Caboxyl tertiary amine derivatives, methods of making, and use in the preparation of botanical fungicides

Abstract

The present application provides a carvacrol tertiary amine derivative, a preparation method and an application in preparing a plant source fungicide, the carvacrol tertiary amine derivative contains a cyclic secondary amine substituent, the preparation method of the derivative is that an intermediate II is dissolved in ethanol, then a cyclic secondary amine is added, heating and stirring are carried out, reflux is kept, after TLC detection reaction is completed, ethanol is removed under reduced pressure, then extraction, water washing and purification are carried out, and the carvacrol tertiary amine derivative is prepared. The carvacrol tertiary amine derivative prepared by the present application exhibits obvious bacteriostatic activity on cucumber anthracnose fungus, tomato botrytis blight fungus, wheat scab fungus and wheat take-all fungus. The preliminary bacteriostatic activity of carvacrol tertiary amine derivatives 1, 3, 10 and 14 on wheat take-all fungus is 60.8%-83.2%, which is significantly better than that of positive control hymexazol (48.6%) and reference control carvacrol (45.0%).

Description

TECHNICAL FIELD

[0001] The present application relates to a cardanol tertiary amine derivative, a preparation method and application in preparing a plant source fungicide, and belongs to the technical field of heterocyclic compounds. BACKGROUND

[0002] Cardanol is a natural antifungal product extracted and separated from the rhizome of Asarum sieboldii

Lee JY, Moon SS, Hwang BK. Isolation and antifungal activity of kakuol, a propiophenone derivative from Asarum sieboldii rhizome. Pest Management Science 2005, 61, 821-825.

[0003] Recent research results show that kakuol and its various derivatives have broad-spectrum antifungal activity against plant pathogenic fungi

Lee JY, Moon SS, Hwang BK. Isolation and antifungal activity of kakuol, a propiophenone derivative from Asarum sieboldii rhizome. Pest Management Science 2005, 61, 821-825; Musso L, Dallavalle S, Merlini L, Farina, G. Synthesis and Antifungal Activity of 2-Hydroxy-4,5-methylenedioxyaryl Ketones as Analogues of Kakuol. Chemistry & Biodiversity 2010, 7(4), 887-897; Shi W, Nandinsuren T, Zhou WM, Gao JM. Natural products as sources of new fungicides (I): synthesis and antifungal activity of Kakuol derivatives againstphytopathogenic fungi. Mongolian Journal of Chemistry 2014, 15 (41), 94-100.

Sui GQ, Shu LL, Zhang AL, Li D, Cao SH. Sulfonate Derivatives Containing a Kakuol Moiety as Potential Fungicidal Candidates: Design, Synthesis and Antifungal Activity Evaluation. Chem. Pharm. Bull., 2024, 72, 186-189.

[0004] The tertiary amine fungicides are a kind of broad-spectrum fungicides, and are widely used in the field of plant protection due to the significant antifungal activity on various plant pathogenic fungi

Debieu D, Bach J, Arnold A, Brousset S, Gredt M, Taton M, Rahier A, Malosse C, Leroux P. Inhibition of ergosterol biosynthesis by morpholine, piperidine, and spiroketalamine fungicides in Microdochium nivale: Effect on sterol composition and sterol Delta8->Delta7-isomerase activity. Pestic. Biochem. Phys., 2000, 67, 85-94; Sui GQ, Zhang W, Zhou K, Li YL, Zhang BY, Xu D, Zou Y, Zhou WM. Trialkylamine Derivatives Containing a Triazole Moiety as Promising Ergosterol Biosynthesis Inhibitor: Design, Synthesis, and Antifungal Activity. Chem. Pharm. Bull., 2017, 65, 82-89.

[0005] In view of the problems in the prior art, the carpanachol tertiary amine derivative, the preparation method and the application in the preparation of the plant source fungicide are provided, and the following purposes are achieved: the prepared carpanachol tertiary amine derivative has high-efficiency and broad-spectrum antifungal activity on plant pathogenic fungi, and can be used for preparing the plant source fungicide.

[0006] To solve the above technical problems, the technical scheme is adopted in the present application.

[0007] The carpanachol tertiary amine derivative has the following general structure:

[0008] .

[0009] The substituent group of the cyclic secondary amine is The substituent group of the cyclic secondary amine is

[0010] The specific structure of the cyclic secondary amine substituent group is as follows:

[0011] .

[0012] The corresponding carbolin tertiary amine derivatives of the cyclic secondary amine substituents 1-17 are compounds 1-17.

[0013] The preparation method of the carbolin tertiary amine derivative is that the intermediate II is dissolved in ethanol, then the cyclic secondary amine is added, heated and stirred, reflux is maintained, after the reaction is completed by TLC detection, ethanol is removed under reduced pressure, and then extraction, water washing and purification are performed to obtain the carbolin tertiary amine derivative.

[0014] The structural formula of the intermediate II is as follows:

[0015] .

[0016] The structural formula of the cyclic secondary amine is as follows:

[0017] .

[0018] The molar ratio of the intermediate II and the cyclic secondary amine is 1:1.5-2.0, preferably 1:1.5.

[0019] The preparation method of the intermediate II is that anhydrous aluminum chloride is added to a flask, then dry dichloromethane is added and stirred under an ice water bath at 0 DEG C, after uniform stirring, methyl acryloyl chloride is added dropwise, then the sesamol I is dissolved in an appropriate amount of dichloromethane and added dropwise into the reaction system by a syringe, after the addition is completed, the ice water bath is removed and the reaction is gradually restored to room temperature, after the reaction is completed by TLC detection, an appropriate amount of ice water mixture is slowly added into the reaction system, then a small amount of concentrated hydrochloric acid is added to quench the reaction, the reaction is continuously stirred for about 10 minutes, then water is added for dilution, and extraction, water washing, drying, concentration and purification are performed to obtain a yellow needle-shaped solid.

[0020] The molar ratio of the sesamol I, anhydrous aluminum chloride and methyl acryloyl chloride is 1.0:1.3:1.3.

[0021] The carbolin tertiary amine derivative is applied to the preparation of a plant source fungicide.

[0022] The carbolin tertiary amine derivative has inhibitory activity on cucumber anthracnose fungus, tomato botrytis cinerea, wheat scab fungus and wheat full-erosion fungus.

[0023] Compared with the prior art, the application has the following beneficial effects:

[0024] In 50 μThe prepared cardol tertiary amine derivatives 1-17 of the application exhibit obvious bacteriostatic activity on cucumber anthracnose fungus (Colletotrichum orbiculare) C. orbiculare ), tomato gray mold (Botrytis cinerea) B. cinerea ), wheat scab (Gibberella zeae) F. graminearum ), and wheat full-etching fungus (Gaeumannomyces graminis) G. graminis ) at a concentration of 50 μ g / mL. The inhibition rate of derivative 14 on cucumber anthracnose fungus is 60.9%, which is better than the positive control hymexazol (57.0%) and the reference control cardol (33.1%). The preliminary bacteriostatic activity of derivatives 1, 3, 10, and 14 on wheat full-etching fungus is 60.8%-83.2%, which is significantly better than the positive control hymexazol (48.6%) and the reference control cardol (45.0%). In addition, the inhibition rates of derivative 14 on cucumber anthracnose fungus and wheat full-etching fungus are both higher than 60% at a concentration of 50 μ g / mL, indicating that derivative 14 has a certain broad-spectrum antifungal activity. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 is the nuclear magnetic resonance hydrogen spectrum of compound 1;

[0026] Figure 2 is the HRMS spectrum of compound 1;

[0027] Figure 3 is the nuclear magnetic resonance hydrogen spectrum of compound 3;

[0028] Figure 4 is the HRMS spectrum of compound 3;

[0029] Figure 5 is the nuclear magnetic resonance hydrogen spectrum of compound 10;

[0030] Figure 6 is the HRMS spectrum of compound 10;

[0031] Figure 7 is the nuclear magnetic resonance hydrogen spectrum of compound 14;

[0032] Figure 8 is the HRMS spectrum of compound 14. DETAILED DESCRIPTION

[0033] In the application, a series of new cardol tertiary amine derivatives 17 are synthesized, and it is found that the 17 newly synthesized cardol tertiary amine derivatives have obvious fungicidal activity on four common plant pathogenic fungi, and some derivatives have the potential to be developed into agricultural fungicides.

[0034] Example 1: Preparation of cardol tertiary amine derivatives 1-17

[0035] The reaction formula is as follows:

[0036]

[0037] The structural formula of the above cyclic secondary amine is as follows:

[0038]

[0039] The cyclic secondary amines a-q, in order, correspond to the carbidol tertiary amine derivatives 1-17, referred to as compounds 1-17, respectively.

[0040] Reference for preparation of intermediate II: Chem. Pharm. Bull., 2024, 72, 186-189.

[0041] Preparation of intermediate II: In a dry flask, anhydrous aluminum chloride (2.5 g, 18.8 mmol) was added, followed by the addition of dry dichloromethane 40 mL and stirring under an ice water bath at 0°C, stirring for about 5 minutes, and then methyl acryloyl chloride (1.8 mL, 18.8 mmol) was added dropwise with a syringe. Then sesamol I (2.0 g, 14.5 mmol) was dissolved in dry dichloromethane (5 mL) and added dropwise to the reaction system, and after the addition was completed, the ice water bath was removed and the reaction was gradually restored to room temperature. The reaction was carried out at room temperature for about 4 hours, and the reaction was detected by TLC. After the reaction was completed, 5 mL of an ice water mixture and 0.5 mL of concentrated hydrochloric acid were slowly added to the reaction system, and stirring was continued for 10 minutes. Then water (40 mL) was added for dilution, and dichloromethane was extracted twice, and the combined organic phase was washed with saturated Na2CO3 solution and saturated brine, dried over anhydrous NaSO4, concentrated, and then purified by column chromatography (PE:EA = 30:1) to obtain a yellow needle-like solid (1.05 g, 5.09 mmol, 35%).

[0042] Preparation of carbidol tertiary amine derivatives 1-17: An appropriate amount of intermediate II (412 mg, 2.0 mmol) was dissolved in 15 mL of ethanol, and then 3.0 mmol of the corresponding cyclic secondary amine was added, and the oil bath was heated and stirred at 80°C to maintain reflux. The reaction was detected by TLC, and after the reaction was completed, the ethanol was removed under reduced pressure, and then water (15 mL) was added for dilution, and ethyl acetate (15 mL) was extracted twice, and the combined organic phase was washed with saturated brine. After removing the solvent, the target product 1-17 was purified by column chromatography.

[0043] The physicochemical properties of carbidol tertiary amine derivatives 1-17 are as follows:

[0044] The physicochemical properties of compound 1 are as follows:

[0045] 1) yellow oily liquid (two configurations, molar ratio 1:1), yield 36%;

[0046] 2) the compound's nuclear magnetic resonance hydrogen spectrum, high resolution mass spectrum characteristics:

[0047] 1 H-NMR (400 MHz, CDCl3) δ : 13.42 (br, s, 1H), 7.17 (s, 1H), 6.45 (s,1H), 5.98 (s, 2H), 3.63–3.50 (m, 1H), 2.88–2.75 (m, 2H), 2.75–2.65 (m,1H),2.38 (dd, J = 12.8, 5.5 Hz, 1H), 1.97–1.83 (m, 1H), 1.80–1.52 (m, 5H), 1.52–1.37 (m, 1H), 1.18 (d, J = 6.9 Hz, 3H), 0.83 (d, J = 5.6 Hz, 1.5H), 0.80 (d, J =5.9 Hz, 1.5H). HRMS (ESI):calcd for C 17 H 24 NO4[M+H] + , 306.1705, found 306.1709.

[0048] The compound 2 physical and chemical properties are as follows:

[0049] 1) yellow oily liquid, yield 45%;

[0050] 2) the compound's nuclear magnetic resonance hydrogen spectrum, high resolution mass spectrum characteristics:

[0051] 1 H-NMR (400 MHz, CDCl3) δ : 13.40 (br, s, 1H), 7.16 (s, 1H), 6.45 (s,1H), 5.99 (s, 2H), 3.68–3.50 (m, 1H), 2.88 (d, J = 11.5 Hz, 2H), 2.78 (d, J =11.4 Hz, 1H), 2.42 (dd, J= 12.8, 5.3 Hz, 1H), 2.09–1.92 (m, 2H), 1.56 (t, J =11.8 Hz, 2H), 1.36–1.21 (m, 3H), 1.18 (d, J = 6.9 Hz, 3H). HRMS (ESI):calcd for C J = 6.2 Hz,3H). HRMS (ESI):calcd for C 17 H 24 NO4[M+H] + , 306.1705, found 306.1709.

[0052] The physicochemical properties of compound 3 are as follows:

[0053] 1) Brown-yellow solid, yield 26%;

[0054] 2) The compound's nuclear magnetic resonance hydrogen spectrum, high-resolution mass spectrum characteristics:

[0055] 1 H-NMR (400 MHz, CDCl3) δ : 13.36 (br, s, 1H), 7.32–7.25 (m, 2H), 7.24–7.12 (m, 4H), 6.48 (s, 1H), 5.99 (s, 2H), 3.70–3.52 (m, 1H), 3.05 (d, J = 11.4Hz, 1H), 2.99–2.85 (m, 2H), 2.53–2.40 (m, 2H), 2.25–2.04 (m, 2H), 1.86–1.64(m, 4H), 1.23 (d, J = 6.9 Hz, 3H). HRMS (ESI):calcd for C 22 H 26 NO4[M+H] + ,368.1862, found 368.1866.

[0056] The physicochemical properties of compound 4 are as follows:

[0057] 1) Light yellow oily liquid, yield 37%;

[0058] 2) The compound's nuclear magnetic resonance hydrogen spectrum, high-resolution mass spectrum characteristics:

[0059] 1 H-NMR (400 MHz, CDCl3)δ : 13.35 (br, s, 1H), 7.14 (s, 1H), 6.45 (s,1H), 5.98 (s, 2H), 3.65 (s, 3H), 3.60–3.47 (m, 1H), 2.92–2.72 (m, 3H), 2.39(dd, J = 12.7, 5.7 Hz, 1H), 2.30–2.19 (m, 1H), 2.14–1.99 (m, 2H), 1.90–1.77 (m,2H), 1.75–1.59 (m, 2H), 1.18 (d, J = 6.9 Hz, 3H). HRMS (ESI):calcd for C 18 H 24 NO6[M+H] + , 350.1604, found 350.1607.

[0060] The physical and chemical properties of compound 5 are as follows:

[0061] 1) Brown yellow oily liquid, yield 23%;

[0062] 2) The nuclear magnetic resonance hydrogen spectrum, high resolution mass spectrum characteristics of the compound:

[0063] 1 H-NMR (400 MHz, CDCl3) δ : 13.31 (br, s, 1H), 7.13 (s, 1H), 6.43 (s,1H), 5.96 (s, 2H), 3.72–3.64 (m, 4H), 3.60–3.48 (m, 1H), 2.95 (d, J = 11.5 Hz,1H), 2.90–2.80 (m, 2H), 2.54–2.46 (m, 4H), 2.40 (dd, J = 12.7, 5.4 Hz, 1H),2.18–2.09 (m, 1H), 2.08–1.91 (m, 2H), 1.82–1.70 (m, 2H), 1.53–1.41 (m, 2H),1.16 (d, J = 6.9 Hz, 3H). HRMS (ESI):calcd for C 20 H 29 N2O5[M+H] + , 377.2076, found377.2081.

[0064] The physicochemical properties of compound 6 are as follows:

[0065] 1) light yellow solid, yield 27%;

[0066] 2) the nuclear magnetic resonance hydrogen spectrum, high resolution mass spectrum characteristics of the compound:

[0067] 1 H-NMR (400 MHz, CDCl3) δ : 13.33 (br, s, 1H), 7.13 (s, 1H), 6.46 (s,1H), 5.99 (s, 2H), 3.58–3.47 (m, 1H), 2.94–2.87 (m, 1H), 2.68–2.46 (m, 7H),2.43 (dd, J = 12.6, 5.6 Hz, 2H), 2.31 (s, 3H), 1.19 (d, J = 7.0 Hz, 3H). HRMS(ESI):calcd for C 16 H 23 N2O4[M+H] + , 307.1658, found 307.1661.

[0068] The physicochemical properties of compound 7 are as follows:

[0069] 1) light yellow solid, yield 30%;

[0070] 2) the nuclear magnetic resonance hydrogen spectrum, high resolution mass spectrum characteristics of the compound:

[0071] 1 H-NMR (400 MHz, CDCl3) δ : 13.34 (br, s, 1H), 7.13 (s, 1H), 6.45 (s,1H), 5.98 (s, 2H), 3.58–3.47 (m, 1H), 2.92–2.83 (m, 1H), 2.68–2.47 (m, 6H),2.45–2.36 (m, 5H), 1.18 (d, J = 6.7 Hz, 3H), 1.07 (t, J = 7.2 Hz, 3H). HRMS(ESI):calcd for C 17 H 25 N2O4[M+H] +, 321.1814, found 321.1815.

[0072] Physical and chemical properties of compound 8 are as follows:

[0073] 1) Brown-yellow solid, yield 33%;

[0074] 2) The compound's nuclear magnetic resonance hydrogen spectrum, high resolution mass spectrum characteristics:

[0075] 1 H-NMR (400 MHz, CDCl3) δ : 13.35 (br, s, 1H), 7.13 (s, 1H), 6.45 (s,1H), 5.98 (s, 2H), 3.57–3.47 (m, 1H), 2.91–2.82 (m, 1H), 2.66–2.60 (m, 1H),2.57–2.44 (m, 8H), 2.40 (dd, J = 12.7, 5.8 Hz, 1H), 1.18 (d, J = 6.8 Hz, 3H),1.02 (d, J = 6.7 Hz, 6H). HRMS (ESI):calcd for C 18 H 27 N2O4[M+H] + , 335.1971, found335.1976.

[0076] Physical and chemical properties of compound 9 are as follows:

[0077] 1) Yellow oily liquid, yield 22%;

[0078] 2) The compound's nuclear magnetic resonance hydrogen spectrum, high resolution mass spectrum characteristics:

[0079] 1 H-NMR (400 MHz, CDCl3) δ : 13.35 (br, s, 1H), 7.13 (s, 1H), 6.45 (s,1H), 5.98 (s, 2H), 3.57–3.46 (m, 1H), 2.91–2.81 (m, 1H), 2.71–2.62 (m, 1H),2.60–2.45 (m, 8H), 2.40 (dd, J= 12.7, 5.7 Hz, 1H), 2.26–2.17 (m, 1H), 1.89–1.82 (m, 2H), 1.80–1.73 (m, 2H), 1.60 (d, J = 12.4 Hz, 1H), 1.27–1.21 (m, 2H),1.18 (d, J = 7.5 Hz, 3H), 1.15–1.03 (m, 2H). HRMS (ESI):calcd for C 21 H 31 N2O4[M+H] + , 375.2284, found 375.2285.

[0080] The physicochemical properties of compound 10 are as follows:

[0081] 1) light yellow solid, yield 26%;

[0082] 2) the compound's nuclear magnetic resonance hydrogen spectrum, high resolution mass spectrum characteristics:

[0083] 1 H-NMR (400 MHz, CDCl3) δ : 13.33 (br, s, 1H), 7.27–7.22 (m, 2H), 7.18(s, 1H), 6.90 (d, J = 8.1 Hz, 2H), 6.85 (t, J = 7.3 Hz, 1H), 6.47 (s, 1H), 5.99(s, 2H), 3.78–3.47 (m, 1H), 3.16 (s, 4H),3.05–2.90 (m, 1H), 2.65 (br, s, 4H),2.57–2.43 (m, 1H), 1.24 (d, J = 7.0 Hz, 3H). HRMS (ESI):calcd for C 21 H 25 N2O4[M+H] + , 369.1814, found 369.1817.

[0084] The physicochemical properties of compound 11 are as follows:

[0085] 1) light yellow oily liquid, yield 30%;

[0086] 2) the compound's nuclear magnetic resonance hydrogen spectrum, high resolution mass spectrum characteristics:

[0087] 1 H-NMR (400 MHz, CDCl3) δ : 13.29 (br, s, 1H), 7.10 (s, 1H), 6.44 (s,1H), 5.97 (s, 2H), 3.65 (s, 3H), 3.57–3.48 (m, 1H), 3.45–3.31 (m, 4H), 2.90–2.81 (m, 1H), 2.50–2.30 (m, 5H), 1.17 (d, J = 6.9 Hz, 3H). HRMS (ESI):calcd for C 17 H 23 N2O6[M+H] + , 351.1556, found 351.1552.

[0088] The physicochemical properties of compound 12 are as follows:

[0089] 1) yellowish oil liquid, yield 28%;

[0090] 2) the nuclear magnetic resonance hydrogen spectrum, high resolution mass spectrum characteristics of the compound:

[0091] 1 H-NMR (400 MHz, CDCl3) δ : 13.30 (br, s, 1H), 7.12 (s, 1H), 6.46 (s,1H), 5.98 (s, 2H), 4.10 (q, J = 7.1 Hz, 2H), 3.60–3.49 (m, 1H), 3.47–3.31 (m,4H), 2.95–2.80 (m, 1H), 2.51–2.30 (m, 5H), 1.23 (t, J = 7.0 Hz, 3H), 1.19 (d, J =6.8 Hz, 3H). HRMS (ESI):calcd for C 18 H 25 N2O6[M+H] + , 365.1713, found 365.1711.

[0092] The physicochemical properties of compound 13 are as follows:

[0093] 1) brownish yellow solid, yield 21%;

[0094] 2) The compound's nuclear magnetic resonance hydrogen spectrum, high resolution mass spectrum characteristics:

[0095] 1 H-NMR (400 MHz, CDCl3) δ : 13.30 (br, s, 1H), 7.12 (s, 1H), 6.46 (s,1H), 5.99 (s, 2H), 3.69–3.45 (m, 3H), 3.45–3.30 (m, 2H), 3.04–2.82 (m, 1H),2.56–2.32 (m, 5H), 2.06 (s, 3H), 1.20 (d, J = 7.0 Hz, 3H). HRMS (ESI):calcd for C 17 H 23 N2O5[M+H] + , 335.1607, found 335.1610.

[0096] The physical and chemical properties of compound 14 are as follows:

[0097] 1) White solid, yield 46%;

[0098] 2) The compound's nuclear magnetic resonance hydrogen spectrum, high resolution mass spectrum characteristics:

[0099] 1 H-NMR (400 MHz, CDCl3) δ : 13.30 (br, s, 1H), 7.13 (s, 1H), 6.46 (s,1H), 5.98 (s, 2H), 3.64 (t, J = 4.7 Hz, 4H), 3.58–3.50 (m, 1H), 2.91–2.82 (m,1H), 2.52–2.43 (m, 4H), 2.43–2.37 (m, 1H), 1.20 (d, J = 7.0 Hz, 3H). HRMS(ESI):calcd for C 15 H 20 NO5[M+H] + , 294.1341, found 294.1344.

[0100] The physical and chemical properties of compound 15 are as follows:

[0101] 1) Light yellow solid, yield 35%;

[0102] 2) The compound's nuclear magnetic resonance hydrogen spectrum, high resolution mass spectrum characteristics:

[0103] 1 H-NMR (400 MHz, CDCl3) δ : 13.31 (br, s, 1H), 7.13 (s, 1H), 6.46 (s,1H), 5.99 (s, 2H), 3.63–3.45 (m, 1H), 2.98–2.85 (m, 1H), 2.80–2.67 (m, 4H),2.65–2.52 (m, 4H), 2.42 (dd, J = 13.2, 5.4 Hz, 1H), 1.17 (d, J = 6.9 Hz, 3H). HRMS (ESI):calcd for C 15 H 20 NO4S [M+H] + , 310.1113, found 310.1117.

[0104] The compound 16's physical and chemical properties are as follows:

[0105] 1) Brownish red needle-like solid, yield 32%;

[0106] 2) The compound's nuclear magnetic resonance hydrogen spectrum, high resolution mass spectrum characteristics:

[0107] 1 H-NMR (400 MHz, CDCl3) δ : 7.13 (s, 1H), 6.41 (s, 1H), 5.94 (s, 2H),3.58–3.43 (m, 1H), 2.97–2.85 (m, 1H), 2.56–2.51 (m, 1H), 2.51–2.42 (m, 4H),1.75–1.64 (m, 4H), 1.18 (d, J = 7.0 Hz, 3H). HRMS (ESI):calcd for C 15 H 20 NO4 [M+H] + , 278.1392, found 278.1396.

[0108] The compound 17's physical and chemical properties are as follows:

[0109] 1) Yellow oily liquid (two rotamers, molar ratio 1:1), yield 23%;

[0110] 2) Characteristics of the compound's 1H NMR spectrum and high-resolution mass spectrometry:

[0111] 1 H-NMR (400 MHz, CDCl3) C. orbiculare : 13.24 (br, s, 1H), 13.21 (br, s, 1H), 7.19(dd, J = 9.0, 6.8 Hz, 1H), 7.10 (t, J = 8.0 Hz, 1H), 6.93 (s, 1H), 6.89 (s, 1H), 6.44 (s, 1H), 6.39 (s, 1H), 6.28–6.19 (m, 2H), 6.07 (s, 1H), 6.01 (s, 1H), 5.99 (s, 1H), 5.96 (s, 1H), 5.92–5.83 (m, 2H), 3.90–3.75 (m, 4H), 3.38–3.26 (m, 2H), 2.98–2.93 (m, 1H), 2.92–2.79 (m, 5H), 2.77–2.66 (m, 2H), 2.49–2.41(m, 3H), 2.40–2.27 (m, 5H), 1.86–1.78 (m, 2H), 1.76–1.67 (m, 2H), 0.97 (d, J =6.9 Hz, 3H), 0.93 (d, J = 6.8 Hz, 3H). HRMS (ESI):calcd for C 22 H 24 N₂O₅Na [M+Na] + ,419.1583, found 419.1586.

[0112] Example 2: Bioassay Experiment on Resistance to Plant Pathogenic Fungi

[0113] 1. Pathogenic fungus: Cucumber anthracnose fungus ( B. cinerea ), tomato gray mold ( F. graminearum ), wheat scab ( G. graminis ), wheat take-all pathogen ( μ The aforementioned pathogenic fungi were passaged and cultured in our laboratory.

[0114] 2. Test samples: Compounds 1-17, calcubin, and oxamyl (97%, Shanghai Titan Technology Co., Ltd.). Oxamyl was used as a positive control.

[0115] 3. Preparation of Potato Dextrose Agar (PDA) medium

[0116] The peeled, cored and washed potato was cut into small pieces. 1000 mL of distilled water was boiled in a pot. 200 g of the cut potato was added to the pot and boiled for 30 min. After filtering with double-layered gauze, the filtrate was poured into a clean pot. 20 g of glucose and 17 g of agar were added and heated until completely dissolved. Finally, the volume was adjusted to 1000 mL, and then it was divided and sterilized by high-pressure steam.

[0117] 4. Bioassay method (mycelial growth rate method)

[0118] The mycelial growth rate method was used to determine the fungicidal activity of the test compounds 1-17, carvyl alcohol and the positive control, hymexazol. In a clean bench, each test compound (5.0 mg) was completely dissolved in 0.5 mL of DMSO, and then 9.5 mL of sterile water was added to prepare a test solution. The solution was then added to 90 mL of sterilized and melted PDA medium, mixed thoroughly, and poured into a sterile Petri dish to prepare a drug-containing flat medium. The final actual concentration of the test compound was 50 μ g / mL, and the DMSO content was 0.5% (v / v). The medium containing only 0.5% DMSO was used as a blank control, and the same method was used to prepare a medium containing 50 μ g / mL of carvyl alcohol and hymexazol as a reference control and a positive control, respectively, and the fungicidal activity was determined in the same way.

[0119] When the medium was semi-solid, a puncher with a diameter of 5.0 mm was used to select the mycelium growing vigorously at the edge of the colony. Then, the mycelial cake was moved into the above-mentioned Petri dish with the inoculation needle, and the mycelium was in contact with the medium. Each treatment was repeated three times. The inoculated Petri dish was placed in a constant temperature incubator at 26°C for incubation. After 72 h, the colony diameter was measured using the cross method, and the mycelial growth inhibition rate was calculated according to the following formula. Finally, the average value ± standard deviation (S.D.) of the inhibition rate was used to represent the final result.

[0120] Mycelial growth inhibition rate (%) = ( d c - d s ) / ( d c -5) x 100%

[0121] wherein d c is the average value of the colony diameter in the blank control, d s is the average value of the diameter of the colonies growing in the medium containing the compound.

[0122] Table 1. Preliminary in vitro antibacterial activity of calcubin tertiary amine derivatives 1-17 (50 μL / L). μ g / mL, 72h)

[0123]

[0124] Conclusion: The results show that at 50 C. orbiculare At a concentration of g / mL, the above-mentioned calcubin tertiary amine derivatives 1-17 were effective against cucumber anthracnose ( B. cinerea ), tomato gray mold ( F. graminearum ), wheat scab ( G. graminis ), wheat take-all pathogen ( μ It exhibited significant antibacterial activity. At 50... μ At a concentration of g / mL, derivative 14 showed an inhibition rate of 60.9% against cucumber anthracnose, superior to the positive control oxadixyl (57.0%) and the reference control calciferol (33.1%). Derivatives 1, 3, 10, and 14 exhibited preliminary inhibitory activities of 60.8%-83.2% against wheat take-all, significantly superior to the positive control oxadixyl (48.6%) and the reference control calciferol (45.0%). Furthermore, at a concentration of 50 g / mL... ​ At a concentration of g / mL, derivative 14 showed inhibition rates of over 60% against both cucumber anthracnose fungus and wheat take-all fungus, indicating that derivative 14 possesses certain broad-spectrum antifungal activity. These results suggest that tertiary amine derivatives 1, 3, 10, and 14 hold promise for the preparation of highly efficient, environmentally friendly, and low-toxicity plant-derived fungicides.

Claims

1. A tertiary amine derivative of calcullol, characterized in that: The general structural formula of the derivative is: ； The It is a cyclic secondary amine substituent; The specific structure of the cyclic secondary amine substituent is as follows: 。 2. The method for preparing the calcullol tertiary amine derivative according to claim 1, characterized in that: The preparation method involves dissolving intermediate II in ethanol, then adding a cyclic secondary amine, heating and stirring under reflux, and after the reaction is completed by TLC detection, removing the ethanol under reduced pressure, followed by extraction, washing with water, and purification to obtain the calcullol tertiary amine derivative. The structural formula of intermediate II is shown below: 。 3. The preparation method according to claim 2, characterized in that: The structural formula of the cyclic secondary amine is shown below: 。 4. The preparation method according to claim 3, characterized in that: The molar ratio of intermediate II to the cyclic secondary amine is 1:1.5-2.

0.

5. The application of the calcullol tertiary amine derivative according to claim 1 in the preparation of plant-derived fungicides.

6. The application according to claim 5, characterized in that: The plant-derived fungicide has inhibitory activity against cucumber anthracnose fungus, tomato gray mold fungus, wheat scab fungus, and wheat take-all fungus.

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