Application of feijoine derivative

Abstract

The invention relates to the technical field of veterinary drugs, and particularly discloses application of a fiji alkali derivative. The invention provides application of a feijoine derivative in preparation of a medicine for treating cow mastitis, the feijoine derivative belongs to a derivative of a natural traditional Chinese medicine, a pentacyclic mother nucleus of the feijoine is reserved, different substituent groups are introduced to the mother nucleus, and a pharmaceutical preliminary test proves that the feijoine derivative has a good application prospect in preparation of a medicine for treating cow mastitis. The compound has the characteristics of stable chemical property, strong antibacterial activity, wide antibacterial spectrum, high bioavailability, low residue, no drug resistance and small toxic and side effects, and is expected to be used as a new veterinary drug for treating the cow mastitis. The dairy cow mammary gland injectant provided by the invention is a novel preparation modified by natural effective components, and compared with antibiotics, the dairy cow mammary gland injectant is not easy to generate drug resistance, fast in-vivo metabolism, high in efficiency, low in toxicity, free of residues, definite in curative effect, safe and reliable, and can replace antibiotic drugs to prevent and treat dairy cow mastitis.

Description

Technical Field

[0001] The invention relates to the technical field of veterinary drugs, and in particular to the application of a fijiline derivative. Background Art

[0002] With the continuous improvement of the scale of animal husbandry production, the demand for highly effective, low-toxic and low-residue animal drugs is increasing rapidly.

[0003] Cow mastitis is an inflammation of the cow's breast caused by chemical, physical, microbial and other factors. It can be divided into clinical, subclinical and nonclinical mastitis, and is a common disease in dairy cows. At present, antibiotics are the first choice for the treatment of cow mastitis, mainly through injection of antibacterial drugs and breast perfusion. However, it is not effective for mastitis caused by environmental pathogens such as Escherichia coli. There are also problems such as insufficient antibacterial spectrum of a single drug, drug resistance, accumulation of drug residues, and intestinal flora imbalance.

[0004] The Ireland research group at the University of Utah and the Clardy research group at Cornell University jointly discovered a red active alkaloid Fascaplysin (whose structural formula is shown in Formula 1) extracted and separated from a sponge. This compound has multiple biological activities, including anti-tumor, anti-bacterial, anti-fungal, anti-viral, anti-malarial, and cholinesterase inhibition, and its three-dimensional structure is a planar five-membered ring. Preclinical pharmacology studies have shown that Fascaplysin can be used as a CDK4 inhibitor. Although it has a significant effect in inhibiting tumor cells, Fascaplysin is not very selective in anti-cancer effects. It will be very difficult to use this type of compound as an anti-cancer drug candidate, so it is very important to find a new use for this type of drug.

[0005]

[0006] Formula 1. Summary of the invention

[0007] In view of the above problems, the present invention provides an application of a fijianine derivative, which has obvious antibacterial effect and can be used as a new veterinary drug for treating bovine mastitis.

[0008] In order to solve the above technical problems, the present invention adopts the following technical solutions: In a first aspect, the present invention provides a use of a fijianine derivative in the preparation of a drug for treating bovine mastitis, wherein the structural formula of the fijianine derivative is shown in Formula I:

[0009] Formula I.

[0010] Compared with the prior art, the application of the fijianine derivatives provided by the present invention, the fijianine derivatives shown in Formula I are derivatives of natural traditional Chinese medicine, retain the five-ring mother nucleus of fijianine, and introduce different substituents on the mother nucleus. Preliminary pharmaceutical experiments have proved that the compound has the characteristics of stable chemical properties, strong antibacterial activity, broad antibacterial spectrum, high bioavailability, low residue, no drug resistance, and small toxic and side effects, and is expected to be used as a new type of veterinary drug for the treatment of bovine mastitis.

[0011] Preferably, the Fijian alkali derivative is prepared by acylation reaction of 1,2,3,4-tetrahydro-β-carboline-1-carboxylic acid and p-methoxybenzoyl chloride as raw materials.

[0012] It should be noted that the present invention does not limit the sources of 1,2,3,4-tetrahydro-β-carboline-1-carboxylic acid and p-methoxybenzoyl chloride, and they can be prepared by conventional methods. The reaction formula for synthesizing the Fijian alkali derivative is shown in Formula 2:

[0013] Formula 2.

[0014] Preferably, the preparation method of the fijianine derivative comprises the following steps: 1,2,3,4-tetrahydro-β-carboline-1-carboxylic acid and a base are added to 1,4-dioxane, and p-methoxybenzoyl chloride is added at -22°C to -17°C, and the temperature is raised to 10°C to 35°C for acylation reaction, and then an acid is added for crystallization and recrystallization to obtain a Fijian alkali derivative.

[0015] Further preferably, the molar ratio of the 1,2,3,4-tetrahydro-β-carboline-1-carboxylic acid to the p-methoxybenzoyl chloride is 1:(1-1.4), more preferably 1:1.2.

[0016] Further preferably, the base comprises 1.8 mol / L to 2.2 mol / L sodium hydroxide aqueous solution, and the pH of the acylation reaction is 7 to 9, more preferably pH 7.5 to 8.5.

[0017] More preferably, the molar volume ratio of the 1,2,3,4-tetrahydro-β-carboline-1-carboxylic acid to the 1,4-dioxane is 1 mmol:(4-6) mL.

[0018] More preferably, the acylation reaction temperature is 15° C. to 25° C., and the acylation reaction time is 6 h to 20 h (more preferably 10 h to 14 h).

[0019] Further preferably, the acid comprises a 4.8 mol / L to 5.2 mol / L aqueous hydrochloric acid solution, and the molar volume ratio of the 1,2,3,4-tetrahydro-β-carboline-1-carboxylic acid to the acid is 1 mmol:(4 to 6) mL.

[0020] Further preferably, the crystallization temperature is 1°C to 3°C, and the crystallization time is 4h to 6h.

[0021] For example, after the crystallization is completed, suction filtration and water washing are also included.

[0022] Further preferably, ethyl acetate is added for the recrystallization, the recrystallization temperature is 1° C. to 3° C., and the recrystallization time is 2 h to 4 h.

[0023] More preferably, the molar volume ratio of the 1,2,3,4-tetrahydro-β-carboline-1-carboxylic acid to the ethyl acetate is 1 mmol:(1.8-2.2) mL.

[0024] The present invention has found through a large number of experiments that the yield and purity of the Fijian alkali derivative can be further improved by controlling the reaction conditions of 1,2,3,4-tetrahydro-β-carboline-1-carboxylic acid and p-methoxybenzoyl chloride.

[0025] In a second aspect, the present invention provides a cow mammary gland injection, comprising a fijianine derivative as shown in formula I;

[0026] Formula I.

[0027] Preferably, the cow mammary gland injection further comprises a pharmaceutically acceptable excipient, and the mass content of the fijianine derivative is 95% to 99.5%.

[0028] Preferably, the injection amount of the cow mammary gland injection is 1 g / head / time to 3 g / head / time. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is the hydrogen spectrum of the Fijian alkali derivative of Example 1 of the present invention; Figure 2 This is the carbon spectrum of the fijianine derivative of Example 1 of the present invention. DETAILED DESCRIPTION

[0030] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0031] The specific components and contents of the culture medium used in the embodiments of the present invention are: Beef extract peptone solid culture medium (referred to as solid culture medium): 3g beef extract, 5g peptone, 5g NaCl, 1000mL distilled water, 15g~20g agar, pH=7.4.

[0032] Beef extract peptone liquid medium (referred to as liquid medium): 5 g peptone, 3 g beef extract, 1000 mL triple distilled water, 5 g NaCl, pH = 7.4; provided by Henan PuLe Education Technology Co., Ltd. Nutrient agar medium, nutrient broth medium, MH nutrient broth, MH nutrient agar, MacConkey agar medium, provided by Guangdong Huankai Microbiology Technology Co., Ltd. LB medium, Lennox, SIGMA.

[0033] In order to better illustrate the present invention, further examples are given below.

[0034] Example 1 This embodiment provides a method for preparing a fijianine derivative, comprising the following steps: S1. At room temperature, add 200 mL of water, 12 g of tryptamine and 6.72 g of hydrated glyoxylic acid to a 250 mL three-necked flask, drop 10% NaOH to adjust the pH of the mixed system to 3-4, stir and react for 6 h, let stand to precipitate a solid, wash it three times with water, and recrystallize it with methanol to obtain 1,2,3,4-tetrahydro-β-carboline-1-carboxylic acid.

[0035] S2, add 20 mL of dry THF, 1 g of p-methoxybenzoic acid, and 1 mL of SOCl to a 100 mL round-bottom flask. 2 and 0.2 mL of pyridine, reflux at 60°C for 3 h, and then evaporate the solvent to obtain p-methoxybenzoyl chloride.

[0036] It should be noted that this embodiment does not limit the sequence of S1 and S2.

[0037] S3, add 4mmol 1,2,3,4-tetrahydro-β-carboline-1-carboxylic acid, 4.2mL 2mol / L NaOH aqueous solution and 10mL 1,4-dioxane into a 100mL round-bottom flask, and stir until all are dissolved; control the temperature at -20°C with an ice-salt bath, dropwise add 4mmol p-methoxybenzoyl chloride, stir and react at room temperature for 12h, add 10mL 5mol / L hydrochloric acid aqueous solution, crystallize at 2°C for 5h, filter and wash with water three times; recrystallize with 8mL ethyl acetate at 2°C for 3h to obtain a Fijian alkali derivative with a yield of 89.4% and a purity of 95%.

[0038] The Fijian alkali derivative synthesized in Example 1 was subjected to nuclear magnetic resonance and mass spectrometry detection, and the detection results are as follows: Figure 1~2 shown.

[0039] mp: 180.6℃~1801.6℃. IR (KBr) v (cm -1 ): 3214(NH),1755(CC=O), 1622(NC=O), 1275(Ar-O-CH 3 ) ; 1 H-NMR (400 MHz, DMSO-d 6 ) δ11.08(s, 1H), 7.46(td, J =17.5, 16.5, 7.7Hz,4H), 7.07(ddt, J =22.5, 14.9, 7.4Hz, 4H),5.99(s, 1H), 4.05(d, J =12.0Hz, 1H),3.83(d, J =19.1Hz, 3H), 3.46-3.67(m, 1H), 2.67-2.98(m, 2H). 13 C-NMR (100 MHz, DMSO-d 6 , TMS, ppm) δ 171.2, 169.7, 161.2,137.0, 129.6,127.5, 127.3, 126.2, 122.3, 119.4, 118.4, 114.5, 112.2, 109.1, 65.4, 55.9,53.3, 47.1, 21.7. ESI-MS: Calcd for C 20 H 18 N 2 O 4 [M+Na]+373.1411, found 373.1381. Example 2 This example provides an antibacterial test of a fijianine derivative.

[0040] 1. Test strains Staphylococcus aureus (Escherichia coli, ATCC 26003), Escherichia coli (Staphylococcus aureus, ATCC BL-21), and Pseudomonas aeruginosa (Pseudomonas aeruginosa, ATCC10104) are all standard bacteria preserved by the Lanzhou Institute of Animal Husbandry and Veterinary Drugs, Chinese Academy of Agricultural Sciences.

[0041] 2. Sterilization of culture medium and activation of bacterial strains 1000mL beef extract peptone culture medium and experimental vessels were sterilized by high pressure wet heat at 121℃ for 30min, the solid culture medium was made into a slant, and a sterility test was performed in a 37℃ incubator. The more thoroughly sterilized slant culture medium was used to activate the strains, and the liquid culture medium was used for group experiments.

[0042] The liquid culture medium was divided into a control group and a test group, and 10 concentration gradients were set for the control group and the test group respectively: 0.0100 g / mL, 0.0500 g / mL, 0.1000 g / mL, 0.2000 g / mL, 0.2200 g / mL, 0.2400 g / mL, 0.2500 g / mL, 0.2600 g / mL, 0.2800 g / mL and 0.3000 g / mL (these concentrations were all converted according to the amount of crude drugs). Kanamycin sulfate, Fijian alkali, Fijian alkali derivative a as shown in formula II, Fijian alkali derivative b as shown in formula III, Fijian alkali derivative c as shown in formula IV, Fijian alkali derivative d as shown in formula V and the Fijian alkali derivative prepared in Example 1 were added according to the gradient requirements, and then 0.1 mL of the activated three test bacterial solutions were added to the container containing the culture medium, and cultured in a constant temperature oscillator at 37°C.

[0043]

[0044] Formula II

[0045] Formula III

[0046] Formula IV

[0047] Formula V.

[0048] Wherein, the preparation method of the Fijian alkali derivative a as shown in formula II comprises the following steps: The Fijian alkali derivative prepared in Example 1 was dissolved in 6 mL of methanol, mixed evenly, and SOCl was added dropwise at -20 °C. 2 , stirred vigorously for 1 h, and continued to react at room temperature for 8 h; after TLC detection, the solvent was dried, ethyl acetate was added to dissolve, and NaHCO 3 The solution was washed three times, and then washed three times with saturated brine. The organic layer was dried over anhydrous sodium sulfate, filtered, the filtrate was spin-dried, and separated by column chromatography to obtain Fiji alkali derivative a.

[0049] The preparation method of the Fijian alkali derivative b shown in Formula III is similar to the preparation method of S3 in Example 1, except that p-methoxybenzoyl chloride is replaced by an equal molar amount of p-iodobenzoyl chloride.

[0050] The preparation method of the Fijian alkali derivative c shown in Formula IV is similar to the preparation method of S3 in Example 1, except that p-methoxybenzoyl chloride is replaced by an equal molar amount of p-fluorobenzoyl chloride.

[0051] The preparation method of the fijianine derivative d as shown in formula V is similar to the preparation method of S3 in Example 1, except that p-methoxybenzoyl chloride is replaced by an equal molar amount of p-trifluoromethylbenzoyl chloride.

[0052] Take the refrigerated Escherichia coli strain, streak culture on the agar plate, place it in a constant temperature incubator, and culture it at 37°C for 24 hours. Pick out a single colony and inoculate it into liquid culture medium, place it in a constant temperature oscillator, and culture it at 37°C for 24 hours to activate the strain.

[0053] 3. Determination of MIC Kanamycin sulfate, Fijian alkali, Fijian alkali derivative a as shown in formula II, Fijian alkali derivative b as shown in formula III, Fijian alkali derivative c as shown in formula IV, Fijian alkali derivative d as shown in formula V and Fijian alkali derivative prepared in Example 1 were diluted with sterile distilled water in multiple ratios to prepare dilutions of the required series of concentrations; then the dilutions were quantitatively transferred to sterile culture dishes, fully mixed with the quantitatively poured culture medium, cooled and solidified at room temperature, and a plate containing drug solution was prepared; then, according to the required dosage of the culture dish, a quantitative test bacterial solution was respectively transferred, applied to the plate, placed in a constant temperature box at 37°C for 1d~2d, and the growth of the colonies was checked, and the number of colonies was calculated by the plate colony counting method. In addition, a sterile negative and a drug-free positive with a 1:5 ratio of each drug-containing solution were taken as controls. The above tests were repeated 3 times. The lowest drug concentration when there was no colony growth at all by naked eye counting was the minimum inhibitory concentration (MIC) of the drug, and the test results are shown in Table 1.

[0054] Table 1. Results of the determination of antibacterial effect (diameter of the inhibition zone / mm)

[0055] In the table, * indicates that there are significant differences between the values ​​in the same column, P < 0.05.

[0056] As shown in Table 1, when fiji alkali, the fiji alkali derivative of Example 1, the fiji alkali derivative a, the fiji alkali derivative b, the fiji alkali derivative c, the fiji alkali derivative d and kanamycin sulfate were respectively acted on Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa, all showed antibacterial effects. Among them, the antibacterial effect of the fiji alkali derivative of Example 1 was significantly stronger than that of the fiji alkali derivatives a to d, fiji alkali and kanamycin sulfate, so the fiji alkali derivative provided by the present invention is used as a drug compound in veterinary clinics and has practical value.

[0057] Example 3 This example provides an in vitro antibacterial test for common pathogenic bacteria of bovine mastitis. The minimum inhibitory concentration (MIC) of the fijianine derivatives is determined by the test tube double dilution method combined with the agar plate method. The in vitro antibacterial activity of the fijianine derivatives provided in Example 1 is observed by the tube-plate method. The diluent is MH broth.

[0058] 1. Test strains Streptococcus agalactiae and Streptococcus uberis were the dominant strains isolated and identified from the milk of cows with mastitis; Streptococcus dysgalactiae (ATCC 35666), Staphylococcus aureus (CMCC(B) 26003), Escherichia coli (CMCC(B) 44102), Proteusbacillus vulgaris (CMCC(B) 49027), Streptococcus pyogenes (ATCC 19615), Pseudomonas aeruginosa (CICC10419), and Bacillus subtilis (CMCC(B) 63501) were purchased from China General Microbiological Culture Collection and China Industrial Microbiological Culture Collection and Management Center, respectively, with official authorization from ATCC.

[0059] 2. Preparation of inoculum Take the broth culture of each bacteria, and prepare the test bacterial suspension with a concentration equivalent to 0.5 McFarland standard turbidimetric tube using 0.9% sterile saline solution according to the 10-fold dilution method, and then dilute it with MH broth to a bacterial count of 1×10 7 CFU / mL, for future use.

[0060] 3. Test tube double dilution method combined with agar plate method The dilution method for the experimental group and the control group was the same. Take each 5mL sterile centrifuge tube, 11 for the experimental group and 11 for the control group, numbered 1#~11#. In each group, except for tube 1#, 2mL MH broth (MH broth containing 2% calf serum was added to streptococci). Add 2mL of sterilized drug stock solution to tube 1# and tube 2# respectively, and tube 1# is the antibacterial drug stock solution (drug concentration is 500mg / mL); mix the drug solution in tube 2# with nutrient broth, aspirate 2mL and add it to tube 3#, and so on, dilute it to tube 10# by multiple times, aspirate 2mL from tube 10# and discard it, add 100μL of diluted bacterial solution to each tube 1#~11# of the experimental group, and do not add diluted bacterial solution to the 11 tubes of the control group. The 11# tube of the test group is a growth control without drugs; the 1# tube of the control group only adds 2mL of drug solution, without MH broth and bacteria, which is a drug solution control; the 11# tube of the control group only adds 2mL of MH broth, without drug solution and bacteria, which is a blank control. The drug concentrations in tubes 1# to 10# are 500mg / mL, 250mg / mL, 125mg / mL, 62.5mg / mL, 31.25mg / mL, 15.625mg / mL, 7.81mg / mL, 2.91mg / mL, 1.95mg / mL, and 0.98mg / mL, respectively.

[0061] Incubate in a 37°C constant temperature incubator for 18h~24h. Preliminary results: compare each tube in the test group with each tube of the same drug concentration in the control group (if the test tube is turbid, it means bacterial growth; if the test tube is clear and transparent, it means no bacterial growth).

[0062] The drugs and bacterial cultures in each tube of the test group and the control group were streaked onto the blood agar plate and placed in a 37°C constant temperature incubator for 18h~24h. The results were determined by observing whether there was bacterial growth on the blood agar plate. The corresponding test tube with no bacterial growth on the agar plate was the minimum inhibitory concentration (MIC) of the test drug to the bacteria. Each bacterium was repeated 3 times and the average value was calculated. The test results are shown in Table 2.

[0063] 4. Tube-plate method: The antibacterial activity is determined by the in vitro antibacterial method using an Oxford cup filled with liquid medicine.

[0064] Preparation of culture medium plates: sterilize hydrolyzed casein (Mueller-Hinton, MH) agar medium at 121℃ for 15min, wait until the temperature drops to 50℃~60℃, pour into sterile plates to make the agar thickness 4mm. For streptococci, use MH agar plates containing 5% defibrinated sheep blood (v / v). MH agar plates should be incubated at 35℃ for 30min before use to dry the surface.

[0065] Determination of the optimal coating concentration: Use nutrient broth (bacteria) to activate each bacterial solution for 10-1 ~10 -5 Dilute the solution 10 times in increments, take 100 μL of each dilution, and spread on the corresponding agar plate. Repeat 3 times for each dilution concentration. Incubate at 37℃ for 18h~24h, observe the growth of the bacterial lawn, and determine the best coating concentration for each bacteria.

[0066] Oxford cup filling solution method: dilute the prepared bacterial solution with sterile saline to the optimal coating concentration, dip the bacterial solution with a sterile cotton swab, and evenly spread it on the MH agar plate. Finally, spread it along the inner edge of the plate for a week to achieve uniform inoculation.

[0067] The test results of the antibacterial strength are shown in Table 2.

[0068] Table 2 Test results of MIC and inhibition zone diameter of fiji alkaloid derivatives

[0069] As can be seen from Table 2, the stock solution of fiji alkali derivatives (500 mg / mL) has good antibacterial effects on various bacteria, among which it has a strong antibacterial effect on Streptococcus and Bacillus subtilis, and has a moderate antibacterial effect on common pathogenic Enterobacter and Staphylococcus aureus. The antibacterial strength is in the following order: Streptococcus > Bacillus subtilis > Staphylococcus aureus > Enterobacter. Therefore, as a new preparation of natural drug structure modification, fiji alkali derivative breast injection has the advantages of mild adverse reactions, not easy to produce drug resistance and broad-spectrum antibacterial effect, which makes it have strong development potential and high development and utilization value in the development of new antibacterial drugs.

[0070] Example 4 This embodiment provides a clinical trial of a cow mammary gland injection for treating cow mastitis.

[0071] Experimental animals: Cows with mastitis were selected from a private dairy farm in Gansu. The main symptoms of the affected cows were abnormal milk in the udder area (the presence of flakes, clots or watery milk), different degrees of redness, swelling, heat and pain in the udder, decreased milk production, but no obvious systemic symptoms.

[0072] Test drugs: cow mammary gland injection X containing the fijianine derivative prepared in Example 1 (the content of fijianine derivative is 99wt%); cow mammary gland injection Y containing the fijianine derivative a shown in Formula II (the content of fijianine derivative a is 99wt%); cow mammary gland injection Z containing fijianine (the content of fijianine is 99wt%); Shuangding injection (trade name: Rulining), specification is 10mL / tube (equivalent to 18g of original drug), produced by Hebei Yuanzheng Pharmaceutical Co., Ltd.

[0073] The cows clinically diagnosed with mastitis were randomly divided into a high-dose group of dairy cow breast injection agent X (3g, referred to as the high-dose group X), a medium-dose group of dairy cow breast injection agent X (2g, referred to as the medium-dose group X), a low-dose group of dairy cow breast injection agent X (1g, referred to as the low-dose group X), a high-dose group of dairy cow breast injection agent Y (3g, referred to as the high-dose group Y), a medium-dose group of dairy cow breast injection agent Y (2g, referred to as the medium-dose group Y), a low-dose group of dairy cow breast injection agent Y (1g, referred to as the low-dose group Y), a high-dose group of dairy cow breast injection agent Z (3g, referred to as the high-dose group Z), a medium-dose group of dairy cow breast injection agent Z (2g, referred to as the medium-dose group Z), a low-dose group of dairy cow breast injection agent Z (1g, referred to as the low-dose group Z) and a double-butan injection group (double-butan control group), 50 cows in each group.

[0074] The statistical results of the number of cows with subclinical mastitis and clinical mastitis are shown in Table 3.

[0075] Table 3 Statistical results of the number of cows with latent mastitis and clinical mastitis

[0076] Usage and dosage: Disinfect the udder before administration, squeeze out 1-2 handfuls of milk, and inject the drug from the milk duct, 2 times / d (once in the morning and once in the evening), 6 days as a course of treatment. After 6 days of clinical treatment, the clinical treatment effect of mastitis cows is shown in Table 4.

[0077] Table 4 Clinical treatment effect of mastitis in dairy cows Healing effect Number of cured / head Effective number / head Invalid number / header Effectiveness / % X high dose group 43 7 0 100 X Medium dose group 38 12 0 100 X low dose group 29 15 6 88 Y high dose group 6 8 36 28 Y medium dose group 5 6 39 22 Y low dose group 3 5 42 16 Highest dose group 8 18 24 52 Medium dose group 5 11 34 32 Z low dose group 2 8 40 20 Double Ding control group 11 29 10 80 The criteria for judging the efficacy of the treatment for bovine mastitis are: "cured" - clinical symptoms disappear, milk appearance is normal, milk production is restored, somatic cell count drops to the normal range (below 500,000 / mL), and milk pathogen culture is negative; "markedly effective" - ​​clinical symptoms are alleviated, milk appearance is normal, milk production is restored, and somatic cell count and milk pathogen count decrease; "ineffective" - ​​all indicators have not improved, or the condition has worsened.

[0078] Detection of milk sample somatic cell count (SCC): Before testing the milk sample, heat it in a water bath and maintain the temperature between 38℃ and 40℃. Use a portable somatic cell counter to detect the somatic cell count. The test results are shown in Table 5.

[0079] Table 5 SCC test results (10,000 / mL) Healing effect Day 0 Day 3 Day 6 X high dose group 238.90±22.51 83.56±21.78** 27.78±4.98** X Medium dose group 267.06±18.11 91.43±22.44** 32.09±6.41** X low dose group 233.46±19.11 120.71±34.21* 60.63±9.81** Y high dose group 246.21±21.47 241.09±20.31 222.76±18.76 Y medium dose group 250.01±19.34 248.99±17.89 235.22±17.45 Y low dose group 245.37±18.39 244.11±16.57 236.02±17.01 Highest dose group 238.21±23.16 193.69±22.96* 165.32±10.11 Medium dose group 241.73±19.32 233.56±25.58 195.15±8.53 Z low dose group 240.34±20.24 238.73±26.62 200.76±11.32 Double Ding control group 281.21±13.44 173.22±25.30* 98.82±5.81* In the table, * indicates that there are significant differences between the values ​​in the same column, P < 0.05; ** indicates that there are extremely significant differences between the values ​​in the same column, P < 0.01.

[0080] As shown in Table 5, before medication (day 0), there was no significant difference in the number of somatic cells in the milk of the affected cows among the groups (P>0.05); on the 6th day after treatment, compared with before medication, the X high-dose group and the X medium-dose group could significantly reduce the number of somatic cells, and the X low-dose group also had a reducing effect on the number of somatic cells, but the effect was not as obvious as that of the X high-dose group and the X medium-dose group; compared with the double-butan control group, the X high-dose group and the X medium-dose group could significantly reduce the number of somatic cells (P<0.01), and the difference between the X low-dose group and the double-butan control group was significant (P<0.05); the effect of fijianine and fijianine derivative a was not obvious.

[0081] In summary, mastitis is an inflammatory reaction caused by stimulation of the mammary gland by many factors, but pathogen infection is the main cause of mastitis, and pathogens are often mainly Staphylococcus aureus, Streptococcus and Escherichia coli. In recent years, antibiotics have played an important role in preventing and treating mastitis in dairy cows, but there are disadvantages in antibiotic treatment, which easily causes antibiotic residues, thereby affecting the quality of dairy products and the health of consumers. The dairy cow mammary gland injection provided by the present invention is a new preparation modified with natural effective ingredients. Compared with antibiotics, it is not easy to develop drug resistance, has faster metabolism in the body, high efficiency and low toxicity, no residue, and has a definite therapeutic effect, is safe and reliable, and can replace antibiotic drugs to prevent and treat dairy cow mastitis.

[0082] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modification, equivalent substitution or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. Use of a fijiline derivative in the preparation of a drug for treating bovine mastitis, characterized in that: The structural formula of the Fijian alkali derivative is shown in Formula I: Formula I.

2. The use of the fijipine derivative according to claim 1 in the preparation of a drug for treating bovine mastitis, characterized in that: The Fijian alkali derivative is prepared by using 1,2,3,4-tetrahydro-β-carboline-1-carboxylic acid and p-methoxybenzoyl chloride as raw materials through an acylation reaction.

3. Use of the fijipine derivative according to claim 1 in preparing a drug for treating bovine mastitis, characterized in that: The preparation method of the Fijian alkali derivative comprises the following steps: 1,2,3,4-tetrahydro-β-carboline-1-carboxylic acid and a base are added to 1,4-dioxane, and p-methoxybenzoyl chloride is added at -22°C to -17°C, and the temperature is raised to 10°C to 35°C for acylation reaction, and then an acid is added for crystallization and recrystallization to obtain a Fijian alkali derivative.

4. Use of the fijianine derivative according to claim 2 or 3 in the preparation of a drug for treating bovine mastitis, characterized in that: The molar ratio of the 1,2,3,4-tetrahydro-β-carboline-1-carboxylic acid to the p-methoxybenzoyl chloride is 1:(1-1.4); The temperature of the acylation reaction is 15°C to 25°C, and the acylation reaction time is 6h to 20h.

5. Use of the fijipine derivative according to claim 3 in the preparation of a drug for treating bovine mastitis, characterized in that: The base includes 1.8 mol / L to 2.2 mol / L sodium hydroxide aqueous solution, and the pH of the acylation reaction is 7 to 9.

6. Use of the fijipine derivative according to claim 3 in the preparation of a drug for treating bovine mastitis, characterized in that: The molar volume ratio of the 1,2,3,4-tetrahydro-β-carboline-1-carboxylic acid to the 1,4-dioxane is 1 mmol:(4-6) mL.

7. Use of the fijipine derivative according to claim 3 in the preparation of a drug for treating bovine mastitis, characterized in that: The acid includes a 4.8 mol / L to 5.2 mol / L hydrochloric acid aqueous solution, and the molar volume ratio of the 1,2,3,4-tetrahydro-β-carboline-1-carboxylic acid to the acid is 1 mmol:(4 to 6) mL.

8. A cow mammary gland injection, characterized in that: Including the Fijian alkaloid derivatives shown in formula I; Formula I.

9. The cow mammary gland injection according to claim 8, characterized in that The cow mammary gland injection also includes pharmaceutically acceptable excipients, and the mass content of the fijianine derivative is 95% to 99.5%.

10. The cow mammary gland injection according to claim 8, characterized in that: The injection amount of the cow mammary gland injection is 1 g / head / time to 3 g / head / time.

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