阅读说明：本技术 含酰腙结构的pityriacitrin生物碱衍生物及其制备方法和用途 (Pityriacitrin alkaloid derivative containing acylhydrazone structure and preparation method and application thereof ) 是由 卢爱党 王禄 王铁男 李红岩 王兹稳 陈建新 于 2021-06-02 设计创作，主要内容包括：本发明含酰腙结构的pityriacitrin生物碱衍生物及其制备方法和用途,涉及含有β-咔啉且1位含有酰腙结构的pityriacitrin生物碱衍生物的杀生剂,含酰腙结构的pityriacitrin生物碱衍生物,具有如下化学结构式通式Ⅰ,其制备方法,具体步骤如下化学反应方程式所示,其用途为用作抗植物病毒剂和用作杀植物病原真菌剂。(The invention relates to pityriacitrin alkaloid derivatives containing acylhydrazone structures, a preparation method and application thereof, and relates to a biocide of pityriacitrin alkaloid derivatives containing beta-carboline and acylhydrazone structures at 1-position, the pityriacitrin alkaloid derivatives containing acylhydrazone structures have the following chemical structural formula I, the preparation method comprises the following steps of as shown in a chemical reaction equation,)

1. Pityriacitrin alkaloid derivative containing acylhydrazone structure is characterized in that: has the following chemical structural formula I,

in the general formula I, R represents methyl, propyl, isopropyl, n-butyl, tert-butyl, benzyl, cyclopentyl, cyclohexyl, phenyl, 2-fluorophenyl, 3, 4-difluorophenyl, 2, 6-difluorophenyl, 2, 4-difluorophenyl, 2, 3-difluorophenyl, 4-fluorophenyl, 2-chlorophenyl, 3, 4-dichlorophenyl, 2, 6-dichlorophenyl, 2, 4-dichlorophenyl, 2, 3-dichlorophenyl, 2, 5-dichlorophenyl, 4-chlorophenyl, 5-chloro-2-fluorophenyl, 2-bromophenyl, 3-bromophenyl, 4-iodophenyl, 2-nitrophenyl, 3-nitrophenyl, tert-butyl, benzyl, cyclopentyl, cyclohexyl, phenyl, 2-fluorophenyl, 4-iodophenyl, 2-nitrophenyl, etc, 4-nitrophenyl, 2-methylphenyl, 3-methylphenyl, 4-methylphenyl, 2-trifluoromethylphenyl, 3-trifluoromethylphenyl, 4-trifluoromethylphenyl, 3, 5-bis (trifluoromethyl) phenyl, 3-methoxyphenyl, 3,4, 5-tris (methoxy) phenyl, 4-methoxyphenyl, 2-thienyl, 2-furyl, 2-pyridyl or 4-pyridyl.

2. The pityriacitrin alkaloid derivative containing an acylhydrazone structure according to claim 1, wherein: are compounds having the following chemical structural formulas I-1 to I-11:

3. the method for preparing pityriacitrin alkaloid derivative containing acylhydrazone structure as claimed in claim 1, wherein: the specific steps are shown in the following chemical reaction equation,

tryptamine shown in a chemical structural formula 1 is used as a raw material to react with a glyoxylic acid aqueous solution to generate a2, 3,4, 9-tetrahydro-pyrido [3,4-b ] indole-1-formic acid intermediate product shown in a chemical structural formula 2; dissolving the intermediate product in methanol, adding thionyl chloride for reaction, and then esterifying to obtain 2,3,4, 9-tetrahydro-pyrido [3,4-b ] indole-1-methyl formate shown as a chemical structural formula 3; then oxidizing under the action of sulfur powder to prepare pyrido [3,4-b ] indole-1-methyl formate shown as a chemical structural formula 4; then, carrying out hydrazinolysis reaction to generate pyrido [3,4-b ] indole-1-formhydrazide shown as a chemical structural formula 5; finally, the pyrido [3,4-b ] indole-1-formylhydrazine reacts with different substituted aldehyde compounds to prepare the N' -methylene-pyrido [3,4-b ] indole-1-formylhydrazine shown as the chemical structural formula I.

4. The use of pityriacitrin alkaloid derivatives containing acylhydrazone structures as claimed in claim 1, wherein: the pityriacitrin alkaloid derivative containing the acylhydrazone structure is used as an anti-plant virus agent, and is a compound shown in the following chemical structural formulas I-1 to I-11:

the plant virus is tobacco mosaic virus, pepper virus, rice virus, tomato virus, sweet potato virus, melon virus and maize dwarf mosaic virus.

5. The use of pityriacitrin alkaloid derivatives containing acylhydrazone structures as claimed in claim 1, wherein: the pityriacitrin alkaloid derivative containing the acylhydrazone structure is a compound represented by the following chemical structural formulas I-1 to I-11 and is used as a plant pathogenic fungicide:

the plant pathogenic fungi are cucumber fusarium wilt, peanut brown spot fungus, apple ring spot fungus, tomato early blight, wheat gibberella, rice bakanae fungus, rape sclerotinia, pepper phytophthora capsici, wheat sheath blight fungus, corn microsporum, watermelon anthracnose fungus, potato late blight fungus, rice blast fungus, rice sheath blight fungus and cucumber gray mold.

Technical Field

The technical scheme of the invention relates to a biocide of pityriacitrin alkaloid derivatives containing beta-carboline and acylhydrazone structure at position 1, in particular to pityriacitrin alkaloid derivatives containing acylhydrazone structure and a preparation method and application thereof.

Background

The Pityriacitrins alkaloid is a beta-carboline alkaloid with a novel structure, and an indole structure is connected at the C-1 position of beta-carboline through a carbonyl functional group. The compounds containing the core skeleton extracted from nature include pityriacitrin, pityriacitrin B, hysiolawesine, 7-MeO-pityriacitrin, 7, 2' - (MeO)₂Pityriacitrin alkaloids (Mini. Rev. Med. chem.,2021,21, 233-one 244), such as pityriacitrin and 2 '-MeO-6' -OH-pityriacitrin, are shown in the structural formula I.

Pityriacitrin alkaloids have been first reported by the Japanese research group in 1999 to be isolated from the marine bacterium Paracoccus (strain F-1547) (JP 11269175A 2,1999), with subsequent confirmation that pityriacitrin and its derivatives are also present in other bacteria and fungi (Arch. Dermatol. Res.,2002,294(3), 131. cndot. 134; J. Nat. Prod.,2002, 65(8), 1173. cndot. 20156; Helv. Chim. acta,2005,8, 1472. cndot. 1485; Mycoses,2006,49(5), 388. cndot. J. Invest. Dermatol. 2008,128(7), 1620. cna 1625; Eur. J. Org. chem.,2010, 2084. cndot. 0; bioorg. chett. chet., 22(6), 2296. cna., 2296, 2299, 20199. cnd. cnotch, 133, 2014. cnotch, 20199, 11. cnotr, 133, 132, 11. cnotr, 11. cnotr, 133, 11. cnotr, 12, 11. cnotr. Pityriacitrin alkaloids have good ultraviolet absorption performance, and also have the function of activating an AhR signal path (Clin. Microbiol. Rev.,2012,25(1), 106-; gaitanis et al (Mycoses,2019,62(7),597-the black yeast exothia dermatitidis), 13 molds including 7 Aspergillus spp, 2 Fusarium spp and 2 Rhizopus oryzae. Lan et al (mar. drugs,2017,15(11),339.) reported that pityriacitrin has moderate cytotoxicity to human colon cancer cells, but did not have any inhibitory effect on four bacteria, Staphylococcus aureus (Staphylococcus aureus ATCC29213), Escherichia coli (Escherichia coli ATCC25922), Pseudomonas aeruginosa (Pseudomonas aeruginosa ATCC27853) and acinetobacter baumannii (Bauman's acinetobacter ATCC 19606). Li et al found pityriacitrin derivatives 7-MeO-pityriacitrin, 7, 2' - (MeO) in 2012₂Examination of the inhibition of human cancer cells (HL-60, SMMC-7721, A-549, MCF-and SW480) by-pityriacitrin, 7-MeO-6 '-OH-pityriacitrin and 2' -MeO-6 '-OH-pityriacitrin revealed that 7, 2' - (MeO)₂Pityriacitrin shows strong cytotoxicity to 4 human cancer cells, SMMC-7721, A-549, MCF-7 and SW 480. However, Tan et al (Helv. Chim. acta,2015,98,72-77.) studied pityriacitrin derivatives 3-OH-7, 6' - (MeO)₂Pityriacitrin did not show significant cytotoxicity against human cancer cells HL-60 and A-549.

Pityriacitrin alkaloids attract a lot of attention due to their unique chemical structures, and various research groups use pityriacitrin as a lead compound for structural modification and biological activity research. Jiang et al (Sun Xiaofei, Synthesis and antitumor Activity evaluation of Marine alkaloid pityriacitrin and analogs, emodin sugar derivatives, China oceanic university [ D ]]2008; Eur.J.Med.chem.,2011,46(12),6089-₅₀12.94,6.35 and 16.37 μ M, respectively). Liew (Tetrahedron,2014,70,4910-And Xu (Eur.J.Med.chem., 2019,168,293-300) et al also report pityriacitrin derivatives on the study of inhibiting cancer cells.

Through literature research, due to the low natural content of pityriacitrins alkaloids, low synthesis yield and high cost, researchers mainly focus on the application research of pityriacitrins alkaloids in the aspect of medicines, particularly in the aspect of tumor resistance, and few reports are made in the aspect of controlling plant diseases and insect pests. With the continuous exploration of new methods for synthesizing pityriacitrins alkaloids, Wu and Bharate et al (Mini.Rev.Med.chem.,2021,21, 233-244.) report a new method for synthesizing pityriacitrin and derivatives thereof by a one-pot method, which greatly reduces the difficulty and cost of synthesis. Therefore, the pityriacitrin and the derivative thereof are applied to controlling plant diseases and insect pests in agricultural production by modifying and modifying the structure of the pityriacitrin, so that the biological activity spectrum of the pityriacitrin and the derivative thereof can be widened, and the pityriacitrin and the derivative thereof have very important application value to the protection of crops.

Disclosure of Invention

The technical problem to be solved by the invention is as follows: the pityriacitrin alkaloid derivative containing the acylhydrazone structure is a novel beta-carboline derivative, and has good activity of resisting plant viruses and germs for the first time.

The technical scheme adopted by the invention for solving the technical problem is as follows: the pityriacitrin alkaloid derivative containing the acylhydrazone structure has the following chemical structural formula I,

in the general formula I, R represents methyl, propyl, isopropyl, n-butyl, tert-butyl, benzyl, cyclopentyl, cyclohexyl, phenyl, 2-fluorophenyl, 3, 4-difluorophenyl, 2, 6-difluorophenyl, 2, 4-difluorophenyl, 2, 3-difluorophenyl, 4-fluorophenyl, 2-chlorophenyl, 3, 4-dichlorophenyl, 2, 6-dichlorophenyl, 2, 4-dichlorophenyl, 2, 3-dichlorophenyl, 2, 5-dichlorophenyl, 4-chlorophenyl, 5-chloro-2-fluorophenyl, 2-bromophenyl, 3-bromophenyl, 4-iodophenyl, 2-nitrophenyl, 3-nitrophenyl, 4-nitrophenyl, 2-methylphenyl, 3-methylphenyl, 4-methylphenyl, 2-trifluoromethylphenyl, 3-trifluoromethylphenyl, 4-trifluoromethylphenyl, 3, 5-bis (trifluoromethyl) phenyl, 3-methoxyphenyl, 3,4, 5-tris (methoxy) phenyl, 4-methoxyphenyl, 2-thienyl, 2-furyl, 2-pyridyl, or 4-pyridyl.

The pityriacitrin alkaloid derivative containing the acylhydrazone structure is preferably a compound shown in the following chemical structural formulas I-1 to I-10:

the preparation method of the pityriacitrin alkaloid derivative containing the acylhydrazone structure comprises the following specific steps of:

tryptamine shown in a chemical structural formula 1 is used as a raw material to react with a glyoxylic acid aqueous solution to generate a2, 3,4, 9-tetrahydro-pyrido [3,4-b ] indole-1-formic acid intermediate product shown in a chemical structural formula 2; dissolving the intermediate product in methanol, adding thionyl chloride for reaction, and then esterifying to obtain 2,3,4, 9-tetrahydro-pyrido [3,4-b ] indole-1-methyl formate shown as a chemical structural formula 3; then oxidizing under the action of sulfur powder to prepare pyrido [3,4-b ] indole-1-methyl formate shown as a chemical structural formula 4; then, carrying out hydrazinolysis reaction to generate pyrido [3,4-b ] indole-1-formhydrazide shown as a chemical structural formula 5; finally, the pyrido [3,4-b ] indole-1-formylhydrazine reacts with different substituted aldehyde compounds to prepare the N' -methylene-pyrido [3,4-b ] indole-1-formylhydrazine shown as the chemical structural formula I.

The pityriacitrin alkaloid derivative containing the acylhydrazone structure is a compound shown in the following chemical structural formulas I-1-I-11 and can be used as an anti-plant virus agent:

the plant virus is tobacco mosaic virus, pepper virus, rice virus, tomato virus, sweet potato virus, melon virus and maize dwarf mosaic virus.

The pityriacitrin alkaloid derivative containing the acylhydrazone structure is a compound shown as the following chemical structural formulas I-1 to I-11 and is used as a plant pathogenic fungicide:

the plant pathogenic fungi include 15 plant pathogenic fungi including cucumber fusarium wilt, peanut brown spot fungus, apple ring spot fungus, tomato early blight fungus, wheat gibberella, rice bakanae fungus, rape sclerotinia, pepper phytophthora capsici, wheat sheath blight fungus, corn microsporum, watermelon anthracnose fungus, potato late blight fungus, rice blast fungus, rice sheath blight fungus and cucumber gray mold fungus.

The preparation method of the pityriacitrin alkaloid derivative containing the acylhydrazone structure comprises the steps of using the related raw materials of tryptamine, acetic aldehyde aqueous solution and SO₂Cl₂Sulfur powder, hydrazine hydrate, various kinds of aldehyde compounds, and the like are commercially available, and the chemical reaction process is within the grasp of those skilled in the art.

The invention has the beneficial effects that: compared with the prior art, the invention has the prominent substantive characteristics and remarkable progress as follows:

(1) the invention synthesizes the pityriacitrin alkaloid derivatives with acylhydrazone-containing structures shown in the chemical structural formula I for the first time, particularly the pityriacitrin alkaloid derivatives with acylhydrazone-containing structures shown in the chemical structural formulas I-1-I-11, provides the preparation methods of the compounds for the first time, and discovers that the pityriacitrin alkaloid derivatives have good activity of resisting plant viruses and germs for the first time.

Detailed Description

Example 1

The method for preparing the pityriacitrin alkaloid derivative containing the acylhydrazone structure with the chemical structural formula I comprises the following steps:

the prepared pityriacitrin alkaloid derivative containing the acylhydrazone structure has the following chemical structural formula I:

in the general formula I, R represents methyl, propyl, isopropyl, n-butyl, tert-butyl, benzyl, cyclopentyl, cyclohexyl, phenyl, 2-fluorophenyl, 3, 4-difluorophenyl, 2, 6-difluorophenyl, 2, 4-difluorophenyl, 2, 3-difluorophenyl, 4-fluorophenyl, 2-chlorophenyl, 3, 4-dichlorophenyl, 2, 6-dichlorophenyl, 2, 4-dichlorophenyl, 2, 3-dichlorophenyl, 2, 5-dichlorophenyl, 4-chlorophenyl, 5-chloro-2-fluorophenyl, 2-bromophenyl, 3-bromophenyl, 4-iodophenyl, 2-nitrophenyl, 3-nitrophenyl, 4-nitrophenyl, 2-methylphenyl, 3-methylphenyl, 4-methylphenyl, 2-trifluoromethylphenyl, 3-trifluoromethylphenyl, 4-trifluoromethylphenyl, 3, 5-bis (trifluoromethyl) phenyl, 3-methoxyphenyl, 3,4, 5-tris (methoxy) phenyl, 4-methoxyphenyl, 2-thienyl, 2-furyl, 2-pyridyl, or 4-pyridyl.

The preparation method of the pityriacitrin alkaloid derivative containing the acylhydrazone structure with the chemical structural formula I comprises the following specific steps of:

the tryptamine shown in a chemical structural formula 1 is used as a raw material to react with a glyoxylic acid aqueous solution to generate a 1,2,3, 4-tetrahydro-beta-carboline-1-carboxylic acid intermediate product shown in a chemical structural formula 2; dissolving the intermediate product in methanol, adding thionyl chloride for reaction, and then esterifying to obtain 1,2,3, 4-tetrahydro-beta-carboline-1-methyl carboxylate shown as a chemical structural formula 3; then oxidizing the mixture under the action of sulfur powder to prepare beta-carboline-1-methyl carboxylate shown as a chemical structural formula 4; then the beta-carboline-1-hydrazide shown as a chemical structural formula 5 is generated through hydrazinolysis reaction; and finally reacting the beta-carboline-1-hydrazide with different substituted aldehyde compounds to obtain the 1-acylhydrazone-beta-carboline derivative shown as the chemical structural formula I.

Example 2

The preparation method of the N' -benzylidene-pyrido [3,4-b ] indole-1-formylhydrazine shown in the chemical structural formula I-1 comprises the following steps:

the chemical structural formula of the N' -benzylidene-pyrido [3,4-b ] indole-1-formylhydrazine I-1 is

The preparation method comprises the following specific steps:

in the first step, aqueous glyoxylic acid monohydrate (10.37g, 50% by mass) was mixed with ethyl acetate (145mL), and K was added dropwise₂CO₃After adjusting the pH of the aqueous solution (20mL, 10% by mass) to 5 and slowly adding tryptamine (9.60 g, 60mmol) with stirring to dissolve it sufficiently, the reaction was stirred at room temperature for 24 hours. After the reaction is finished, filtering, washing the solid product by using ethyl acetate and a small amount of ethanol, and drying to obtain 10.702g of yellow solid compound 1 with the yield of 83%, wherein the yellow solid substance is determined to have the following relevant parameters:¹H NMR(400MHz,DMSO-d₆)δ2.81–2.85(m,1H,CH₂), 2.88–2.95(m,1H,CH₂),3.27–3.46(m,2H,CH₂),4.66(s,1H,CH),6.97–7.03(m,2H,ArH), 7.38-7.47(m,2H,ArH),8.91(brs,1H,NH),10.64(s,1H,COOH)；¹³C NMR(100MHz, DMSO-d₆) δ 18.7,41.2,55.9,104.3,112.6,117.7,118.7,121.1,126.5,129.3,136.6,165.8; the intermediate product was identified as 2,3,4, 9-tetrahydro-pyrido [3,4-b]Indole-1-carboxylic acid.

In the second step, 2,3,4, 9-tetrahydro-pyridine is addedAnd [3,4-b ]]Indole-1-carboxylic acid (10.70g, 49.5mmol) dissolved in CH₃In OH (110mL), SOCl was slowly added dropwise while controlling the temperature to 0 deg.C₂(10.78 mL). After the dropwise addition, the temperature is raised to room temperature, the temperature is controlled to 70 ℃ for reflux, and the solvent is removed in vacuum after the detection reaction is completed. To the residue was added 40mL of water, and saturated Na was added dropwise₂CO₃The solution was adjusted to neutral pH, extracted three times with dichloromethane (40 mL. times.3), the combined organic phases were dried over anhydrous sodium sulfate, filtered with suction and the solvent removed in vacuo to give a reddish brown solid which was directly taken to the next step without purification.

Step three, adding dimethylbenzene (200mL) and sulfur powder (2.88g) into the crude product obtained in the step two, refluxing for 5 hours, cooling to room temperature, removing the solvent in vacuum, and purifying the residual liquid by column chromatography (the eluent is petroleum ether: ethyl acetate: 4:1) to obtain 7.55g of brown solid product 4 with the yield of 89%; relevant parameters of the brown solid matter were determined to be:¹H NMR(CDCl₃,400MHz)δ9.91(s,1H,NH),8.59(d,J＝5.0Hz,1H,ArH),8.17–8.5 (m,2H,ArH),7.62–7.59(m,2H,ArH),7.36–7.32(m,1H,ArH),4.13(s,3H,OCH₃)；¹³C NMR(DMSO-d₆100MHz) delta 166.17,139.71,137.79,136.06,130.43,128.41,128.32,120.86, 119.72,119.66,117.63,110.80, 51.76; the product was identified as pyrido [3,4-b ]]Indole-1-carboxylic acid methyl ester.

In the fourth step, pyrido [3,4-b ] is reacted with]Indole-1-methyl formate (5.79g, 25.59mmol) is added into a mixed system of ethanol (100mL) and hydrazine hydrate (19.84mL, mass fraction 80%), heating and refluxing are carried out for 4h, the raw materials are directly filtered after complete reaction, and then 5.21g of white solid product 5 can be obtained, wherein the yield is 90%; the white solid matter was determined to have the following relevant parameters:¹H NMR(DMSO-d₆,400MHz):δ11.70(s,1H,NH),10.00(s,1H,NH),8.37-8.25(m,3H,ArH), 7.81(d,J＝8.2Hz,1H,ArH),7.57(t,J＝7.4Hz,1H,ArH),(t,J＝7.2Hz,1H,ArH),4.65(s, 2H,NH₂)；¹³C NMR(DMSO-d₆100MHz) δ 164.04,141.49,136.71,134.10,132.32,130.44, 128.69,121.70,119.87,119.68,117.57,112.98; the product was identified as pyrido [3,4-b ]]Indole-1-carboxylic acid hydrazide.

The fifth step is toPyrido [3,4-b]Dissolving indole-1-formylhydrazine (0.68g, 3mmol) and benzaldehyde (0.32g, 3mmol) in tetrahydrofuran (30mL), refluxing for 4 hours, cooling to room temperature after the raw materials react completely, removing the solvent in vacuum, and washing with diethyl ether to obtain a white solid product I-1 with a yield of 95%; the white solid matter was determined to have the following relevant parameters:¹H NMR(400MHz,CDCl₃)δ11.21(s,1H,CONH),10.42(s,1H,NH),8.40(d,J＝ 4.9Hz,1H,ArH),8.33(s,1H,N＝CH),8.16–8.14(m,2H,ArH),7.85–7.84(m,2H,ArH), 7.65–7.59(m,2H,ArH),7.43(brs,3H,ArH),7.33(t,J＝6.9Hz,1H,ArH)；¹³C NMR(100 MHz,DMSO-d₆)δ161.8,148.6,141.7,136.7,134.8,134.5,131.8,130.1,128.93,128.85,127.1, 121.8,119.9,118.4,113.0；HRMS(ESI):calcd for C₁₉H₁₅N₄O,[M+H]⁺315.1240; 315.1237 for found; the product was identified as N' -benzylidene-pyrido [3,4-b]Indole-1-carboxylic acid hydrazide.

Example 3

The preparation method of the N' - (4-methylbenzylidene) -pyrido [3,4-b ] indole-1-formylhydrazine shown as the chemical structural formula I-2 comprises the following steps:

the chemical structural formula of the N' - (4-methylbenzylidene) -pyrido [3,4-b ] indole-1-formylhydrazine I-2 is

The preparation method comprises the following specific steps:

the first to fourth steps, the same as example 1;

step five, except using 4-methyl benzaldehyde, other examples 1, diethyl ether recrystallization purification light yellow solid, yield 92%; the relevant parameters of the yellowish solid material were determined to be:¹H NMR(400MHz,DMSO-d₆)δ11.24 (s,1H,CONH),11.83(s,1H,NH),8.69(s,1H,N＝CH),8.44(d,J＝4.3and 22.4Hz,2H,ArH), 7.84(d,J＝8.1Hz,1H,ArH),7.84(d,J＝8.1Hz,1H,ArH),7.65(d,J＝7.5Hz,2H,ArH),7.60 (t,J＝7.7Hz,1H,ArH),7.30–7.28(m,2H,ArH),2.35(s,3H,CH₃)；¹³C NMR(100MHz, DMSO-d₆)δ161.7,148.7,141.6,139.9,136.7,134.8,131.9,131.8,130.8,129.5,128.9,127.1, 121.8,119.9,118.3,113.0,21.0；HRMS(ESI):calcd for C₂₀H₁₇N₄O,[M+H]⁺329.1397; 329.1396 for found; the product was identified as N' - (4-methylbenzylidene) -pyrido [3,4-b]Indole-1-carboxylic acid hydrazide.

Example 4

The preparation method of the N' - (4-methoxybenzylidene) -pyrido [3,4-b ] indole-1-formylhydrazine shown in the chemical structural formula I-3 comprises the following steps:

the chemical structural formula of the N' - (4-methoxybenzylidene) -pyrido [3,4-b ] indole-1-formylhydrazine I-3 is

The preparation method comprises the following specific steps:

the first to fourth steps, the same as example 1;

step five, except using 4-methoxy benzaldehyde, other examples 1, diethyl ether recrystallization purification get light yellow solid, the yield 93%; the relevant parameters of the yellowish solid material were determined to be:¹H NMR(400MHz,DMSO-d₆)δ11.24 (s,1H,CONH),11.83(s,1H,NH),8.69(s,1H,N＝CH),8.45(dd,J＝5.0and 22.7Hz,2H,ArH), 8.31(d,J＝7.8Hz,1H,ArH),7.84(d,J＝8.2Hz,1H,ArH),7.70(d,J＝8.6Hz,2H,ArH),7.60 (t,J＝7.4Hz,1H,ArH),7.30(t,J＝7.3Hz,1H,ArH),7.04(t,J＝8.6Hz,1H,ArH),3.82(s,3H, OCH₃)；¹³C NMR(100MHz,DMSO-d₆)δ161.6,160.8,148.5,141.6,136.7,134.8,132.0, 130.8,128.9,128.7,127.0,121.8,119.9,118.3,114.4,113.0,55.3；HRMS(ESI):calcd for C₂₀H₁₇N₄O,[M+H]⁺345.1346; 345.1349 for found; the product was identified as N' - (4-methoxybenzylidene) -pyrido [3,4-b]Indole-1-carboxylic acid hydrazide.

Example 5

The preparation method of the N' - (4-fluorobenzylidene) -pyrido [3,4-b ] indole-1-formylhydrazine shown as the chemical structural formula I-4 comprises the following steps:

the chemical structural formula of the N' - (4-fluorobenzylidene) -pyrido [3,4-b ] indole-1-formylhydrazine I-4 is

The preparation method comprises the following specific steps:

the first to fourth steps, the same as example 1;

step five, except using 4-fluorobenzaldehyde, other examples 1, diethyl ether recrystallization purification get light yellow solid, the yield 89%; the relevant parameters of the yellowish solid material were determined to be:¹H NMR(400MHz,DMSO-d₆)δ12.32 (s,1H,CONH),11.84(s,1H,NH),8.72(s,1H,N＝CH),8.45(dd,J＝4.6and 21.0Hz,2H,ArH), 8.31(d,J＝7.7Hz,1H,ArH),7.84–7.81(m,3H,ArH),7.77(d,J＝8.2Hz,2H,ArH),7.61(t,J ＝7.1Hz,1H,ArH),7.35–7.30(m,3H,ArH)；¹³C NMR(100MHz,DMSO-d₆)δ164.3,161.8, 147.5,141.7,136.7,134.8,131.8,131.1,130.8,129.3,129.2,128.9,121.8,119.92,119.89, 118.4,116.1,115.8,113.0；HRMS(ESI):calcd for C₁₉H₁₄FN₄O,[M+H]⁺333.1146; 333.1144 for found; the product was identified as N' - (4-fluorobenzylidene) -pyrido [3,4-b]Indole-1-carboxylic acid hydrazide.

Example 6

The preparation method of the N' - (4-chlorobenzylidene) -pyrido [3,4-b ] indole-1-formhydrazide shown in the chemical structural formula I-5 comprises the following steps:

the chemical structural formula of the N' - (4-chlorobenzylidene) -pyrido [3,4-b ] indole-1-formylhydrazine I-5 is

The preparation method comprises the following specific steps:

the first to fourth steps, the same as example 1;

the fifth step, same as example 1 except that 4-chlorobenzaldehyde was used, diethyl ether was recrystallizedPurifying to obtain white solid with the yield of 85 percent; the white solid matter was determined to have the following relevant parameters:¹H NMR(400MHz,DMSO-d₆)δ12.38(s, 1H,CONH),11.84(s,1H,NH),8.72(s,1H,N＝CH),8.45(dd,J＝4.6and 20.7Hz,2H,ArH), 8.31(d,J＝7.8Hz,1H,ArH),7.84(d,J＝8.2Hz,1H,ArH),7.77(d,J＝8.2Hz,2H,ArH),7.60 (t,J＝7.3Hz,1H,ArH),7.31(t,J＝7.4Hz,1H,ArH)；¹³C NMR(100MHz,DMSO-d₆)δ161.8, 147.3,141.7,136.7,134.8,134.5,133.4,131.7,130.9,129.0,128.7,121.8,119.93,119.89, 118.4,113.0；HRMS(ESI):calcd for C₁₉H₁₄ClN₄O,[M+H]⁺349.0851; 349.0850 for found; the product was identified as N' - (4-chlorobenzylidene) -pyrido [3,4-b]Indole-1-carboxylic acid hydrazide.

Example 7

The preparation method of the N' - (4-bromobenzylidene) -pyrido [3,4-b ] indole-1-formylhydrazine shown as the chemical structural formula I-6 comprises the following steps:

the chemical structural formula of the N' - (4-bromobenzylidene) -pyrido [3,4-b ] indole-1-formylhydrazine I-6 is

The preparation method comprises the following specific steps:

the first to fourth steps, the same as example 1;

step five, except using 4-bromobenzaldehyde, other examples 1, diethyl ether recrystallization purification get light yellow solid, the yield 95%; the relevant parameters of the yellowish solid material were determined to be:¹H NMR(400MHz,DMSO-d₆)δ12.38 (s,1H,CONH),11.83(s,1H,NH),8.70(s,1H,N＝CH),8.46(dd,J＝4.9and 20.6Hz,2H,ArH), 8.31(d,J＝7.6Hz,1H,ArH),7.83(d,J＝8.1Hz,1H,ArH),7.72–7.69(m,3H,ArH),7.61(t,J ＝7.3Hz,1H,ArH),7.30(t,J＝7.4Hz,1H,ArH)；¹³C NMR(100MHz,DMSO-d₆)δ161.8, 14.4,141.7,136.7,134.8,133.8,131.9,131.7,130.9,123.3,121.8,119.94,119.89,118.4,113.0； HRMS(ESI):calcd for C₁₉H₁₄BrN₄O,[M+H]⁺,3493.0346and 395.0325, respectively; found 3493.0349 and 395.0326; the product was identified as N' - (4-bromobenzylidene) -pyrido [3,4-b]Indole-1-carboxylic acid hydrazide.

Example 8

The preparation method of the N' - (4-trifluoromethyl benzylidene) -pyrido [3,4-b ] indole-1-formhydrazide shown in the chemical structural formula I-7 comprises the following steps:

the chemical structural formula I-7 of the N' - (4-trifluoromethyl benzylidene) -pyrido [3,4-b ] indole-1-formhydrazide is

The preparation method comprises the following specific steps:

the first to fourth steps, the same as example 1;

step five, except using 4-trifluoromethyl benzaldehyde, other examples 1, diethyl ether recrystallization purification to get white solid, yield 93%; the white solid matter was determined to have the following relevant parameters:¹H NMR(400MHz,DMSO-d₆) δ12.53(s,1H,CONH),11.86(s,1H,NH),8.80(s,1H,N＝CH),8.47(dd,J＝4.9and 20.1Hz, 2H,ArH),8.32(d,J＝7.7Hz,1H,ArH),7.96(d,J＝7.7Hz,1H,ArH),7.85–7.84(m,3H,ArH), 7.62(t,J＝7.3Hz,1H,ArH),7.31(t,J＝7.3Hz,1H,ArH)；¹³C NMR(100MHz,DMSO-d₆)δ162.5,147.4,142.2,139.0,137.2,135.4,132.1,131.4,130.0,129.5,128.1,126.3,126.2,123.2, 122.3,120.5,120.4,119.0,113.5；HRMS(ESI):calcd for C₂₀H₁₄F₃N₄O,[M+H]⁺383.1114; 383.1115 for found; the product was identified as N' - (4-trifluoromethylbenzylidene) -pyrido [3,4-b]Indole-1-carboxylic acid hydrazide.

Example 9

The preparation method of the N' - (furan-2-methylene) -pyrido [3,4-b ] indole-1-formylhydrazine shown as the chemical structural formula I-8 comprises the following steps:

the chemical structural formula of the N' - (furan-2-methylene) -pyrido [3,4-b ] indole-1-formylhydrazine is I-8

The preparation method comprises the following specific steps:

the first to fourth steps, the same as example 1;

step five, except using furan-2-formaldehyde, other examples 1, diethyl ether recrystallization purification to obtain yellow solid, yield 91%; the relevant parameters of the yellow solid material were determined to be:¹H NMR(400MHz,DMSO-d₆)δ12.36(s,1H, CONH),11.80(s,1H,NH),8.62(s,1H,N＝CH),8.45(dd,J＝4.7and 20.4Hz,2H,ArH),8.30 (d,J＝7.7Hz,1H,ArH),8.89(s,1H,ArH),7.61(t,J＝7.3Hz,1H,ArH),6.93(d,J＝2.6Hz, 1H,ArH),6.67–6.66(m 1H,ArH)；¹³C NMR(100MHz,DMSO-d₆)δ162.2,150.2,145.7, 142.1,138.9,137.1,135.3,132.3,131.4,129.4,122.3,120.4,120.4,118.8,114.0,113.6,112.7； HRMS(ESI):calcd for C₁₇H₁₃N₄O₂,[M+H]⁺305.1033; 305.1035 for found; the product was identified as N' - (furan-2-methylene) -pyrido [3,4-b]Indole-1-carboxylic acid hydrazide.

Example 10

The preparation method of the N' - (thiophene-2-methylene) -pyrido [3,4-b ] indole-1-formhydrazide shown in the chemical structural formula I-9 comprises the following steps:

the chemical structural formula of the N' - (thiophene-2-methylene) -pyrido [3,4-b ] indole-1-formylhydrazine I-9 is

The preparation method comprises the following specific steps:

the first to fourth steps, the same as example 1;

step five, except using thiophene-2-formaldehyde, other examples 1, diethyl ether recrystallization purification get light yellow solid, the yield 82%; the relevant parameters of the yellowish solid material were determined to be:¹H NMR(400MHz,DMSO-d₆)δ12.34 (s,1H,CONH),11.80(s,1H,NH),8.90(s,1H,N＝CH),8.44(dd,J＝4.8and 21.9Hz,2H,ArH), 8.31(d,J＝7.8Hz,1H,ArH),7.83(d,J＝8.2Hz,1H,ArH),7.69(d,J＝4.7Hz,1H,ArH),7.61 (t,J＝7.4Hz,1H,ArH),7.44(d,J＝2.8Hz,1H,ArH),7.30(t,J＝7.5Hz,1H,ArH),7.17(t,J ＝4.1Hz,1H,ArH)；¹³C NMR(100MHz,DMSO-d₆)δ162.1,144.1,142.1,139.9,137.2,135.3, 132.4,131.3,131.2,129.42,129.38,128.4,122.3,120.41,120.39,118.8,113.5；HRMS(ESI): calcd for C₁₇H₁₃N₄OS,[M+H]⁺321.0805; 321.0803 for found; the product was identified as N' - (furan-2-methylene) -pyrido [3,4-b]Indole-1-carboxylic acid hydrazide.

Example 11

The preparation method of the N' - (pyridine-2-methylene) -pyrido [3,4-b ] indole-1-formylhydrazine shown as the chemical structural formula I-10 comprises the following steps:

the chemical structural formula of the N' - (pyridine-2-methylene) -pyrido [3,4-b ] indole-1-formylhydrazine I-10 is

The preparation method comprises the following specific steps:

the first to fourth steps, the same as example 1;

step five, except using pyridine-2-formaldehyde, other examples 1, diethyl ether recrystallization purification to get white solid, yield 78%; the white solid matter was determined to have the following relevant parameters:¹H NMR(400MHz,DMSO-d₆)δ12.70(s,1H, CONH),11.88(s,1H,NH),8.78(s,1H,N＝CH),8.65(d,J＝3.6Hz,1H,ArH),8.49(dd,J＝4.7 and 22.0Hz,2H,ArH),8.33(d,J＝7.8Hz,1H,ArH),8.06(d,J＝7.7Hz,1H,ArH),7.94(d,J ＝7.2Hz,1H,ArH),7.85(d,J＝8.1Hz,1H,ArH),7.63(t,J＝7.2Hz,1H,ArH),7.46(t,J＝5.2 Hz,1H,ArH),7.33(t,J＝7.2Hz,1H,ArH)；¹³C NMR(100MHz,DMSO-d₆)δ162.2,153.5, 149.5,148.8,141.7,139.4,136.8,136.7,134.9,131.7,130.9,129.0,126.3,124.3,121.8,120.0, 119.9,118.5,113.0；HRMS(ESI):calcd for C₁₈H₁₃N₅O,[M+H]⁺315.1120; 315.1124 for found; the product was determined to be N' - (pyridine-2-methylene)) -pyrido [3,4-b]Indole-1-carboxylic acid hydrazide.

Example 12

The preparation method of the N' - (cyclohexylmethylene) -pyrido [3,4-b ] indole-1-formylhydrazine shown in the chemical structural formula I-11 comprises the following steps:

the chemical structural formula of the N' - (cyclohexylmethylene) -pyrido [3,4-b ] indole-1-formylhydrazine is I-11

The preparation method comprises the following specific steps:

the first to fourth steps, the same as example 1;

fifthly, the method is the same as the example 1 except that the cyclohexyl formaldehyde is used, and the yellow solid is obtained by recrystallization and purification of the diethyl ether, and the yield is 75 percent; the relevant parameters of the yellow solid material were determined to be:¹H NMR(400MHz,DMSO-d₆)δ11.79(s,1H, CONH),11.75(s,1H,NH),8.41(dd,J＝4.9and 15.9Hz,2H,ArH),8.23(d,J＝7.8Hz,1H, ArH),7.89(d,J＝5.6Hz,1H,ArH),7.82(d,J＝8.2Hz,1H,N＝CH),7.59(t,J＝7.4Hz,1H, ArH),7.29(t,J＝7.4Hz,1H,ArH),2.36–2.29(m,1H,CH₂CH),1.81–1.62(m,5H,CH₂), 1.33–1.22(m,5H,CH₂)；¹³C NMR(100MHz,DMSO-d₆)δ161.6,156.9,141.7,136.7,134.8, 132.1,130.9,129.0,121.9,120.0,118.2,113.1,40.4,29.8,29.8,25.6,25.0；HRMS(ESI):calcd for C₁₉H₂₁N₄O,[M+H]⁺321.1710; 321.1711 for found; the product was identified as N' - (cyclohexylmethylene) -pyrido [3,4-b]Indole-1-carboxylic acid hydrazide.

Example 13

The determination of the anti-tobacco mosaic virus activity of the individual compounds shown in the chemical structural formulas I-1 to I-11 in the pityriacitrin alkaloid derivative I containing the acylhydrazone structure is carried out by the following steps:

the first step, tobacco mosaic virus purification and concentration determination:

the purification and concentration determination of the tobacco mosaic virus are carried out according to the specification of tobacco mosaic virus SOP compiled by the institute of elements, institute of southern development university, the virus crude extract is subjected to 2-time polyethylene glycol centrifugation treatment, the concentration is determined to be 20 mug/mL, and the virus crude extract is refrigerated at 4 ℃ for standby;

secondly, preparing individual compound medicament solution shown in chemical structural formulas I-1 to I-11 in the pityriacitrin alkaloid derivative I containing the acylhydrazone structure:

weighing 40mg of individual compounds shown in chemical structural formulas I-1-I-11 in the pityriacitrin alkaloid derivative I containing the acylhydrazone structure as raw medicines respectively, and adding 0.4mL of DMF (dimethyl formamide) into each raw medicine for dissolving to obtain 1 × 10⁵Diluting the mother solution with mu g/mL by using a Tween 80 aqueous solution with the mass percentage concentration of 1 per thousand to the test concentration of 500 mu g/mL or 100 mu g/mL, thus preparing individual compound medicament solutions shown in chemical structural formulas I-1 to I-11 in the pityriacitrin alkaloid derivative I containing the acylhydrazone structure, and taking a ningnanmycin preparation to be directly diluted by adding water as a contrast;

step three, in vitro action:

preparing ten rubbingly inoculated 3-5 leaf-period Sanxi tobacco leaves, respectively washing with running water, wherein the virus concentration is 10 mug/mL, cutting off after drying, splitting along the leaf vein, respectively soaking the left and right half leaves in a Tween 80 aqueous solution with the mass percentage concentration of 1 thousandth and an individual compound medicament solution shown by chemical structural formulas I-1-I-11 in the pityriacitrinin alkaloid derivative I containing the acylhydrazone structure prepared in the second step, taking out after 30min, performing moisture preservation culture under the condition of normal-temperature illumination, repeating for 1 time for every 3 leaves, repeating for 3 times, recording the number of disease spots after 3 days, and calculating the prevention effect;

step four, protecting the living body:

respectively selecting ten parts of 3-5-leaf-period Sanxi tobaccos with uniform growth vigor, respectively spraying individual compound medicament solution shown in chemical structural formulas I-1-I-11 in the pityriacitrin alkaloid derivative I containing the acylhydrazone structure prepared in the second step on the whole plant, repeating the treatment for 3 times, setting a Tween 80 aqueous solution with the mass percentage concentration of 1 thousandth for comparison, spreading 500-mesh carborundum on the leaf surface after 24 hours, dipping the carborundum solution in a writing brush, lightly rubbing the whole leaf surface for 2 times along the branch vein direction, supporting the lower part of the leaf with a palm, keeping the virus concentration at 10 mu g/mL, washing with running water after inoculation, recording the number of disease spots after 3 days, and calculating the prevention effect;

the fifth step, the in vivo therapeutic action:

respectively selecting ten 3-5 leaf-period Sanxi tobaccos with uniform growth vigor, respectively inoculating viruses by using whole leaves of a writing brush, wherein the virus concentration is 10 mu g/mL, washing the inoculated leaves by using running water, drying the leaf surfaces, spraying the individual compound medicament solution shown in the chemical structural formula I-1-I-11 in the pityriacitrin alkaloid derivative I containing the acylhydrazone structure prepared in the second step by using the whole plant, repeating the treatment for 3 times, setting a Tween 80 aqueous solution with the mass percentage concentration of 1 thousandth for comparison, recording the number of lesions after 3 days, and calculating the prevention effect;

sixthly, in-vivo passivation:

respectively selecting ten parts of 3-5-leaf-period coral tobacco with uniform growth vigor, respectively mixing and passivating individual compound medicament solution shown in chemical structural formulas I-1-I-11 in the pityriacitrin alkaloid derivative I containing the acylhydrazone structure prepared in the second step with virus juice with the same volume for 30min, performing friction inoculation, wherein the virus concentration is 20 mu g/mL, flushing with running water after inoculation, repeating for 3 times, setting the Tween 80 aqueous solution with the mass percentage concentration of 1 thousandth for comparison, and counting the number of disease spots after 3 days to calculate the result;

the results of the determination of the anti-tobacco mosaic virus activity of individual compounds represented by the chemical structural formulas I-1 to I-11 in the pityriacitrin alkaloid derivative I containing the acylhydrazone structure are shown in Table 1.

TABLE 1 anti-TMV Activity test results of individual compounds represented by chemical structural formulas I-1 to I-11 in pityriacitrin alkaloid derivative I containing acylhydrazone structure:

from Table 1, the pityriacitrin alkaloid derivative I containing the acylhydrazone structure shows good anti-TMV activity, and the anti-TMV of the compounds I-2 to I-5, I-7 and I-11 exceeds that of the commercial varieties of ribavirin in living passivation, living treatment and living protection; the compound I-7 is obviously superior to commercial Ningnanmycin in vivo tests including in vivo inactivation, in vivo treatment and in vivo protection, and has great development value.

Example 14

The individual compounds shown in chemical structural formulas I-1-I-11 in the pityriacitrin alkaloid derivative I containing the acylhydrazone structure are subjected to an antibacterial activity test and an in-vitro sterilization test, and the determination procedures are as follows:

cell growth rate measurement method, i.e. plate method: dissolving 3mg of individual compounds shown in chemical structural formulas I-1-I-11 in the pityriacitrin alkaloid derivative I containing the acylhydrazone structure in 0.03mL of acetone respectively, diluting with an aqueous solution containing 200 mu g/mL of Tween 80 until the test concentration is 50mg/kg, sucking 1mL of liquid medicine respectively, injecting into a culture dish corresponding to the liquid medicine, adding 9mL of culture medium respectively, shaking uniformly to prepare a medicine-containing flat plate with the concentration of 50 mu g/mL, taking a flat plate added with 1mL of sterilized purified water as blank control, cutting a bacterial disc along the outer edge of hyphae by using a puncher with the diameter of 4mm, transferring to the medicine-containing flat plate, repeating the treatment for three times every treatment, placing the culture dish in a constant temperature incubator at 24 +/-1 ℃ for culturing, investigating the expansion diameter of each treated bacterial disc after 48 hours, calculating the average value, and comparing with the blank control to calculate the relative bacteriostasis rate.

The results of the in vitro fungicidal activity of the individual compounds represented by the chemical structural formulae I-1 to I-11 in the pityriacitrin alkaloid derivative I containing the acylhydrazone structure are shown in Table 2.

TABLE 2 in vitro bactericidal activity test results of individual compounds represented by chemical structural formulas I-1 to I-11 in pityriacitrin alkaloid derivative I containing acylhydrazone structure

As can be seen from the data in Table 2, individual compounds shown in chemical structural formulas I-1 to I-11 in pityriacitrin alkaloid derivative I containing an acylhydrazone structure show broad-spectrum activity against plant germs, and part of the compounds show over 50 percent of inhibition rate and have broad-spectrum bactericidal activity.

The percentages in the above examples are percentages by mass.

The raw materials and reagents involved in the above examples are commercially available, and the chemical reaction process is within the skill of those in the art.