Application of patchoulone in the preparation of drugs for preventing and treating porcine reproductive and respiratory syndrome
A technology for respiratory syndrome and patchouli ketone, which is applied in the direction of pharmaceutical formulations, medical preparations containing active ingredients, antiviral agents, etc., can solve the problem of unpatented and unseen patchouli ketone anti-pig reproductive and respiratory syndrome To achieve good antiviral effect, clear medicinal ingredients and mechanism of action, and high biological safety
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Embodiment 1
[0024] Example 1: Extraction of Patchoulone
[0025] Patchouli whole herb 50kg, steam distillation to obtain 1.78kg of pale yellow oil, rectification under reduced pressure, under 1.3KPa, to obtain 159-163 ℃ of fractions 345g, carry out silica gel column chromatography, with petroleum ether ( 30-60 ℃)-diethyl ether gradient elution, combined 10% diethyl ether-petroleum ether eluent, and then subjected to silica gel column chromatography to obtain colorless needle crystals (260mg), melting point 32.5-33 ℃, by mass spectrometry and nuclear magnetic resonance Hydrogen spectrum analysis identified patchoulone.
Embodiment 2
[0026] Example 2: Cytotoxicity of patchoulone on Marc-145 determined by CCK-8 method
[0027] After the Marc-145 cells grow to a monolayer in the 96-well cell culture plate, the medium is discarded, washed twice with PBS (phosphate buffered saline), and 100 μL of DMEM cell maintenance solution containing 2% fetal bovine serum is added to dilute it. different concentrations of patchoulone, and three replicate wells were made for each concentration. After continuing to incubate at 37°C for 48h, add 10 μL of LCCCK-8 (Japan Dojin Chemical) reagent to each well, incubate at 37°C for about 2h, and detect the absorbance value at a wavelength of 450nm. from figure 1 It can be seen that patchoulone does not inhibit the proliferation of Marc-145 cells at a concentration of 10 μg / mL.
Embodiment 3
[0028] Example 3: Determination of the inhibitory effect of patchoulone on the proliferation of porcine reproductive and respiratory syndrome virus RNA in Marc-145 cells by real-time fluorescence quantitative PCR
[0029] After the Marc-145 cells grow to a monolayer in a 6-well cell culture plate, the medium is discarded, washed twice with PBS (phosphate buffered saline), and diluted with DMEM cell maintenance solution containing 2% fetal bovine serum. Including 100TCID 50 of porcine reproductive and respiratory syndrome virus (PRRSV), 600 μL / well, incubated at 37°C for 2 h, discarded the virus supernatant, washed twice with PBS, and added 12, 6, and 3 μg / mL of patchoulone, 2 mL / well, respectively. . In the experiment, a normal control group (without test drug, without PRRSV) and a PRRSV control group (without test drug) were set at the same time, and three parallels were set for each test concentration. The cells were cultured at 37°C until 48 h after infection. After obse...
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