CN1166637C - Antibacterial Activity, Induction, Isolation and Synthesis of N-Acyltryptamine Phytoalexin Derivatives - Google Patents

Abstract

Antibacterial activity, induction, isolation and synthesis of N-acyltryptamine phytoalexin derivatives. It involves the induction, separation and structure determination of two N-acyltryptamine phytoalexins. Using these two acyltryptamine phytoalexins as lead compounds, 25 derivatives were synthesized, involving compounds of the formula [I], wherein: R ₁ represents hydrogen and a group optionally substituted on the indole ring; R ₂ represents hydrogen And a group optionally substituted on the aromatic ring; R ₃ and R ₄ represent an optionally substituted group. The compound has obvious inhibitory effect on various plant pathogenic bacteria and Candida albicans, and is used as a new type of antibacterial drug for plant disease prevention and treatment and clinically.

Description

Antibacterial Activity, Induction, Isolation and Synthesis of N-Acyltryptamine Phytoalexin Derivatives

technical field

The invention relates to the induction and separation of phytoalexin with antibacterial activity, the synthesis method of phytoalexin derivatives, and the application as a high-efficiency fungicide, belonging to the technical field of pesticides and medicines.

Background technique

Plant-derived antibacterial substances are one of the main mechanisms for plants to resist pathogen infection during the long-term co-evolution process with pathogens. Phytoalexin is an inducible antibacterial compound. So far, at least 400 phytoalexins have been isolated and identified, and they are mainly distributed in 35 families of plants, among which the Gramineae, Leguminosae and Solanaceae crops are the plants with the most phytoalexins found so far. From the chemical structure point of view, phytoalexin can be roughly divided into phenols, flavonoids, terpenes, alkaloids and so on. Phytoalexin is easy to degrade in plants, and it is possible to develop a new efficient and safe fungicide by chemically studying it and synthesizing its derivatives. Although the chemical structure of many phytoalexins has been elucidated, there are not many successful examples of fungicides developed on this basis. The main reasons are: (1) The government and enterprises have not paid enough attention to plant-derived antibacterial compounds, and lack of attention to plant protection. (2) The in vitro antibacterial activity of most phytoalexins is not high, and the synthesis of their derivatives, evaluation of antibacterial and disease control effects are lacking.

Rice is a major food crop, and rice blast has been one of the serious obstacles to rice production for a long time. In order to control the harm of rice blast, people have carried out in-depth research on the mechanism of rice blast resistance. So far, more than 10 phytoprotectins have been isolated from rice, which are structurally divided into diterpenoids, flavonoids and oxidized fatty acids. The extraction and isolation of N-acyltryptamine from rice has not been reported yet.

It has been found that N-acyltryptamine compounds have various physiological activities and can be used clinically as an anxiolytic, sedative and hypnotic agent (Chemical Abstract, 1997, 123: 350261); they can also be used as endocrine drugs, immune system drugs, Ophthalmic disease drugs and animal feed additives, etc. (United States Patent, 1995, 5403851). However, there is no report on N-acyltryptamine compounds as agricultural and medical antibacterial agents.

purpose of invention

In order to protect the environment, protect the sustainable development of human beings and agriculture, it is urgent to develop environmentally friendly pesticides with high efficiency, low toxicity and low residue. One of the purposes of the present invention is to provide a method for inducing and extracting N-acyltryptamines phytoalexin; another object is to provide a synthetic method for plantalexins derivatives; the third purpose is to provide N-acyltryptamines as Examples of high-efficiency antimicrobial applications.

Contents of the invention

In order to achieve the above object, the present invention provides the following technical methods:

The present invention relates to rice (Oryza sativa cv. Aichiasahi) N-acyltryptamine phytoalexin and its derivatives, including antibacterial activity, induction, separation method and synthesis method, discovering highly active compounds, used as antibacterial drugs for plant disease control and clinical .

Two N-acyltryptamine phytoalexins were obtained by separating and purifying from rice leaves: N-benzoyltryptamine and N-cinnamoyltryptamine. Taking N-benzoyl tryptamine and N-cinnamoyl tryptamine as lead compounds, another 25 kinds of N-acyl tryptamine compounds were synthesized by chemical synthesis method (Example 3-Example 27). Rice phytoalexin N-benzoyl tryptamine and N-cinnamoyl tryptamine and their derivatives have significant antibacterial activity.

The present invention relates to N-acyltryptamine derivatives represented by general formula [I], general formula [I] is as follows:

in:

_R represents hydrogen and a group optionally substituted on the indole ring (such as hydroxyl, halogen, alkoxy, nitro, aryl, cyano, alkylthio, amino, heterocyclic);

_R represents hydrogen and a group optionally substituted on the aromatic ring (such as hydroxyl, halogen, alkoxy, nitro, aryl, cyano, alkylthio, amino, heterocyclic);

R ₃ represents an optionally substituted group {such as -(CH=CH)n-, -(CH ₂ )n-, -(C≡C)n-};

R ₄ represents an optionally substituted group {such as -(CH═CH)n-, -(CH ₂ )n-, -(C≡C)n-}.

The invention provides an induction method, an extraction and separation method, an analysis method, a synthesis method and an identification method for the formation of rice N-benzoyl tryptamine and N-cinnamoyl tryptamine phytoalexin.

The method for inducing rice N-acyltryptamine phytoalexin involved in the present invention is to spray and inoculate rice seedlings with a conidia suspension of Magnaporthe grisea, and then cut and inoculate them after a period of time at 100% relative humidity and away from light. Leaves are reserved for extraction and separation of phytoalexin.

The method for extracting and separating the rice N-acyl-tryptamine phytoalexin involved in the patent of the present invention is as follows: the fat-soluble substances of rice leaves are separated by phytochemical separation methods (such as silica gel column chromatography, preparative thin layer chromatography and high performance liquid chromatography) ), to obtain N-benzoyl tryptamine and N-cinnamoyl tryptamine two kinds of plant hormones.

See accompanying drawing 1 for the high performance liquid chromatography (HPLC) of the whole fat extract of rice leaves inoculated with Magnaporthe grisea for 48 hours. At the same retention time, two strong absorption peaks P1 and P2 appeared in the leaf extract inoculated with Magnaporthe grisea. P1 is N-benzoyl tryptamine, and P2 is N-cinnamoyl tryptamine.

The invention provides a method for synthesizing N-benzoyl tryptamine and N-cinnamoyl tryptamine. Take a certain amount of tryptamine and dissolve it in an appropriate volume of pyridine, and add dropwise benzoyl chloride (for the preparation of N-benzoyl tryptamine) or dropwise addition of cinnamoyl chloride (for the preparation of N-cinnamoyl tryptamine) under cooling in an ice-water bath ), stirred for a period of time (such as 1 hour) under cooling in an ice-water bath, then stirred for a period of time (such as 3 hours) at room temperature, the reaction mixture was poured into ice water, extracted three times with an appropriate volume of chloroform, and the chloroform part was diluted with water and Wash with hydrochloric acid until pyridine is removed, dry the chloroform part with a desiccant (such as anhydrous sodium sulfate), and concentrate in vacuo to obtain the reaction product, whose structural formula is as follows. The hydrogen spectrum ( ¹ H-NMR) and carbon spectrum ( ¹³ C-NMR) data of the reaction product are shown in Table 2 and Table 3. Example-1 is N-benzoyltryptamine, and Example-2 is N- Cinnamoyltryptamine.

N-Benzoyltryptamine (Example-1)

N-cinnamoyl tryptamine (Example-2)

The present invention is based on the synthesis of a series of N-acyltryptamine compounds, through resistance to plant pathogenic bacteria (including blast fungus, Fusarium wilt of watermelon, Fusarium wilt of cucumber, Fusarium wilt of cotton, Gibberella wheat, Rhizoctonia solani, Rhizoctonia oryzae) The structure-activity relationship research with the activity of human pathogenic bacteria (including Candida albicans) found highly active substances as a new type of antibacterial drug. The idea of its compound structure design is to introduce oxygen-containing, halogen, hydroxyl and other substituents with p-π conjugation with the aromatic ring in the aromatic ring of indole ring, benzoyl or cinnamoyl to improve the antibacterial activity of the compound . In order to achieve the above object, the present invention further provides the following technical methods.

Method A: Mix tryptamine and excess triethylamine in a reaction vessel, add an appropriate amount of chloroform, cool and stir in an ice-water bath, add dropwise an acid chloride solution with an equimolar number of triethylamine, and react in the dark. Allow the reaction mixture to rise to room temperature naturally, stir overnight, add an appropriate volume of water, separate the aqueous layer from the chloroform layer, wash the aqueous layer once with an equal amount of chloroform, combine the chloroform layers, and successively wash with acid solution (such as 10% hydrochloric acid ), lye (such as 2M sodium hydroxide solution) and water, and after drying with a desiccant (such as anhydrous sodium sulfate), the solvent is recovered to obtain the acylated product.

Method B: Mix 5-hydroxytryptamine hydrochloride with water and excess alkali aqueous solution (such as 2M sodium hydroxide) in a reaction vessel, add an appropriate amount of chloroform, and add dropwise the equimolar alkali aqueous solution under cooling and stirring in an ice-water bath. A few acid chloride solutions, reacted in the dark, let the reaction mixture rise to room temperature naturally, stirred overnight, a white precipitate appeared, filtered, washed the precipitate three times with an appropriate volume of chloroform, and then dissolved the precipitate in ethyl acetate, followed by Acid solution (such as 10% hydrochloric acid), alkaline solution (such as 2M sodium hydroxide solution) and water washing, and after drying with desiccant (such as anhydrous sodium sulfate), the solvent is recovered to obtain the acylated product.

The compound of general formula [I] synthesized by the technical method of the present invention has antibacterial activity. For example, N-benzoyl-5-hydroxytryptamine has a half-inhibitory concentration of 0.15 (μg/ml) for the growth of the blastomyces oryzae strain P131 germ tubes, and a half-inhibitory concentration of 0.44 (μg/ml) for the growth of the blastomyces oryzae strain R1528 germ tubes. ), and their action strengths are 110 times (16.6 μg/ml) and 46 times (20.2 μg/ml) respectively that of the lead N-benzoyltryptamine.

Examples of synthesis of N-acyl-tryptamine and N-acyl-5-hydroxytryptamine derivatives by methods A and B are as follows. The reaction products, melting points (MP, °C) and yields (Yield, %) are shown in Table 1. The hydrogen spectrum ( ¹ H-NMR) and carbon spectrum ( ¹³ C-NMR) data of the reaction product are shown in Table 2 and Table 3, respectively.

                                             

Compound R ₁ R ₂ R ₃ R ₄ Synthetic Method Yield (%) MP (℃)

Example-1 - - - -CH ₂ -CH ₂ -A - -

Example-2 - - -CH ²⁰ =CH ²¹ - -CH ₂ -CH ₂ -A 96.7 135-136

Example-3 5-HO- - - -CH ₂ -CH ₂ -B 72.4 195-197

Example-4 5-HO- o-Cl- - -CH ₂ -CH ₂ - B 91.2 62-64

Example-5 5-HO-p-Cl--- _CH2 - _CH2 -B 91.2 142-144

Example-6 5-HO- 2,4-2Cl- - -CH ₂ -CH ₂ - B 79.3 -

Example-7 5-HO-3,4-2Cl- - -CH ₂ -CH ₂ -B 80.0 179-181

Example-8 5-HO-p-OCH ₃ ²⁰ - - -CH ₂ -CH ₂ -B 89.1 209-210

Example-9 5-HO- - -CH ²⁰ =CH ²¹ - -CH ₂ -CH ₂ - B 100.0 80-82

Example-10 5-HO- - -CH ₂ ²⁰ - -CH ₂ -CH ₂ - B 91.8 -

Example-11 - - -CH ₂ ²⁰ - -CH ₂ -CH ₂ -A 94.2 139-140

Example-12 - o-Cl - -CH ₂ -CH ₂ - A 90.2 -

Example-13 - p-Cl - -CH ₂ -CH ₂ - A 92.2 -

Example-14 - 2,4-2Cl- - -CH ₂ -CH ₂ -A 91.5 -

Example-15 - 3,4-2Cl- - -CH ₂ -CH ₂ -A 91.5 -

Example-16 - pF- - -CH ₂ -CH ₂ -A 77.8 138-140

Example-17 - oF- - -CH ₂ -CH ₂ - A 78.9 -

Example-18 5-HO- pF- - -CH ₂ -CH ₂ -B 58.6 172-173

Example-19 5-HO- oF- - -CH ₂ -CH ₂ - B 58.0 -

Example-20-2,4-2F--- _CH2 - _CH2 -A 85.0-

Example-21-3,4-2F--- _{CH2- CH2} -A 94.1 79-81

Example -22 - 2,4,5-3F- - -CH ₂ -CH ₂ -A 58.3 -

Example-23 5-HO- 2,4-2F- - -CH ₂ -CH ₂ - B 93.9 -

Example-24 5-HO- 3,4-2F- - -CH ₂ -CH ₂ - B 61.8 -

Example-25 5-HO-2,4,5-3F- - -CH ₂ -CH ₂ -B 68.6 177-179

Example-26 5-HO- o-NH ₂ - - -CH ₂ -CH ₂ - B 63.7 -

Example-27 5-MeO- o-NH ₂ - - -CH ₂ -CH ₂ -B 84.4 86-89

The hydrogen spectrum data of table 2 reaction product N-acyltryptamine ^*

Compound 1-H 5-OH Arom.-H 10-H 11-H 12-H 20-H 21-H NH ₂ CH ₃

Example-1 8.2s - 7.07- 3.10t 3.80d 6.20s - - - - - - -

...

Example-2 11.64s - 7.24- 3.39t, 4.11q, 9.08s 7.06s 7.10s - -

8.16m 7.2Hz 6.8Hz

Example-3 11.55s 9.41t, 7.19- 3.31t, 4.04q, 8.71s - - - - - - -

5.3Hz 8.44m 7.4Hz 6.8Hz

Example-4 11.54s 9.19t, 7.21- 3.39t, 4.09m 8.71s - - - - -

5.6Hz 8.14m 7.2Hz

Example-5 11.56s 9.30s 7.20- 3.30t, 4.01q, 8.71s - - - - - - -

8.29m 7.2Hz 6.7Hz

Example-6 11.56s 9.32t, 7.20- 3.31t, 4.01q, 8.71s - - - - - - -

5.5Hz 8.71m 7.0Hz 7.0Hz

Example-7 11.52s 9.42t, 7.22- 3.33t, 4.03q, 8.71s - - - - - - -

5.4Hz 8.41m 7.2Hz 6.9Hz

Example-8 11.54s 9.06t, 6.99- 3.32t, 4.06q, 8.71s - - - - - - -

5.8Hz 8.30m 7.4Hz 6.8Hz

Example-9 11.53s 8.89t, 7.20- 3.28t, 4.03q, 8.71s 6.93d, 6.97d, - - -

5.6Hz 8.08m 7.2Hz 6.8Hz 10.8Hz 10.4Hz

Example-10 11.40s 8.71s 7.17- 3.16t, 4.10m 8.58s 3.78m - - - - -

7.66m 7.2Hz

Example-11 11.81s - 7.19- 3.19t, 4.07q, 8.71s 3.77m - - - - -

8.67m 7.2Hz 7.1Hz

Example-12 7.91s - 6.92- 2.95t, 3.66q, 6.06s - - - - - - -

7.48m 7.0Hz 7.0Hz

Example-13 8.13s - 7.10- 3.13t, 3.82q, 6.19s - - - - - - -

...

Example-14 8.13s - 7.10- 3.04t, 3.85q, 6.29s - - - - - - -

...

Example-15 8.13s - 7.10- 3.13t, 3.81q, 6.18s - - - - - - -

                                                                                           

Example-16 11.85s - 7.15-8.30m 3.37t, 4.05q, 8.73s - - - - - - -

7.2Hz 6.4Hz

Example-17 11.85s - 7.21-8.82m 3.36t, 4.07q, 8.71s - - - - - - -

7.2Hz 6.4Hz

Example-18 11.56s 9.23d, 7.11-8.25m 3.30t, 4.01q, 8.69s - - - - - - -

5.2Hz 7.2Hz 6.8Hz

Example-19 11.60s 8.82d 7.08 3.33t, 4.07q, 8.71s - - - - - - -

5.2Hz 8.13m 7.2Hz 6.8Hz

Example-20 - - - - - - - - 8.71s - - - - - - -

Example-21 11.86s - 7.19-8.54m 3.34t, 4.10q, 8.71s - - - - - - -

7.2Hz 6.4Hz

Example-22 11.88s - 7.18-8.83m 3.35t, 4.04q, 8.71s - - - - - - -

5.8Hz 5.2Hz

Example-23 12.09s 8.86d 6.95-8.25m 33.3t, 4.02q, 8.71s - - - - - - -

5.2Hz 7.2Hz 6.7Hz

Example-24 11.58s 9.39d 7.00-8.54m 3.31t, 4.00q, 8.71s - - - - - - -

6.0Hz 7.2Hz 6.8Hz

Example-25 1164s 8.87d 717-8.01m 3.31t, 4.00m 8.71s - - - - - - -

5.2Hz 7.2Hz

Example-26 11.59s 9.00t 6.82-8.39m 3.38m 4.00m 8.70s - - - 6.61m -

5.4Hz

Example-27 11.69s - 7.01-8.11m 3.33t, 4.02q, 8.71s - - 6.62q, 3.78s

7.2Hz 6.4Hz 7.2Hz

^* Chemical shift value (δ) is in ppm

Table 3 The carbon spectrum data of the reaction product N-acyltryptamine ^*

C- Example-1 Example-2 Example-3 Example-4 Example-5 Example-6 Example-7 Example-8 Example-9

2 122.07d 123.2d 124.0d 124.0d 124.1d 124.1d 124.0d 124.1d 124.2d

3 113.05s 113.4s 112.6s 113.0s 112.7s 112.7s 112.5s 112.7s 112.5s

4 118.75d 119.5d 112.9d 113.1d 113.0d 113.0d 113.0d 112.7d 113.0d

5 119.37d 119.4d 152.3s 152.4s 152.4s 152.3s 152.4s 152.3s 152.3s

6 122.29d 122.1d 104.1d 104.2d 104.1d 104.0d 104.1d 104.7d 104.7d

7 111.27d 112.3d 112.6d 112.4d 112.5d 112.7d 112.8d 112.7d 112.7d

8 136.44s 137.9s 132.5s 132.6s 132.6s 132.4s 132.6s 132.6s 132.6s

9 127.32s 128.7s 129.6s 129.7s 129.7s 129.6s 129.7s 129.6s 129.6s

10 25.29t 26.5t 26.6t 26.6t 26.5t 26.3t 26.4t 26.7t 26.6t

11 40.22t 41.3t 41.5t 41.5t 41.6t 41.7t 41.7t 41.5t 41.0t

13 167.44s 166.8s 167.8s 168.0s 166.8s 165.5s 165.8s 167.4s 166.4s

14 134.69s 140.2s 136.4s 136.4s 132.6s 135.8s 135.6s 128.6s 139.9s

15 126.81d 128.7d 130.3d 138.4s 134.7s 136.9s 131.0s 132.6s 128.6d

16 128.48d 124.0d 128.7d 129.7d 129.0d 130.8d 129.7s 114.1d 124.2d

17 131.30d 129.8d 132.8d 133.5d 137.0s 136.9s 135.1s 162.4s 129.7d

18 128.48d 123.8d 127.9d 124.5d 128.9d 134.3d 136.3d 112.9d 124.2d

19 126.81d 128.3d 131.2d 132.6d 132.6d 135.4d 132.6d 132.6d 128.1d

20 - 112.3d - - - - - 55.4q ^** 113.0d

21 - 136.1d - - - - - - - 136.0d

^* Chemical shift value (δ) is in ppm; ^-CH ₂ -be assigned. ^** -O-CH ₃ be assigned

Continued Table 3 The carbon spectrum data of the reaction product N-acyltryptamine ^*

C- Example-10 Example Example-12 Example-13 Example-14 Example-15 Example-16 Example-17 Example-18

-11

2 124.1d 123.3d 122.2d 122.1d 122.2d 122.1d 123.8d 123.8d 123.9d

3 112.4s 113.2s 112.8s 113.0s 112.7s 112.9s 113.3s 113.1s 113.0s

4 113.0d 119.2d 118.7d 118.7d 118.7d 118.6d 119.2d 119.3d 104.1d

5 152.4s 119.2d 119.5d 119.6d 119.6d 119.7d 121.9d 121.9d 152.4d

6 104.1d 121.8d 122.3d 122.4d 122.3d 122.4d 119.2d 119.3d 104.1d

7 112.8d 112.0d 112.2d 111.3d 111.3d 111.4d 112.0d 112.1d 112.6d

8 132.6s 137.7s 130.6s 133.0s 133.5s 133.0s 137.7d 137.8d 132.6d

9 129.6s 128.5s 127.2s 127.3s 127.2s 127.3s 132.6s 134.3s 130.5s

10 26.5t 26.2t 25.2t 25.2t 25.1t 25.1t 26.3t 26.2t 26.6t

11 41.6t 41.4t 40.2t 40.4t 40.3t 40.6t 41.6t 41.4t 41.7t

13 171.4s 171.0s 166.5s 166.3s 165.4s 165.2s 166.7s 164.3s 166.9s

14 135.3s 135.3s 135.2s 136.4s 136.4s 136.4s 128.6s 128.5s 129.7s

15 129.6d 129.8d 136.4s 128.7d 131.4s 126.0d 130.3d 161.8s 130.5d

16 129.0d 128.9d 127.0d 128.3d 127.4d 134.5s 115.4d 116.2d 115.5d

17 127.1d 127.3d 131.1d 137.5s 136.6s 135.7s 166.0s 132.9d 166.1s

18 129.0d 128.8d 130.2d 128.3d 130.0d 129.1d 115.6d 124.8d 115.7d

19 129.1d 128.9d 130.0d 128.7d 131.2d 130.5d 130.4d 131.5d 130.5d

20 61.0t ^60.8d ^- - - - - - - - - - - -

twenty one - - - - - - - - -

^* Chemical shift values (δ) are in ppm; ^-CH ₂ -be assigned.

Continued Table 3 The carbon spectrum data of the reaction product N-acyltryptamine ^*

C- Example-19 Example-20 Example-21 Example-22 Example-23 Example-24 Example-25 Example-26 Example

-27

2 124.1d 123.5d 123.6d 123.8d 123.8d 124.0d 124.1d 124.1d 124.1d

3 112.2s 112.8s 113.0s 112.9s 113.6s 112.4s 112.1s 112.4s 113.3s

4 104.1d 119.0d 120.0d 119.3d 106.1d 104.0d 104.0d 104.4d 101.3d

5 152.3s 121.7d 121.7d 122.0d 150.2s 152.3s 152.4s 152.4s 154.5s

6 104.1d 119.0d 120.1d 119.2d 105.9d 104.0d 104.0d 104.1d 101.3d

7 112.6d 111.7d 111.8d 112.2d 112.5d 112.6d 112.6d 112.7d 112.4d

8 132.7s 137.5s 137.5s 137.8s 133.2s 133.5s 132.5s 132.6s 132.9s

9 131.5s 134.5s 133.3s 128.5s 128.8s 129.6s 129.5s 129.6s 129.0s

10 26.4t 25.9t 26.0t 26.1t 26.0t 26.3t 26.1t 26.7t 26.5t

11 41.3t 41.2t 41.5t 41.6t 41.5t 41.7t 41.5t 41.1t 41.2t

13 164.3s 165.4s 165.3s 165.5s 167.5s 166.6s 162.2s 163.7s 170.4s

14 129.5s 120.6s 133.3s 120.6s 121.0s 133.4s 119.4s 117.3s 116.9s

15 161.8s 163.1s 125.6s 162.3s 163.5s 124.7d 162.2s 128.8d 128.7d

16 116.4d 104.5d 118.5d 106.6d 104.6d 117.5d 106.6d 116.8d 117.2d

17 132.6d 165.3s 153.2s 150.1s 165.6s 152.3s 150.1s 132.2d 132.2d

18 124.8d 111.8d 148.8d 137.8d 111.9d 149.1s 135.8s 115.8d 115.6d

19 131.5d 132.9d 118.3d 119.3d 133.1d 117.4d 119.2d 152.4s 150.8s

20 - - - - - - - - - - - - - - -

twenty one - - - - - - - - -

CH ₃ O - - - - - - - - 55.8q

^* Chemical shift values (δ) are in ppm; ^-CH ₂ -be assigned.

Description of drawings

Fig. 1 is the high-performance liquid chromatography (HPLC) figure of the rice leaf full-fat extract and the simulated inoculation rice leaf full-fat extract of inoculating Magnaporthe grisea for 48 hours, and Fig. 1A is the high performance liquid chromatography (HPLC) figure of the simulated inoculation rice leaf full-fat extract HPLC graph, Fig. 1B is the HPLC graph of rice leaf whole fat extract inoculated with Magnaporthe grisea. The chromatographic column adopted is a C18 reverse phase liquid chromatographic column; the mobile phase is methanol:water=52:48 (v/v); the flow rate: 1ml/min; the absorption wavelength is 220nm. Under the same extraction conditions and chromatographic conditions, two strong absorption peaks P1 and P2 appeared in the extract of leaves inoculated with Magnaporthe grisea (Fig. 1B), and the retention times were 21.31min and 28.42min respectively; The leaf (Fig. 1A) extract showed only very small absorption peaks P1 and P2 at the same retention time. P1 is N-benzoyl tryptamine, and P2 is N-cinnamoyl tryptamine.

Implementation

In order to better understand the present invention, the substantive content of the present invention will be further described below in conjunction with the examples, but the content of the present invention is not limited thereto.

Example-1: Dissolve 0.6g of tryptamine in 15ml of pyridine, add dropwise 0.58g of benzoyl chloride under cooling in an ice-water bath, stir for 1 hour under cooling in an ice-water bath, then stir at room temperature for 3 hours, and pour the reaction mixture into Put into ice water, extract three times with 15ml of chloroform, wash the chloroform part with water and dilute hydrochloric acid until pyridine is removed, dry the chloroform part with anhydrous sodium sulfate, and concentrate the solvent in vacuo to obtain Product Example-2, the structural formula is as follows, the molecular formula is For C ₁₇ H ₁₆ N ₂ O, see Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Example-1 Structural formula

Example-2: Dissolve 0.6g of tryptamine in 15ml of pyridine, add dropwise 0.62g of cinnamoyl chloride under cooling in an ice-water bath, stir for 1 hour under cooling in an ice-water bath, then stir at room temperature for 3 hours, and pour the reaction mixture into In ice water, extract three times with 15ml of chloroform, wash the chloroform part with water and dilute hydrochloric acid until pyridine is removed, dry the chloroform part with anhydrous sodium sulfate, and concentrate the solvent in vacuo to obtain Product Example-2, the structural formula is as follows, and the molecular formula is C ₁₉ H ₁₈ N ₂ O, EIMS 290 (M ⁺ , 13), 159 (C ₁₀ H ₉ NO ⁺ , 8), 143 (C ₉ H ₈ NO ⁺ , 77), 131 (C ₉ H ₇ O ⁺ , 29 ), 130 (C ₉ H ₈ N ⁺ , 65), 117 (C ₈ H ₇ N ⁺ , 3), 115 (C ₉ H ₇ ⁺ , 4), 103 (C ₉ H ₆ ⁺ , 22), 91 ( C ₆ H ₅ CH ₂ ⁺ , 8), 77 (C ₆ H ₅ ⁺ , 17).IR (KBr), cm ^-1 3311 (NH), 263, 2840 (CH), 1653 (C=O), 1607 , 1558 (C=C), 1457, 1348, 1379 (CH), 1219, 1076, 1060, 1006 (CO), 978, 819, 744, 686 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Example-2 Structural formula

Example-3: In a reaction vessel, 5-hydroxytryptamine hydrochloride (100mg, 0.4702mM) was mixed with water (5ml) and an excess of 150-200% 2M sodium hydroxide solution, added 5ml of chloroform, cooled in an ice-water bath and Under stirring, add dropwise a solution of benzoyl chloride (dissolved in 1ml chloroform) with an equimolar number of sodium hydroxide, react in the dark, allow the reaction mixture to rise to room temperature naturally, stir overnight, a white precipitate appears, filter, and use 30ml Wash the precipitate three times with chloroform, then dissolve the precipitate in ethyl acetate, wash with 10% hydrochloric acid, 2M sodium hydroxide solution and water successively and dry with anhydrous sodium sulfate, then concentrate the solvent in vacuo to obtain the product Example-3, the structural formula is as follows , the molecular formula is C ₁₇ H ₁₆ N ₂ O ₂ , EIMS 280 (M ⁺ , 12), 263 (M ⁺ -OH, 2), 159 (C ₁₀ H ₉ NO ⁺ , 100), 146 (C ₉ H ₈ NO ⁺ , 62), 130 (C ₉ H ₈ N ⁺ , 2), 117 (C ₈ H ₇ N ⁺ , 7), 105 (C ₆ H ₅ CO ⁺ , 28), 91 (C ₆ H ₅ CH ₂ ⁺ , 7), 77(C ₆ H ₅ ⁺ , 29).IR(KBr), cm ^-1 3426, 3294(OH, NH), 1701(C=O), 1578, 1538(C=C), 1454, 1376 (CH), 1186, 1290, 1066 (CO), 939, 850, 708, 624 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Example-3 Structural formula

Example-4: Example-4 was synthesized according to the method described in Synthesis Example-3, but the acid chloride was 2-chlorobenzoyl chloride. The structural formula of Example-4 is as follows, the molecular formula is C ₁₇ H ₁₅ ClN ₂ O ₂ , EIMS 314 (M ⁺ , 11), 297 (M ⁺ -OH, 0.2), 263 (M + -Cl-OH, 0.2), 159 ( C ₁₀ H ₉ NO ⁺ , 100), 146 (C ₉ H ₈ NO ⁺ , 70), 130 (C ₉ H ₈ N ⁺ , 2), 117 (C ₈ H ₇ N ⁺ , 7), 105 (C ₆ H ₅ CO ⁺ , 4), 91(C ₆ H ₅ CH ₂ ⁺ , 7), 77(C ₆ H ₅ ⁺ , 5).IR(KBr), cm ^-1 3310(OH, NH), 1692(C =O), 1593, 1538 (C=C), 1471, 1315 (CH), 1276, 1187, 1050 (CO), 933, 746 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Example-4 Structural formula

Example-5: Example-5 was synthesized according to the method described in Synthesis Example-3, but the acid chloride was 4-chlorobenzoyl chloride. The structural formula of Example-5 is as follows, the molecular formula is C ₁₇ H ₁₅ ClN ₂ O ₂ , EIMS 314 (M ⁺ , 11), 297 (M ⁺ -OH, 0.2), 263 (M + -Cl-OH, 0.2), 159 ( C ₁₀ H ₉ NO ⁺ , 100), 146 (C ₉ H ₈ NO ⁺ , 70), 130 (C ₉ H ₈ N ⁺ , 2), 117 (C ₈ H ₇ N ⁺ , 7), 105 (C ₆ H ₅ CO ⁺ , 4), 91 (C ₆ H ₅ CH ₂ ⁺ , 7), 77 (C ₆ H ₅ ⁺ , 5).IR(KBr), cm ^-1 3412, 3344 (OH, NH), 1624 (C=O), 1567, 1546 (C=C), 1486, 1443 (CH), 1285, 1184, 1094 (CO), 937, 844, 754 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Example-5 Structural formula

Example-6: Example-6 was synthesized according to the method described in Synthesis Example-3, but the acid chloride was 2,4-dichlorobenzoyl chloride. Example-6 The structural formula is as follows, the molecular formula is C ₁₇ H ₁₄ Cl ₂ N ₂ O ₂ , EIMS 349 (M ⁺ , 2), 348 (M ⁺ -1, 12), 331 (M ⁺ -OH, 1), 314 (M ⁺ -Cl, 0.5), 278 (0.2), 190 (M + -C ₁₀ H ₉ NO, 2), 173 (C ₆ H ₂ Cl ₂ CO ⁺ , 20), 174 (C ₆ H ₃ Cl ₂ CO ⁺ , 12), 159 (C ₁₀ H ₉ NO ⁺ , 100), 146 (C ₉ H ₈ NO ⁺ , 91), 130 (C ₉ H ₈ N ⁺ , 3), 117 (C ₈ H ₇ N ⁺ , 10), 109 (C ₅ H ₅ NO ⁺ , 6), 105 (C ₆ H ₅ CO ⁺ , 5), 91 (C ₆ H ₅ CH ₂ ⁺ , 11), 77 (C ₆ H ₅ ⁺ , 6) .IR (KBr), cm ^-1 3375 (OH, NH), 1642 (C=O), 1588, 1542 (C=C), 1460, 1378 (CH), 1217, 1190, 1063 (CO), 938, 801, 762, 624 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Embodiment-6 Structural formula

Example-7: Example-7 was synthesized according to the method described in Synthesis Example-3, but the acid chloride was 3,4-dichlorobenzoyl chloride. Example-7 The structural formula is as follows, the molecular formula is C ₁₇ H ₁₄ Cl ₂ N ₂ O ₂ , EIMS 349 (M ⁺ , 5), 348 (M ⁺ -1, 29), 331 (M ⁺ -OH, 1), 314 (M ⁺ -Cl, 3), 190 (M + -C ₁₀ H ₉ NO, 2), 173 (C ₆ H ₂ Cl ₂ CO ⁺ , 42), 174 (C ₆ H ₃ Cl ₂ CO ⁺ , 28), 159 (C ₁₀ H ₉ NO ⁺ , 99), 146 (C ₉ H ₈ NO ⁺ , 100), 130 (C ₉ H ₈ N ⁺ , 8), 117 (C ₈ H ₇ N ⁺ , 24), 109 ( C ₅ H ₅ NO ⁺ , 6), 105 (C ₆ H ₅ CO ⁺ , 5), 91 (C ₆ H ₅ CH ₂ ⁺ , 11), 77 (C ₆ H ₅ ⁺ , 6).IR(KBr) , cm ^-1 3333 (OH, NH), 1640 (C=O), 1589, 1542 (C=C), 1466, 1375, 1313 (CH), 1221, 1183, 1030 (CO), 936, 839, 754 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Example-7 Structural formula

Example-8: Example-8 was synthesized according to the method described in Synthesis Example-3, but the acid chloride was 4-methoxybenzoyl chloride. Example-8 The structural formula is as follows, the molecular formula is C ₁₈ H ₁₈ N ₂ O ₃ , EIMS 310 (M ⁺ , 19), 293 (M ⁺ -OH, 0.2), 175 (C ₁₀ H ₁₁ N ₂ O ⁺ , 2) , 159 (C ₁₀ H ₉ NO ⁺ , 100), 146 (C ₉ H ₈ NO ⁺ , 77), 135 (C ₈ H ₇ O ₂ ⁺ , 61), 130 (C ₉ H ₈ N ⁺ , 4), 117 (C ₈ H ₇ N ⁺ , 10), 109 (C ₅ H ₅ NO ⁺ , 2), 105 (C ₆ H ₅ CO ⁺ , 3), 91 (C ₆ H ₅ CH ₂ ⁺ , 11), 77 (C ₆ H ₅ ⁺ , 25).IR(KBr), cm ^-1 3411, 3313 (OH, NH), 2932, 2849 (CH ₃ ), 1615, 1603 (C=O), 1543, 1504 (C= C), 1485, 1320 (CH), 1259, 1180, 1063, 1024 (CO), 936, 847, 803, 664 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Embodiment-8 Structural formula

Example-9: Example-9 was synthesized according to the method described in Synthesis Example-3, but the acid chloride was cinnamoyl chloride. The structural formula of Example-9 is as follows, the molecular formula is C ₁₉ H ₁₈ N ₂ O ₂ , EIMS 306 (M ⁺ , 19), 175 (C ₁₀ H ₁₁ N ₂ O ⁺ , 2), 159 (C ₁₀ H ₉ NO ⁺ , 100), 146 (C ₉ H ₈ NO ⁺ , 90), 131 (C ₉ H ₇ O ⁺ , 48), 117 (C ₈ H ₇ N ⁺ , 15), 109 (C ₅ H ₅ NO ⁺ , 2) , 105 (C ₆ H ₅ CO ⁺ , 2), 103 (C ₈ H ₇ ⁺ , 46), 91 (C ₆ H ₅ CH ₂ ⁺ , 16), 77 (C ₆ H ₅ ⁺ , 40).IR( KBr), cm ^-1 3383 (OH, NH), 1662 (C=O), 1611, 1540 (C=C), 1454, 1372 (CH), 1215, 1187, 1069 (CO), 965, 937, 833 , 766 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Embodiment-9 Structural formula

Example-10: Example-10 was synthesized according to the method described in Synthesis Example-3, but the acid chloride was phenylacetyl chloride. The structural formula of Example-10 is as follows, the molecular formula is C ₁₈ H ₁₈ N ₂ O ₂ , EIMS 294 (M ⁺ , 24), 159 (C ₁₀ H ₉ NO ⁺ , 100), 146 (C ₉ H ₈ NO ⁺ , 77) .130 (C ₉ H ₈ N ⁺ , 6), 117 (C ₈ H ₇ N ⁺ , 11), 105 (C ₆ H ₅ CO ⁺ , 2), 91 (C ₆ H ₅ CH ₂ ⁺ , 56), 77( _{C6H5 +} ^, 5).IR(KBr), cm ^-1 3335(OH, NH), 2727(CH), 1703(C=O), 1539, 1496, 1456(C=C), 1342 , 1190, 1031 (CH), 1285, 1184, 1094 (CO), 932, 798, 700, 678 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Embodiment-10 Structural formula

Example-11: Mix tryptamine (100mg, 0.4702mM) with an excess of 150-200% triethylamine in a reaction vessel, add 10ml of chloroform, cool and stir in an ice-water bath, add dropwise an equimolar amount of triethylamine Several phenylacetyl chloride (dissolved in 1ml chloroform) solution, reacted in the dark, let the reaction mixture rise to room temperature naturally, stirred overnight, added 10ml water, separated the water layer from the chloroform layer, and then used an equal amount of Wash once with chloroform, combine the chloroform layers, wash with 10% hydrochloric acid, 2M sodium hydroxide solution and water successively and dry with anhydrous sodium sulfate, then concentrate the solvent in vacuo to obtain product Example-11, the structural formula is as follows, and the molecular formula is C ₁₈ H ₁₈ N ₂ O, EIMS 278 (M ⁺ , 42), 159 (C ₁₀ H ₉ NO ⁺ , 12), 143 (C ₉ H ₈ NO ⁺ , 100), 131 (C ₉ H ₇ O ⁺ , 36), 130 (C ₉ H ₈ N ⁺ , 95), 117 (C ₈ H ₇ N ⁺ , 15), 115 (C ₉ H ₇ ⁺ , 25), 103 (C ₉ H ₆ ⁺ , 32), 91 (C ₆ H ₅ CH ₂ ⁺ , 80), 77 (C ₆ H ₅ ⁺ , 42).IR (KBr), cm ^-1 3395 (NH), 3252 (CH), 3077 (CH), 1655 (C=O), 1635 , 1566 (C=C), 1457, 1355, 1379 (CH), 1275, 1095, 1071 (CO), 930, 800, 744, 695 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Example-11 Structural formula

Example-12: Example-12 was synthesized according to the method described in Synthesis Example-11, but the acid chloride was 2-chlorobenzoyl chloride. The structural formula of Example-12 is as follows, the molecular formula is C ₁₇ H ₁₅ ClN ₂ O, EIMS 298 (M ⁺ , 15), 143 (C ₉ H ₈ NO ⁺ , 78), 130 (C ₉ H ₈ N ⁺ , 100), 117(C ₈ H ₇ N ⁺ , 7), 105(C ₆ H ₅ CO ⁺ , 5), 77(C ₆ H ₅ ⁺ , 28).IR(KBr), cm ^-1 3261(NH), 1624( C=O), 1594, 1567 (C=C), 1455, 1433 (CH), 1221, 1172, 1106 (CO), 733; 727 (=CH). ¹ H-NMR (Table 2), ¹³ C-NMR (Table 3). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Embodiment-12 Structural formula

Example-13: Example-13 was synthesized according to the method described in Synthesis Example-11, but the acid chloride was 4-chlorobenzoyl chloride. The structural formula of Example-13 is as follows, the molecular formula is C ₁₇ H ₁₅ ClN ₂ O, EIMS 298 (M ⁺ , 16), 143 (C ₉ H ₈ NO ⁺ , 100), 130 (C ₉ H ₈ N ⁺ , 92), 117 (C ₈ H ₇ N ⁺ , 3), 105 (C ₆ H ₅ CO ⁺ , 2), 77 (C ₆ H ₅ ⁺ , 15). IR(KBr), cm ^-1 3390, 3230(NH), 1632(C=O), 1596, 1550(C=C), 1487, 1454(CH), 1227, 1201, 1180(CO), 849, 740 , 683 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Embodiment-13 Structural formula

Example-14: Example-14 was synthesized according to the method described in Synthesis Example-11, but the acid chloride was 2,4-dichlorobenzoyl chloride. Example-14 The structural formula is as follows, the molecular formula is C ₁₇ H ₁₄ Cl ₂ N ₂ O, EIMS 332 (M ⁺ , 12), 173 (C ₆ H ₂ Cl ₂ CO ⁺ , 20), 143 (C ₉ H ₈ NO ⁺ , 100), 130 (C ₉ H ₈ N ⁺ , 88), 117 (C ₈ H ₇ N ⁺ , 3), 105 (C ₆ H ₅ CO ⁺ , 1), 77 (C ₆ H ₅ ⁺ , 11) .IR(KBr), cm ^-1 3362, 3294(NH), 1628(C=O), 1587, 1540(C=C), 1466(CH), 1222, 1106(CO), 877, 866, 826, 723, (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Embodiment-14 Structural Formula

Example-15: Example-15 was synthesized according to the method described in Synthesis Example-11, but the acid chloride was 3,4-dichlorobenzoyl chloride. The structural formula of Example-15 is as follows, the molecular formula is C ₁₇ H ₁₄ Cl ₂ N ₂ O, EIMS 332 (M ⁺ , 20), 173 (C ₆ H ₂ Cl ₂ CO ⁺ , 19), 143 (C ₉ H ₈ NO ⁺ , 98), 130 (C ₉ H ₈ N ⁺ , 100), 117 (C ₈ H ₇ N ⁺ , 2), 105 (C ₆ H ₅ CO ⁺ , 0.5), 77 (C ₆ H ₅ ⁺ , 8) .IR(KBr), cm ^-1 3392, 3285(NH), 1630(C=O), 1589, 1545(C=C), 1462(CH), 1348, 1229, 1132(CO), 886, 845, 757, 738 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Example-15 Structural Formula

Example-16: Example-16 was synthesized according to the method described in Synthesis Example-11, but the acid chloride was 4-fluorobenzoyl chloride. The structural formula of Example-16 is as follows, the molecular formula is C ₁₇ H ₁₅ FN ₂ O, EIMS m/z: 283 (M ⁺ +1, 12), 282 (M ⁺ , 41), 264 (M ⁺ +1-F, 0.5 ), 155 (C ₈ H ₁₀ FNO ⁺ , 1), 143 (C ₁₀ H ₉ N ⁺ , 100), 130 (C ₉ H ₈ N ⁺ , 93), 123 (C ₇ H ₇ FO ⁺ , 55), 115 (C ₈ H ₅ N ⁺ , 13), 103 (C ₆ H ₃ CO ⁺ , 22), 95 (C ₆ H ₄ F ⁺ , 33), 77 (C ₆ H ₅ ⁺ , 26). IR(KBr)cm ^-1 : 3391, 3283(NH), 1628(C=O), 2933, 1564, 1506(C=C), 1454, 1328(CH), 1242, 1162, 1096(CO), 855 , 741, 669 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Embodiment-16 Structural Formula

Example-17: Example-17 was synthesized according to the method described in Synthesis Example-11, but the acid chloride was 2-fluorobenzoyl chloride. The structural formula of Example-17 is as follows, the molecular formula is C ₁₇ H ₁₅ FN ₂ O, EIMS m/z: 283 (M ⁺ +1, 10), 282 (M ⁺ , 34), 264 (M ⁺ +1-F, 0.5 ), 155 (C ₈ H ₁₀ FNO ⁺ , 2), 143 (C ₁₀ H ₉ N ⁺ , 100), 130 (C ₉ H ₈ N ⁺ , 89), 123 (C ₇ H ₇ FO ⁺ , 50), 115 (C ₈ H ₅ N ⁺ , 13), 103 (C ₆ H ₃ CO ⁺ , 22), 95 (C ₆ H ₄ F ⁺ , 42), 77 (C ₆ H ₅ ⁺ , 26). IR (KBr), cm ^-1 3276 (NH), 1621 (C=O), 1565 (C=C), 1454, 1333 (CH), 1221, 1166, 1105 (CO), 845, 763, 746 (= CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Embodiment-17 Structural formula

Example-18: Example-18 was synthesized according to the method described in Synthesis Example-3, but the acid chloride was 4-fluorobenzoyl chloride. Example-18 The structural formula is as follows, the molecular formula is C ₁₇ H ₁₅ FN ₂ O ₂ , EIMS m/z: 299 (M ⁺ +1, 8), 298 (M ⁺ , 34), 281 (M ⁺ -OH, 2) , 159 (C ₁₀ H ₉ NO ⁺ , 100), 146 (C ₉ H ₈ NO ⁺ , 90), 130 (C ₉ H ₈ N ⁺ , 2), 117 (C ₈ H ₇ N ⁺ , 20), 105 (C ₆ H ₅ CO ⁺ , 2), 91 (C ₆ H ₅ CH ₂ ⁺ , 22), 77 (C ₆ H ₅ ⁺ , 7). IR (KBr), cm ^-1 3408, 3324 (OH, NH), 2929, 1629 (C=O), 1584, 1548 (C=C), 1500, 1443, 1370 (CH), 1287, 1185, 1160 ( CO), 937, 851, 759 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Embodiment-18 Structural formula

Example-19: Example-19 was synthesized according to the method described in Synthesis Example-3, but the acid chloride was 2-fluorobenzoyl chloride. Example-19 The structural formula is as follows, the molecular formula is C ₁₇ H ₁₅ FN ₂ O ₂ , EIMS m/z: 299 (M ⁺ +1, 6), 298 (M ⁺ , 28), 281 (M ⁺ -OH, 2) , 159 (C ₁₀ H ₉ NO ⁺ , 100), 146 (C ₉ H ₈ NO ⁺ , 91), 130 (C ₉ H ₈ N ⁺ , 7), 117 (C ₈ H ₇ N ⁺ , 22), 105 (C ₆ H ₅ CO ⁺ , 2), 91 (C ₆ H ₅ CH ₂ ⁺ , 21), 77 (C ₆ H ₅ ⁺ , 7). IR(KBr), cm ^-1 3419, 3311(OH, NH), 2927, 2862, 1641(C=O), 1530(C=C), 1479, 1433(CH), 1294, 1218, 1099(CO) , 937, 839, 755 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Embodiment-19 Structural Formula

Example-20: Example-20 was synthesized according to the method described in Synthesis Example-11, but the acid chloride was 2,4-difluorobenzoyl chloride. The structural formula of Example-20 is as follows, the molecular formula is C ₁₇ H ₁₄ F ₂ N ₂ O, EIMS m/z: 301 (M ⁺ +1, 8), 300 (M ⁺ , 31), 155 (C ₈ H ₁₀ FNO ⁺ , 2), 143 (C ₁₀ H ₉ N ⁺ , 97), 130 (C ₉ H ₈ N ⁺ , 100), 115 (C ₈ H ₅ N ⁺ , 16), 113 (C ₆ H ₃ F ₂ ⁺ , 36), 103 (C ₆ H ₃ CO ⁺ , 30), 91 (C ₆ H ₅ CH ₂ ⁺ , 2), 77 (C ₆ H ₅ ⁺ , 26). IR(KBr), cm ^-1 3248(NH), 1654(C=O), 1530(C=C), 1488, 1338(CH), 1296, 1262, 1229, 1091(CO), 968, 843, 746 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Example-20 Structural Formula

Example-21: Example-21 was synthesized according to the method described in Synthesis Example-11, but the acid chloride was 3,4-difluorobenzoyl chloride. Example-21 The structural formula is as follows, the molecular formula is C ₁₇ H ₁₄ F ₂ N ₂ O, EIMS m/z: 301 (M ⁺ +1, 3), 300 (M ⁺ , 15), 155 (C ₈ H ₁₀ FNO ⁺ , 2), 143 (C ₁₀ H ₉ N ⁺ , 100), 130 (C ₉ H ₈ N ⁺ , 95), 115 (C ₈ H ₅ N ⁺ , 5), 113 (C ₆ H ₃ F ₂ ⁺ , 29), 103 (C ₆ H ₃ CO ⁺ , 10), 91 (C ₆ H ₅ CH ₂ ⁺ , 2), 77 (C ₆ H ₅ ⁺ , 14). IR(KBr)cm ^-1 3347(NH), 3067, 1636(C=O), 1547, 1513(C=C), 1447, 1322(CH), 1284, 1204, 1109(CO), 934,736( =CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Example-21 Structural Formula

Example-22: Example-22 was synthesized according to the method described in Synthesis Example-11, but the acid chloride was 2,4,5-trifluorobenzoyl chloride. The structural formula of Example-22 is as follows, the molecular formula is C ₁₇ H ₁₃ F ₃ N ₂ O, EIMS m/z: 319 (M ⁺ +1, 5), 318 (M ⁺ , 23), 190 (C ₈ H ₅ F ₃ NO ⁺ , 5), 176 (C ₇ H ₅ F ₃ NO ⁺ , 28), 159 (C ₇ H ₂ F ₃ O ⁺ , 100), 143 (C ₁₀ H ₉ N ⁺ , 61), 131 (C ₆ H ₂ F ₃ ⁺ , 47), 130 (C ₉ H ₈ N ⁺ , 68), 115 (C ₈ H ₅ N ⁺ , 4), 113 (C ₆ H ₂ F ₂ ⁺ , 5), 103 (C ₆ H ₃ CO ⁺ , 6), 91 (C ₆ H ₅ CH ₂ ⁺ , 2), 77 (C ₆ H ₅ ⁺ , 11). IR(KBr)cm ^-1 3267(NH), 3064, 1662(C=O), 1611, 1540(C=C), 1425, 1342(CH), 1196, 1141, 1069(CO), 901, 862, 839, 791, 751 (=CH), 607. See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Example-22 Structural Formula

Example-23: Example-23 was synthesized according to the method described in Synthesis Example-3, but the acid chloride was 2,4-difluorobenzoyl chloride. The structural formula of Example-23 is as follows, the molecular formula is C ₁₇ H ₁₄ F ₂ N ₂ O ₂ , EIMS m/z: 316 (M ⁺ , 2), 299 (C ₁₇ H ₁₃ F ₂ N ₂ O, 70), 286 ( 45), 159 (C ₁₀ H ₉ NO ⁺ , 8), 146 (C ₉ H ₈ NO ⁺ , 13), 141 (C ₇ H ₃ F ₂ O ⁺ , 100), 130 (C ₉ H ₈ N ⁺ , 6), 113 (C ₆ H ₃ F ₂ ⁺ , 24), 105 (C ₆ H ₅ CO ⁺ , 2), 91 (C ₆ H ₅ CH ₂ ⁺ , 3), 77 (C ₆ H ₅ ⁺ , 2 ). IR(KBr)cm ^-1 3396, 3293(OH, NH), 3065(CH), 1654(C=O), 1540, 1493, 1432(C=C), 1266(CH), 1184, 1113(CO) , 970, 855, 762 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Example-23 Structural Formula

Example-24: Example-24 was synthesized according to the method described in Synthesis Example-3, but the acid chloride was 3,4-difluorobenzoyl chloride. The structural formula of Example-24 is as follows, the molecular formula is C ₁₇ H ₁₄ F ₂ N ₂ O ₂ , EIMS m/z: 316 (M ⁺ , 60), 299 (C ₁₇ H ₁₃ F ₂ N ₂ O, 17), 286 ( 10), 160 (C ₁₀ H ₁₀ NO ⁺ , 35), 159 (C ₁₀ H ₉ NO ⁺ , 100), 146 (C ₉ H ₈ NO ⁺ , 100), 141 (C ₇ H ₃ F ₂ O ⁺ , 65), 130 (C ₉ H ₈ N ⁺ , 6), 117 (C ₈ H ₇ N ⁺ , 17), 113 (C ₆ H ₃ F ₂ ⁺ , 50), 105 (C ₆ H ₅ CO ⁺ , 2 ), 91 (C ₆ H ₅ CH ₂ ⁺ , 16), 77 (C ₆ H ₅ ⁺ , 5). IR(KBr), cm ^-1 IR(KBr)cm ^-1 3405, 3162(OH, NH), 2952, 2882(CH), 1643(C=O), 1550, 1503, 1468(C=C), 1294 (CH), 1202, 1105, 1070 (CO), 932, 852, 776 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Example-24 Structural Formula

Example-25: Example-25 was synthesized according to the method described in Synthesis Example-3, but the acid chloride was 2,4,5-trifluorobenzoyl chloride. The structural formula of Example-25 is as follows, the molecular formula is C ₁₇ H ₁₃ F ₃ N ₂ O ₂ , EIMS m/z: 334 (M ⁺ , 8), 298 (C ₁₇ H ₁₂ F ₂ N ₂ O, 1), 176 ( C ₇ H ₅ F ₃ NO ⁺ , 5), 160 (C ₁₀ H ₁₀ NO ⁺ , 8), 159 (C ₁₀ H ₉ NO ⁺ , 75), 146 (C ₉ H ₈ NO ⁺ , 48), 141 ( C ₇ H ₃ F ₂ O ⁺ , 4), 13 (C ₉ H ₉ N ⁺ , 15), 117 (C ₈ H ₇ N ⁺ , 5), 113 (C ₆ H ₃ F ₂ ⁺ , 2), 105 (C ₆ H ₅ CO ⁺ , 5), 91 (C ₆ H ₅ CH ₂ ⁺ , 16), 77 (C ₆ H ₅ ⁺ , 5). IR(KBr)cm ^-1 3316(OH, NH), 1651(C=O), 1542, 1426(C=C), 1327, 1276, 1192(CH), 1141, 1103, 1044(CO), 931, 845,798 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Example-25 Structural Formula

Example-26: Example-26 was synthesized according to the method described in Synthesis Example-3, but the acid chloride was 2-aminobenzoyl chloride. The structural formula of Example-26 is as follows, the molecular formula is C ₁₇ H ₁₇ N ₃ O ₂ , EIMS m/z: 296 (M ⁺ +1, 10), 295 (M ⁺ , 40), 163 (C ₉ H ₁₁ N ₂ O ⁺ , 9), 159 (C ₁₀ H ₉ NO ⁺ , 8), 146 (C ₉ H ₈ NO ⁺ , 100), 130 (C ₉ H ₈ N ⁺ , 11), 105 (C ₆ H ₅ CO ⁺ , 5), 92 (C ₆ H ₆ N, 65), 77 (C ₆ H ₅ ⁺ , 14). IR(KBr)cm ^-1 3369(OH, NH), 2918, 1625(C=O), 1579, 1542(C=C), 1485, 1366(CH), 1283, 1159, 1095(CO), 935, 755 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Example-26 Structural Formula

Example-27: Mix 5-methoxytryptamine hydrochloride (100mg, 0.4702mM) with water (5ml) and an excess of 150-200% 2M sodium hydroxide solution in a reaction vessel, add 5ml of chloroform, and Under water bath cooling and stirring, add dropwise a solution of 2-aminobenzoyl chloride (dissolved in 1ml of chloroform) with an equimolar number of sodium hydroxide, and react in the dark, let the reaction mixture rise to room temperature naturally, stir overnight, and white Precipitate, filter, wash the precipitate three times with 30ml chloroform, then dissolve the precipitate in ethyl acetate, wash with 10% hydrochloric acid, 2M sodium hydroxide solution and water successively and dry with anhydrous sodium sulfate, then concentrate the solvent in vacuo to obtain the product. Example-27, the structural formula is as follows, the molecular formula is C ₁₈ H ₁₉ N ₃ O ₂ , EIMS m/z: 309 (M ⁺ , 5), 163 (C ₉ H ₁₁ N ₂ O ⁺ , 55), 146 (C ₉ H ₈ NO ⁺ , 7), 137 (C ₇ H ₇ N ₂ ⁺ O, 15), 105 (C ₆ H ₅ CO ⁺ , 1), 92 (C ₆ H ₆ N, 92), 77 (C ₆ H ₅ ⁺ , 5). IR(KBr)cm ^-1 3358(NH), 2990, 2930, 1629(C=O), 1582, 1536, 1485(C=C), 1325, 1260, 1214(CH), 1157, 1034, 1016(CO ), 922, 825, 807, 755 (=CH). See Table 1 for yield and melting point data, Table 2 for hydrogen spectrum ( ¹ H-NMR) data, and Table 3 for carbon spectrum ( ¹³ C-NMR) data.

Example-27 Structural Formula

Example-28: A research example of the inhibitory effect of the compounds synthesized by the present invention on the mycelial growth of plant pathogenic fungi

1. Strains: Fusarium oxysporum f.sp.vasinfectum, Verticillium dahliae, Fusarium oxysporum f.sp.niveum, Fusarium oxysporum f.sp. cucumerimum), Rhizoctonia solani from rice, Rhizoctonia solani from cotton, Fusarium graminearum.

2. Culture medium:

①PDA (Potato Dextrose Agar) medium: used for the subculture and strain preservation of the tested fungi. Each liter of medium contains 200g of potato boiled filtrate (the specific operation is to take 200g of potatoes, peel them, cut them into small pieces or small pieces, boil them in boiling water for 30min, filter them with four layers of gauze), 20g of glucose, 20g of agar, adjust the pH value to 5.5, Sterilize with damp heat at 121°C for 15 minutes.

②Czapek-Dox medium: used for the subculture and strain preservation of Verticillium dahliae, Rhizoctonia solani and Rhizoctonia oryzae. Each liter of medium contains KCl 0.5g, KH ₂ PO ₄ 1g, MgSO ₄ 7H ₂ O 0.5g (sterilized separately from other components), NaNO ₃ 2g, FeSO ₄ 7H ₂ O 10mg, sucrose 30g, agar 20g ( Liquid culture medium does not add agar), adjust the pH value to 6.0, and sterilize with damp heat at 121°C for 15 minutes.

③Armstrong Fusarium medium: used for liquid culture of Fusarium fungi (including shaker suspension culture and microplate culture). Each liter of medium contains KCl 1.6g, KH ₂ PO ₄ 1.1g, MgSO ₄ 7H ₂ O 0.4g (sterilized separately from other components), Ca(NO ₃ ) ₂ 4H ₂ O 8.49g, MnSO ₄ H ₂ O 0.2 μg, FeCl ₃ ·6H ₂ O 0.2 μg, ZnSO ₄ ·7H ₂ O 0.2 μg, glucose 20 g, adjust the pH value to 4.5, and sterilize at 121°C for 15 minutes.

3. Preparation of compound concentration: Take 1mg sample, dissolve it completely with absolute ethanol, and dilute to 1ml to make a mother solution of 1000μg/ml, and store at -20°C. Before the experiment, the compound mother solution was temporarily diluted to different concentrations, and the ethanol concentration of the solution was fixed at 10%.

4. Fungal liquid suspension culture: Add 20 ml of sterilized liquid medium into a sterile Erlenmeyer flask with a volume of 50 ml. Pick a little mycelium growing on the edge of the fungal colony on the solid medium and put it in 20ml of culture solution, culture it in suspension on a shaker (175±5rpm), 25°C, dark, every 48-72 hours of cultivation (different strains have different cultivation times) ), replace the culture medium once (i.e. take 200 μl of the culture medium containing the bacteria and put it in 20ml of fresh culture medium, and continue to cultivate).

5. Preparation of bacteria solution for antibacterial activity determination: take the bacteria suspension cultured for 48-72 hours, crush the mycelium mass with tweezers, filter with a single layer of gauze, add fresh culture solution with 2 times the concentration to dilute the short mycelia into a certain concentration for the determination of antibacterial activity.

6. Specific operation: take a microwell plate with 96 wells, add 50 μl of compound solutions of different concentrations to each well, repeat 5 wells for each concentration, add 50 μl of the prepared bacterial solution to each well, mix evenly on a shaker, and then The light absorbance value of each well was measured at a wavelength of 655nm, and then incubated in the dark, and the light absorbance value of each well was repeatedly measured after 48 hours or longer.

7. Calculation of inhibition rate and growth half-inhibition rate (IC ₅₀ ): 3 times standard deviation test method was used to eliminate abnormal data. The difference between the average value (ΔC) of the change in the light absorption value of the control and the average value (ΔT) of the change in the light absorption value of the compound at a certain concentration is divided by the average value (ΔC) of the change in the light absorption value of the control, and then multiplied by 100%, that is is the inhibition rate of the compound at a certain concentration.

8, to the mensuration (table 4) of phytopathogenic fungus mycelia growth inhibition

          The compounds in Table 4 inhibit the growth of mycelium of plant pathogenic fungi

Half-inhibitory concentration of mycelial growth (IC ₅₀ , μg/ml)

compound

Fusarium wilt of watermelon Fusarium wilt of cucumber Fusarium wilt of cotton Gibberella wheat Rhizoctonia solani Rhizoctonia solani Rhizoctonia oryzae

Example-1 0.26 1.00 1.05 - - - -

Example-2 2.21 0.11 1.10 - - - -

Example-12 0.46 8.20 0.44 - - - -

Example-13 0.30 - 0.83 3.5 19.20 -

Example-14 0.76 15.30 0.22 0.60 - -

Example-15 0.70 8.50 0.90 0.49 - 0.86

Example-29: A research example of the inhibitory effect of the compounds synthesized by the present invention on the spore germination of Magnaporthe grisea

1. Strains: Magnaporthe grisea P131 and S1528 (Magnaporthe grisea P131 and S1528).

2. Culture medium: Oat tomato Agar is used for the cultivation of blast fungus. Each liter of medium contains 150ml of tomato juice (take 250g of tomato, squeeze the juice with a juicer, and then filter it with double-layer gauze), 750ml of oat juice (30g of oatmeal, add 1000ml of water, boil for 15min, and then filter with double-layer gauze) , agar 20g, 121 ℃ damp heat sterilization for 15min.

3. Preparation of compound concentration: Take 1mg sample, dissolve it completely with absolute ethanol, and dilute to 1ml to make a mother solution of 1000μg/ml, and store at -20°C. Before the experiment, the compound mother solution was diluted to different concentrations, and the ethanol concentration of the solution was fixed at 20%.

4, the cultivation of rice blast bacterium and the preparation of spore suspension: 1) Bacteria coating: wash down the mycelia of rice blast fungus with 700 μ l sterile water, move to the culture medium, coat evenly, and dry naturally (about 30 Minutes), cultivate under light at 25°C; 2) Wash the aerial hyphae: After the aerial mycelium grows (usually 2 days), add 1ml of sterile water, and gently interrupt the aerial mycelium with a sterilized cotton ball , upside down to dry, covered with sterilized gauze, cultured under light at 25°C; 3) preparation of spore suspension: after growing gray spores (generally 1 day), wash the spores with sterile water, filter with four layers of gauze, 2640g (rotating speed) 5000rpm) centrifugation for 15 minutes, remove the supernatant, the spores are made into a spore suspension with sterile water, then filter with four layers of gauze, count with a Neubauer hemocytometer, and then dilute the spore suspension to 1×10 ⁶ ml.

5. Specific operation: Take 30 μl of the compound solution and 30 μl of the spore suspension, mix them evenly in an eppendorf tube, take 10 μl on a glass slide to form water droplets, and incubate at a relative humidity of 100% at 25°C in the dark for 16 hours, then observe under a microscope Spore germination and germ tube growth status. Under a low-magnification microscope, 100-200 spores were randomly inspected. Those with germ tubes were regarded as germinated, and those without germ tubes were considered not to germinate.

First calculate the inhibition rate of spore germination and germ tube growth at each concentration, and then use the interpolation formula to calculate the half inhibitory concentration (IC ₅₀ ) of the compound. The calculation formulas of spore germination inhibition rate and germ tube growth inhibition rate are as follows:

6. Determination of inhibitory effect on spore germination of Magnaporthe grisea (Table 5).

The inhibitory effect of table 5 compounds on Magnaporthe grisea strain P131 and S1528 compound Magnaporthe grisea strain P131 Magnaporthe grisea strain S1528 Germ tube growth half-inhibitory concentration (μg/ml) Half inhibitory concentration of spore germination (μg/ml) Germ tube growth half-inhibitory concentration (μg/ml) Half inhibitory concentration of spore germination (μg/ml) Example-1 16.60 - 20.02 - Example-2 13.55 - 17.38 - Example-3 0.15 5.83 0.44 6.25 Example-4 0.45 5.31 2.49 15.63 Example-5 0.44 2.32 0.55 15.63 Example-6 1.32 11.36 1.55 7.50 Example-7 0.27 5.00 1.99 9.06 Example-8 1.98 7.32 2.05 5.00 Example-9 0.12 4.57 1.01 3.59 Example-10 0.63 6.50 0.99 5.83 Example-11 0.36 7.75 0.40 1.83 Example-12 4.94 - 28.54 - Example-13 12.44 - 1.91 - Example-14 1.98 7.88 2.04 14.63 Example-15 1.64 7.72 1.77 8.22 Example-16 41.30 - / / Example-17 23.54 42.75 / / Example-18 - - / / Example-19 - - / / Example-20 27.84 35.74 / / Example-21 38.50 41.79 / / Example-22 65.17 63.94 / / Example-23 50.77 - / / Example-24 - - / / Example-25 - - / / Example-26 - - / / Example-27 73.52 98.16 / /

/: Indicates that antibacterial activity has not been determined.

Embodiment-30: The research example of anti-Candida albicans activity of the compound synthesized by the present invention

1. Strains: Candida albicans Berkh ATCC10231.

2. Candida albicans culture medium: used for plate culture and liquid culture of Candida albicans. Each liter of medium contains KH ₂ PO ₄ 0.5g, MgSO ₄ 7H ₂ O 0.5g (sterilized separately from other components), CaCl ₂ 2H ₂ O 0.3g, (NH ₄ ) ₂ SO ₄ 2g, KI 0.1g , asparagine (L-asparagine) 2g, glucose 20g, a small amount of trace elements (B, Mn, Zn, Cu, Mo, Fe) (the specific formulation is: from MnCl ₂ 4H ₂ O 450.7mg, ( NH ₄ ) ₆ Mo ₇ O ₂₄ 4H ₂ O 23mg, H ₃ BO ₃ 282.2mg, ZnSO ₄ 7H ₂ O 11mg, CuSO ₄ 5H ₂ O 9.8mg, FeC ₆ H ₅ O ₇ 5H ₂ O 2.998g Take 1ml from 500ml of mother liquor), D-biotin (vitamin H) 2mg, agar 20g (do not add in liquid medium), adjust the pH value to 5.5, and sterilize with damp heat at 121°C for 15min.

3. Liquid suspension culture of Candida albicans: Add 20 ml of sterilized liquid culture medium into a sterile Erlenmeyer flask with a volume of 50 ml. Pick a little mycelium growing on the edge of the fungal colony on the solid medium and put it in 20ml of culture solution, culture it in suspension on a shaker (175±5rpm), at 25°C, in the dark, and replace the culture solution once every 48-72 hours of cultivation ( That is, take 200 μl of the culture medium containing the bacteria and put it in 20ml of fresh culture medium, and continue to cultivate).

4. Preparation of compound concentration: take 1mg of sample, dissolve it completely with absolute ethanol, and dilute to 1ml to make a mother solution of 1000μg/ml, and store at -20°C. Before the experiment, the compound mother solution was temporarily diluted to different concentrations, and the ethanol concentration of the solution was fixed at 10%.

5. Specific operation: take a microwell plate with 96 wells, add 50 μl of compound solutions of different concentrations to each well, repeat 5 wells for each concentration, add 50 μl of the prepared bacterial solution to each well, mix evenly on a shaker, and The light absorbance value of each well was measured at a wavelength of 655nm, and then incubated in the dark, and the light absorbance value of each well was repeatedly measured after 48 hours or longer.

6. Calculation of inhibition rate and growth half-inhibition rate (IC ₅₀ ): 3 times standard deviation test method was used to eliminate abnormal data. The difference between the average value (ΔC) of the change in the light absorption value of the control and the average value (ΔT) of the change in the light absorption value of the compound at a certain concentration is divided by the average value (ΔC) of the change in the light absorption value of the control, and then multiplied by 100%, that is is the inhibition rate of the compound at a certain concentration.

7. Determination of the inhibitory effect on the growth of Candida albicans

Among the tested compounds, 6 compounds showed inhibitory effect on the growth of Candida albicans at the tested concentrations (the results are shown in Table 6).

         The compounds in Table 6 have an inhibitory effect on Candida albicans

Compound IC ₅₀ (μg/ml) Compound IC ₅₀ (μg/ml)

Example-1 6.15 Example-17 14.56

Example-2 8.31 Example-21 10.59

Example-5 16.46 Example-22 19.89

Example-6 14.38 Example-23 12.45

Example-10 63.15 Example-25 10.14

Example-14 8.74 Example-24 12.69

Example-15 11.98 Example-26 5.08

Example-16 12.95 Example-27 3.80

Example-31: Example of research on the control effect of some compounds synthesized by the present invention on cotton wilt

1. Strains: Fusarium oxysporum f.sp.vasinfectum

2. Culture medium: 1) PDA (Potato Dextrose Agar) medium: used for subculture and strain preservation of the tested fungi. Each liter of medium contains 200g of potato boiled filtrate (the specific operation is to take 200g of potatoes, peel them, cut them into small pieces or small pieces, boil them in boiling water for 30min, filter them with four layers of gauze), 20g of glucose, 20g of agar, adjust the pH value to 5.5, Sterilize with damp heat at 121°C for 15 minutes. 2) Wheat grain sand medium: Take wheat grains in a beaker, add water and boil for 30 minutes, then mix with sieved and washed fine sand at a volume ratio of 1:2, and sterilize with moist heat at 121°C for 60 minutes.

3. Preparation of the inoculum: Cut the agar block containing mycelia from the edge of the colony of the newly cultured Fusarium wilt, inoculate it in a triangular flask filled with wheat sand medium, mix well, and cultivate it at 25°C for about 10 days , Pay attention to shake well every day, and use it for the preparation of diseased soil.

4. Pre-treatment of seeds before sowing: develvet cotton seeds in concentrated sulfuric acid at 100±5°C (about 10ml of concentrated sulfuric acid is needed for 500 seeds), then rinse with tap water until neutral, remove grains, and dry in the air. Before accelerating germination, soak the seeds in warm water at 58°C for 30 minutes, then place them on wet gauze, and accelerate germination in a constant temperature box at 37°C (usually 24 hours).

5. Compound preparation: the preparation concentration is 20 μg/g. Based on the calculation of 500g of soil for each treatment (that is, 125g of soil per pot, 4 small pots in total), weigh 10mg of the compound, dissolve it with 3ml of ethanol, add 5g of quartz sand, mix well, and wait for the ethanol to volatilize for later use.

6. Preparation of diseased soil: take the sterilized cultivation soil in the pot, add wheat grain sand containing Fusarium wilt in a certain proportion (cultivation soil: wheat grain sand=10:1, w/w; or cultivation soil : Dried barley sand = 15:1, w/w), mix well. Take by weighing 500g diseased soil, add the quartz sand containing compound, make the concentration of compound be 20ug/g. In addition, the soil containing bacteria but no compound, soil containing compound but no bacteria, and neither bacteria nor compound were taken as controls.

7. Sowing: Sow the germinated seeds into cups, sow 5 seeds per cup, soak them with sterile water from the bottom of the cups, and then put them in a growth room for cultivation at 22°C. At the first week and the seventh week, the observation results showed that the symptoms of cotton wilt in this study were mainly bacterial wilt, and the diseased plants were dwarf. If it withers and dries up, it is considered withered. The expression method of control effect is:

8, the control effect measurement of cotton fusarium wilt (table 7)

Compounds Example-7, Example-14 and Example-15 were selected, and the control effect of the compounds on cotton fusarium wilt was determined by a pot test method. The concentration of the compound in the soil was 20mg/kg, and the results are shown in Table 7. Example-7, Example-14 and Example-15 all have certain control effects on cotton fusarium wilt, and the disease prevention effects of Example-14 and Example-15 are better than Example-7.

Control effect of table 7 compounds on cotton fusarium wilt deal with Observation results of 3 weeks of cultivation Observation results of 7 weeks of cultivation Withering rate (%) Control effect (%) Withering rate (%) Control effect (%) Control 1 (do not add fungus) control 2 (add fungus) embodiment 7 (add fungus) embodiment 14 (add fungus) embodiment 15 (add fungus) embodiment 7 (do not add fungus) embodiment 14 (do not add fungus) Embodiment 15 (do not add fungus) 060501120000 --16.6781.6766.67--- 090754450000 --16.6751.1144.44---

-: Indicates that this column is blank.

Claims (8)

1, the N-acyl-trypamine compound shown in the general formula [I], general formula [I] is:

Wherein:

R ₁Expression hydrogen or hydroxyl, halogen, alkoxyl group;

R ₂Expression hydrogen or hydroxyl, halogen, alkoxyl group, alkylthio;

R ₃Expression-CH=CH-,-CH ₂-or do not exist;

R ₄Expression-(CH ₂) ₂-.

2, the method for a kind of discovery and isolation identification paddy rice N-acyl-trypamine plant protecting chemical: with rice blast pathogen conidiospore suspension spray inoculation paddy rice seedling; then at 100% relative humidity, lucifuge clip inoculation blade after for some time; with the vegetable chemistry separation method of rice leaf lipid-soluble substance by silica gel column chromatography, preparation of lamina chromatography or high performance liquid chromatography; the described N-acyl-trypamine of claim 1 compound is arrived in separation and purification: N-benzoyl tryptamines and two kinds of plant protecting chemicals of N-cinnamoyl tryptamines; pass through wave spectrum analysis; carried out structure determination, its structural formula is as follows:

N-benzoyl tryptamines

N-cinnamoyl tryptamines

3, a kind of method of the N-of preparation benzoyl tryptamines: get in the pyridine that a certain amount of tryptamines is dissolved in proper volume; under the ice-water bath cooling, drip Benzoyl chloride; under the ice-water bath cooling, stir for some time; at room temperature stir for some time then, reaction mixture is poured in the frozen water, divides three extractions with the chloroform of proper volume; chloroform part water and dilute hydrochloric acid washing; be removed until pyridine, chloroform is partly used the siccative drying, and vacuum concentration promptly gets N-benzoyl tryptamines.

4, a kind of method of the N-of preparation cinnamoyl tryptamines: get in the pyridine that a certain amount of tryptamines is dissolved in proper volume; under the ice-water bath cooling, drip cinnamyl chloride; under the ice-water bath cooling, stir for some time; at room temperature stir for some time then, reaction mixture is poured in the frozen water, divides three extractions with the chloroform of proper volume; chloroform part water and dilute hydrochloric acid washing; be removed until pyridine, chloroform is partly used the siccative drying, and vacuum concentration promptly gets N-cinnamoyl tryptamines.

5; a kind of not method of the N-acyl-trypamine acylate of hydroxyl of 5-position for preparing: in reaction vessel that tryptamines and excess of triethylamine is mixed; add an amount of chloroform; in ice-water bath cooling and stir under; the solution of acid chloride of mole numbers such as dropping and triethylamine; under lucifuge, react; allow reaction mixture rise to room temperature naturally; stirring is spent the night; the water that adds proper volume, with after chloroform layer separates, water layer is again with the equal amounts of chloroform washing once with water layer; the combined chloroform layer; use acid solution successively; alkali lye and water washing, and with after the siccative drying are reclaimed solvent and are promptly obtained the 5-position acylate of hydroxyl not in the claim 1.

6; a kind of method for preparing the N-acyl-trypamine acylate of 5-position hydroxyl: in reaction vessel that serotonin hydrochloride and water and excessive alkali aqueous solution is mixed; add an amount of chloroform; under the ice-water bath cooling and stirring; the solution of acid chloride of mole numbers such as dropping and alkali aqueous solution; under lucifuge, react; allow reaction mixture rise to room temperature naturally; stirring is spent the night; white precipitate occurs, filter, divide three washing precipitations with the chloroform of proper volume; then precipitation is dissolved in ethyl acetate; use acid solution successively; alkali lye and water washing, and, reclaim the acylate that solvent promptly obtains the 5-position hydroxyl in the claim 1 with after the siccative drying.

7, the N-acyl-trypamine compound in the claim 1 is used to prepare the medicine of anti-phytopathogen, and affiliated phytopathogen comprises: rice blast fungus, watermelon Fusarium oxysporum, cucumber Fusarium oxysporum, cotton wilt fusarium, gibberella saubinetii, cotton dry thread Pyrenomycetes, the withered bacterium of rice line.

8, the N-acyl-trypamine compound in the claim 1 is used to prepare the medicine of anti-human body pathogenic bacteria, and affiliated human body pathogenic bacteria comprises Candida albicans.