CN113499337A - Application of kurarinone in preparation of aquatic pathogen resisting medicine or feed additive - Google Patents
Application of kurarinone in preparation of aquatic pathogen resisting medicine or feed additive Download PDFInfo
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- CN113499337A CN113499337A CN202110919995.1A CN202110919995A CN113499337A CN 113499337 A CN113499337 A CN 113499337A CN 202110919995 A CN202110919995 A CN 202110919995A CN 113499337 A CN113499337 A CN 113499337A
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- kurarinone
- aquatic
- preparation
- feed additive
- vibrio
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/116—Heterocyclic compounds
- A23K20/121—Heterocyclic compounds containing oxygen or sulfur as hetero atom
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/489—Sophora, e.g. necklacepod or mamani
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/04—Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
- Y02A40/818—Alternative feeds for fish, e.g. in aquacultures
Abstract
The invention discloses an application of Kurarinone in preparing drugs or feed additives for resisting aquatic pathogenic bacteria, in particular to an application in preventing or treating diseases caused by aquatic pathogenic bacteria, especially in preparing vibrio harveyi (Vibrio harveyi) for inhibiting aquatic pathogenVibrio harveyi) Vibrio parahaemolyticus: (Vibrio parahaemolyticus) Aeromonas hydrophila (b) ((b))Aeromonas hydrophila) And Edwardsiella tarda: (Edwardsiella tarda) The invention has good inhibition effect on aquatic pathogen Vibrio harveyi, Vibrio parahaemolyticus, Aeromonas hydrophila and Edwardsiella tarda through biological activity test, and the minimum inhibitory concentration is in the range of 0.0625-0.125 mg/mL, which shows that the kushenone can be used as an antibiotic substitute for preventing or treating aquatic pathogen infection.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to application of kurarinone in preparation of an anti-aquatic pathogen medicament or a feed additive.
Background
In recent thirty years, the aquatic product aquaculture industry in China develops rapidly, the yield of aquatic products is at the top of the world, and with the rapid development of the intensive aquaculture industry, many problems related to aquatic animal diseases and water body environmental pollution are gradually exposed, so that the stable development of the aquaculture industry is severely restricted. The main pathogens of aquatic animals include bacteria, viruses, parasites and the like, and the annual loss of China caused by aquatic animal diseases is estimated to exceed 100 billion yuan. Wherein, the disease loss of the aquatic animals caused by bacterial diseases accounts for 58 percent, and is the most serious factor causing the economic loss of aquaculture; the pathogeny of bacterial diseases in aquaculture mainly comprises vibrio harveyi, vibrio parahaemolyticus, aeromonas hydrophila, edwardsiella tarda and the like, at present, antibacterial drugs applied to aquaculture are mainly transplanted from medical and veterinary antibiotics, the types of special antibacterial raw material drugs for aquaculture are few, the drug resistance of pathogenic bacteria is continuously increased due to long-term application or abuse of the antibiotics in aquaculture, the curative effect of the antibiotics is more and more unsatisfactory, and in addition, the antibiotics remained in the aquaculture also pose a certain threat to the health and safety of human beings. Along with the gradual understanding of negative effects of antibiotics and food safetyThe importance of the whole is increasing day by day, and the development of 'green' antibiotic substitutes becomes urgent; the Chinese herbal medicine resources have the excellent characteristics of abundant, cheap and easily obtained Chinese herbal medicine resources, extremely small toxicity, few side effects, relatively few residues and almost no pollution, so that the Chinese herbal medicine resources have great potential and wide application prospect in the aspect of developing green and environment-friendly antibiotic substitutes; the Chinese herbal medicine radix Sophorae Flavescentis is Leguminosae plant radix Sophorae Flavescentis (Sophora flavescenesAit), the compound species of which are rich, including alkaloid, flavone, triterpenoid saponin, lignin, phenolic acid and the like, and the lightyellow sophora root which is used as a traditional Chinese medicinal material has the medicinal history of thousands of years, is widely used clinically, and has the effects of resisting tumor, virus, pathogenic microorganism, oxidation, inflammation and the like; at present, the sophora flavescens is approved by the ministry of agriculture of China to be used for preventing and treating aquatic animal diseases, but the antibacterial active substance of the sophora flavescens is not clear and the whole curative effect is unstable, so that the identification of the active molecule of the sophora flavescens resisting aquatic pathogenic bacteria has important significance for developing green and environment-friendly antibiotic substitutes.
Disclosure of Invention
The invention discloses an application of Kurarinone in preparing an anti-aquatic pathogen medicament or a feed additive, wherein the Kurarinone has the following structural formula:
further, the kushenone has activity against aquatic pathogens Vibrio harveyi, Vibrio parahaemolyticus, Aeromonas hydrophila single cell and Edwardsiella tarda.
Further, the kushenone is applied to vibriosis, hemorrhagic disease and Edwardsiellosis disease caused by aquatic pathogenic bacteria.
Further, the kurarinone can be prepared by various methods or modified methods known in the art, such as separation, extraction, synthesis or semi-synthesis from plants, or can be directly obtained commercially.
Further, the kurarinone is obtained by extracting and separating kurarinone from radix sophorae flavescentis, and the preparation method of the kurarinone comprises the following steps:
(1) mixing anhydrous ethanol and radix Sophorae Flavescentis at a liquid-to-material ratio of 30:1 mL/g, heating and extracting at 79 deg.C for 146 min to obtain a mixed solution;
(2) carrying out suction filtration separation on the mixed solution, removing residues and reserving an extracting solution;
(3) evaporating the extractive solution under reduced pressure to recover solvent to obtain solid radix Sophorae Flavescentis extract;
(4) using Agilent 1260 high performance liquid chromatograph, using CLC-ODS (10.0 mm × 250 mm, 5 μm, 1.5 mL/min) as chromatographic column, and 295 nm as detection wavelength; the mobile phase is 0.1 percent of phosphoric acid water and methanol; gradient elution procedure (v/v) 0 min 30% methanol, 20 min 80% methanol, 45 min 90% methanol, 60 min 100% methanol; collecting the eluting solution of kurarinone peak, concentrating under reduced pressure, and drying to obtain kurarinone.
Compared with the prior art, the invention has the following advantages:
biological activity tests prove that the kushenone has good inhibition effect on aquatic pathogen Vibrio harveyi, Vibrio parahaemolyticus, Aeromonas hydrophila and Edwardsiella tarda, and the minimum inhibitory concentrations are 0.0625 mg/mL, 0.125 mg/mL and 0.0625 mg/mL respectively, which shows that the kushenone can be used as an antibiotic substitute for preventing or treating aquatic pathogen infection.
Drawings
FIG. 1 is a high performance liquid chromatogram of a kurarinone standard (a).
FIG. 2 is a high performance liquid chromatogram of Sophora flavescens Aiton extract (b).
Detailed Description
The invention is described in detail below with reference to the accompanying drawings: the application of Kurarinone in the preparation of medicines or feed additives for resisting aquatic pathogenic bacteria is disclosed, wherein the Kurarinone has the following structural formula:
preferably, the kushenone has activity against aquatic pathogens Vibrio harveyi, Vibrio parahaemolyticus, Aeromonas hydrophila, and Edwardsiella tarda.
As a preferred scheme, the kushenone is applied to vibriosis, hemorrhagic disease and Edwardsiellosis disease caused by aquatic pathogenic bacteria.
Preferably, the kurarinone can be prepared by various methods or variations known in the art from plants by isolation, extraction, synthesis or semisynthesis, or can be obtained commercially directly.
Preferably, the kurarinone is extracted and separated from radix sophorae flavescentis, and the preparation method of the kurarinone comprises the following steps:
(1) mixing anhydrous ethanol and radix Sophorae Flavescentis at a liquid-to-material ratio of 30:1 mL/g, heating and extracting at 79 deg.C for 146 min to obtain a mixed solution;
(2) carrying out suction filtration separation on the mixed solution, removing residues and reserving an extracting solution;
(3) evaporating the extractive solution under reduced pressure to recover solvent to obtain solid radix Sophorae Flavescentis extract;
(4) using Agilent 1260 high performance liquid chromatograph, using CLC-ODS (10.0 mm × 250 mm, 5 μm, 1.5 mL/min) as chromatographic column, and 295 nm as detection wavelength; the mobile phase is 0.1 percent of phosphoric acid water and methanol; gradient elution procedure (v/v) 0 min 30% methanol, 20 min 80% methanol, 45 min 90% methanol, 60 min 100% methanol; collecting the eluting solution of kurarinone peak, concentrating under reduced pressure, and drying to obtain kurarinone.
Example 1: preparing a sophora flavescens extract: taking a certain amount of Chinese medicinal material radix Sophorae Flavescentis, taking absolute ethanol as solvent, mixing absolute ethanol and radix Sophorae Flavescentis according to liquid-material ratio of 30:1 mL/g, heating and extracting at 79 deg.C for 146 min, and preparing mixed solution; carrying out suction filtration separation on the mixed solution, removing residues and reserving an extracting solution; evaporating the extractive solution under reduced pressure to recover solvent to obtain solid radix Sophorae Flavescentis extract.
Example 2: identification and preparation of kurarinone in sophora flavescens extracts: adopting kurarinone standard substance as reference compound, and comparing chromatogram of the reference compound and radix Sophorae Flavescentis extract by High Performance Liquid Chromatography (HPLC) to identify and prepare kurarinone. An Agilent 1260 high performance liquid chromatograph with CLC-ODS (10.0 mm. times.250 mm, 5 μm, 1.5 mL/min) column and 295 nm detection wavelength was used. The mobile phase is 0.1 percent of phosphoric acid water and methanol; the gradient elution procedure was as follows (v/v) 0 min 30% methanol, 20 min 80% methanol, 45 min 90% methanol, 60 min 100% methanol.
Referring to FIGS. 1-2, FIG. 1 is an HPLC chart of kurarinone, its retention time (t)R) 45.097 min, FIG. 2 shows HPLC chart of radix Sophorae Flavescentis extract containing various chemical components including kurarinone under the same conditions, and retention time (t) in radix Sophorae Flavescentis extract is identified by comparing absorption spectra of chromatographic peaksR) The peak at 44.239 min is kurarinone.
Repeatedly collecting eluting solution of kurarinone peak, concentrating under reduced pressure, and drying to obtain kurarinone.
Example 3: and (3) determining the activity of the kurarinone against aquatic pathogenic bacteria: determining the activity of the sophora flavescens extract and the kurarinone against aquatic pathogenic bacteria by adopting an oxford cup method: dissolving radix Sophorae Flavescentis extract and kurarinone in methanol respectively, and preparing into sample solutions with series concentrations by two-fold dilution method; the test strains are Vibrio harveyi, Vibrio parahaemolyticus, Aeromonas hydrophila and Edwardsiella tarda; 20 mL of MH broth was poured into the plate, and after coagulation, 100. mu.L of each of the 4 bacterial suspensions (1X 10)6cfu/mL) was spread evenly on the medium; the Oxford cup is placed on the culture medium, 200 mu L of sample solution is added into the Oxford cup, the mixture is cultured for 24 h at 37 ℃, and the diameter of the inhibition zone is measured. The lowest concentration of the compound that produced an inhibitory zone was defined as the Minimum Inhibitory Concentration (MIC) as shown in table 1 below:
TABLE 1 Minimum Inhibitory Concentration (MIC) of Sophora flavescens extract and kurarinone against aquatic pathogens
Through biological activity tests, the sophora flavescens extract and the kurarinone thereof have good inhibition effects on aquatic pathogens vibrio harveyi, vibrio parahaemolyticus, aeromonas hydrophila and edwardsiella tarda, wherein the minimum inhibitory concentrations of the kurarinone on the vibrio harveyi, the vibrio parahaemolyticus, the aeromonas hydrophila and the edwardsiella tarda are respectively 0.0625 mg/mL, 0.125 mg/mL and 0.0625 mg/mL, which indicates that the kurarinone can be used as an antibiotic substitute for preventing or treating aquatic pathogen infection.
The above examples of the present invention are merely examples for clearly illustrating the present invention and are not intended to limit the embodiments of the present invention. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. Not all embodiments are exhaustive. All obvious changes or modifications of the technical solution of the present invention are within the protection scope of the present invention.
Claims (5)
1. The application of the kurarinone in the preparation of the medicament or the feed additive for resisting aquatic pathogenic bacteria is characterized in that: the kurarinone has the following structural formula:
2. the use of kurarinone according to claim 1 in the preparation of a medicament or feed additive against aquatic pathogens, wherein: the kushenone has activity against aquatic pathogens Vibrio harveyi, Vibrio parahaemolyticus, Aeromonas hydrophila unicell and Edwardsiella tarda.
3. The use of kurarinone according to claim 2 in the preparation of a medicament or feed additive against aquatic pathogens, characterized in that: the kushenone can be used for treating vibriosis, hemorrhagic disease and Edwardsiellosis caused by aquatic pathogenic bacteria.
4. The use of kurarinone according to claim 2 in the preparation of a medicament or feed additive against aquatic pathogens, characterized in that: the kurarinone is obtained by extraction and separation from plants or a synthetic or semi-synthetic method.
5. The use of kurarinone according to claim 4 in the preparation of a medicament or feed additive against aquatic pathogens, wherein: the kurarinone is obtained by extracting and separating kurarinone from radix sophorae flavescentis.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110919995.1A CN113499337B (en) | 2021-08-11 | 2021-08-11 | Application of kurarinone in preparation of aquatic pathogenic bacteria resistant medicines or feed additives |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202110919995.1A CN113499337B (en) | 2021-08-11 | 2021-08-11 | Application of kurarinone in preparation of aquatic pathogenic bacteria resistant medicines or feed additives |
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| CN113499337A true CN113499337A (en) | 2021-10-15 |
| CN113499337B CN113499337B (en) | 2023-04-28 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115785049A (en) * | 2022-12-17 | 2023-03-14 | 山东方舟生物科技有限公司 | Extraction method of kurarinone and application of kurarinone in resisting crop pathogenic fungi |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102125062A (en) * | 2011-03-28 | 2011-07-20 | 山东大学威海分校 | Compound lightyellow sophora root preparation and application thereof |
| CN105249025A (en) * | 2015-11-13 | 2016-01-20 | 广西壮族自治区中国科学院广西植物研究所 | Application of sophora flavescens extract or sophora flavescens ultra-fine powder in preparing fodder additives |
| WO2019146826A1 (en) * | 2018-01-29 | 2019-08-01 | 전진바이오팜 주식회사 | Composition for preventing or treating parasitic infections in fish, comprising sophora flavescens aiton extract or fraction thereof |
-
2021
- 2021-08-11 CN CN202110919995.1A patent/CN113499337B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102125062A (en) * | 2011-03-28 | 2011-07-20 | 山东大学威海分校 | Compound lightyellow sophora root preparation and application thereof |
| CN105249025A (en) * | 2015-11-13 | 2016-01-20 | 广西壮族自治区中国科学院广西植物研究所 | Application of sophora flavescens extract or sophora flavescens ultra-fine powder in preparing fodder additives |
| WO2019146826A1 (en) * | 2018-01-29 | 2019-08-01 | 전진바이오팜 주식회사 | Composition for preventing or treating parasitic infections in fish, comprising sophora flavescens aiton extract or fraction thereof |
Non-Patent Citations (2)
| Title |
|---|
| 薛飞等: "苦参生物碱及其复方制剂对嗜水气单胞菌的抑制作用" * |
| 黄琦等: "苦参生物碱和黄酮体外抑菌活性比较" * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115785049A (en) * | 2022-12-17 | 2023-03-14 | 山东方舟生物科技有限公司 | Extraction method of kurarinone and application of kurarinone in resisting crop pathogenic fungi |
| CN115785049B (en) * | 2022-12-17 | 2024-01-09 | 山东方舟生物科技有限公司 | Extraction method of kurarinone and application of kurarinone in resisting crop pathogenic fungi |
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| CN113499337B (en) | 2023-04-28 |
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