CN117209428A - Anthraquinone compound with tobacco black shank resistance activity and preparation method and application thereof - Google Patents
Anthraquinone compound with tobacco black shank resistance activity and preparation method and application thereof Download PDFInfo
- Publication number
- CN117209428A CN117209428A CN202311180155.3A CN202311180155A CN117209428A CN 117209428 A CN117209428 A CN 117209428A CN 202311180155 A CN202311180155 A CN 202311180155A CN 117209428 A CN117209428 A CN 117209428A
- Authority
- CN
- China
- Prior art keywords
- compound
- black shank
- solvent
- tobacco black
- resisting
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 241000208125 Nicotiana Species 0.000 title claims abstract description 43
- 235000002637 Nicotiana tabacum Nutrition 0.000 title claims abstract description 43
- 241000233647 Phytophthora nicotianae var. parasitica Species 0.000 title claims abstract description 41
- PYKYMHQGRFAEBM-UHFFFAOYSA-N anthraquinone Natural products CCC(=O)c1c(O)c2C(=O)C3C(C=CC=C3O)C(=O)c2cc1CC(=O)OC PYKYMHQGRFAEBM-UHFFFAOYSA-N 0.000 title claims abstract description 37
- -1 Anthraquinone compound Chemical class 0.000 title claims abstract description 32
- 230000000694 effects Effects 0.000 title claims abstract description 32
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 150000001875 compounds Chemical class 0.000 claims abstract description 62
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 48
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 26
- 239000000284 extract Substances 0.000 claims abstract description 24
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims abstract description 22
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 18
- 239000000741 silica gel Substances 0.000 claims abstract description 14
- 229910002027 silica gel Inorganic materials 0.000 claims abstract description 14
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 11
- 238000000034 method Methods 0.000 claims abstract description 10
- OAIVIYSBZFEOIU-UHFFFAOYSA-N chloroform;propan-2-one Chemical compound CC(C)=O.ClC(Cl)Cl OAIVIYSBZFEOIU-UHFFFAOYSA-N 0.000 claims abstract description 9
- 239000002994 raw material Substances 0.000 claims abstract description 9
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical group CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 39
- 239000002904 solvent Substances 0.000 claims description 31
- 239000000243 solution Substances 0.000 claims description 26
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 16
- 241000196324 Embryophyta Species 0.000 claims description 14
- 238000010828 elution Methods 0.000 claims description 11
- 229920005654 Sephadex Polymers 0.000 claims description 10
- 239000012507 Sephadex™ Substances 0.000 claims description 10
- 238000002156 mixing Methods 0.000 claims description 10
- 239000012043 crude product Substances 0.000 claims description 8
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 8
- 239000007864 aqueous solution Substances 0.000 claims description 7
- 238000001035 drying Methods 0.000 claims description 7
- 238000004587 chromatography analysis Methods 0.000 claims description 6
- 238000001514 detection method Methods 0.000 claims description 6
- 239000003814 drug Substances 0.000 claims description 6
- 235000015110 jellies Nutrition 0.000 claims description 6
- 239000008274 jelly Substances 0.000 claims description 6
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 claims description 6
- 238000010298 pulverizing process Methods 0.000 claims description 6
- 239000012634 fragment Substances 0.000 claims description 5
- 238000009738 saturating Methods 0.000 claims description 4
- 238000000605 extraction Methods 0.000 abstract description 10
- 230000002401 inhibitory effect Effects 0.000 abstract description 5
- 150000002611 lead compounds Chemical class 0.000 abstract description 4
- 239000000575 pesticide Substances 0.000 abstract description 4
- 239000003480 eluent Substances 0.000 abstract 1
- 238000010898 silica gel chromatography Methods 0.000 abstract 1
- 239000012071 phase Substances 0.000 description 20
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 12
- 150000004056 anthraquinones Chemical class 0.000 description 11
- 238000005481 NMR spectroscopy Methods 0.000 description 8
- 125000003118 aryl group Chemical group 0.000 description 8
- 238000001914 filtration Methods 0.000 description 8
- IOLCXVTUBQKXJR-UHFFFAOYSA-M potassium bromide Chemical compound [K+].[Br-] IOLCXVTUBQKXJR-UHFFFAOYSA-M 0.000 description 8
- 238000010992 reflux Methods 0.000 description 7
- 229960005091 chloramphenicol Drugs 0.000 description 6
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 6
- 201000010099 disease Diseases 0.000 description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 6
- 238000004896 high resolution mass spectrometry Methods 0.000 description 6
- 241000233645 Phytophthora nicotianae Species 0.000 description 5
- 244000037364 Cinnamomum aromaticum Species 0.000 description 4
- 235000014489 Cinnamomum aromaticum Nutrition 0.000 description 4
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 4
- 239000008346 aqueous phase Substances 0.000 description 4
- 239000011521 glass Substances 0.000 description 4
- 150000002500 ions Chemical class 0.000 description 4
- 229930014626 natural product Natural products 0.000 description 4
- 238000000643 oven drying Methods 0.000 description 4
- 229920006395 saturated elastomer Polymers 0.000 description 4
- 239000011734 sodium Substances 0.000 description 4
- 235000002906 tartaric acid Nutrition 0.000 description 4
- 239000011975 tartaric acid Substances 0.000 description 4
- 241000220485 Fabaceae Species 0.000 description 3
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000004440 column chromatography Methods 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 3
- 238000001704 evaporation Methods 0.000 description 3
- 238000003919 heteronuclear multiple bond coherence Methods 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 230000003595 spectral effect Effects 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- AZQWKYJCGOJGHM-UHFFFAOYSA-N 1,4-benzoquinone Chemical compound O=C1C=CC(=O)C=C1 AZQWKYJCGOJGHM-UHFFFAOYSA-N 0.000 description 2
- MUVQKFGNPGZBII-UHFFFAOYSA-N 1-anthrol Chemical class C1=CC=C2C=C3C(O)=CC=CC3=CC2=C1 MUVQKFGNPGZBII-UHFFFAOYSA-N 0.000 description 2
- XILIYVSXLSWUAI-UHFFFAOYSA-N 2-(diethylamino)ethyl n'-phenylcarbamimidothioate;dihydrobromide Chemical compound Br.Br.CCN(CC)CCSC(N)=NC1=CC=CC=C1 XILIYVSXLSWUAI-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 238000000862 absorption spectrum Methods 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 230000000853 biopesticidal effect Effects 0.000 description 2
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 229940125782 compound 2 Drugs 0.000 description 2
- 125000000524 functional group Chemical group 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 125000000623 heterocyclic group Chemical group 0.000 description 2
- 238000001052 heteronuclear multiple bond coherence spectrum Methods 0.000 description 2
- 238000005570 heteronuclear single quantum coherence Methods 0.000 description 2
- 238000002329 infrared spectrum Methods 0.000 description 2
- 238000003973 irrigation Methods 0.000 description 2
- 230000002262 irrigation Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- 125000000325 methylidene group Chemical group [H]C([H])=* 0.000 description 2
- 239000000779 smoke Substances 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 125000001424 substituent group Chemical group 0.000 description 2
- 241000173529 Aconitum napellus Species 0.000 description 1
- 206010007247 Carbuncle Diseases 0.000 description 1
- 206010051625 Conjunctival hyperaemia Diseases 0.000 description 1
- 206010010726 Conjunctival oedema Diseases 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 206010017553 Furuncle Diseases 0.000 description 1
- 206010019280 Heart failures Diseases 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 241000546273 Lindera <angiosperm> Species 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 208000004880 Polyuria Diseases 0.000 description 1
- 229940023019 aconite Drugs 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- XAIKOVRFTSBNNU-UHFFFAOYSA-N anthracene-9,10-dione Chemical compound C1=CC=C2C(=O)C3=CC=CC=C3C(=O)C2=C1.C1=CC=C2C(=O)C3=CC=CC=C3C(=O)C2=C1 XAIKOVRFTSBNNU-UHFFFAOYSA-N 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 150000004777 chromones Chemical class 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 230000001882 diuretic effect Effects 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 230000002439 hemostatic effect Effects 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 239000006877 oatmeal agar Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000010451 perlite Substances 0.000 description 1
- 235000019362 perlite Nutrition 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
Landscapes
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention discloses an anthraquinone compound with tobacco black shank resistance activity, a preparation method and application thereof, wherein the compound takes concha haliotidis as a raw material, high-concentration methanol, high-concentration acetone/water or high-concentration ethanol/water are used for extraction, and extracting solutions are combined, filtered and decompressed and concentrated into extractum; loading the extract into a column by a silica gel dry method, and performing silica gel column chromatography; gradient eluting with chloroform-acetone solution; separating and purifying the 8:2 part of the eluent by high pressure liquid chromatography to obtain the required anthraquinone compound. The invention also discloses application of the compound, and an activity test shows that the compound has a good inhibiting effect on tobacco black shank. The compound disclosed by the invention is novel in structure, has good black shank resistance activity, can be used as a lead compound for resisting the tobacco black shank, and is used for researching and developing biological pesticides for resisting the tobacco black shank.
Description
Technical Field
The invention belongs to the technical field of natural product chemistry, and particularly relates to a compound with tobacco black shank resistance activity extracted from Dai medicine ear leaf shell of the genus Cassia of the family Leguminosae, and application thereof in preparing a medicine for resisting tobacco black shank.
Background
Tobacco black shank is one of the most destructive diseases in tobacco production, also called tobacco epidemic disease, and tobacco growers are called black stem crazy, black root and aconite. The main smoke producing areas in China all occur to different degrees, wherein Anhui, shandong and Henan provinces are historic severe areas; southern smoke areas such as Yunnan, guizhou, sichuan, hunan, guangdong, guangxi and Fujian are also quite common. At present, the prevention and control of the black shank are mainly realized by methods of rotation cultivation, variety gene improvement, biological pesticide and the like. Among them, control with biopesticides is the most commonly used and readily achievable method. Therefore, searching for more efficient biopesticide lead compounds has important significance for preventing and controlling tobacco black shank.
Radix seu folium Juglandis ImmaturusCassia auriculataL.) is a perennial shrub shawl herb of the genus Cassia of the family Leguminosae, is a common Chinese medicinal material in Yunnan province, has the effects of promoting heat and removing dampness, eliminating dampness and toxin, and treating main heat syndrome and heart failure, and is often used for treating cold-dampness diarrhea, wind-heat cough, conjunctival congestion and swelling and pain, carbuncle, sore furuncle and other diseases. Previous studyThe main active ingredients in the concha haliotidis are anthraquinone and chromone compounds. Of these two main active ingredients, anthraquinone (Anthraquinone) is a quinone type chemical. Anthraquinone derivatives are widely found in nature or can be synthesized artificially. Anthraquinones include products and dimers of varying degrees of reduction, such as anthracenols, oxidized anthracenols, anthracnones, and the like, and in addition, glycosides of these compounds. In natural products, anthraquinone is often present in the metabolites of higher and lower plant lichens and fungi, and has hemostatic, antibacterial, toxic materials removing, purgation, and diuretic effects. The invention discovers a novel anthraquinone compound from the cassia ear of the genus Cassia of the family Leguminosae, and is notable that the compound contains [2,3- ] in rare anthraquinone structurec]The novel skeleton molecule of the aza Zhuo Huanjie structural fragment has higher novelty in structure, and the compound has remarkable tobacco black shank resistance activity, has a control effect (72.6+/-3.8)% on tobacco black shank, is superior to a control effect (63.2+/-3.5) of control agricultural chloramphenicol, and can be used as a lead compound for resisting tobacco black shank for researching and developing biological pesticide for resisting tobacco black shank.
Disclosure of Invention
The first object of the present invention is to provide an anthraquinone compound having an activity against tobacco black shank; the second aim is to provide a preparation method of the anthraquinone compound with the activity of resisting tobacco black shank; the third aim is to provide the application of the anthraquinone compound with the activity of resisting tobacco black shank.
The first object of the present invention is achieved in that the anthraquinone compound with activity of resisting tobacco black shank is a light red jelly, and the molecular formula is: c (C) 21 H 17 NO 4 Designated as 2',3' -dihydro-5, 4' -dimethyl-6-methoxy-1HAnthracene [2,3 ]c]Azepine-1',9,10-trione, english name: 6-methoxy-5,4'-dimethyl-2',3'-dihydro-1'H-anthra[2,3-c]Azepine-1',9,10-trione. Characterized in that the compound contains [2,3 ]c]An aza Zhuo Huanjie fragment, the structure of the compound is as follows:
。
the second object of the invention is realized by taking concha haliotidis as a raw material, which comprises the following steps:
1) Sun-drying and pulverizing the whole plant of ear leaf Concha Haliotidis, extracting with a first solvent, concentrating, extracting with a second solvent, saturating the water phase with sodium carbonate, extracting with the second solvent, mixing, and concentrating to obtain extract a; the first solvent is an ethanol aqueous solution with the concentration of 90-96%; the second solvent is ethyl acetate;
2) Dissolving the extract a with a third solvent, separating by a silica gel column, performing gradient elution with chloroform-acetone solution, and collecting a part with the volume ratio of chloroform-acetone solution of 8:2 to obtain an elution component b; the third solvent is methanol, ethanol or acetone;
3) Purifying the eluting component b by a sephadex column to obtain a crude product of the target compound; and separating the crude target compound by HPLC chromatography to obtain the pure target compound.
The anthraquinone compound structure with the tobacco black shank resistance prepared by the steps can be identified by the following method:
TABLE 1 Compounds 1 H NMR 13 C NMR data (CDCl) 3 )
Appearance observation finds that: the compound of the invention is a pale red jelly; the ultraviolet-visible absorption spectrum shows that the compound has maximum absorption at 215, 268 and 415 and nm, and the aromatic ring structure is proved to exist in the compound; infrared spectra (potassium bromide tablets) showed-NH (3342 cm) -1 ，3138 cm -1 ) Kenocarbonyl (1668 cm) -1 ) Aromatic ring (1620, 1542, 1463 cm) -1 ) A characteristic functional group; high resolution mass spectrometry (HRESIMS) gives an excimer ion peak 370.1064 [ M+Na ]] + The molecular formula of the compound can be determined to be C 21 H 17 NO 4 。
For it 1 H NMR、 13 C NSpectroscopic data of MR, DEPT and HSQC were analyzed in detail and found to have 3 carbonyl groups, 9 aromatic aprotic carbons, 5 tertiary aromatic carbons, 1 methylene, 1 methoxy and 2 methyl groups present. HMBC spectra show that H-1 is associated with C-4a/C-3 and C-9, H-4 is associated with C-1a/C-2 and C-10, H-7 is associated with C-5 and C-9a, and H-8 is associated with C-10a/C-6 and C-9, indicating the presence of the 2,3,5, 6-tetrasubstituted-9, 10-anthraquinone substructure. In addition, H-NH is associated with C-1 '/C-2/C-3 ' and C-4 ', H 2 -3 ' is associated with C-1 '/C-4 ' and C-5 ', H-5 ' is associated with C-2/C-3/C-3 ' and C-4 ', and H 3 -6 'is related to C-3'/C-4 'and C-5', indicating the presence of a methyl-substituted nitrogen-containing seven-membered heterocycle. Whereas the HMBC correlation of H-1 with C-1 ', H-4 with C-5 ', and H-5 ' with C-4 indicates that the two subunits are located at the C-2 and C-3 positions. The remaining substituents being through H 3 -7' to C-5, C-6 and C-10a, and C-6 to C-6. To this end, the structure of the compound can be confirmed and is named: 2',3' -dihydro-5, 4' -dimethyl-, 6-methoxy-1HAnthracene [2,3 ]c]Azepine-1',9,10-trione, english name: 6-methoxy-5,4'-dimethyl-2',3'-dihydro-1'H-anthra[2,3-c]- azepine-1',9,10-trione。
Infrared, ultraviolet, mass spectral data for the compounds: UV (methanol)l max (loge) 215 (4.36), 268 (3.68), 415 (3.58) nm nm; IR (KBr)n max 3342, 3138, 2946, 1668, 1655, 1620, 1542, 1463, 1348, 1274, 1162, 1059, 842 cm -1 ; 1 H and 13 C NMR data are shown in Table 1, HRESIMS (positive ion mode)m/ z370.1064 [M+Na] + (C 21 H 17 NNaO 4 Calculated 370.1055).
The third object of the invention is realized in such a way that the anthraquinone compound is applied to the preparation of the tobacco black shank resistant medicine.
The beneficial effects of the invention are as follows:
1) The invention extracts anthraquinone compound with novel structure from concha haliotidis for the first time, and is worth focusing on: the compound is an anthraquinone new skeleton molecule containing [2,3-c ] aza Zhuo Huanjie structural fragment in a rare structure in a natural product, has high novelty, has good black shank resistance activity, and can be used as a lead compound for resisting the black shank of tobacco for researching and developing biological pesticides for resisting the black shank of tobacco.
2) The invention takes the traditional Chinese medicine concha haliotidis as the raw material for the first time, and provides a new research direction for the development of natural products of plant sources with the plant disease control effect.
3) The raw materials for preparing the compound have wide sources and low cost, can provide sufficient and continuous raw material support for the preparation of the compound, and are easy to realize industrialized production.
Drawings
FIG. 1 is a nuclear magnetic resonance spectrum of an anthraquinone compound prepared in example 1;
FIG. 2 is a nuclear magnetic resonance hydrogen spectrum of the anthraquinone compound prepared in example 1;
FIG. 3 is a main HMBC-related diagram of the anthraquinone compound prepared in example 1.
Detailed Description
The invention is further described below with reference to examples and figures, but is not limited in any way, and any alterations or substitutions based on the teachings of the invention are within the scope of the invention.
The anthraquinone compound with the activity of resisting the tobacco black shank is a light red jelly, and the molecular formula is as follows: c (C) 21 H 17 NO 4 Designated as 2',3' -dihydro-5, 4' -dimethyl-6-methoxy-1HAnthracene [2,3 ]c]Azepine-1',9,10-trione, english name: 6-methoxy-5,4'-dimethyl-2',3'-dihydro-1'H-anthra[2,3-c]Azepine-1',9,10-trione. Characterized in that the compound contains [2,3 ]c]An aza Zhuo Huanjie fragment, the structure of the compound is as follows:
。
the anthraquinone compound with the tobacco black shank resistance activity is prepared from concha haliotidis serving as a raw material, and specifically comprises the following components:
1) Sun-drying and pulverizing the whole plant of ear leaf Concha Haliotidis, extracting with a first solvent, concentrating, extracting with a second solvent, saturating the water phase with sodium carbonate, extracting with the second solvent, mixing, and concentrating to obtain extract a; the first solvent is an ethanol aqueous solution with the concentration of 90-96%; the second solvent is ethyl acetate;
2) Dissolving the extract a with a third solvent, separating by a silica gel column, performing gradient elution with chloroform-acetone solution, and collecting a part with the volume ratio of chloroform-acetone solution of 8:2 to obtain an elution component b; the third solvent is methanol, ethanol or acetone;
3) Purifying the eluting component b by a sephadex column to obtain a crude product of the target compound; and separating the crude target compound by HPLC chromatography to obtain the pure target compound.
The volume ratio of the chloroform-acetone solution is sequentially 20:1, 9:1, 8:2, 7:3, 6:4 and 5:5.
The mobile phase purified by the sephadex column in step 3) is methanol.
HPLC chromatography in step 3) was performed using 21.2 mm X25 cm,5μC of m 18 The chromatographic column takes 58% methanol water solution as a mobile phase, the flow rate is 12 mL/min, the detection wavelength of the ultraviolet detector is 415 nm, and the chromatographic peak is collected for 33.6 min.
The application of the anthraquinone compound disclosed by the invention is the application of the anthraquinone compound with the activity of resisting tobacco black shank in preparing a medicine for resisting tobacco black shank.
The preparation method comprises the following specific operations:
(1) Extracting extract: sun-drying and pulverizing the whole plant of ear leaf Concha Haliotidis, extracting with a first solvent, concentrating, extracting with a second solvent, saturating the water phase with sodium carbonate, extracting with the second solvent, mixing, and concentrating to obtain extract;
(2) Dissolving the extract with a third solvent, separating by a silica gel column, eluting with chloroform/acetone gradient, and collecting chloroform: an eluting component having an acetone volume ratio of 8:2;
(3) Purifying the elution component obtained in the step 2 by using a sephadex column to obtain a crude product of the target compound, and finally separating the crude product of the target compound by using HPLC (high performance liquid chromatography) chromatography to obtain a pure product of the target compound.
The first solvent is a 95% ethanol aqueous solution.
In the step 1, the first solvent is adopted to carry out 2-3 times of reflux extraction, and the time of each reflux extraction is 35-60 min.
Concentrating the extractive solution obtained by reflux extraction, diluting with 3% tartaric acid, and extracting for 2-3 times.
The aqueous phase saturated with sodium carbonate is extracted 2-3 more times with a second solvent.
In the step 2, the volume ratio of chloroform to acetone for gradient elution is 20:1, 9:1, 8:2, 7:3, 6:4 and 5:5 in sequence.
In the step 2, a 160-200 mesh silica gel column is adopted for separation, and after the extract is dissolved by a second solvent with the amount of 1.5-3 times, the extract is stirred with 80-120 mesh crude silica gel.
In the process of mixing samples, the weight of the sample mixing silica gel is 0.8 to 2.5 times of that of the extract.
In the step 3, the high pressure liquid chromatography is performed by using a Zorbax PrepHT GF (21.2 mm ×25 cm) reverse phase column, 58% methanol water solution is used as a mobile phase, the flow rate is 12 mL/min, the detection wavelength of an ultraviolet detector is 415 nm, and a chromatographic peak of 33.6 min is collected.
In the step 3, the sephadex column purification is carried out by taking methanol as a mobile phase.
The second solvent is ethyl acetate.
The third solvent is methanol, ethanol or acetone.
The invention also provides application of the anthraquinone compound in preparing a preparation for resisting tobacco black shank. Through an activity test for resisting tobacco black shank, the compound shown in the formula (I) has a remarkable inhibiting effect on tobacco black shank.
The invention can be realized by adopting the concha haliotidis raw materials without being limited by regions and varieties, and the invention is further described below by adopting the concha haliotidis raw materials from Yunnan university.
Example 1
The ear leaf concha haliotidis sample of the embodiment is derived from the mouth of a Yunnan red river. Sun drying the whole plant of ear leaf Concha Haliotidis, and pulverizing to about 40 mesh. Weighing a 3.5-kg crushed sample, placing the sample in a 20-L glass reaction kettle, adding a 95% ethanol aqueous solution 6-L, carrying out reflux extraction for 50 min, and filtering out an extracting solution; and adding 95% ethanol water solution 6L into the residue again, reflux-extracting for 50 min, and filtering to remove the extractive solution. The two extracts were combined and concentrated to a small volume, then diluted with 3% tartaric acid solution 6L and extracted 2 times with 6L ethyl acetate. After extraction, the aqueous phase was saturated with sodium carbonate, extracted 2 times with ethyl acetate 6. 6L again, the extracted ethyl acetate phases were combined and concentrated under reduced pressure to give anthraquinone fraction extract 53.6 g. Mixing the extract with 80 g (80-120 mesh) crude silica gel, oven drying, subjecting to 200 g silica gel (160-200 mesh) column chromatography, chloroform: acetone (20:1, 9:1, 8:2, 7:3, 6:4, 5:5) in a gradient, split into 6 fractions. Selecting an 8:2 elution part, concentrating, dissolving with methanol, taking methanol as a mobile phase, and purifying with a sephadex column to obtain a crude compound. The crude product is further separated by HPLC: the method comprises the steps of using Zorbax PrepHT GF (21.2 mm multiplied by 25 cm) reverse phase column of Agilent company, using 58% methanol water solution as mobile phase, using the flow rate of 12 mL/min, using an ultraviolet detector to detect the wavelength of 415 nm, collecting chromatographic peaks of 33.6 min, accumulating for multiple times, and evaporating to dryness to obtain a light red jelly, namely a pure product of the target compound.
Example 2
The ear leaf concha haliotidis sample is from Lindera yunnanensis Meng Ding. Sun drying the whole plant of ear leaf Concha Haliotidis, and pulverizing to about 40 mesh. Weighing a 4.0 and kg crushed sample, placing the sample in a 20L glass reaction kettle, adding a 95% ethanol water solution 8L, carrying out reflux extraction for 40min, and filtering out an extracting solution; and adding 95% ethanol water solution 8L into the residue again, reflux-extracting for 40min, and filtering to remove the extractive solution. The two extracts were combined and concentrated to a small volume, then diluted with 3% tartaric acid solution 8L and extracted 2 times with ethyl acetate 8L. After extraction, the aqueous phase was saturated with sodium carbonate, extracted 2 times with ethyl acetate 8. 8L again, the extracted ethyl acetate phases were combined and concentrated under reduced pressure to give anthraquinone fraction extract 68.5. 68.5 g. Mixing the extract with 100 g (80-120 mesh) crude silica gel, oven drying, separating by column chromatography with 220 g silica gel (160-200 mesh), and separating chloroform: acetone (20:1, 9:1, 8:2, 7:3, 6:4, 5:5) in a gradient, split into 6 fractions. Selecting an 8:2 elution part, concentrating, dissolving with methanol, taking methanol as a mobile phase, and purifying with a sephadex column to obtain a crude compound. The crude product was further subjected to HPLC for further separation: the pure compound is obtained by using Zorbax PrepHT GF (21.2 mm multiplied by 25 cm) reverse phase column of Agilent company, using 58% methanol water solution as mobile phase, the flow rate is 12 mL/min, the detection wavelength of ultraviolet detector is 415 nm, collecting chromatographic peaks of 33.6 min, accumulating for multiple times, and evaporating.
The structural identification of the compound obtained in this example was the same as that of example 1, and it was confirmed that the compound prepared in this example was the anthraquinone compound-2 ',3' -dihydro-5, 4' -dimethyl-6-methoxy-1HAnthracene [2,3-c ]]Azepine-1',9,10-trione.
Example 3
The ear leaf Concha Haliotidis sample is derived from Menghai, xishuangbanna, yunnan. Taking the whole plant of the concha haliotidis, sun-drying, and crushing to about 50 meshes. Weighing a crushed sample of 4.6 and kg, placing the crushed sample in a glass reaction kettle of 20L, adding a 95% ethanol aqueous solution of 10L, carrying out reflux extraction for 30 min, and filtering out an extracting solution; and adding a 95% ethanol aqueous solution 10L into the residue again, extracting under reflux for 50 min, and filtering to remove the extract. The two extracts were combined and concentrated to a small volume, then diluted with 3% tartaric acid solution 10L and extracted 2 times with 10L ethyl acetate. After extraction, the aqueous phase was saturated with sodium carbonate, extracted 2 times with ethyl acetate 8. 8L again, the extracted ethyl acetate phases were combined and concentrated under reduced pressure to give anthraquinone fraction extract 88.5. 88.5 g. Mixing the extract with 130 g (80-120 mesh) crude silica gel, oven drying, separating by column chromatography with 300 g silica gel (160-200 mesh), and separating chloroform: acetone (20:1, 9:1, 8:2, 7:3, 6:4, 5:5) in a gradient, split into 6 fractions. Selecting an 8:2 elution part, concentrating, dissolving with methanol, taking methanol as a mobile phase, and purifying with a sephadex column to obtain a crude compound. The crude product was further subjected to HPLC for further separation: the pure compound is obtained by using Zorbax PrepHT GF (21.2 mm multiplied by 25 cm) reverse phase column of Agilent company, using 58% methanol water solution as mobile phase, the flow rate is 12 mL/min, the detection wavelength of ultraviolet detector is 415 nm, collecting chromatographic peaks of 33.6 min, accumulating for multiple times, and evaporating.
The chemistry obtained in this exampleStructure identification of the Compound As in example 1, it was confirmed that the compound prepared in this example was the anthraquinone compound-2 ',3' -dihydro-5, 4' -dimethyl-6-methoxy-1HAnthracene [2,3-c ]]Azepine-1',9,10-trione.
Example 4
The structure of the anthraquinone compound prepared by the method is identified by the following method:
the compound of the invention is found to be a pale red jelly by visual observation; the ultraviolet-visible absorption spectrum shows that the compound has maximum absorption at 215, 268 and 415 and nm, and the aromatic ring structure is proved to exist in the compound; infrared spectra (potassium bromide tablets) showed-NH (3342 cm) -1 ，3138 cm -1 ) Kenocarbonyl (1668 cm) -1 ) Aromatic ring (1620, 1542, 1463 cm) -1 ) A characteristic functional group; high resolution mass spectrometry (HRESIMS) gives an excimer ion peak 370.1064 [ M+Na ]] + The molecular formula of the compound can be determined to be C 21 H 17 NO 4 。
For it 1 H NMR、 13 The spectral data of C NMR, DEPT and HSQC were analyzed in detail and found to have 3 carbonyl groups, 9 aromatic aprotic carbons, 5 tertiary aromatic carbons, 1 methylene, 1 methoxy and 2 methyl groups present. HMBC spectra show that H-1 is associated with C-4a/C-3 and C-9, H-4 is associated with C-1a/C-2 and C-10, H-7 is associated with C-5 and C-9a, and H-8 is associated with C-10a/C-6 and C-9, indicating the presence of the 2,3,5, 6-tetrasubstituted-9, 10-anthraquinone substructure. In addition, H-NH is associated with C-1 '/C-2/C-3 ' and C-4 ', H 2 -3 ' is associated with C-1 '/C-4 ' and C-5 ', H-5 ' is associated with C-2/C-3/C-3 ' and C-4 ', and H 3 -6 'is related to C-3'/C-4 'and C-5', indicating the presence of a methyl-substituted nitrogen-containing seven-membered heterocycle. Whereas the HMBC correlation of H-1 with C-1 ', H-4 with C-5 ', and H-5 ' with C-4 indicates that the two subunits are located at the C-2 and C-3 positions. The remaining substituents being through H 3 -7' to C-5, C-6 and C-10a, and C-6 to C-6. To this end, the structure of the compound can be confirmed and is named: 2',3' -dihydro-5, 4' -dimethyl-, 6-methoxy-1HAnthracene [2,3 ]c]Azepine-1',9,10-trione, english name:6-methoxy-5,4'-dimethyl-2',3'-dihydro-1'H-anthra[2,3-c]- azepine-1',9,10-trione。
infrared, ultraviolet, mass spectral data for the compounds: UV (methanol)l max (loge) 215 (4.36), 268 (3.68), 415 (3.58) nm nm; IR (KBr)n max 3342, 3138, 2946, 1668, 1655, 1620, 1542, 1463, 1348, 1274, 1162, 1059, 842 cm -1 ; 1 H and 13 C NMR data are shown in Table 1, HRESIMS (positive ion mode)m/ z370.1064 [M+Na] + (C 21 H 17 NNaO 4 Calculated 370.1055).
Example 5
Test of tobacco Black shank resistance Activity of the Compounds of the invention
1. Test for inhibiting Phytophthora nicotianae Activity
Adding water 1000 and mL to oatmeal, heating on boiling water bath for 1 hr, filtering with gauze, adding water to complement 1000 mL, adding sugar and agar, heating to melt agar completely, filtering with gauze (absorbent cotton is added in the middle) in a triangular flask while hot, sterilizing at 121deg.C for 20min (min), taking out, cooling to about 45deg.C, adding ampicillin (5 mg/100 mL) on aseptic console, mixing, pouring into a dish, culturing at 28deg.C for 48 hr, checking for sterility, and keeping stand-by.
Taking round filter paper with diameter of 5 mm, placing into a culture dish, sterilizing at 15 lbs. under high pressure for 30 min, oven drying, and soaking into 20 respectivelyµM in the compound prepared in example 1, 75% ethanol in water and sterile distilled water. On a sterile operating table, respectively sucking fresh bacterial liquid of 0.2 mL phytophthora nicotianae on a oatmeal agar medium plate by using a sterile suction tube. Uniformly coating with triangular glass coating rod, respectively lightly attaching filter paper sheets to corresponding flat plates with tweezers, culturing at 28deg.C, observing experimental results, and determining the size of the inhibition zone. Meanwhile, chloramphenicol for agricultural use was used as a positive control.
The test results show that: the diameter of the antibacterial ring of the anthraquinone compound is 15.26+/-2.2 mm, and the diameter of the antibacterial ring of the positive control agricultural chloramphenicol is 12.8+/-2.0 mm. The result shows that the effect of the compound for inhibiting phytophthora nicotianae is obviously better than that of positive control agricultural chloramphenicol, and the compound has outstanding activity for inhibiting black shank.
2. Effect detection for preventing and controlling tobacco black shank
Tobacco seedlings are transplanted into flowerpots with the diameter of 10 cm and the height of 10 cm, and the cultivation substrates are as follows: sterilized soil, turf, perlite (2:2:1), 1 plant per pot. After transplanting and seedling-recovering, adding fungus and cereal 10 g per plant into root, placing tobacco seedling into artificial climate chamber for culturing, at 30 ℃ in daytime and 28 ℃ at night, and illuminating: darkness (12 h: 12 h), 95% relative humidity, caused tobacco seedling onset. And (3) root irrigation treatment is carried out on tobacco seedlings by using the compound disclosed by the invention with the particle size of 20 mu M before the occurrence of black shank, wherein 10 per plant is irrigated mL, and the total amount of irrigation is 2 times. After repeating 10 plants per treatment for 3 times and 14 to d, the disease condition was investigated and the disease index was calculated.
Results: the control effect of the compound on tobacco black shank is (72.6+/-3.8)%, and the control effect of the compound on control agricultural chloramphenicol is (63.2+/-3.5), and the control effect of the compound is superior to that of the compound on control agricultural chloramphenicol, so that the anthraquinone compound has remarkable tobacco black shank control effect.
Claims (6)
1. The anthraquinone compound with the activity of resisting the tobacco black shank is a light red jelly, and the molecular formula is as follows: c (C) 21 H 17 NO 4 Designated as 2',3' -dihydro-5, 4' -dimethyl-6-methoxy-1HAnthracene [2,3 ]c]Azepine-1',9,10-trione, english name: 6-methoxy-5,4'-dimethyl-2',3'-dihydro-1'H-anthra[2,3-c]Azepine-1',9,10-trione. Characterized in that the compound contains [2,3 ]c]An aza Zhuo Huanjie fragment, the structure of the compound is as follows:
。
2. an anthraquinone compound with tobacco black shank resistance according to claim 1, which is prepared from concha haliotidis as a raw material, and specifically comprises the following steps:
1) Sun-drying and pulverizing the whole plant of ear leaf Concha Haliotidis, extracting with a first solvent, concentrating, extracting with a second solvent, saturating the water phase with sodium carbonate, extracting with the second solvent, mixing, and concentrating to obtain extract a; the first solvent is an ethanol aqueous solution with the concentration of 90-96%; the second solvent is ethyl acetate;
2) Dissolving the extract a with a third solvent, separating by a silica gel column, performing gradient elution with chloroform-acetone solution, and collecting a part with the volume ratio of chloroform-acetone solution of 8:2 to obtain an elution component b; the third solvent is methanol, ethanol or acetone;
3) Purifying the eluting component b by a sephadex column to obtain a crude product of the target compound; and separating the crude target compound by HPLC chromatography to obtain the pure target compound.
3. The preparation method according to claim 2, wherein the volume ratio of the chloroform-acetone solution is 20:1, 9:1, 8:2, 7:3, 6:4 and 5:5 in sequence.
4. The process according to claim 2, wherein the mobile phase purified by the sephadex column in step 3) is methanol.
5. The process according to claim 2, wherein the HPLC chromatography in step 3) is carried out using 21.2 mm X25 cm,5μC of m 18 The chromatographic column takes 58% methanol water solution as a mobile phase, the flow rate is 12 mL/min, the detection wavelength of the ultraviolet detector is 415 nm, and the chromatographic peak is collected for 33.6 min.
6. The application of the anthraquinone compound with the activity of resisting the tobacco black shank according to claim 1, which is characterized in that the application of the anthraquinone compound with the activity of resisting the tobacco black shank in preparing a medicine for resisting the tobacco black shank.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311180155.3A CN117209428A (en) | 2023-09-13 | 2023-09-13 | Anthraquinone compound with tobacco black shank resistance activity and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311180155.3A CN117209428A (en) | 2023-09-13 | 2023-09-13 | Anthraquinone compound with tobacco black shank resistance activity and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117209428A true CN117209428A (en) | 2023-12-12 |
Family
ID=89041996
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311180155.3A Pending CN117209428A (en) | 2023-09-13 | 2023-09-13 | Anthraquinone compound with tobacco black shank resistance activity and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117209428A (en) |
-
2023
- 2023-09-13 CN CN202311180155.3A patent/CN117209428A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110483396B (en) | Isopentenyl isoquinoline alkaloid compound and preparation method and application thereof | |
| CN110483535B (en) | Isoquinoline tricyclic alkaloid compound and preparation method and application thereof | |
| CN110452170B (en) | Isoquinoline alkaloid compound and preparation method and application thereof | |
| CN103524472B (en) | Phenolic compound, and preparation method and application thereof | |
| CN113087664B (en) | Quinoline alkaloid compound and preparation method and application thereof | |
| CN113200989B (en) | Preparation method and application of chromone alkaloid compound | |
| CN113214152B (en) | Active compound for resisting tobacco black shank in thalictrum delavayi Franch, preparation method and application thereof | |
| CN114456102A (en) | Indole alkaloid compound and preparation method and application thereof | |
| CN111072616B (en) | Compound for resisting tobacco black shank and preparation method and application thereof | |
| CN110357894B (en) | Tricyclic alkaloid compound and preparation method and application thereof | |
| CN110590723B (en) | Sterol compound in tobacco as well as preparation method and application thereof | |
| CN115286561B (en) | Indole alkaloid compound in gene editing tobacco, and preparation method and application thereof | |
| CN113200911B (en) | Quinoline alkaloid compound and preparation method and application thereof | |
| CN114409661B (en) | Indole alkaloid compound and preparation method and application thereof | |
| CN113717184B (en) | Quinoline alkaloid with tobacco mosaic virus resisting activity in cigar and preparation method and application thereof | |
| CN110467623B (en) | Benzoisofuran compound and preparation method and application thereof | |
| CN117209428A (en) | Anthraquinone compound with tobacco black shank resistance activity and preparation method and application thereof | |
| CN107501225B (en) | Flavonoid compound and preparation method and application thereof | |
| CN113173843B (en) | Tobacco black shank resistant active compound and preparation method and application thereof | |
| CN109438406B (en) | Chromone derivative extracted from anshansenia glauca and preparation method and application thereof | |
| CN107445934B (en) | Flavonoid compound and preparation method and application thereof | |
| CN110372707B (en) | Benzazepine alkaloid compound and preparation method and application thereof | |
| CN117185980A (en) | Azacyclic anthraquinone compound with tobacco mosaic virus resistance activity, and preparation method and application thereof | |
| CN111574492B (en) | Compound for resisting tobacco mosaic virus, preparation method and application thereof, and tobacco mosaic virus inhibitor containing compound | |
| CN115557960B (en) | Isopentenyl indole alkaloid compound and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |