CN107033134B - Bisamide compound containing pyridinium and 1,3, 4-oxadiazolyl and preparation method and application thereof - Google Patents

Abstract

The invention discloses a bisamide compound containing pyridinium and 1,3, 4-oxadiazolyl, a preparation method and application thereof, wherein the compound has a structure shown as a general formula (I):

Description

Bisamide compound containing pyridinium and 1,3, 4-oxadiazolyl and preparation method and application thereof

Technical Field

The invention relates to the technical field of pharmaceutical chemistry, in particular to a bisamide compound containing pyridinium and 1,3, 4-oxadiazolyl, and a preparation method and application thereof.

Background

In recent years, with the continuous change of ecological conditions, varieties and cultivation systems, plant bacterial diseases (such as bacterial leaf blight of rice, citrus canker and the like) and virus diseases (such as tobacco mosaic disease and the like) are generated and popularized in large scale in China, and have a further spreading and spreading trend in China, so that the disease range is wide, the control is difficult, the control efficiency of the existing commercial antibacterial agents (such as bismerthiazol, thiediazole and the like) and antiviral agents (ningnanmycin, ribavirin and the like) is low, and efficient plant disease bacteria or virus resistant active compounds are urgently needed to be found, so that green new pesticides from main intellectual property rights are created on the basis, and candidate drugs are provided for the control of bacterial and viral diseases of crops.

Bisamide-based compounds have a wide range of biological activities against bacteria and viruses, and thus have been one of the hot spots of research researchers, and commercially available bisamide-based compounds include pesticides (such as chlorantraniliprole (chlorantraniliprole), flubendiamide (flubendiamide), etc.) and fungicides (such as pyraflufen (fenpyrazamine), etc.). Meanwhile, the thioether compound containing a pyridinium group or a 1,3,4-oxadiazole group also shows wide biological activity in the aspect of pesticides. In order to search for active compounds with high-efficiency sterilization and antivirus, the invention takes bisamide compounds as the basis, introduces pyridinium groups and 1,3,4-oxadiazole groups which can improve the biological activity of target compounds into the system, synthesizes a series of bisamide compounds containing pyridinium and 1,3,4-oxadiazole groups, inspects the biological activity of the bisamide compounds, and provides important scientific basis for the research and development of new pesticides.

The research on the biological activity of the bisamide compounds progresses as follows:

wu et al [ Wu, J.; kang, s.h.; luo, l.j.; shi, q.c.; ma, j.; yin, j.; song, b.a.; hu, d.y.; a series of bisamide derivatives containing 1,3, 4-oxadiazolyl are synthesized by Yang, S.Synthesis and anti-inflammatory derivatives of novel amino derivatives connecting 1,3,4-oxadiazole.chem.Cent.J.2013,7,64 ]. The compound shows certain inhibition activity on wheat scab, pepper fusarium wilt and apple canker.

Yang et al Z.B.Yang, D.Y.Hu, S.Zeng, B.A.Song, Novel hydrogel associating pyridine amide viscosity Design, synthesis, ligation activity biology, Med.chem.Lett.2016,26,1161-]A series of bisamide compounds containing acylhydrazone structures are synthesized, and biological activity test results show that the target compound has insecticidal activity on brown planthoppers, diamondback moths and culex pipiens, wherein the compound 5j has the strongest insecticidal activity on the culex pipiens, and the LC of the compound₅₀And LC₉₅2.44 and 5.76mg/L, respectively.

Good et al (J.A.good, J.silver, C) 2016.

W.Bahnan,K.S.Krishnan,O.Salin,P.

R.Svensson,P.Artursson,

Gylfe,S.F.Almqvist,Thiazolino 2-Pyridone Amide Inhibitors of Chlamydia trachomatisInfectivity.J.Med.Chem.2016,59,2094-2108]A series of thiazolo 2-pyridone amide compounds are reported, and biological activity test results show that part of target compounds show better inhibitory activity on Chlamydia trachomatis.

The research on the biological activity of the derivatives containing 1,3,4-oxadiazole group has progressed as follows:

in 2013, thioether and sulfone compounds containing 1,3,4-oxadiazole group were designed and synthesized by Xu et al [ W.M.Xu, S.Z.Li, M.He, S.Yang, X.Y.Li, P.Li.Synthesis and selectivity of novel thio ether/sulfo derivatives connecting 1,2,3-thiadiazole and 1,3,4-oxadiazole/thiadiazole mobility. bioorg.Med.chem.Lett.,2013,23,5821 and 5824 ]. Preliminary antiviral bioactivity tests show that the compounds show better inhibitory activity to tobacco mosaic virus at a concentration of 500 mug/mL, wherein the compounds 7a, 7c, 7d, 8a and 9a show higher inhibitory activity.

2015, Wu et al [ W.N.Wu, Q.Chen, A.Q.Tai, G.Q.Jiang, G.P.Ouyang, Synthesis and activity of 2-substistuted methyl-5- (4-amino-2-methyl pyrimidin-5-yl) -1,3, 4-oxadiazine derivatives.bioorg.Med.chem.Le2015, 25,2243-]A series of pyrimidine-bis-1, 3,4-oxadiazole compounds are synthesized. Antiviral activity test shows that the target compound has better inhibitory activity to tobacco mosaic virus, wherein the compound 8i has higher therapeutic activity to tobacco mosaic virus and EC thereof₅₀It was 246.48. mu.g/mL.

Some benzodioxan derivatives with 1,3,4-oxadiazole group were reported in 2016, khalilulah et al [ h.khalilulah, s.khan, m.s.nomani, b.ahmed, arab.j.chem.2016,9, S1029-S1035 ]. Biological activity tests show that part of target compounds show good inhibitory activity on bacteria (such as staphylococcus aureus, escherichia coli, bacillus subtilis and the like) and fungi (aspergillus niger, aspergillus flavus, candida albicans and the like).

The study of the biological activity of amphiphiles based on pyridinium salts has progressed as follows:

a series of pyridinium-containing Ionic Liquids were synthesized in 2015 by Messali et al [ M.Messali, Eco-Friendly Synthesis of a New Class of pyridine-Based Ionic Liquids with active Ionic activities, Molecules 2015,20,14936-14949 ]. Such compounds exhibit certain inhibitory activity against Gram- (+) such as Streptococcus pneumoniae (Streptococcus pneumoniae) and Bacillus subtilis, and Gram- (-) such as Pseudomonas aeruginosa (Pseudomonas aeruginosa) and Escherichia coli (Escherichia coli).

In 2016, Jia et al [ R.X.Jia, Y.F.Duan, Q.Fang, X.Y.Wang, J.Y.Huang, Pyridine-grafted chitosan derivative as an anti-chitosan agent, food chem.2016,196,381-387] grafted pyridinium onto chitosan to synthesize a pyridinium-containing chitosan derivative. The minimum inhibitory concentration and the minimum fungicidal concentration of the compound on the botrytis cinerea are respectively 0.13 and 1.0mg/mL, and the minimum inhibitory concentration and the minimum fungicidal concentration on the botrytis cinerea are respectively 0.13 and 4.0 mg/mL.

Disclosure of Invention

The purpose of the invention is: provides a bisamide compound containing pyridinium and 1,3, 4-oxadiazolyl, a preparation method and application thereof, which has good inhibition effect on pathogenic bacteria and viruses, and the synthesis method is very economical and simple.

The invention is realized by the following steps: a bisamide compound containing pyridinium and 1,3, 4-oxadiazolyl is characterized in that: the compound has a structure shown as a general formula (I):

in the formula, R on the benzene ring₁Is hydrogen, C_1-4Alkyl, trifluoromethyl, C_1-3Alkoxy, trifluoromethoxy, propenyl, acrylamido, C_1-3Alkylamino, nitro or halogen atoms; r₂Is hydrogen, C_1-4Alkyl or different substituted benzyl, wherein the substituent on the different substituted benzyl is more than one, and the substituent is hydrogen or C_1-4Alkyl, trifluoromethyl, C_1-3Alkoxy, trifluoromethoxy, propenyl, acrylamido, C_1-3Alkylamino, nitro or halogen atoms; r on pyridinium cation₃Is hydrogen, C_1-4Alkyl, trifluoromethyl, hydroxy, C_1-3Alkoxy, trifluoromethoxy, propenyl, acrylamido, amino, C_1-3Alkylamino, nitro or halogen atoms; x is oxygen atom or sulfur atom; n is a natural number greater than 1.

2. A process for producing a bisamide compound containing a pyridinium salt and 1,3, 4-oxadiazolyl according to claim 1, wherein: the synthetic route is as follows;

in the formula, R on the benzene ring₁Is hydrogen, C_1-4Alkyl, trifluoromethyl, C_1-3Alkoxy, trifluoromethoxy, propenyl, acrylamido, C_1-3Alkylamino, nitro or halogen atoms; r₂Is hydrogen, C_1-4Alkyl or different substituted benzyl, wherein the substituent on the different substituted benzyl is more than one, and the substituent is hydrogen or C_1-4Alkyl, trifluoromethyl, C_1-3Alkoxy, trifluoromethoxy, propenyl, acrylamido, C_1-3Alkylamino, nitro or halogen atoms; r on pyridinium cation₃Is hydrogen, C_1-4Alkyl, trifluoromethyl, hydroxy, C_1-3Alkoxy, trifluoromethoxy, propenyl, acrylamido, amino, C_1-3Alkylamino, nitro or halogen atoms;x is oxygen atom or sulfur atom; n is a natural number greater than 1.

Application of bisamide compounds containing pyridinium and 1,3, 4-oxadiazolyl in preparation of medicines for resisting pathogenic bacteria and viruses.

By adopting the technical scheme, the invention takes the aryl bisamide compound as the base, introduces pyridinium cation and 1,3, 4-oxadiazolyl which can improve the biological activity of a target compound into the system, synthesizes a series of bisamide compounds containing pyridinium and 1,3, 4-oxadiazolyl, and finds that the compounds have good inhibition effect on pathogenic bacteria and viruses, have good inhibition effect on pathogenic bacteria [ such as Xanthomonas oryzae pv. oryzae, Xoo), Citrus canker (Xanthomonas axaoxonoporus pv. citri, Xac) and the like ] and plant viruses [ such as Tobacco Mosaic Virus (Tobacco Mosaic Virus, TMV) and the like ], and provide important scientific basis for the research and development of new pesticides.

Detailed Description

Example 1 of the invention: preparation of target compound 5- (2- (2-methyl-5- (methylthioether) -1,3, 4-oxadiazole) benzamido) amino) -5-pentanoyl) -1-bromopyridine salt

Adding 5-bromovaleryl chloride (0.45mmol) into 10mL dry dichloromethane solution dissolved with 2-amino-N- (2-methyl-5- (methyl sulfide group) -1,3, 4-oxadiazole) benzamide (0.45mmol) and triethylamine (0.91mmol), reacting at normal temperature for 5 hr, stopping reaction, desolventizing, adding 50mL ethyl acetate, washing with water 20mL × 3 times, drying with anhydrous sodium sulfate, desolventizing, dissolving in 5mL pyridine, reacting at 50 deg.C for 12 hr, desolventizing, and performing column chromatography (CH eluent)₂Cl₂:CH₃OH 6:1) gave a yellow oil in 41.2% yield.

Synthesis of other target compounds reference example 1.

The structure, nuclear magnetic resonance hydrogen spectrum and carbon spectrum data of the synthesized part of the bisamide compound containing the pyridinium and the 1,3, 4-oxadiazolyl are shown in table 1, and the physicochemical properties are shown in table 2.

TABLE 1 NMR hydrogen and carbon spectra data for some of the compounds

Table 2 physicochemical properties of the target compounds

Pharmacological example 1:

the inhibition rate of the target compound on plant pathogenic bacteria is tested by a turbidity method, and the test objects are rice bacterial blight (Xoo) and citrus canker pathogen (Xac). DMSO was dissolved in the medium as a blank control. Placing rice bacterial leaf blight bacteria (in M210 solid culture medium) in NB culture medium, and shake culturing in constant temperature shaking table at 28 deg.C and 180rpm to logarithmic phase for use; the citrus canker pathogen (on M210 solid medium) was placed in NB medium and shake-cultured in a constant temperature shaker at 28 ℃ and 180rpm until logarithmic phase for use. 5mL of toxin-containing NB liquid culture medium prepared by the drug (compound) into different concentrations (example: 100, 50. mu.g/mL) is added into a test tube, 40. mu.L of NB liquid culture medium containing plant-borne bacteria is respectively added, shaking is carried out in a constant temperature shaking table at 28-30 ℃ and 180rpm, the bacterial blight pathogenic bacteria of rice are cultured for 36h, and the citrus canker pathogenic bacteria are cultured for 48 h. The OD was measured on a spectrophotometer using the bacterial solutions of the respective concentrations₅₉₅Value, and additionally determining the OD of the corresponding concentration of the sterilized NB-containing liquid medium₅₉₅The value is obtained.

EC₅₀(mean effective concentration) is an important means for evaluating the sensitivity of plant pathogenic bacteria to chemical compoundsThe target is also an important parameter for setting the concentration of the compound when researching the action mechanism of the target compound. In the concentration gradient experiment, proper 5 concentrations are set by a double dilution method, finally the inhibition rate of the medicament on plant pathogenic bacteria and the medicament concentration are converted into paired numerical values, a toxicity curve is obtained through SPSS software regression analysis, and EC is calculated₅₀。

Testing the effective medium concentration EC of target compound on plant pathogenic bacteria by adopting turbidity method₅₀The test subjects were rice bacterial blight (Xoo) and citrus canker (Xac). DMSO was dissolved in the medium as a blank control. Placing rice bacterial leaf blight bacteria (in M210 solid culture medium) in NB culture medium, and shake culturing in constant temperature shaking table at 28 deg.C and 180rpm to logarithmic phase for use; the citrus canker pathogen (on M210 solid medium) was placed in NB medium and shake-cultured in a constant temperature shaker at 28 ℃ and 180rpm until logarithmic phase for use. 5mL of toxic NB-containing liquid medium prepared by the drug (compound) into different concentrations (for example: 80,40,20,10, 5. mu.g/mL) is added into a test tube, 40. mu.L of NB liquid medium containing plant-borne bacteria is respectively added, shaking is carried out in a constant temperature shaking table at 180rpm at 28-30 ℃, the bacterial blight pathogenic bacteria of rice is cultured for 36h, and the citrus canker pathogenic bacteria are cultured for 48 h. The OD was measured on a spectrophotometer using the bacterial solutions of the respective concentrations₅₉₅Value, and additionally determining the OD of the corresponding concentration of the sterilized NB-containing liquid medium₅₉₅The value is obtained.

Corrected OD value-bacteria-containing medium OD value-sterile medium OD value

Percent inhibition ratio [ (corrected contrast culture medium bacterial liquid OD value-corrected toxin-containing culture medium OD value)

OD value of the control medium bacterial liquid after correction is multiplied by 100

The examples of the present invention are given to illustrate the technical solution of the present invention, but the contents of the examples are not limited thereto, and some experimental results of the target compounds are shown in tables 3 and 4.

TABLE 3 inhibitory Activity of bisamide-based Compounds containing pyridinium salt and 1,3, 4-oxadiazolyl against plant pathogenic bacteria

"NT" means not tested

TABLE 4 EC of bisamide containing pyridinium salt and 1,3, 4-oxadiazolyl compound against plant pathogenic bacteria₅₀

As can be seen from tables 3 and 4, the target compounds showed good inhibitory activity against plant pathogenic bacteria (e.g., bacterial blight of rice and canker citrus) in the in vitro test. Wherein, the inhibitory activity of the compounds 7, 8, 9 and 10 on the rice bacterial blight under the concentration of 100 mug/mL is 90.5, 74.0, 99.7 and 86.8 percent respectively, and the inhibitory activity of the compound 9 on the rice bacterial blight under the concentration of 50 mug/mL is 98.3 percent; the inhibition rates of the compounds 1,3, 8 and 10 on the citrus canker pathogenic bacteria at the concentration of 100 mu g/mL are 67.6, 70.0, 71.5 and 99.8 percent respectively, and the inhibition activity of the compound 9 on the citrus canker pathogenic bacteria at the concentration of 50 mu g/mL is 99.8 percent. Further test results show that compound 9 has EC against these two pathogens₅₀Respectively 0.7 and 5.9 mug/mL, can be used for preparing pesticides for resisting plant pathogenic bacteria.

Pharmacological example 2: test for antiviral Activity of part of target Compound (anti-tobacco mosaic Virus)

The anti-plant virus activity of the compound is determined by a hemiphyllous spot method. 3mg of the test compound was accurately weighed in a weighing bottle, and 60. mu.L of DMSO as a solvent was added to dissolve it sufficiently. The solution was prepared into 500mg/L compound solution with 1% Tween 20 in double distilled water. And adding a solvent DMSO of 60 mu L into 250 mu L of 2% ningnanmycin aqueous solution, and preparing 500mg/L ningnanmycin solution by 10mL of secondary distilled water containing 1% Tween 20.

In vivo therapeutic activity of the agent against TMV infection. Selecting heart-leaf tobacco with consistent growth vigor, dipping virus solution (concentration of 6 × 10) with a row ratio^-3mg/mL) is manually rubbed and inoculated on the leaf surface (whole leaf) along the branch vein direction of the leaf surface (whole leaf) on the leaf blade scattered with carborundum, the inoculation force of the left and right leaf blades is kept consistent as much as possible, and the lower part of the leaf blade is leveledAnd supporting the whole wood plate. And after the virus liquid is dried, washing away carborundum on the blade by using running water. After the leaves are dry, the left half is coated with the drug and the right half is coated with sterile water as a control. 3 plants are set for each medicament treatment, each plant has 3-4 leaves, then the plants are placed in a light incubator for moisture preservation and culture, the temperature is controlled at 23 ℃, the illumination is 10000LuX, and the number of the generated scorched spots is observed and recorded after 2-4 days. The inhibition rate was calculated as 3 replicates per dose as described above.

In vivo protective activity of the agent against TMV infection. Selecting the heart-leaf tobacco with consistent growth vigor, lightly applying the medicament on the left half leaf and applying the sterilized water on the right half leaf by using a writing brush as a control, and inoculating the virus after 24 hours. Dipping the virus liquid with a spread pen (concentration 6X 10)^{- 3}mg/mL), the leaf surface (whole leaf) is manually rubbed along the branch direction to be inoculated on the leaf blade scattered with carborundum, the inoculation force of the left and right leaf blades is kept consistent as much as possible, and the lower part of the leaf blade is supported by a flat wood plate. And after the virus liquid is dried, washing away carborundum on the blade by using running water. 3 plants are set for each medicament treatment, each plant has 3-4 leaves, then the plants are put into a light incubator for moisture preservation and culture, the temperature is controlled at 23 ℃, and after the light irradiation is carried out for 10000Lux and 2-4d, the number of the generated scorched spots is observed and recorded. The inhibition ratio Y (%) ═ R-L)/rx100% was calculated by repeating 3 times per drug in the above manner.

Wherein: y is the inhibition rate of the compound on the tobacco mosaic virus; r is the number of the control group (right half leaf) dry spots, and L is the number of the treatment group (left half leaf) dry spots. The embodiment of the present invention is provided to illustrate the technical solution of the present invention, but the content of the embodiment is not limited thereto, and the experimental results are shown in table 5.

TABLE 5 inhibitory Activity of bisamide-based Compounds containing pyridinium salt and 1,3, 4-oxadiazolyl against plant viruses

As can be seen from Table 5, the in vivo test results show that the series of compounds have certain antiviral activity at the concentration of 500 mug/mL, wherein the therapeutic activity of the compounds 3,5 and 9 on tobacco mosaic virus is 50.6, 60.0 and 51.9 percent respectively; the protective activity of the compound 1 on tobacco mosaic virus is 51.2%, so that the compound can be used for preparing pesticide for resisting plant virus.

Claims (3)

1. A bisamide compound containing pyridinium and 1,3, 4-oxadiazolyl is characterized in that: the compound has a structure shown as a general formula (I):

in the formula, R on the benzene ring₁Is hydrogen, C_1-4Alkyl, trifluoromethyl, C_1-3Alkoxy, trifluoromethoxy, propenyl, acrylamido, C_1-3Alkylamino, nitro or halogen atoms; r₂Is hydrogen, C_1-4Alkyl or different substituted benzyl, wherein the substituent on the different substituted benzyl is more than one, and the substituent is hydrogen or C_1-4Alkyl, trifluoromethyl, C_1-3Alkoxy, trifluoromethoxy, propenyl, acrylamido, C_1-3Alkylamino, nitro or halogen atoms; r on pyridinium cation₃Is hydrogen, C_1-4Alkyl, trifluoromethyl, hydroxy, C_1-3Alkoxy, trifluoromethoxy, propenyl, acrylamido, amino, C_1-3Alkylamino, nitro or halogen atoms; x is oxygen atom or sulfur atom; n is a natural number greater than 1.

2. A process for producing a bisamide compound containing a pyridinium salt and 1,3, 4-oxadiazolyl according to claim 1, wherein: the synthetic route is as follows:

in the formula, R on the benzene ring₁Is hydrogen, C_1-4Alkyl, trifluoromethyl, C_1-3Alkoxy, trifluoromethoxy, propenyl, acrylamido, C_1-3Alkylamino, nitro or halogen atoms; r₂Is hydrogen, C_1-4Alkyl or different substituted benzyl, wherein the substituent on the different substituted benzyl is more than one, and the substituent is hydrogen or C_1-4Alkyl, trifluoromethyl, C_1-3Alkoxy, trifluoromethoxy, propenyl, acrylamido, C_1-3Alkylamino, nitro or halogen atoms; r on pyridinium cation₃Is hydrogen, C_1-4Alkyl, trifluoromethyl, hydroxy, C_1-3Alkoxy, trifluoromethoxy, propenyl, acrylamido, amino, C_1-3Alkylamino, nitro or halogen atoms; x is oxygen atom or sulfur atom; n is a natural number greater than 1.

3. The use of the bisamide compounds containing a pyridinium salt and 1,3, 4-oxadiazolyl according to claim 1 for the preparation of pesticides against pathogenic bacteria and viruses.