rot of rapeseed, gray mold of cucumber, sheath blight of rice, wilt of cucumber, brown spot of peanut, ring rot of apple, sheath blight of wheat, small leaf spot of corn, anthracnose of watermelon, and bakanae disease of rice.

[0026] This agricultural fungicide may contain the aforementioned diselenyl ether compounds and one or more agriculturally acceptable carriers. The carriers include diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorbents, lubricants, or synergists conventional in the pesticide field. Its formulation may be an emulsifiable concentrate, wettable powder, soluble powder, water-in-oil emulsion, microemulsion, aqueous solution, suspension concentrate, microcapsule, or water-dispersible granules.

[0027] The present invention also provides an anticancer drug that, at an effective dose, has a good inhibitory effect on lung adenocarcinoma cells, liver cancer cells, pancreatic cancer cells and breast cancer cells.

[0028] This anticancer drug may contain the aforementioned diselenyl ether compounds and one or more pharmaceutically acceptable carriers. The carriers include diluents, excipients, fillers, binders, humectants, disintegrants, absorption enhancers, surfactants, adsorbents, lubricants, or synergists commonly used in the pharmaceutical field. Its dosage form may be an injection, tablet, pill, capsule, suspension, or emulsion. Attached Figure Description

[0029] Figure 1 It is the NMR selenium spectrum of compound SJR-33 ( 77 SeNMR), Figure 2 It is the single-crystal structure of compound SJR-50. Figure 3 This is a high-resolution mass spectrometry (HRMS) spectrometer of compound SJR-51. Detailed Implementation

[0030] The essential features of the present invention can be seen from the following embodiments, but these embodiments are only illustrative and not intended to limit the invention.

[0031] The raw material 1 used in this invention is the following compound (the numbers below are the corresponding CAS numbers), which can be purchased from Aurora Chemicals in the United States and Chemhere Chemicals in Hong Kong, China.

[0032]

[0033]

[0034] When the raw material 1 used in this method is one of the following compounds, it is an unreported intermediate and needs to be prepared.

[0035]

[0036] 1-1 and 1-2 are prepared by the following method

[0037] Compounds 1-3 were prepared by the following method

[0038]

[0039] The raw material 2 used in this invention is the following compound

[0040]

[0041]

[0042] Prepared by the following method

[0043]

[0044] Example 1. Preparation of raw material 1-1

[0045] In a 100 mL single-necked flask, metallic sodium (0.46 g, 20 mmol) was added to isopropanol (60 mL), and the mixture was heated to reflux until the sodium solid completely disappeared. Methyl anthranilate was then added to the reaction system, and the reaction was allowed to proceed overnight. After the reaction was completed as monitored by TLC, the solvent was removed by concentration under reduced pressure. Ethyl acetate and water were added for extraction, and the organic phase was washed with saturated sodium chloride solution (100 mL × 3). The mixture was dried over anhydrous magnesium sulfate overnight, filtered, and concentrated under reduced pressure to obtain the product, with a yield of 93%.

[0046] In a 250 mL single-necked flask, isopropyl anthranilate (3.02 g, 20 mmol) was added. 6M hydrochloric acid solution (12 mL) was slowly added under ice-water bath conditions. After stirring for 15 min, freshly prepared 3M sodium nitrite solution (8 mL) was slowly added dropwise, stirring until the solution was clear and transparent. Sodium acetate was added to adjust the pH to 4-5, and potassium selenocyanate (2.88 g, 20 mmol) was quickly added. Bubbles were rapidly generated in the solution, and an orange-yellow / red solid was formed. Ethyl acetate (100 mL) was added, followed by separation. The solution was extracted with ethyl acetate (50 mL × 2), and the organic phases were combined. The organic phase was washed with saturated sodium chloride solution (100 mL × 3), dried over anhydrous sodium sulfate overnight, filtered, and concentrated under reduced pressure to give a yellow solid with a yield of 75%. 1 The H NMR data are as follows:

[0047] 1 H NMR (400MHz, Acetone-d6) δ8.19 (dd, J=7.8, 1.6Hz, 1H, Ph-H), 8.02 (dd, J=8.1, 1.0Hz, 1H, Ph-H), 7.78 (td, J=7. 8,1.6Hz,1H,Ph-H),7.58(td,J=7.6,1.1Hz,1H,Ph-H),5.29(hept,J=6.2Hz,1H,CH),1.42(d,J=6.3Hz,6H,CH3).

[0048] Similarly, the raw materials 1-2 of the present invention can be prepared.

[0049] Example 2. Preparation of raw materials 1-3

[0050] In a 250 mL single-necked flask, dissolve 6.4 g (46 mmol) of anthranilic acid in 32 mL of 6 M hydrochloric acid solution and stir in an ice bath. Dissolve 5.6 g (811 mmol) of sodium nitrite in 16 mL of water and slowly add it dropwise to the above solution. After adding about 13 mL, stop adding; at this point, the reaction system is a clear, yellow color. Then adjust the pH to alkaline using 2.4 mL of 4 M sodium hydroxide solution, and then place the reaction mixture in an ice bath until ready for use.

[0051] In a 500 mL single-necked flask, sodium borohydride (0.76 g, 20 mmol), sodium hydroxide (3.2 g, 80 mmol), and hexadecyltrimethylammonium bromide (0.1 g) were dissolved in water (50 mL). After stirring for 15 min, selenium powder (4 g, 50 mmol) was added. The mixture was stirred at room temperature for 15 min, at which point the reaction system turned purplish-black. The mixture was then heated to 75 °C and reacted for 1.5 h, at which point the reaction system turned reddish-brown. After the reaction mixture cooled to room temperature, it was transferred to an ice-water bath and allowed to stand for 15 min. The freshly prepared diazonium salt solution was then slowly added dropwise. After the addition was complete, the temperature was raised to 50 °C and the reaction was allowed to proceed for 3 h. After the reaction mixture cooled, it was filtered. The filtrate was acidified with 3 M hydrochloric acid solution and filtered again to obtain a pink filter cake. The filter cake was dissolved in a saturated sodium bicarbonate solution and heated under reflux for 1 hour. After the reaction cooled, it was filtered to obtain a clear yellow solution. The solution was acidified with 3M hydrochloric acid solution, and a large amount of yellow solid was observed to precipitate. The solution was then filtered and the filter cake was dried overnight in a vacuum drying oven to obtain 5.325 g of yellowish-brown solid product, with a yield of 51%.

[0052] In a 100 mL single-necked flask, 2,2'-diselenobisbenzoic acid (1.0 g, 2.5 mmol) was dissolved in 13 mL of thionyl chloride and heated to 80 °C for 2.5 h. After the reaction was completed, the solvent was removed by concentration under reduced pressure. The solid was redissolved in n-hexane, filtered, and the filtrate was concentrated under reduced pressure to obtain a bright yellow solid for later use.

[0053] In a 500 mL single-necked flask, 7-amino-4-methylcoumarin (AMC) (0.968 g, 5.5 mmol) was dissolved in 270 mL of ultra-dry dichloromethane. The solution was stirred in an ice bath for 3 h, and then filtered to obtain a dichloromethane solution of AMC. The obtained 2-chloroselenobenzoyl chloride was dissolved in 20 mL of ultra-dry dichloromethane and slowly added dropwise to the dichloromethane solution of AMC. The system became turbid. After the addition was complete, the mixture was transferred to room temperature and reacted overnight. The filter cake was the product, with a yield of 65%. (The following appears to be a separate, unrelated sentence: Starting materials 1-3...) 1 The H NMR data are as follows:

[0054] 1 H NMR (400MHz, Acetone-d6) δ8.00-7.93(m,2H,Ph-H),7.82-7.70(m,2H,Ph-H),7.61-7.49(m ,2H,Ph-H),7.26(td,J=7.6,1.9Hz,1H,Ph-H),6.23(t,J=1.2Hz,1H,CH),2.49(s,3H,CH3).

[0055] Example 3. Preparation of raw materials 2-14

[0056] In a 250 mL three-necked flask, dissolve 8.16 g (30 mmol) of p-trifluoromethyliodobenzene, 0.24 g (3 mmol) of nano-copper oxide (40 nm, 3 mmol), and 4.74 g (60 mmol) of selenium powder in 100 mL of dimethyl sulfoxide. Then, purge with nitrogen, stir, and heat to 90 °C. After reacting for 15 minutes, add 3.36 g (60 mmol) of potassium hydroxide and continue to maintain the temperature at 90 °C for 5 hours. After the reaction is complete, quench with water (200 mL) and extract. Wash the mixture with ethyl acetate (300 mL × 3) and separate the organic phase. Then wash the organic phase with saturated brine (300 mL × 3), combine the organic phases, dry with anhydrous magnesium sulfate (6 g), filter, and concentrate under reduced pressure. Column chromatography (eluent: petroleum ether) is used to purify the solid, di-p-trifluoromethylphenyl diselenyl ether, in a golden-yellow powder, with a yield of 67.5%.

[0057] Take a 50 mL two-necked flask, add the intermediate di-p-trifluoromethylphenyl diselenyl ether (0.2694 g, 0.6 mmol), add 15 mL of ethanol, stir until dissolved, purge with nitrogen, and after 15 minutes, add sodium borohydride (0.068 g, 1.8 mmol), continue stirring for 10 minutes, add citric acid (0.576 g, 3.0 mmol), and stir for 15 minutes. After the reaction is complete, extract, wash the organic phase with diethyl ether (80 mL), collect the organic phase, dry with anhydrous magnesium sulfate (6 g), filter, concentrate the organic phase under reduced pressure to obtain a yellow oily substance 2-5, which can be used directly in the next step without purification (because this type of substance is easily oxidized by air, it must be used immediately for the next reaction).

[0058] Similarly, raw materials 2-1, 2-2, 2-3, 2-4, 2-5, 2-6, 2-7, 2-8, 2-9, 2-10, 2-11, 2-12, 2-13, 2-15, 2-16, 2-17, 2-18, 2-19 and 2-20 can be prepared.

[0059] Example 4. Preparation of compound SJR-1

[0060] Selenol 2-1 (0.3096 g, 1.8 mmol) was dissolved in 15 mL of acetone and stirred. Then, ebuselenoline (CAS No. 60940-34-3, 0.2475 g, 0.9 mmol) was quickly added, and the mixture was stirred at room temperature for 5 minutes. After the reaction was complete, the organic phase was concentrated under reduced pressure. The mixture was then purified by column chromatography (eluent: n-hexane / ethyl acetate (v / v = 15 / 1)). The purified organic phases were combined and concentrated under reduced pressure to 1 / 10 of the original volume. 20 mL of n-hexane was added, and the mixture was shaken to precipitate the product. The product was filtered, and the filter cake was washed with a large amount of n-hexane to obtain a white powdery solid with a yield of 5%.

[0061] Similarly, SJR-2 up to SJR-16 of the present invention can be prepared.

[0062] Example 5. Preparation of compound SJR-17

[0063] Selenol 2-7 (0.3744 g, 1.8 mmol) and trace amounts of alumina were dissolved in 15 mL of acetone. After stirring, o-cyanobenzoate (CAS No. 78337-05-6, 0.241 g, 1.0 mmol) was quickly added, and the mixture was stirred at room temperature for 5 minutes. The residue was purified by silica gel column chromatography (n-hexane → n-hexane: ethyl acetate = 200:1). The purified organic phases were combined and concentrated under reduced pressure to give a yellow solid in 6% yield.

[0064] Similarly, SJR-18 up to SJR-52 of the present invention can be prepared.

[0065] Example 6. Preparation of compound SJR-53

[0066] Raw material 1-3 (0.3577 g, 1 mmol) was dissolved in acetone (15 mL) and dimethyl sulfoxide (2 mL). Raw material selenophenol 2-19 (0.2826 g, 1.8 mmol) was added to the solution. The system was observed to change from turbid to clear and transparent. The reaction was stopped, and the solvent was removed by concentration under reduced pressure. The residue was dissolved in ethyl acetate (60 mL) and washed with saturated sodium chloride solution (50 mL × 5). The organic phase was collected, dried over anhydrous sodium sulfate for 20 min, filtered, and concentrated under reduced pressure to 1 / 10 of the original volume. 20 mL of n-hexane was added, and the mixture was shaken to precipitate the product. The product was then filtered, and the filter cake was washed with a large amount of n-hexane to obtain a white powder, with a yield of 57%.

[0067] Similarly, SJR-54 up to SJR-68 of the present invention can be prepared.

[0068] Properties, yields, melting points, and other characteristics of SJR-1 to SJR-68 1 HNMR, 13 CNMR, 77 SeNMR, 19F NMR, and HRMS data are shown in Table 1.

[0069] Table 1. Physicochemical characterization and spectral data of SJR-1 to SJR-68

[0070]

[0071]

[0072]

[0073]

[0074]

[0075]

[0076]

[0077]

[0078]

[0079]

[0080]

[0081]

[0082]

[0083]

[0084]

[0085]

[0086]

[0087]

[0088]

[0089]

[0090] Example 7. Indoor assay of the compound's activity against tobacco mosaic virus

[0091] Virus purification: Using the Gooding method, upper and middle leaves of systemically infected host plants inoculated for more than 3 weeks were selected, homogenized in phosphate buffer, filtered, centrifuged, subjected to two polyethylene glycol precipitation treatments, centrifuged again, and the precipitate was washed with phosphate buffer to suspend it, thus obtaining the crude extract of tobacco mosaic virus (hereinafter referred to as TMV). The entire experiment was carried out under a sterile environment at 4°C.

[0092] The viral mass concentration in the crude viral extract was calculated using ultraviolet spectrophotometry.

[0093] Virus mass concentration (mg / L) = (dilution factor × A) 260 ) / 3.1

[0094] Test reagent preparation: Weigh the test compound, dissolve it in DMF, and prepare a solution of 1×10⁻⁶. 5The mother liquor was prepared at a concentration of mg / L and then quickly diluted with a series of aqueous solutions containing 1‰ Tween 80 to the required mass concentration.

[0095] In vivo inactivation: Select 3-5 leaf stage *Nicotiana sambac* plants with similar growth. Mix the agent with an equal volume of virus solution, achieving a final virus concentration of 10 mg / L. After 30 minutes of inactivation, apply the mixture to the entire leaf of the *Nicotiana sambac* plant sprinkled with carborundum. A control group with the same concentration of virus-water mixture was included, with each group containing three replicates. Incubate in a greenhouse for 2-3 days until the virus is fully infected, then check the results and calculate the control efficacy.

[0096] Efficacy (%) = (Disease index of control group - Disease index of treatment group) / Disease index of control group × 100

[0097] The results of in vivo inactivation of the compound and the control drug ribavirin against TMV are shown in Table 2.

[0098] Table 2. In vivo passivation and prevention effects of the target compound at a concentration of 500 mg / L on TMV.

[0099]

[0100]

[0101] It can be seen that the compounds of the present invention exhibit good in vivo passivation protection against TMV, among which compounds SJR-28, SJR-45, SJR-48, SJR-50, SJR-51, SJR-56 and SJR-62 are superior to ribavirin.

[0102] Example 8. Determination of the bactericidal activity of the compound

[0103] The tested fungi were *Alternaria solani* Sorauer. (early blight of tomato), *Fusarium graminearum* Schw. (wheat sclerosis), *Pyricularia oryzae* Cav. (rice blast fungus), *Phytophthora capsica.* (pepper bud blight), *Sclerotinia sclerotiorum.* (rapeseed sclerotinia sclerotiorum.), *Botrytis cinerea.* (cucumber gray mold), *Thanatephorus cucumeris* (Frank) Donk. (rice sheath blight), *Fusarium oxysporum* (Schl.) F.sp. cucumerinum Owen. (cucumber wilt), *Cercospora arachidicola.* (peanut brown spot), *Physalospora piricola.* (apple ring rot), *Rhizoctonia cerealis.* (wheat sheath blight), and *Helminthosporium* (corn leaf spot). The fungi causing watermelon anthracnose (Colletotrichum orbiculare.) and rice bakanae disease (Fusarium moniliforme Sheld.) are also mentioned.

[0104] The in vitro antibacterial activity of the target compound was determined, with chlorothalonil as a control, and the room temperature was maintained at 25±1℃.

[0105] In vitro antibacterial activity was determined using the mycelial growth rate method: the compound was prepared at a concentration of 3.0 × 10⁻⁶ using DMSO as the solvent. 4 Dilute the test solution (mg / L) with Tween solution to a concentration of 50 mg / L. Under aseptic conditions, add 1.0 mL of the test solution to 9 mL of PDA medium and inoculate with the test strain. The blank control group is medium supplemented with 1 mL of sterile water. Incubate the above medium at 25 ± 1 °C for 72 hours. Measure the diameter (D) of the colonies and calculate the inhibition rate using the following formula:

[0106] Inhibition rate = (D 空白 -D 药剂处理 ) / D 空白 ×100%

[0107] The bactericidal activities of the compounds are shown in Table 3.

[0108] Table 3. Inhibition rate (%) of the target compound against 14 agricultural pathogenic fungi at a concentration of 50 mg / L.

[0109]

[0110]

[0111] In the table, A represents early blight pathogen of tomato, B represents Fusarium head blight pathogen of wheat, C represents rice blast pathogen of rice, D represents Phytophthora infestation of pepper, E represents Sclerotinia sclerotiorum var. ...

[0112] It can be seen that most of the compounds of this invention have good inhibitory effects on crop pathogenic fungi. Some compounds have an antibacterial effect of more than 90% at a concentration of 50 mg / L. Among them, SJR-15 and SJR-35 have an inhibitory effect of 100% on rice blast fungus, which is significantly better than that of chlorothalonil (76.0%). SJR-3, SJR-5, SJR-9, SJR-15, SJR-18, SJR-20, SJR-21, SJR-22, SJR-24, SJR-26, SJR-27, SJR-28, SJR-30, SJR-33, SJR-34, SJR-35, SJR-37 and SJR-42 have an inhibitory effect of significantly better than that of apple scab fungus (68.1%).

[0113] Example 9. Anticancer bioactivity of the compound in a cell model

[0114] The cells used in the experiment were obtained from the Shanghai Cell Bank of the Chinese Academy of Sciences. The cells used in this experiment were lung adenocarcinoma H820, liver cancer Huh7, pancreatic cancer PANC1, breast cancer MCF-7, and normal cells 3T3.

[0115] Preparation of complete culture medium: 45 mL DMEM / RPMI 1640, 5 mL FBS, and 0.55 mL Penicillin-Streptomycin.

[0116] 10×MTT:

[0117] Weigh 2.5g of MTT powder and dissolve it in 450mL of 10mM PBS buffer. After mixing thoroughly, add 10mM PBS buffer to bring the volume to 500mL. Then filter the solution through a 0.22μm filter membrane for sterilization. Aliquot the filtrate and store it at -20℃.

[0118] Cell resuscitation:

[0119] (1) Place the complete culture medium, PBS, etc. required for the experiment into the clean bench and turn on the ultraviolet lamp to sterilize for 15 minutes.

[0120] (2) Quickly remove the cells, put them in warm water, and shake them to dissolve them quickly.

[0121] (3) Centrifuge at 800 rpm for 5 min and discard the supernatant.

[0122] (4) Add an appropriate amount of culture medium to a cell culture dish of appropriate size, resuspend the cell pellet with 1 mL of culture medium, add it evenly to the cell culture dish, and gently shake it to distribute it evenly.

[0123] (5) Place in a cell culture incubator for culture.

[0124] Cell passage:

[0125] (1) Observe the cell density. When the cell density reaches about 80% and the culture medium turns yellow, remove the culture medium and wash with PBS.

[0126] (2) Add an appropriate amount of pancreatic enzyme and place it in a cell culture incubator for 2-5 minutes.

[0127] (3) Remove cells that start to detach from the bottom of the culture dish when observed with the naked eye. Stop digestion, add complete culture medium, and pipette until the cells are dispersed and fully dissolved in the culture medium. Transfer to a 10 mL tube, centrifuge, discard the supernatant, and wash plasma cells with PBS 1-3 times.

[0128] (4) Finally, resuspend the cell pellet and passage it into a culture dish at a ratio of 1:3 to 1:5 and place it in a cell culture incubator for culture.

[0129] Cell cryopreservation:

[0130] (1) Add 4 mL of fetal bovine serum and 1 mL of DMSO to 5 mL of complete culture medium, mix thoroughly, and store in a refrigerator at 4 °C.

[0131] (2) Before freezing cells, a gradient cooling box needs to be prepared in advance, and the level of isopropanol in the box should reach the marked line on the surface of the cooling box to restore it to room temperature.

[0132] (3) The cells to be frozen should be processed according to the cell digestion method in cell passage, centrifuged, and then the supernatant should be discarded and washed with PBS 1-3 times.

[0133] (4) Based on the number of cells, add an appropriate amount of cell cryopreservation solution to the cell pellet, resuspend it thoroughly, and then dispense it into cryovials.

[0134] (5) Place the cryovials into a gradient cooling box and put them in a -80℃ freezer. After 24 hours, they can be transferred to liquid nitrogen for long-term storage.

[0135] MTT assay for drug activity:

[0136] (1) Digest the cells according to the method of cell digestion in cell passage, centrifuge, wash with PBS 1-3 times, and resuspend in an appropriate amount of complete culture medium.

[0137] (2) Use a hemocytometer to count the cells. If there are many cells, a small amount of cell suspension should be diluted before counting.

[0138] (3) According to the formula 5000 cells / well × well plate = required number of cells, 200 μL / well × number of wells = total volume of cell suspension, fully resuspend the cell suspension in sufficient culture medium, and then use a multichannel pipette to aspirate 200 μL of cell suspension into each well of the 96-well plate.

[0139] (4) After 24 hours of plate preparation, the compound to be tested is serially diluted to the final concentration, and then the corresponding concentration of the compound is added to each well.

[0140] (5) 72 hours after drug administration, add MTT solution to each cell well to make the final concentration 500 μg / mL, and place in a cell culture incubator to stand for 3-5 hours.

[0141] (6) Quickly remove the culture medium from the well plate and add an appropriate amount of dimethyl sulfoxide to completely dissolve the formazan precipitate.

[0142] (7) The 96-well plate to be tested was placed flat in the microplate reader, and its absorbance was measured at a wavelength of 570 nm. The data was exported and analyzed. Diphenyldiselenoether was used as a control drug in the cell model experiment.

[0143] Table 4 lists the activities of the target compounds against lung adenocarcinoma cells and normal cells. Table 5 lists the broad-spectrum anticancer effects of the highly active compounds against liver cancer cells, pancreatic cancer cells, and breast cancer cells.

[0144] Table 4. Experimental results of the compounds on lung adenocarcinoma cells and normal cells in cell models.

[0145]

[0146]

[0147] It can be seen that the compounds of this invention have good activity against H820 lung adenocarcinoma cells in cell models. Many compounds have better activity than diphenyldiselenoether, among which SJR-33 has the strongest activity, with an IC50 value of [missing value]. 50The concentration was 5.38 μM. The ratio of the toxicity of diphenyldiselelenide to normal cells to that to lung adenocarcinoma cells was 0.39 (17.11 / 44.16), while the ratio of the toxicity of the compound SJR-33 to normal cells to that to lung adenocarcinoma cells was 1.93 (10.39 / 5.38), and the ratio of the toxicity of SJR-37 to normal cells to that to lung adenocarcinoma cells was 2.83 (18.74 / 6.63), indicating better cell selectivity.

[0148] Table 5. Broad-spectrum anticancer effects of highly active compounds in cell models

[0149]

[0150] It can be seen that the highly active compounds exhibited higher anticancer activity than diphenyldiselelenide against a variety of cancer cells. Among them, SJR-33 showed higher IC50 activity against pancreatic cancer cells. 50 The value is 4.33 μM.

Claims

1. A diseleno ether compound, characterized by This diselenyl ether compound is and salts of the above compounds.

2. The use of the diselenyl ether compound of claim 1 in the preparation of an antiviral agent for plants.

3. The use of the diselenyl ether compound of claim 1 in the preparation of agricultural fungicides.

4. Use of the diselenyl ether compound of claim 1 in the preparation of anticancer drugs.

5. Use according to claim 2, characterized in that The aforementioned antiviral agent is effective against tobacco mosaic virus, pepper virus, rice virus, tomato virus, sweet potato virus, potato virus, cucurbit virus, and maize dwarf mosaic virus. It can effectively prevent and control viral diseases in tobacco, pepper, rice, tomato, cucurbits, corn, wheat, grains, vegetables, and legumes, and is especially suitable for the prevention and control of tobacco mosaic virus.

6. Use according to claim 3, characterized in that The aforementioned agricultural fungicide is an agricultural fungicide that targets early blight of tomato, scab of wheat, rice blast, Phytophthora blight of pepper, sclerotinia stem rot of rapeseed, gray mold of cucumber, sheath blight of rice, wilt of cucumber, brown spot of peanut, ring rot of apple, sheath blight of wheat, small leaf spot of corn, anthracnose of watermelon, and bakanae disease of rice.

7. Use according to claim 4, characterized in that The aforementioned anticancer drugs are for lung adenocarcinoma, liver cancer, pancreatic cancer, and breast cancer.

8. An agent against plant viruses, characterized in that It contains a diselenide compound as described in claim 1 and one or more agriculturally acceptable carriers; the carriers include diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorbents, lubricants, or synergists commonly used in the pesticide field.

9. An agricultural fungicide characterized by It contains a diselenide compound as described in claim 1 and one or more agriculturally acceptable carriers; the carriers include diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorbents, lubricants, or synergists commonly used in the pesticide field.

10. An anticancer drug, characterized by It contains a diselenide compound as described in claim 1 and one or more pharmaceutically acceptable carriers; the carriers include diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorbents, lubricants, or synergists commonly used in the pharmaceutical field.

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