CN110467623B - Benzoisofuran compound and preparation method and application thereof - Google Patents
Benzoisofuran compound and preparation method and application thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
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- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
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Abstract
The invention discloses a benzisothiafuran compound and a preparation method and application thereof. The benzisofuran compound is separated from the ansu adenophora and has a molecular formula of C15H16O4A yellow gum having the following structural formula:
. The preparation method comprises the steps of extract extraction, silica gel column chromatography and high-pressure liquid chromatography separation and purification. The application is the application of the benzisofuran compound in preparing bacteriostatic agents or the application of the benzisofuran compound in preparing biological pesticides for resisting tobacco black shank.
Description
Technical Field
The invention belongs to the technical field of phytochemistry, and particularly relates to a benzisofuran compound and a preparation method and application thereof.
Background
Tuberose senna (latin name:Cassia pumilalam.) is a perennial shrub-like, draping herbaceous plant of the genus Cassia of the family Leguminosae, which is grown in shrubs or grasses in mountainous and open areas. The method is mainly distributed in subtropical regions of Guangdong, Yunnan, Guangxi and other provinces in China. Distribution is also found in india, central and south peninsula, malaysia, australia, etc. The cassia plant is a traditional medicinal plant in China, and has very wide application and long medicinal history in China. The literature reports that the chemical components of cassia plants are mainly flavonoids and anthraquinones, and in addition, terpenes, steroids, alkaloids, chromones and the like. At present, few chemical components of the plants of Cassia genus, namely the anshansenna canna are reported in domestic and foreign documents, and a large number of compounds with novel structures and good biological activity are found in various plants of Cassia genus.
The tobacco black shank is one of the most devastating diseases in tobacco production, and is also called tobacco epidemic disease, and the tobacco growers are called black stalk crazy, black root and aconite disease. The main tobacco producing areas in China occur in different degrees, wherein Anhui, Shandong and Henan provinces are historically serious disease areas; the occurrence of tobacco in southern areas such as Yunnan, Guizhou, Sichuan, Hunan, Guangdong, Guangxi and Fujian is also quite common. At present, the prevention and the treatment of the black-stem disease are mainly realized by methods such as crop rotation cultivation, variety gene improvement, biological pesticide and the like. Where control with biopesticides is the most common and most easily achieved method.
Based on the above consideration, we studied the chemical composition of the whole dry strain of Dolichos canephora produced in Yunnan, and isolated a new benzoisofuran compound from the same, which has not been reported yet. It is worth mentioning that the compound has remarkable activity of resisting the tobacco black stem disease.
Disclosure of Invention
The first purpose of the invention is to provide a benzisofuran compound; the second purpose is to provide a preparation method of the benzisothiafuran compound; the third purpose is to provide the application of the benzisothiafuran compound.
The first purpose of the invention is realized by that the benzisothians compound is separated from the dolichos lablab and has the molecular formula C15H16O4A yellow gum having the following structural formula:
the second purpose of the invention is realized by taking the anshan senna as a raw material and carrying out the steps of extract extraction, silica gel column chromatography and high-pressure liquid chromatography separation and purification, and the method specifically comprises the following steps:
A. extracting the extractum: crushing and sieving the pedunculate hydrangea, ultrasonically extracting for 3-5 times by using an organic solvent with the weight 3-10 times that of the pedunculate hydrangea for 30-60 min each time, combining the extracting solutions, and concentrating to obtain a flowable viscous extract a;
B. silica gel column chromatography: performing silica gel column chromatography on the extract a by using 160-300-mesh silica gel dry method which is 2-8 times of the weight of the extract a, performing gradient elution by using chloroform-acetone solutions with volume ratios of 1:0, 20:1, 9:1, 8:2, 7:3, 6:4, 1:1 and 1:2, collecting gradient eluent, concentrating, and combining the same parts;
C. high-pressure liquid chromatography separation: part 9:1 of the eluate from step B is subjected to reverse phase C18Separating and purifying by medium pressure liquid chromatography to obtain eluate corresponding to chromatographic peak of 31.6min, and further separating and purifying by high pressure liquid chromatography to obtainTarget benzisothiafuran compounds.
The structures of the benzisothiafuran compounds prepared by the method are identified by the following method:
the compounds of the invention are yellow gums; HRESI-MS shows that the peak of the excimer ion is 283.0952 [ M + Na ]]+(calculated 283.0946), combined1H NMR and DEPT spectra confirm that the molecular formula is C15H16O4The unsaturation degree is 8. The infrared spectrum shows hydroxyl groups (3415 cm)-1) Carbonyl group (1682 cm)-1) And aromatic rings (1614, 1556 and 1442 cm)-1) The resonance absorption peak of (1). The existence of aromatic ring structures in the compound is also indicated by the maximum absorption of the ultraviolet spectrum at 312, 272 and 210 nm. Process for preparing compounds1H、13C and DEPT NMR spectra (Table-1) showed that it contained 15 carbons and 16 hydrogens, 1,2,3,4, 5-pentasubstituted benzene rings (C-4 to C-7, C-4a and C-7a, H-7), one hydroxyethyl group (-CH)2CH2OH, C-6 'and C-7', H26' and H2-7'), one harmonic dimethylchromene ring (-CH = CH-C (CH)3）2-O-, C-1 '-C-5', H-1', H-2' and H6-4', 5'), one ester carbonyl (C-1), one oxidized methylene (C-3, H)2-3). According to the structural fragment of the compound, except for the unsaturation degree of a benzene ring of 4, the unsaturation degree of a synthylchromene ring of 2 and the unsaturation degree of an ester carbonyl of 1, the compound should be provided with a ring so as to meet the requirement that the unsaturation degree of the compound is 8. According to the presence of H in the compound2HMBC between-3 and C-1, C-4, C-4a, C-7a, and H-7 and C-1, C-4a, C-7a (FIG. 2) can confirm that the compound forms a 5-membered ring between C-1 and C-3 through an ester bond, and is an isobenzofuranolactone compound.
After the basic skeleton and the groups in the compound are determined, the positions of the substituents can be further determined according to the HMBC correlation of the compound. According to H2HMBC phases of-6 'and C-4, C-5, C-4a, and H-7' and C-4(ii) off, the hydroxyethyl substitution can be determined at the C-4 position; according to H-1' and C-5, C-6, C-7; h-2' and C-6, and H-7 and C-1' are related to HMBC, presumably gem-dimethyl chromene ring substitution at the C-5 and C-6 positions of the compound, and the C-1' carbon is attached at the C-6 position of the phenyl ring. The structure of the compound is now determined.
Infrared, ultraviolet and mass spectral data of compounds: the UV (methanol) light is emitted from the UV,λ max（logε) 312 (3.26), 272 (3.64), 210 (3.93) nm; IR (potassium bromide pellet):ν max 3415、3057、2948、1682、1614、1556、1442、1368、1149、1032、932、853cm-1；1h and13c NMR data (500 and 125MHz, CDCl)3) See Table-1; positive ion mode ESIMSm/ z 283 [M+Na]+(ii) a Positive ion mode HRESIMSm/z 283.0952 [M+Na]+(calculation value C)15H16O4，283.0946）。
The third purpose of the invention is realized by the application of the benzisothiafuran compound in preparing bacteriostatic agent.
The application of the benzisothiafuran compound in preparing the biological pesticide for resisting tobacco black shank is provided.
The tobacco black-stem disease is caused by phytophthora nicotianae infection, and the activity of the compound for inhibiting phytophthora nicotianae is firstly measured in the invention. The method comprises the following steps:
(1) preparation of oatmeal agar medium: adding 1000mL of water into oatmeal, heating the oatmeal in a boiling water bath for 1 hour, filtering the oatmeal by using gauze, adding the water for supplementing 1000mL, then adding sugar and agar, heating the oatmeal to completely melt the agar, filtering the oatmeal in a triangular flask by using the gauze (absorbent cotton is added in the middle) while the oatmeal is hot, sterilizing the oatmeal for 20min (minutes) at 121 ℃ and 15 pounds, taking out the oatmeal and cooling the oatmeal to about 45 ℃, adding ampicillin (5 mg/100 mL) into an aseptic operation table, uniformly mixing the ampicillin and the ampicillin, pouring the ampicillin into the flat dish, culturing the ampicillin in the flat dish for 48 hours at 28 ℃, and checking the ampicillin for later use after sterility is checked.
(2) Bacteriostatic experiments: placing circular filter paper with diameter of 5mm into a culture dish, placing in a 15 pound autoclave for 30min, oven drying, and respectively soaking in 20µM compound, 75% ethanol solution andbacteria distilled water. Respectively sucking 0.2mL of fresh bacterial liquid of phytophthora nicotianae on oatmeal agar medium plates by using a sterile suction pipe on a sterile operating table. Coating with triangular glass coating rod, lightly sticking the filter paper sheets onto corresponding flat plates with tweezers, culturing at 28 deg.C, observing experimental results, and measuring the size of antibacterial zone. Meanwhile, agricultural chloramphenicol was used as a positive control.
The test result shows that: the diameter of the inhibition zone of the benzisothiafuran compound is 16.5 +/-1.2 mm, and the diameter of the inhibition zone of the positive control agricultural chloramphenicol is 12.2 +/-1.0 mm. The compound has the effect of inhibiting phytophthora nicotianae remarkably superior to that of positive control agricultural chloramphenicol, and has outstanding activity of inhibiting black stem diseases.
The compound has the effects of preventing and treating the black stem disease of tobacco: transplanting tobacco seedlings into flowerpots with the diameter of 10cm and the height of 10cm, wherein the culture medium is as follows: sterilizing soil, peat and perlite (2: 2: 1), and culturing 1 strain per pot. After transplanting and seedling slowing, 10g of bacterial grains are added into roots, tobacco seedlings are placed in an artificial climate room for cultivation, the temperature is 30 ℃ in the daytime, the temperature is 28 ℃ in the night, the illumination is dark (12 h:12 h), and the relative humidity is 95%, so that the tobacco seedlings are attacked. Before the black shank disease occurs, the tobacco seedlings are subjected to root irrigation treatment by using the compound of the invention with the particle size of 20 mu M, and 10mL of the compound is poured on each tobacco seedling; the pouring is carried out for 2 times. The disease index was calculated by repeating the treatment of 10 strains each for 3 times and examining the disease after 14 days. The results show that: the compound has a control effect on tobacco black shank of (77.2 +/-3.4)%, and has an obvious control effect on tobacco black stalk diseases.
Compared with the prior art, the invention has the following outstanding advantages:
(1) the compound is separated from the plant of Cassia plant Amur Dolichos stalk, which is a perennial subalpine mantle herb, and has the advantages of large biological yield, wide raw material release, low compound separation and preparation raw material cost.
(2) And the compound has simple preparation process, easy realization of industrial production and large-scale popularization and application conditions. And the compound has simple structure and is easy to realize by adopting an artificial synthesis process.
(3) The compound shows good activity of resisting the tobacco black-stem disease, the diameter of a bacteriostasis zone of the benzisofuran compound is 16.5 +/-1.2 mm, and the effect of inhibiting the tobacco phytophthora is obviously better than that of positive control agricultural chloramphenicol. The actual control effect also shows that the control effect of the compound on the tobacco black shank is (77.2 +/-3.4)%, and the compound has a remarkable control effect on the tobacco black stalk disease. The tobacco black shank is one of the most destructive diseases in tobacco production, and the application of the compound of the invention provides a novel efficient and safe biological pesticide molecular structure for preventing and treating the tobacco black shank.
Drawings
FIG. 1 is a nuclear magnetic resonance carbon spectrum of a benzisothiafuran compound of the invention;
FIG. 2 is a nuclear magnetic resonance hydrogen spectrum of the benzisothiafuran compounds of the invention;
FIG. 3 is a nuclear magnetic resonance HSQC spectrum of the benzisothiafuran compounds of the invention;
FIG. 4 is a nuclear magnetic resonance HMBC spectrum of the benzisothiafuran compounds of the invention;
FIG. 5 is a graph of the main HMBC correlation of the benzisothiafurans of the present invention.
Detailed Description
The present invention is further illustrated by the following examples and the accompanying drawings, but the present invention is not limited thereto in any way, and any modifications or alterations based on the teaching of the present invention are within the scope of the present invention.
The benzisofuran compound is separated from the anshana senna and has the molecular formula of C15H16O4A yellow gum having the following structural formula:
the preparation method of the benzisothiafuran compounds takes the anshansenia as a raw material, and comprises the steps of extract extraction, silica gel column chromatography and high-pressure liquid chromatography separation and purification, and specifically comprises the following steps:
A. extracting the extractum: crushing and sieving the pedunculate hydrangea, ultrasonically extracting for 3-5 times by using an organic solvent with the weight 3-10 times that of the pedunculate hydrangea for 30-60 min each time, combining the extracting solutions, and concentrating to obtain a flowable viscous extract a;
B. silica gel column chromatography: performing silica gel column chromatography on the extract a by using 160-300-mesh silica gel dry method which is 2-8 times of the weight of the extract a, performing gradient elution by using chloroform-acetone solutions with volume ratios of 1:0, 20:1, 9:1, 8:2, 7:3, 6:4, 1:1 and 1:2, collecting gradient eluent, concentrating, and combining the same parts;
C. high-pressure liquid chromatography separation: part 9:1 of the eluate from step B is subjected to reverse phase C18Separating and purifying by medium pressure liquid chromatography to obtain eluate corresponding to 31.6min chromatographic peak, and further separating and purifying by high pressure liquid chromatography to obtain target compound of benzoisofuran.
And the crushing and sieving in the step A is to crush and sieve the mixture through a sieve with 30-50 meshes.
The organic solvent in the step A is acetone with the volume percentage concentration of 60-90%, ethanol with the volume percentage concentration of 80-100% or methanol with the volume percentage concentration of 80-100%.
Dissolving the extract a with an organic solvent which is 1.5-3 times of the weight of the extract a before the extract a is subjected to silica gel column chromatography, and then mixing the sample with 80-100 mesh silica gel which is 0.8-1.5 times of the weight of the extract d.
The organic solvent is pure methanol, pure ethanol or pure acetone.
Reverse phase C in step C18The medium-pressure liquid chromatography separation and purification is to separate and purify C with the size of 21.2mm multiplied by 250mm and 5 mu m18The chromatographic column is stationary phase, 68% methanol is mobile phase, flow rate is 20mL/min, wavelength detected by ultraviolet detector is 282nm, each sample injection is 500 μ L, chromatographic peak of 31.6min is collected, and evaporation is carried out after multiple accumulation.
And dissolving the compound separated and purified by the high pressure liquid chromatography again by using pure methanol, and separating by using gel column chromatography by using the pure methanol as a mobile phase so as to further separate and purify.
The application of the benzisothiafuran compounds is the application of the benzisothiafuran compounds in preparing bacteriostatic agents.
The application of the benzisothiafuran compounds is the application of the benzisothiafuran compounds in preparing the biological pesticide for resisting tobacco black shank.
The invention is further illustrated by the following specific examples:
EXAMPLE 1 preparation of the Compounds
The petasites ananatis sample is from Yunnan red river. 2.5kg of dried anshansa herb is sampled, crushed and extracted with 95% methanol for 5 times, each time for 45 minutes, the extracting solutions are combined, filtered and concentrated under reduced pressure to obtain 165g of extract. Dissolving the extract with 2.0 times of pure methanol, mixing with 180g of 100 mesh crude silica gel, loading 1.2kg of 160 mesh silica gel into column, performing silica gel column chromatography, gradient eluting with chloroform-acetone at volume ratio of 1:0, 20:1, 9:1, 8:2, 7:3, 6:4, 1:1 and 1:2, monitoring by TLC, mixing the same fractions to obtain 8 fractions, wherein the chloroform-acetone eluted fraction at volume ratio of 9:1 is separated by HPLC with ANJIERAN 1100 semi-preparative HPLC, using 68% methanol as mobile phase, and Zorbax SB-C18 (21.2.2)
250mm, 5 μm) column as stationary phase, flow rate of 20ml/min, wavelength of 282nm detected by ultraviolet detector, 500 μ L per sample injection, collecting chromatographic peak of 31.6min, accumulating for multiple times, and evaporating to dryness; dissolving the obtained product with pure methanol again, taking the pure methanol as a mobile phase, and carrying out Sephadex LH-20 gel column chromatography separation to obtain the new compound.
EXAMPLE 2 preparation of the Compounds
The Dolichos stalk-glandular senna sample is from Guangxi Baishi, the dried Dolichos stalk-glandular senna sample is crushed to 30 meshes, 4.2kg of the sample is sampled, the 4 times of extraction are carried out by 95 percent ethanol for 40 minutes each time, the extracting solutions are combined, filtered and concentrated under reduced pressure to obtain 302g of extract. Dissolving the extract with 2.0 times of pure methanol, mixing with 320g of 80 mesh crude silica gel, loading 2.2kg of 200 mesh silica gel into a column, performing silica gel column chromatography, performing gradient elution with chloroform-acetone with volume ratio of 1:0, 20:1, 9:1, 8:2, 7:3, 6:4, 1:1 and 1:2, monitoring by TLC, mixing the same parts to obtain 8 parts, separating the chloroform-acetone eluted part with volume ratio of 9:1 by using an anselan 1100 semi-preparative high performance liquid chromatography,using 68% methanol as mobile phase, Zorbax SB-C18 (21.2)
250mm, 5 μm) column as stationary phase, flow rate of 20ml/min, wavelength of 282nm detected by ultraviolet detector, 500 μ L per sample injection, collecting chromatographic peak of 31.6min, accumulating for multiple times, and evaporating to dryness; dissolving the obtained product with pure methanol again, taking the pure methanol as a mobile phase, and carrying out Sephadex LH-20 gel column chromatography separation to obtain the new compound.
EXAMPLE 3 preparation of the Compound
The Lupinus hancei sample is from Yuanjiang, 5.8kg of Lupinus hancei is sampled and crushed, extracted by ultrasonic for 3 times by 75% of acetone for 50 minutes each time, the extracting solutions are combined, filtered and concentrated under reduced pressure to obtain 427g of extract. Dissolving the extract with 1.6 times of pure methanol, mixing with 480g of 90 mesh crude silica gel, loading 3.2kg of 180 mesh silica gel into a column, performing silica gel column chromatography, performing gradient elution with chloroform-acetone with volume ratio of 1:0, 20:1, 9:1, 8:2, 7:3, 6:4, 1:1 and 1:2, monitoring by TLC, mixing the same parts to obtain 8 parts, wherein the chloroform-acetone elution part with volume ratio of 9:1 is separated by preparative high performance liquid chromatography of Anjiren 1100 half, using 68% methanol as mobile phase, and Zorbax SB-C18 (21.2.2)
250mm, 5 μm) column as stationary phase, flow rate of 20ml/min, wavelength of 332nm detected by ultraviolet detector, collecting chromatographic peak of 31.6min after each sample introduction of 500 μ L, accumulating for multiple times, and evaporating to dryness; dissolving the obtained product with pure methanol again, taking the pure methanol as a mobile phase, and carrying out Sephadex LH-20 gel column chromatography separation to obtain the new compound.
Example 4 structural characterization of Compounds
Taking the compounds prepared in examples 1 to 3, the structures of the compounds were determined by the following methods, and the results are shown in FIGS. 1 to 5:
the compounds of the invention are yellow gums; HRESI-MS shows that the peak of the excimer ion is 283.0952 [ M + Na ]]+(calculated 283.0946), combined1H NMR and DEPT spectra confirm that the molecular formula is C15H16O4The unsaturation degree is 8. The infrared spectrum shows hydroxyl groups (3415 cm)-1) Carbonyl group (1682 cm)-1) And aromatic rings (1614, 1556 and 1442 cm)-1) The resonance absorption peak of (1). The existence of aromatic ring structures in the compound is also indicated by the maximum absorption of the ultraviolet spectrum at 312, 272 and 210 nm. Process for preparing compounds1H、13C and DEPT NMR spectra (Table-1) showed that it contained 15 carbons and 16 hydrogens, 1,2,3,4, 5-pentasubstituted benzene rings (C-4 to C-7, C-4a and C-7a, H-7), one hydroxyethyl group (-CH)2CH2OH, C-6 'and C-7', H26' and H2-7'), one harmonic dimethylchromene ring (-CH = CH-C (CH)3）2-O-, C-1 '-C-5', H-1', H-2' and H6-4', 5'), one ester carbonyl (C-1), one oxidized methylene (C-3, H)2-3). According to the structural fragment of the compound, except for the unsaturation degree of a benzene ring of 4, the unsaturation degree of a synthylchromene ring of 2 and the unsaturation degree of an ester carbonyl of 1, the compound should be provided with a ring so as to meet the requirement that the unsaturation degree of the compound is 8. According to the presence of H in the compound2HMBC between-3 and C-1, C-4, C-4a, C-7a, and H-7 and C-1, C-4a, C-7a (FIG. 2) can confirm that the compound forms a 5-membered ring between C-1 and C-3 through an ester bond, and is an isobenzofuranolactone compound.
After the basic skeleton and the groups in the compound are determined, the positions of the substituents can be further determined according to the HMBC correlation of the compound. According to H26 'is related to HMBC at C-4, C-5, C-4a, and H-7' is related to HMBC at C-4, and the hydroxyethyl substitution can be determined at C-4; according to H-1' and C-5, C-6, C-7; h-2' and C-6, and H-7 and C-1' are related to HMBC, presumably gem-dimethyl chromene ring substitution at the C-5 and C-6 positions of the compound, and the C-1' carbon is attached at the C-6 position of the phenyl ring. The structure of the compound is now determined.
EXAMPLE 5 test of the antibacterial Activity of Compounds
The benzisofuran compounds prepared in the examples 1-3 are taken for the activity test of inhibiting phytophthora, and the method mainly comprises the following steps:
(1) preparation of oatmeal agar medium: adding 1000mL of water into oatmeal, heating the oatmeal in a boiling water bath for 1 hour, filtering the oatmeal by using gauze, adding the water for supplementing 1000mL, then adding sugar and agar, heating the oatmeal to completely melt the agar, filtering the oatmeal in a triangular flask by using the gauze (absorbent cotton is added in the middle) while the oatmeal is hot, sterilizing the oatmeal for 20min (minutes) at 121 ℃ and 15 pounds, taking out the oatmeal and cooling the oatmeal to about 45 ℃, adding ampicillin (5 mg/100 mL) into an aseptic operation table, uniformly mixing the ampicillin and the ampicillin, pouring the ampicillin into the flat dish, culturing the ampicillin in the flat dish for 48 hours at 28 ℃, and checking the ampicillin for later use after sterility is checked.
(2) Bacteriostatic experiments: placing circular filter paper with diameter of 5mm into a culture dish, placing in a 15 pound autoclave for 30min, oven drying, and respectively soaking in 20µM compound, 75% ethanol solution and sterilized distilled water. Respectively sucking 0.2mL of fresh bacterial liquid of phytophthora nicotianae on oatmeal agar medium plates by using a sterile suction pipe on a sterile operating table. Coating with triangular glass coating rod, lightly sticking the filter paper sheets onto corresponding flat plates with tweezers, culturing at 28 deg.C, observing experimental results, and measuring the size of antibacterial zone. Meanwhile, agricultural chloramphenicol was used as a positive control.
The test result shows that: the diameter of the inhibition zone of the benzisothiafuran compound is 16.5 +/-1.2 mm, and the diameter of the inhibition zone of the positive control agricultural chloramphenicol is 12.2 +/-1.0 mm. The compound has the effect of inhibiting phytophthora nicotianae remarkably superior to that of positive control agricultural chloramphenicol, and has outstanding activity of inhibiting black stem diseases.
EXAMPLE 5 Effect test on prevention and treatment of Black Stem disease in tobacco by Compound
The benzisofuran compounds prepared in examples 1-3 were used for the test of the control effect of tobacco black-stem disease, and the method mainly included the following steps:
transplanting tobacco seedlings into flowerpots with the diameter of 10cm and the height of 10cm, wherein the culture medium is as follows: sterilizing soil, peat and perlite (2: 2: 1), and culturing 1 strain per pot. After transplanting and seedling slowing, 10g of bacterial grains are added into roots, tobacco seedlings are placed in an artificial climate room for cultivation, the temperature is 30 ℃ in the daytime, the temperature is 28 ℃ in the night, the illumination is dark (12 h:12 h), and the relative humidity is 95%. To make the tobacco seedling attack. Before the black shank disease occurs, the tobacco seedlings are subjected to root irrigation treatment by using the compound of the invention with the particle size of 20 mu M, and 10mL of the compound is poured on each tobacco seedling; the pouring is carried out for 2 times. Each of 10 treated plants was repeated 3 times for 14 days, and then the disease incidence was investigated to calculate the disease index. The results show that: the compound has a control effect on tobacco black shank of (77.2 +/-3.4)%, and has an obvious control effect on tobacco black stalk diseases.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications or equivalents thereof, which are within the spirit and scope of the present invention, should be construed as being included therein.
Claims (10)
1. A benzisothiafuran compound is characterized in that the benzisothiafuran compound is separated from Dolichos anserina and has a molecular formula of C15H16O4A yellow gum having the following structural formula:
2. the preparation method of the benzisofuran compounds as claimed in claim 1, wherein the method comprises the steps of extract extraction, silica gel column chromatography and high pressure liquid chromatography separation and purification by taking the anshansa as a raw material, and specifically comprises the following steps:
A. extracting the extractum: crushing and sieving the pedunculate hydrangea, ultrasonically extracting for 3-5 times by using an organic solvent with the weight 3-10 times that of the pedunculate hydrangea for 30-60 min each time, combining the extracting solutions, and concentrating to obtain a flowable viscous extract a;
B. silica gel column chromatography: performing silica gel column chromatography on the extract a by using 160-300-mesh silica gel dry method which is 2-8 times of the weight of the extract a, performing gradient elution by using chloroform-acetone solutions with volume ratios of 1:0, 20:1, 9:1, 8:2, 7:3, 6:4, 1:1 and 1:2, collecting gradient eluent, concentrating, and combining the same parts;
C. high-pressure liquid chromatography separation: part 9:1 of the eluate from step B is subjected to reverse phase C18Separating and purifying by medium pressure liquid chromatography to obtain eluate corresponding to chromatographic peak of 31.6min, and further separating and purifying by high pressure liquid chromatography to obtain the final productA target substance of benzoisothiafuran compounds.
3. The method for producing benzisothiafuran compounds according to claim 2, wherein the pulverizing and sieving in step a is pulverizing and sieving with a 30-50 mesh sieve.
4. The method according to claim 2, wherein the organic solvent in step A is acetone, ethanol or methanol with a volume percentage of 60-90%, 80-100% or 80-100%.
5. The method for preparing benzisothiafuran compounds according to claim 2, wherein the extract a is dissolved in an organic solvent 1.5-3 times the weight of the extract a before being subjected to silica gel column chromatography, and then mixed with 80-100 mesh silica gel 0.8-1.5 times the weight of the extract d.
6. The process according to claim 5, wherein the organic solvent is pure methanol, pure ethanol or pure acetone.
7. The process for producing benzisothiafuran-based compounds according to claim 2, wherein the phase inversion C in the step C is18The medium-pressure liquid chromatography separation and purification is to separate and purify C with the size of 21.2mm multiplied by 250mm and 5 mu m18The chromatographic column is stationary phase, 68% methanol is mobile phase, flow rate is 20mL/min, wavelength detected by ultraviolet detector is 282nm, each sample injection is 500 μ L, chromatographic peak of 31.6min is collected, and evaporation is carried out after multiple accumulation.
8. The process according to claim 2, wherein the purified compound is further purified by dissolving in pure methanol and separating by gel column chromatography using pure methanol as a mobile phase.
9. Use of the benzoiso-furan-like compound of claim 1 in the preparation of a phytophthora bacteriostatic agent.
10. Use of the benzoisothiafuran-based compound of claim 1 in the preparation of a biological pesticide against tobacco black shank.
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| CN106916160A (en) * | 2017-02-14 | 2017-07-04 | 云南民族大学 | A kind of isobenzofuran class compounds process for production thereof that can improve cigarette suction comfort in the root of kudzu vine |
| CN108912136A (en) * | 2018-08-07 | 2018-11-30 | 云南中烟工业有限责任公司 | A kind of benzisoxa Furanones compound, preparation method and the usage for having effects that drop thorn and promoting the production of body fluid |
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| CN106916160A (en) * | 2017-02-14 | 2017-07-04 | 云南民族大学 | A kind of isobenzofuran class compounds process for production thereof that can improve cigarette suction comfort in the root of kudzu vine |
| CN108912136A (en) * | 2018-08-07 | 2018-11-30 | 云南中烟工业有限责任公司 | A kind of benzisoxa Furanones compound, preparation method and the usage for having effects that drop thorn and promoting the production of body fluid |
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| Isolation and structure elucidation of new salfredin-type metabolites from Crucibulum laeve DSM 1653 and DSM 8519;Tobias Neumann，等;《Journal of Basic Microbiology》;19991231;第39卷(第5-6期);357-363页 * |
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