Application of Glyantrypine family alkaloid derivatives in the treatment of plant viral and bacterial diseases

Abstract

The invention relates to an application of a Glyantrypine family alkaloid derivative in treating plant viral and pathogenic diseases. The invention first finds that the Glyantrypine family alkaloid derivatives I-1 to I-16 exhibit good anti-plant viral and anti-pathogenic activity and can well inhibit 14 plant pathogens including tobacco mosaic virus (TMV) and cucumber wilt, peanut brown spot, apple ring pattern, wheat sheath blight, corn leaf spot, watermelon anthrax, rice seedling blight, tomato early blight, wheat gibberella, rice blast, pepper phytophthora, rape sclerotia, cucumber gray mold and rice sheath blight.

Description

Technical Field

[0001] The invention relates to application of Glyantrypine family alkaloid derivatives in treating plant virus and bacterial diseases, and belongs to the technical field of agricultural protection. Background Art

[0002] In 2015, Chen Hao's research group at the First Institute of Oceanography of the State Oceanic Administration isolated 14 alkaloids including glyantrypine from the marine fungus Penicillium expansum Y32, and used a zebrafish in vivo model to evaluate the cardiovascular effects of each compound. All compounds had a significant alleviating effect on bradycardia induced by astemizole (ASM) (Mar. Drugs 2015, 13 (10), 6489-6504.). In 2017, Lan Wenjian's research group at Sun Yat-sen University isolated 22 alkaloids from different amino acid culture media of the marine fungus Scedosporium apiospermum F41-1. Scedapin C is the first Fumiquinozoline alkaloid found to contain a sulfonamide group. Scedapin C and scequinadoline D have significant anti-hepatitis C (HCV) activity against recombinant J8CC, EC 50 The values ​​are 110.35 μM and 128.60 μM respectively (Org. Lett. 2017, 19, 4888-4891.). So far, there is no report on the use of Glyantrypine family alkaloid derivatives in the prevention and treatment of plant viral pathogens and fungi. Summary of the invention

[0003] In view of the shortcomings of the prior art, the present invention provides the use of Glyantrypine family alkaloids I-1 to I-16 in preventing and treating plant viral diseases. The Glyantrypine family alkaloid derivatives I-1 to I-16 of the present patent have good anti-plant virus and anti-bacterial activity. The Glyantrypine family alkaloid derivatives of the present invention are compounds shown in I-1 to I-16 (structural formula 1).

[0004]

[0005] Compared with the prior art, the present invention first discovered that the Glyantrypine family alkaloid derivatives I-1 to I-16 exhibited good anti-plant virus and pathogenic disease activity, and could well inhibit 14 plant pathogens including tobacco mosaic virus (TMV) and cucumber wilt, peanut brown spot, apple ring spot, wheat sheath blight, corn spot, watermelon anthrax, rice seedling blight, tomato early blight, wheat fusarium, rice blast, pepper phytophthora, rape sclerotium, cucumber gray mold, and rice sheath blight. Some compounds have reached the activity level of commercial varieties and have good application prospects. DETAILED DESCRIPTION

[0006] I-1 to I-3 of the Glyantrypine family alkaloid derivatives of the present invention are prepared according to the following references: J. Org. Chem. 2000, 65, 1022-1030. I-4 to I-6 are prepared according to the following references: Org. Biomol. Chem. 2018, 16, 1679-1685. I-7 to I-10 are prepared according to the following references: J. Chem. Res. 2016, 41, 18-20. I-11 is prepared according to the following references: Org. Chem. Front. 2020, 7, 1635-1639. I-12 to I-14 are prepared according to the following references: Tetrahedron Lett. 2017, 58, 1071-1074. I-15 to I-16 were prepared according to the following reference: J. Med. Chem. 2016, 59, 8422-8440.

[0007] Example 1: Experimental data of Glyantrypine family alkaloid derivatives I-1 to I-16:

[0008] I-1: white solid, melting point 159-161°C. 1 H NMR (400MHz, CDCl3) δ8.68 (s, 1H), 8.35 (d, J=7.2Hz, 1H), 7.79-7.71 (m, 1H), 7.5 5-7.45 (m, 2H), 7.31 (d, J=8.0Hz, 1H), 7.26-7.20 (m, 1H), 7.06 (t, J=7.5Hz, 1H), 6.98 (d, J=3.6Hz, 1H), 6.84 (t, J=7.5Hz, 1H), 6.67 (d, J=2.0Hz, 1H), 5.63-5.49 ( m, 1H), 3.76 (dd, J=16.9, 4.1Hz, 1H), 3.71-3.52 (m, 2H), 2.80 (d, J=16.9Hz, 1H). 13C NMR (100MHz, CDCl3) δ169.5, 160.6, 148.4, 147.1, 136.0, 135.0, 127.3, 127.1, 1 27.0, 126.7, 123.7, 122.6, 120.2, 120.0, 118.4, 111.3, 109.0, 56.9, 44.6, 27.2.

[0009] I-2: white solid, melting point 119-121°C. 1 H NMR (400MHz, CDCl3) δ8.40-8.30 (m, 2H), 7.76 (t, J=7.6Hz, 1H), 7.59 (d, J=8.1Hz, 1H), 7.52 (t, J=7.5Hz, 1H), 7.38 (d, J=8.0Hz, 1H), 7.28 (d, J=8.1Hz, 1 H), 7.11 (t, J=7.5Hz, 1H), 6.89 (t, J=7.4Hz, 1H), 6.78-6.63 (m, 2H), 5.67 (t , J=4.2Hz, 1H), 3.75-3.60 (m, 2H), 3.19-3.06 (m, 1H), 1.36 (d, J=6.7Hz, 3H). 13 CNMR (100MHz, CDCl3) δ169.3, 160.8, 151.7, 147.1, 136.0, 134.7, 127.3, 127.1, 126.9, 123.6, 122.5, 120.3, 120.0, 118.5, 111.2, 109.4, 57.6, 49.2, 27.1, 19.1.

[0010] I-3: white solid, melting point 122-124°C. 1 H NMR (400MHz, CDCl3) δ8.41-8.31 (m, 2H), 7.76 (t, J=7.5Hz, 1H), 7.59 (d, J=8. 1Hz, 1H), 7.52 (t, J=7.5Hz, 1H), 7.38 (d, J=8.0Hz, 1H), 7.29 (d, J=8.1Hz, 1H), 7.11 (t, J=7.6Hz, 1H), 6.90 (t, J=7.5Hz, 1H), 6.85 (s, 1H), 6.70 (s, 1H), 5.73- 5.62 (m, 1H), 3.73-3.55 (m, 2H), 3.17 (q, J=6.4Hz, 1H), 1.37 (d, J=6.5Hz, 3H). 13C NMR (100MHz, CDCl3) δ169.4, 160.8, 151.7, 147.1, 136.0, 134.7, 127.4, 127.3, 127. 1, 126.8, 123.6, 122.5, 120.2, 120.0, 118.5, 111.3, 109.4, 57.6, 49.2, 27.1, 19.0.C 21 H 19 N4O2[M+H] + 359.1503, found 359.1506.

[0011] I-4: white solid, melting point 243-245°C. 1 H NMR (400MHz, CDCl3) δ8.37 (d, J=7.9Hz, 1H), 8.15 (dd, J=7.7, 1.5Hz, 1H), 7.82 (t, J=6.3Hz, 2H), 7.61 (dd, J=8. 0, 1.3Hz, 1H), 7.57-7.51(m, 1H), 7.50-7.45(m, 1H), 7.42-7.37(m, 1H), 7.36-7.31(m, 2H), 7.31-7.26(m, 2H). 13 CNMR (100MHz, CDCl3) δ161.9, 157.3, 151.5, 147.0, 134.9, 133.0, 131.8, 129.8 , 129.6, 128.8, 127.8, 127.5, 127.4, 127.1, 125.9, 125.1, 121.5, 120.6, 120.1.

[0012] I-5: white solid, melting point 242-244°C. 1 H NMR (400MHz, CDCl3) δ8.36 (d, J=7.8Hz, 1H), 8.03 (d, J=7.9Hz, 1H), 7.83-7.78 (m, 2H), 7.60 (dd, J=8.1 , 1.4Hz, 1H), 7.56-7.49(m, 1H), 7.41-7.31(m, 2H), 7.26-7.21(m, 1H), 7.15-7.08(m, 2H), 2.40(s, 3H). 13 C NMR (100MHz, CDCl3) δ162.0, 161.7, 157.3, 151.5, 147.1, 144.2, 134.8, 131.5, 129.7 , 129.5, 129.0, 127.7, 127.4, 127.1, 126.7, 125.0, 124.1, 121.5, 120.5, 120.4, 21.4.

[0013] I-6: white solid, melting point 246-248°C. 1 H NMR (400MHz, DMSO-d6) δ8.57 (d, J=7.6Hz, 1H), 8.52-8.46 (m, 1H), 8.26 (d, J=7.8Hz, 1H), 7.96 (t, J=7.3Hz, 1H), 7.82 (d, J= 8.1Hz, 1H), 7.73 (d, J=8.0Hz, 1H), 7.66 (t, J=7.3Hz, 1H), 7.60-7.53 (m, 2H), 7.48 (t, J=7.6Hz, 1H), 7.37 (t, J=7.5Hz, 1H). 13 C NMR (100MHz, DMSO-d6) δ164.6, 161.3, 154.7, 151.7, 150.0, 146.8, 142.8, 135.9 , 130.7, 130.5, 128.7, 128.4, 128.0, 127.4, 126.1, 123.5, 122.4, 121.7, 120.7.

[0014] I-7: khaki solid, melting point 288-290°C. 1 H NMR (400MHz, DMSO-d6) δ8.82 (d, J=7.5Hz, 1H), 8.28 (dd, J=8.0, 1.0Hz, 1H), 8.20 (d, J=7 .6Hz, 1H), 8.05-7.93(m, 2H), 7.93-7.89(m, 1H), 7.88-7.84(m, 1H), 7.64-7.56(m, 1H).

[0015] I-8: off-white solid, melting point 327-329°C. 1 H NMR (400MHz, DMSO-d6) δ8.89-8.84(m, 1H), 8.25-8.16(m, 2H), 7.96-7.88(m, 2H) , 7.85-7.73 (m, 2H), 7.69-7.61 (m, 1H), 7.52-7.43 (m, 1H), 2.98 (d, J=4.6Hz, 3H). 13 C NMR (100MHz, DMSO-d6) δ157.8, 149.6, 146.3, 143.7, 134.2, 133.5, 132.7, 129.0, 127.5, 127.1, 126.8, 126.1, 123.3, 122.1, 120.3, 28.8.

[0016] I-9: yellow solid, melting point 224-226°C.1 H NMR (400MHz, DMSO-d6) δ8.94-8.85 (m, 1H), 8.28 (d, J=7.8Hz, 1H), 8.13-8.04 (m, 1H), 8.02-7.92 (m, 2H), 7. 90 (t, J=7.2Hz, 1H), 7.86-7.80 (m, 1H), 7.56 (t, J=7.4Hz, 1H), 3.88 (t, J=4.0Hz, 4H), 3.36 (t, J=4.4Hz, 4H). 13 C NMR (100MHz, DMSO-d6) δ157.5, 153.9, 146.0, 143.6, 134.5, 133.0, 132.7, 129.9, 127.2, 126.8, 126.4, 126.1, 125.6 (s), 122.1, 119.9, 65.8, 51.1.

[0017] I-10: white solid, melting point 219-221°C. 1 H NMR (400MHz, DMSO-d6) δ8.96-8.88 (m, 1H), 8.34-8.29 (m, 1H), 8.27 (d, J=8.0Hz, 1H), 8.03-7.95 (m, 2H), 7.92-7.78 (m, 3H), 7.54 (t, J=7.4Hz, 1H), 3.67 (dd, J=11.3, 5.8Hz, 2H), 2.91-2.76 (m, 2H), 2.42 (s, 6H). 13 C NMR (100MHz, DMSO-d6) δ158.0, 149.3, 146.3, 143.4, 134.4, 133.5, 133.0, 129.0, 127.6, 127.0, 126.7, 126.2, 123.6, 121.9, 120.1, 58.2, 45.4, 45.2.

[0018] I-11: yellow solid, melting point 157-159°C. 1 H NMR (400MHz, CDCl3) δ8.48 (d, J=7.6Hz, 1H), 8.31 (d, J=8.0Hz, 1H), 7.81-7.70 (m, 2H), 7.49-7.39 (m, 3H), 7.27 (d, J=8.2Hz, 1H), 4.41 (t, J=6.3Hz, 2H), 3.09 (t, J=6.2Hz, 2H). 13C NMR (100MHz, CDCl3) δ161.7, 149.4, 147.8, 137.1, 134.2, 131.7, 129.6, 128.0, 127.6, 127.6, 127.5, 126.9, 126.54, 120.8, 39.6, 27.5.

[0019] I-12: khaki solid, melting point 149-151°C. 1 H NMR (400MHz, DMSO-d6) δ8.10 (d, J=7.9Hz, 1H), 7.79 (t, J=7.6Hz, 1H), 7.59 (d, J=8.1 Hz, 1H), 7.49 (t, J=7.5Hz, 1H), 5.58 (s, 1H), 2.77 (s, 2H), 2.28 (s, 3H), 1.04 (s, 6H). 13 C NMR (100MHz, DMSO-d6) δ159.4, 154.3, 146.1, 134.4, 132.9, 128.9, 126.5, 126.4, 126.2, 121.8, 45.9, 29.4, 26.2, 20.0.

[0020] I-13: white solid, melting point 129-131°C. 1 H NMR (400MHz, DMSO-d6) δ7.53 (d, J=8.8Hz, 1H), 7.49 (d, J=2.9Hz, 1H), 7.41-7 .36(m, 1H), 5.57(s, 1H), 3.86(s, 3H), 2.73(s, 2H), 2.28(s, 3H), 1.03(s, 6H). 13 C NMR (100MHz, DMSO-d6) δ159.2, 157.6, 151.9, 140.5, 132.9, 129.0, 128.0, 123.7, 122.5, 106.5, 55.6, 45.7, 29.5, 26.2, 20.1.

[0021] I-14: white solid, melting point 144-146°C. 1 H NMR (400MHz, CDCl3) δ8.35 (s, 1H), 7.77 (d, J=8.6Hz, 1H), 7.46 (d, J=8.6Hz, 1H), 5.48 (s, 1H), 2.77 (s, 2H), 2.34 (s, 3H), 1.09 (s, 6H). 13CNMR (100MHz, CDCl3) δ159.2, 154.9, 145.3, 137.3, 133.6, 129.5, 129.3, 128.3, 123.8, 119.9, 46.9, 29.9, 26.7, 20.5.

[0022] I-15: yellow solid, melting point 300-302°C. 1 H NMR (400MHz, DMSO-d6) δ12.36 (s, 1H), 8.15 (d, J=7.2Hz, 1H), 7.72 (t, J=7.0Hz, 1H), 7.6 3(d, J=2.1Hz, 1H), 7.47 (d, J=8.4Hz, 1H), 7.38 (d, J=2.0Hz, 1H), 7.25 (t, J=7.3Hz, 1H). 13 C NMR (100MHz, DMSO-d6) δ157.5, 146.3, 145.2, 134.6, 127.2, 123.3, 122.0, 121.8, 112.9, 107.2.

[0023] I-16: Yellow solid, melting point>400℃. 1 H NMR (400MHz, DMSO-d6) δ7.87 (dd, J=7.9, 1.3Hz, 1H), 7.70 (s, 1H), 7.58-7.51 (m, 1H), 7.32 (dt, J=8.3, 2.0Hz, 1H), 7.12 (d, J=8.0Hz, 1H), 7.10-7.03 (m, 1H), 5.65-5.54 (m, 1H), 4.03 (dd, J=3.3, 2.2Hz, 2H). 13 C NMR (100MHz, DMSO-d6) δ158.6, 150.3, 148.2, 135.2, 127.2, 124.4, 122.2, 119.3, 116.7, 110.1, 39.9.

[0024] Example 2: Determination of anti-tobacco mosaic virus activity, the determination procedure is as follows:

[0025] 1. Virus purification and concentration determination:

[0026] Virus purification and concentration determination were carried out in accordance with the SOP specification for tobacco mosaic virus compiled by the Laboratory of the Institute of Element Analysis of Nankai University. The crude virus extract was centrifuged twice with polyethylene glycol, the concentration was determined, and the extract was refrigerated at 4°C for later use.

[0027] 2. Preparation of compound solution:

[0028] After weighing, the compound and ribavirin stock were added to DMF and dissolved to prepare 1×10 5 μg / mL stock solution, and then diluted to the required concentration with an aqueous solution containing 1‰ Tween 80.

[0029] 3. In vivo protection:

[0030] Select 3-5 leaf-stage Sansi tobacco plants with uniform growth, spray the whole plant with 3 replicates per treatment, and set a 1‰ Tween 80 aqueous solution as a control. After 24 hours, sprinkle emery (500 mesh) on the leaves, dip a brush in the virus solution, and gently rub the entire leaf surface along the branch vein twice, support the lower part of the leaf with the palm of your hand, the virus concentration is 10μg / mL, and rinse with running water after inoculation. Record the number of lesions after 3 days and calculate the prevention effect.

[0031] 4. In vivo therapeutic effect:

[0032] Select 3-5 leaf-stage Sansi tobacco plants with uniform growth. Use a brush to inoculate the virus on the whole leaf at a virus concentration of 10 μg / mL. Rinse with running water after inoculation. After the leaf surface is dry, spray the whole plant with the drug. Repeat 3 times for each treatment, and set a 1‰ Tween 80 aqueous solution as a control. Record the number of lesions after 3 days and calculate the prevention effect.

[0033] 5. In vivo passivation effect:

[0034] Select Sansi tobacco plants with uniform growth at the 3-5 leaf stage, mix the agent with an equal volume of virus juice and inactivate for 30 minutes, then inoculate by friction, with a virus concentration of 20 μg / mL. Rinse with running water immediately after inoculation, repeat 3 times, and set a 1‰ Tween 80 aqueous solution as a control. Count the number of lesions after 3 days and calculate the results.

[0035] Inhibition rate (%) = [(number of control necrosis spots - number of treated necrosis spots) / number of control necrosis spots] × 100%

[0036] All compounds were tested for their in vivo inactivation activity against tobacco mosaic virus at a treatment dose of 500 μg / mL, and compounds with a relative inhibition rate greater than 40% were further tested for in vivo therapeutic and active protective activity at a treatment dose of 500 μg / mL.

[0037] Table 1 Test results of anti-tobacco mosaic virus (TMV) activity of glyantrypine family alkaloid derivatives I-1 to I-16

[0038]

[0039] From the data in Table 1, it can be seen that the Glyantrypine family alkaloid derivatives I-1 to I-16 all showed good anti-TMV activity. Among them, at 500 μg / mL, the activities of compounds I-1, I-5, I-10, and I-16 were comparable to those of ribavirin, and had great development value.

[0040] Example 3: Antibacterial activity test, the determination procedure is as follows:

[0041] In vitro bactericidal test, bacterial growth rate determination method (plate method):

[0042] Dissolve a certain amount of the drug in an appropriate amount of acetone, then dilute it to the required concentration with an aqueous solution containing 200μg / mL emulsifier, then draw 1mL of the drug solution and inject it into the culture dish, then add 9mL of culture medium respectively, shake well to make a 50μg / mL drug-containing plate, and use the plate with 1mL of sterilized water as a blank control. Use a 4mm diameter puncher to cut the bacterial disc along the outer edge of the hyphae and move it to the drug-containing plate. Repeat each treatment three times. Place the culture dish in a constant temperature incubator at 24±1℃ for culture. After 48 hours, investigate the expansion diameter of the bacterial disc of each treatment, calculate the average value, and calculate the relative inhibition rate compared with the blank control.

[0043]

[0044] Table 2 In vitro bactericidal activity test results of Glyantrypine family alkaloid derivatives I-1 to I-16:

[0045]

[0046] Glyantrypine family alkaloids showed broad-spectrum inhibitory activity against 14 tested bacteria at a test concentration of 50 μg / mL. Compound I-5 had an inhibition rate of 84% against apple ring rot and wheat stripe blight, and compound I-7 had an inhibition rate of more than 80% against wheat stripe blight and rapeseed sclerotinia. In addition, compound I-11 had an inhibition rate of 91% against apple ring rot.

Claims

1. Application of a Glyantrypine family alkaloid derivative I-5 in preventing and treating tobacco mosaic virus disease 2. Use of the Glyantrypine family alkaloid derivative I-5 as claimed in claim 1 in preventing and controlling apple ring rot and wheat sheath blight.

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