Application of isoxanthohumol in sophora flavescens extract in preparation of product for preventing and treating pathogenic bacteria of dairy cow
By using isochlorohumol in the extract of Sophora ginseng, a pharmaceutical composition or feed additive for the treatment of mastitis of dairy cow is prepared, which solves the problem of antibiotic resistance in the prior art and realizes pathogenic bacteria of dairy cow mastitis Effective inhibition and prevention.
Patent Information
- Application Number
- CN202411594543.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-06-27
- Publication Date
- 2025-02-14
AI Technical Summary
In the prior art, the treatment of dairy cow mastitis mainly relies on antibiotics, but due to the spread of drug resistance mechanisms, finding safe and effective alternative antibiotics has become an urgent problem.
Isochohumphenol in Sophora ginseng extract is used as an active ingredient to prepare a pharmaceutical composition or feed additive for the prevention and treatment of dairy cow mastitis caused by dairy cow pathogenic bacteria.
Isochohumol has good antibacterial inhibitory activity against pathogenic bacteria such as Streptococcus alactis and Streptococcus alopecia, and has the advantages of green, high efficiency, low toxicity, low residue and low resistance, and is effective in preventing and treating dairy cow mastitis.
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Abstract
Description
[0001] The present invention is a divisional application of application number 202410840476X, application date June 27, 2024, and invention name "Use of isopentenyl flavonoid compounds in a Sophora flavescens extract in the preparation of products for preventing and treating pathogenic bacteria in dairy cows". Technical Field
[0002] The invention relates to the field of sophora flavescens extracts, and in particular to use of isoxanthohumol in the sophora flavescens extracts in preparing products for preventing and treating pathogenic bacteria in dairy cows. Background Art
[0003] Cow mastitis has a high incidence rate and complex etiology, and is one of the main diseases that affect the development of the world's dairy industry. Nearly 50% of cow mastitis is caused by bacteria, of which Streptococcus agalactiae and Streptococcus dysgalactiae are the main bacterial pathogens that cause cow mastitis. Cow mastitis seriously endangers the health of dairy cows, directly leading to a decrease in milk production, affecting the quality and nutritional content of milk. Cow mastitis has caused huge economic losses to the dairy farming industry and dairy processing industry around the world. In addition, the resulting food safety issues also seriously threaten human health.
[0004] In the prior art, cow mastitis is mainly treated with antibiotics during the infection period. However, bacterial resistance has become an increasingly serious threat as resistance mechanisms are spreading globally. Therefore, finding safe and effective alternative antibiotics to maintain the health of cow mammary glands and using compounds from natural products with strong antibacterial effects and environmental stability to prevent and treat cow mastitis is an important topic in today's green prevention and control of cow mastitis.
[0005] Sophora flavescens is a plant of the genus Sophora in the subfamily Papilionoideae of the family Lenguinosae. Modern medical research has found that Sophora flavescens has anti-inflammatory, antiviral, and anti-tumor effects, and can be used to treat viral hepatitis, arrhythmia, viral myocarditis and other diseases. Sophora flavescens has a relatively rich chemical composition. At present, more than 200 compounds have been isolated from Sophora flavescens, among which the main components have been identified as flavonoids and alkaloids. Flavonoids are a relatively representative class of compounds in Sophora flavescens extracts. Its flavonoids substituted with isopentenyl at the C-8 or C-6 position are one of the taxonomically characteristic components of Sophora flavescens extracts. Among them, isoxanthohumol belongs to the flavonoids in Sophora flavescens extracts. At the same time, according to existing literature reports, flavonoids in Sophora flavescens extracts have anti-inflammatory, anti-allergic, anti-viral, anti-osteoporosis, antioxidant and other activities, with great application potential and value for further research and development.
[0006] Thus, the invention provides a use of isoxanthohumol in a Sophora flavescens extract in the preparation of a product for preventing and treating pathogenic bacteria in dairy cows, and the use of isoxanthohumol in the prevention and / or treatment of related diseases caused by bacterial infection caused by pathogenic bacteria in dairy cows; in particular, the new use of isoxanthohumol in a Sophora flavescens extract in preventing and treating mastitis in dairy cows caused by pathogenic bacteria has important technical significance and value. Summary of the invention
[0007] In order to solve the technical problems existing in the prior art, the present invention provides a use of isoxanthohumol in a Sophora flavescens extract in the preparation of a product for preventing and treating pathogenic bacteria of dairy cows, and the use of the isoxanthohumol in preventing and / or treating related dairy cow diseases caused by bacterial infection caused by pathogenic bacteria of dairy cows; in particular, a new use of isoxanthohumol in a Sophora flavescens extract in preventing and treating cow mastitis caused by pathogenic bacteria of dairy cows.
[0008] In order to solve the above technical problems, the technical solution adopted by the present invention is as follows: Use of isoxanthohumol in a Sophora flavescens extract in preparing a product for preventing and treating pathogenic bacteria in dairy cows, and use of the isoxanthohumol in preparing a product for preventing and / or treating relevant dairy cow diseases caused by pathogenic bacteria in dairy cows; The chemical structural formula of the isoxanthohumol is: ; The dairy cattle pathogenic bacteria are at least one of the following: Streptococcus agalactiae Streptococcus agalactiae , Streptococcus dysgalactiae Streptococcus dysgalactiae ; The related dairy cow diseases include: dairy cow mastitis and bacterial infection.
[0009] Furthermore, the products include: pharmaceutical compositions and feed additives.
[0010] A pharmaceutical composition for treating bovine mastitis caused by bovine pathogenic bacteria uses isoxanthohumol or its salt in the aforementioned Sophora flavescens extract as an effective component or a main effective component and is prepared into a pharmaceutically acceptable dosage form with a pharmaceutically acceptable excipient.
[0011] A feed additive for preventing bovine mastitis caused by bovine pathogenic bacteria uses isoxanthohumol or its salt in the aforementioned Sophora flavescens extract as an effective component or a main effective component, and is prepared into a nutritionally acceptable dosage form with a nutritionally acceptable excipient.
[0012] Compared with the prior art, the present invention has the following beneficial effects: (1) The isoxanthohumol extracted and separated from Sophora flavescens is used in the present invention to target the following pathogenic bacteria, including: Streptococcus agalactiae Streptococcus agalactiae ATCC12386, Streptococcus agalactiae Streptococcus agalactiaATCC27956, Streptococcus dysgalactiae Streptococcus dysgalactiae ATCC27957, and the results showed good antibacterial inhibition activity, providing the use of isoxanthohumol in Sophora flavescens extract in preventing and / or treating related diseases caused by bacterial infection caused by pathogenic bacteria of dairy cows; in particular, the new use of isoxanthohumol in Sophora flavescens extract in preventing and treating bovine mastitis caused by pathogenic bacteria of dairy cows has broad application prospects.
[0013] (2) The isoxanthohumol in the Sophora flavescens extract of the present invention is used to prevent and treat bovine mastitis caused by pathogenic bacteria. This is because the isoxanthohumol in the Sophora flavescens extract is a natural plant-derived antibiotic obtained from common natural plants, and has the advantages of being green, highly efficient, low toxicity, low residue, and not prone to resistance. It has great application value in preventing and treating bovine mastitis caused by pathogenic bacteria, and helps the subsequent research of related drugs.
[0014] (3) The isoxanthohumol in the Sophora flavescens extract of the present invention, at a dosage concentration of 100 μg / mL, can achieve an inhibition rate of 100% against Streptococcus agalactiae ATCC12386; an inhibition rate of 99.2% against Streptococcus agalactiae ATCC27956; and an inhibition rate of 98.28% against Streptococcus dysgalactiae ATCC27957. DETAILED DESCRIPTION
[0015] In order to have a clearer understanding of the technical features, purposes and effects of the present invention, the specific implementation methods of the present invention are now described.
[0016] This embodiment provides a use of isoxanthohumol in a Sophora flavescens extract in preparing a product for preventing and treating pathogenic bacteria in dairy cows, and the use of the isoxanthohumol in preparing a product for preventing and / or treating related dairy cow diseases caused by pathogenic bacteria in dairy cows.
[0017] The chemical structural formula of the isoxanthohumol is: ; Furthermore, the dairy cow pathogenic bacteria is at least one of the following: Streptococcus agalactiae and Streptococcus dysgalactiae.
[0018] Furthermore, the related cow diseases include: cow mastitis and bacterial infection.
[0019] Furthermore, the products include: pharmaceutical compositions and feed additives.
[0020] This embodiment also provides a pharmaceutical composition for treating bovine mastitis caused by bovine pathogenic bacteria, which uses the aforementioned isoxanthohumol or its salt as an effective ingredient or a main effective ingredient and is prepared into a pharmaceutically acceptable dosage form with a pharmaceutically acceptable excipient.
[0021] This embodiment also provides a feed additive for preventing bovine mastitis caused by bovine pathogenic bacteria, which uses the aforementioned isoxanthohumol or its salt as an effective ingredient or a main effective ingredient, and is prepared into a nutritionally acceptable dosage form with a nutritionally acceptable excipient.
[0022] Wherein, the aforementioned compound salt refers to the aforementioned derivative of isoxanthohumol; the parent compound is modified by making an acid salt or an alkaline salt of the parent compound. Examples of compound salts include, but are not limited to: inorganic acid salts or organic acid salts of basic residues (such as amine groups); alkaline salts or organic salts of acidic residues (such as carboxylic acids, etc.). The compound salts of the present invention can be synthesized from a parent compound containing a basic part or an acidic part by conventional chemical methods. In addition, the aforementioned compound salts can be prepared in situ during the final separation and purification of the compound.
[0023] The aforementioned excipients refer to any excipients, solvents, dispersion media, absorption retardants, diluents or adjuvants that do not cause secondary reactions such as allergic reactions in humans or animals; such as preservatives or antioxidants, fillers, binders, disintegrants, wetting agents, emulsifiers, suspending agents, solvents, dispersion media, coating agents, antibacterial agents, isotonic agents and absorption delay agents. Examples of typical excipients include, but are not limited to, mannitol, lactose, magnesium stearate, sodium saccharide, talc, cellulose, cross-linked sodium carboxymethyl cellulose, glucose, gelatin, starch, lactose, dicalcium phosphate, sucrose, kaolin, magnesium carbonate, wetting agents, emulsifiers, solubilizers, sterile water, physiological saline, pH buffers, nonionic surfactants, lubricants, stabilizers, binders and edible oils (such as peanut oil, sesame oil, etc.). At the same time, the use of the aforementioned excipients in the pharmaceutical composition or feed additives is foreseeable.
[0024] Example 1 The activity of isoxanthohumol in Sophora flavescens extract against the pathogenic bacteria of bovine mastitis is determined as follows: 1) Test agents: isoxanthohumol (K3), total flavonoids from Sophora flavescens (control), and norfloxacin (control); The chemical structure of isoxanthohumol is as follows: ; 2) Test bacteria: Pathogenic bacteria of bovine mastitis were used, specifically, Streptococcus agalactiae, deposited with number ATCC12386; Streptococcus agalactiae, deposited with number ATCC27956; Streptococcus dysgalactiae, deposited with number ATCC27957; Escherichia coli, deposited with number ATCC25922; Staphylococcus aureus Newman.
[0025] 3) Test method: The strain used in the test experiment of this embodiment is a strain frozen in 30% glycerol at a temperature of -80°C in the laboratory.
[0026] The frozen strains of Streptococcus agalactiae and Streptococcus dysgalactiae were taken out respectively, streaked on sheep blood agar plates respectively, and cultured at a constant temperature of 37°C until single colonies grew; single colonies were picked out and added to brain heart infusion broth (BHI) liquid culture medium (tryptone: 10.0 g, beef heart extract powder: 17.5 g, sodium chloride: 5.0 g, glucose: 2.0 g, disodium hydrogen phosphate (12H2O): 2.5 g, distilled water: 1 L; sterilized at 121°C for 20 min), and cultured in a constant temperature shaker at 37°C and 140 rpm until the logarithmic growth phase.
[0027] Similarly, the frozen strains of Escherichia coli and Staphylococcus aureus were taken out, streaked on MH solid culture medium (beef powder: 2.0 g, soluble starch: 1.5 g, acid hydrolyzed casein: 17.5 g, agar: 20 g, distilled water: 1 L; sterilized at 121°C for 20 min), and cultured at 37°C until single colonies grew; single colonies were picked up and transferred to MH broth culture medium (beef powder: 2.0 g, soluble starch: 1.5 g, acid hydrolyzed casein: 17.5 g, distilled water: 1 L; sterilized at 121°C for 20 min) liquid culture medium, and cultured in a constant temperature shaker at 37°C and 140 rpm until the logarithmic growth phase.
[0028] Then, each strain in the logarithmic growth phase was diluted to 106 CFU / mL with the corresponding liquid culture medium for later use.
[0029] Dissolve each of the above-mentioned test drugs in DMSO, add them to the liquid culture medium, mix them evenly, and prepare a drug-containing liquid culture medium with a concentration of 200μg / mL. Take 50μL of drug-containing culture medium and the same volume of bacterial culture containing 106CFU / mL and add them to the wells of the 96-well plate to control the final drug concentration to 100μg / mL. And use 100μL of bacterial solution of the same concentration containing an equal amount of DMSO as a control. Incubate the 96-well plate in a 37℃ constant temperature incubator for 12-24h until the bacterial solution in the control group grows, and measure the OD value (OD600) of the bacterial solution in the well on the microplate reader. In addition, measure the OD value of 100μL of liquid culture medium and the drug with a concentration of 100μg / mL, and correct the OD value caused by the culture medium and the drug itself. The calculation formula for corrected OD value and inhibition rate is as follows: Corrected OD value = OD value of bacteria-containing culture medium - OD value of sterile culture medium; Inhibition rate = (corrected OD value of bacterial solution in control culture medium - corrected OD value of drug-containing culture medium) / corrected OD value of bacterial solution in control culture medium × 100%.
[0030] All the above experiments were repeated three times to determine the inhibition rate of each test agent on each bovine mastitis pathogen. The specific test results are shown in the following table:
[0031] It can be seen from the above test results that the isoxanthohumol in the Sophora flavescens extract of the present invention exhibits high antibacterial activity against the tested strains Streptococcus agalactiae ATCC12386, Streptococcus agalactiae ATCC27956, and Streptococcus dysgalactiae ATCC27957.
[0032] Example 2 On the basis of Example 1, the drug-containing liquid culture medium of isoxanthohumol was diluted in a 96-well plate by a two-fold dilution method to obtain 50 μL of drug-containing culture medium of a series of concentrations, and then the inhibition rate corresponding to the series of concentrations was determined according to the same test method in Example 1; the lowest concentration with an inhibition rate greater than 90% was defined as MIC, and the MIC value of each test agent for each bovine mastitis pathogen was determined. The specific measurement results are shown in the following table:
[0033] In summary, the isoxanthohumol in the Sophora flavescens extract of the present invention exhibits good bactericidal activity against Streptococcus agalactiae ATCC12386, Streptococcus agalactiae ATCC27956, and Streptococcus dysgalactiae ATCC27957.
[0034] It can be seen that the isoxanthohumol in the Sophora flavescens extract involved in the present invention can effectively inhibit the pathogenic bacteria of bovine mastitis, and isoxanthohumol or its salt can be used as an effective ingredient or a main effective ingredient to prepare a pharmaceutical composition or feed additive for preventing and treating diseases such as bovine mastitis or bacterial infection caused by the above-mentioned bovine pathogenic bacteria.
[0035] Unless otherwise specified, all percentages used in the present invention are by mass.
[0036] Finally, it should be noted that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.
Claims
1. A use of isoxanthohumol in a Sophora flavescens extract in preparing a product for preventing and treating pathogenic bacteria in dairy cows, characterized in that: Use of the isoxanthohumol in preparing a product for preventing and / or treating relevant cow diseases caused by cow pathogenic bacteria; The chemical structural formula of the isoxanthohumol is: ； The dairy cattle pathogenic bacteria are at least one of the following: Streptococcus agalactiae Streptococcus agalactiae , Streptococcus dysgalactiae Streptococcus dysgalactiae ; The related dairy cow diseases include: dairy cow mastitis and bacterial infection.
2. The use of isoxanthohumol in the Sophora flavescens extract according to claim 1 in preparing a product for preventing and treating pathogenic bacteria in dairy cows, characterized in that: The products include: pharmaceutical compositions and feed additives.
3. A pharmaceutical composition for treating bovine mastitis caused by bovine pathogenic bacteria, characterized in that: The isoxanthohumol or its salt in the Sophora flavescens extract according to claim 1 is used as an effective ingredient or a main effective ingredient, and is prepared into a pharmaceutically acceptable dosage form with a pharmaceutically acceptable excipient.
4. A feed additive for preventing cow mastitis caused by cow pathogenic bacteria, characterized in that: The isoxanthohumol or its salt in the Sophora flavescens extract according to claim 1 is used as an effective ingredient or a main effective ingredient, and is prepared into a nutritionally acceptable dosage form with a nutritionally acceptable excipient.