Use of flavonoids G in a sophora flavescens extract in preparing a product for preventing and treating pathogenic bacteria in dairy cows

Abstract

The present invention provides a use of flavonoids G in a Sophora flavescens extract in preparing a product for preventing and treating pathogenic bacteria of dairy cows, and relates to the field of Sophora flavescens extracts. The use of flavonoids G in a Sophora flavescens extract in preparing a product for preventing and treating pathogenic bacteria of dairy cows, and the use of flavonoids G in preparing a product for preventing and treating related dairy cow diseases caused by pathogenic bacteria of dairy cows. The use of flavonoids G in a Sophora flavescens extract of the present invention in preparing a product for preventing and treating pathogenic bacteria of dairy cows, flavonoids G exhibits good antibacterial activity against pathogenic bacteria of dairy cows, Streptococcus agalactiae, Streptococcus dysgalactiae, Escherichia coli, and Staphylococcus aureus, and can effectively target related diseases caused by pathogenic bacteria of dairy cows, especially in preventing and treating mastitis of dairy cows caused by pathogenic bacteria; the flavonoids G has the advantages of being green and efficient, low toxicity, low residue, and not easy to produce resistance.

Description

Technical Field

[0001] The invention relates to the field of sophora flavescens extracts, and in particular to use of sophora flavescens flavonoids G in a sophora flavescens extract in preparing a product for preventing and treating pathogenic bacteria in dairy cows. Background Art

[0002] Cow mastitis has a high incidence rate and complex etiology, and is one of the major diseases affecting the development of the world's dairy industry. Nearly 50% of cow mastitis is caused by bacteria, among which Streptococcus agalactiae ( Streptococcus agalactiae ) and Streptococcus dysgalactiae ( Streptococcus dysgalactiae ) is the main bacterial pathogen causing mastitis in dairy cows. 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. At present, cow mastitis has caused huge economic losses to the dairy farming industry and dairy processing industry around the world. In addition, the food safety issues caused by it also seriously threaten human health.

[0003] 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.

[0004] Sophora flavescens is a plant of the genus Sophora in the subfamily Papilionoideae of the family Lenguinosae. It is one of the traditional Chinese medicines with a long history in my country. It was first seen in Shennong's Herbal Classic, which records that it has the effects of "removing qi stagnation, lumps, accumulations, and pain and swelling in the heart and abdomen". 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. Sophoraflavanone G is a relatively representative compound in Sophora flavescens and is one of the iconic components of Sophora flavescens.

[0005] According to existing literature reports, flavonoids G in Sophora flavescens extract has multiple biological activities such as anti-oxidation, anti-tumor, anti-inflammatory, anti-angiogenesis, immune response regulation, and anti-allergy. Specifically, studies have shown that the cytotoxicity of various human tumor cells can be inhibited by flavonoids G; at the same time, flavonoids G also has a high inhibitory activity on tyrosinase.

[0006] Therefore, the function of flavonoids G is further developed, the application field of flavonoids G is expanded, and a use of flavonoids G in a Sophora flavescens extract in the preparation of a product for preventing and treating pathogenic bacteria in dairy cows is provided, specifically the use of flavonoids G in preventing and / or treating related diseases caused by bacterial infection caused by pathogenic bacteria in dairy cows; in particular, the use of flavonoids G 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 flavonoids G in a Sophora flavescens extract in the preparation of a product for preventing and treating pathogenic bacteria of dairy cows, and the use of flavonoids G in preventing and / or treating related dairy cow diseases caused by bacterial infection caused by pathogenic bacteria of dairy cows; in particular, the use of flavonoids G 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:

[0009] Disclosed is a use of flavonoids G in a Sophora flavescens extract in preparing a product for preventing and treating pathogenic bacteria in dairy cows, and the flavonoids G is used in preparing a product for preventing and / or treating related dairy cow diseases caused by pathogenic bacteria in dairy cows.

[0010] The chemical structural formula of the sophoranone G is:

[0011] .

[0012] The dairy cattle pathogenic bacteria are at least one of the following: Streptococcus agalactiae Streptococcus agalactiae , Streptococcus dysgalactiae Streptococcus dysgalactiae , Escherichia coli Escherichia coli , Staphylococcus aureus Staphylococcus aureus .

[0013] The related dairy cow diseases include: dairy cow mastitis and bacterial infection.

[0014] Furthermore, the products include: pharmaceutical compositions and feed additives.

[0015] A pharmaceutical composition for treating bovine mastitis caused by bovine pathogenic bacteria uses the sophora flavanone G or its salts, tautomers, stereoisomers, and precursor compounds as active ingredients or main active ingredients, and is prepared into a pharmaceutically acceptable dosage form with a pharmaceutically acceptable excipient.

[0016] A feed additive for preventing bovine mastitis caused by bovine pathogenic bacteria uses the sophora flavanone G or its salts, tautomers, stereoisomers, and precursor compounds as effective ingredients or main effective ingredients, and is prepared into a nutritionally acceptable dosage form with nutritionally acceptable excipients.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] (1) The present invention uses the sophora flavanone G extracted and separated from Sophora flavescens to target the following pathogenic bacteria, including: Streptococcus agalactiae Streptococcus agalactiae ATCC12386, Streptococcus agalactiae Streptococcus agalactia ATCC27956, Streptococcus dysgalactiae Streptococcus dysgalactiae ATCC27957, Escherichia coli Escherichia coli ATCC25922 and Staphylococcus aureus Staphylococcus aureus Newman , and the inhibitory activity screening was carried out, and the results showed good antibacterial inhibitory activity, providing the use of flavonoids G 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 use of flavonoids G in Sophora flavescens extract in preventing and treating bovine mastitis caused by pathogenic bacteria of dairy cows, further expanding the application field of flavonoids G, and having broad application prospects.

[0019] (2) Sophora flavescens extract described in the present invention is used to prevent and treat bovine mastitis caused by pathogenic bacteria. 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 easy to produce resistance. It has great application value in preventing and treating bovine mastitis caused by pathogenic bacteria, and helps the subsequent research of related drugs.

[0020] (3) When the sophora flavescens extract of the present invention is used, the inhibition rate of sophora flavescens G against Streptococcus agalactiae ATCC12386 is 99.92% at a dosage concentration of 100 μg / mL, and the inhibition rate against Streptococcus agalactiae ATCC27956, Streptococcus dysgalactiae ATCC27957, and Staphylococcus aureus Newman can reach 100%; the inhibition rate against Escherichia coli ATCC25922 is 56.17%; when used in related products, it can inhibit the aforementioned dairy cow pathogens and achieve a preventive and / or therapeutic effect on related diseases caused by them; in particular, it is used in the prevention and treatment of dairy cow mastitis caused by the aforementioned dairy cow pathogens. DETAILED DESCRIPTION

[0021] 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.

[0022] This embodiment provides a use of flavonoids G in a Sophora flavescens extract in preparing a product for preventing and treating pathogenic bacteria in dairy cows, and the use of flavonoids G in preparing a product for preventing and / or treating related dairy cow diseases caused by pathogenic bacteria in dairy cows.

[0023] The chemical structural formula of the sophoranone G is:

[0024] .

[0025] The dairy cattle pathogenic bacteria are at least one of the following: Streptococcus agalactiae Streptococcus agalactiae , Streptococcus dysgalactiae Streptococcus dysgalactiae , Escherichia coli Escherichia coli , Staphylococcus aureus Staphylococcus aureus .

[0026] Furthermore, the related cow diseases include: cow mastitis and bacterial infection.

[0027] Furthermore, the products include: pharmaceutical compositions and feed additives.

[0028] This embodiment also provides a pharmaceutical composition for treating bovine mastitis caused by bovine pathogenic bacteria, which uses the sophora flavanone G or its salts, tautomers, stereoisomers, and precursor compounds as active ingredients or main active ingredients, and is prepared into a pharmaceutically acceptable dosage form with pharmaceutically acceptable excipients.

[0029] This embodiment also provides a feed additive for preventing bovine mastitis caused by bovine pathogenic bacteria, which uses the sophora flavanone G or its salts, tautomers, stereoisomers, and precursor compounds as active ingredients or main active ingredients, and is prepared into a nutritionally acceptable dosage form with nutritionally acceptable excipients.

[0030] Wherein, the aforementioned compound salt refers to the derivative of the sophora flavanone G; 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.

[0031] 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.

[0032] Example 1

[0033] The activity of flavonoids G in Sophora flavescens extract against the pathogenic bacteria of bovine mastitis is determined as follows:

[0034] 1) Test agents: Sophora flavanone G, Sophora flavescens (control), Sophora flavescens total flavonoids (control), Norfloxacin (control);

[0035] The chemical structural formula of the sophora flavanone G is as follows:

[0036] .

[0037] 2) Test strains:

[0038] Use the pathogenic bacteria of dairy cow mastitis, specifically, Streptococcus agalactiae Streptococcus agalactiae , deposit number ATCC12386; Streptococcus agalactiae Streptococcus agalactiae , deposit number ATCC27956; Streptococcus dysgalactiae Streptococcus dysgalactiae , deposit number ATCC27957; Escherichia coli Escherichia coli , deposit number ATCC25922; Staphylococcus aureus Staphylococcus aureus Newman .

[0039] 3) Test method:

[0040] 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.

[0041] 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.

[0042] 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.

[0043] Then, each strain in the logarithmic growth phase was diluted to 106 CFU / mL with the corresponding liquid culture medium for later use.

[0044] 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:

[0045] Corrected OD value = OD value of bacterial culture medium - OD value of sterile culture medium;

[0046] 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%.

[0047] 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:

[0048]

[0049] The above test results show that the flavonoids G in the Sophora flavescens extract of the present invention exhibits good antibacterial activity against all tested strains, especially against Streptococcus agalactiae. 、 Streptococcus dysgalactiae 、 Staphylococcus aureus showed high antibacterial activity.

[0050] Example 2

[0051] On the basis of Example 1, the drug-containing liquid culture medium of flavonoid G 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:

[0052]

[0053] In summary, the flavonoids G in the Sophora flavescens extract of the present invention showed good antibacterial activity against the tested strains; among them, against Streptococcus agalactiae 、 Streptococcus dysgalactiae 、 It showed excellent antibacterial activity against Staphylococcus aureus, with MIC values ​​ranging from 1.56 to 3.12 μg / mL.

[0054] It can be seen that the flavonoids G of the Sophora flavescens extract of the present invention can effectively inhibit the pathogenic bacteria of bovine mastitis, and flavonoids G or its salts, tautomers, stereoisomers, and precursor compounds can be used as effective ingredients or main effective ingredients 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.

[0055] Unless otherwise specified, all percentages used in the present invention are by mass.

[0056] 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 flavonoids G in a Sophora flavescens extract in the preparation of a product for preventing and treating pathogenic bacteria in dairy cows, characterized in that: Use of the sophora flavanone G in preparing a product for preventing and / or treating relevant cow diseases caused by cow pathogenic bacteria; The chemical structural formula of the sophoranone G 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, bacterial infection; The products include: pharmaceutical compositions, feed additives; The pharmaceutical composition uses flavonoids G in the Sophora flavescens extract as an effective ingredient or a main effective ingredient, and is prepared into a pharmaceutically acceptable dosage form with a pharmaceutically acceptable excipient; The feed additive uses flavonoids G in the 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.

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