Application of bifunctional iodocarboxylic acid as coxsackie virus inhibitor
A technology of iodocarboxylic acid and coxsackie virus is applied in the application field of benzoic acid as a coxsackie virus inhibitor, which can solve the problem of no report on inhibitory activity, and achieves enhanced cell survival rate, easy synthesis, structure simple effect
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Embodiment 1
[0029] Embodiment 1: Difunctional group iodocarboxylic acid L 4 , L 5 Toxicity to host Hep-2 cells
[0030] Hep-2 cells were plated in 96-well plates at 37°C, 5% CO 2 After the monolayer was grown in the incubator, the cell culture solution was discarded, and the cell maintenance solution containing different concentrations of L4 and L5 was added to continue the culture. After 48 hours, the cytotoxicity was visually observed under the microscope and recorded respectively, and the cell survival rate was determined by the MTT method. The specific steps of the MTT method are as follows: add 30 μL of MTT (5 mg·mL -1 ), after incubation for 3-4 h, the supernatant was removed, and 50 μL of DMSO was added to dissolve the precipitate. Read the corresponding absorbance (OD) at 492 nm with a microplate reader 492 value).
[0031] SPSS 11.5 software was used to calculate the median toxic concentration (Median cyctoxic concentration, CC50) of the drug on the cells.
[0032] Cell via...
Embodiment 2
[0033] Embodiment 2: Difunctional group iodocarboxylic acid L 4 , L 5 Inhibitory activity against CVB3
[0034] Hep-2 cells were plated in 96-well plates at 37°C, 5% CO 2 After the monolayer was grown in the incubator, discard the culture medium, infect the cells with 100TCID50 CVB3 virus solution for 1h, and add different concentrations (2.5μg / mL, 5μg / mL, 10μg / mL, 20μg / mL, 40μg / mL, 80μg / mL) of compound L 4 , L 5 (ribavirin as a positive control drug) to incubate the cells. After continuing to culture for about 48 hours, when about 90% of the CPE lesions appeared in the virus control wells, the cytopathic effect (CPE) was observed under a microscope. Observation and recording method of CPE: No cytopathic disease is marked as -, cytopathic disease of less than 25% is recorded as +, 25%-50% cytopathic disease is recorded as ++, 50%-75% cytopathic disease is recorded as +++, more than 75% Cytopathy was recorded as ++++.
[0035] After the observation of CPE, the inhibitor...
Embodiment 3
[0048] Embodiment 3: Difunctional group iodocarboxylic acid L 4 , L 5 Inhibition of CVB3 Progeny Virus Yield
[0049] Hep-2 cells in the logarithmic growth phase were plated in 24-well plates, and 100TCID after the monolayer was overgrown 50 Infect the cells with CVB3, incubate at 37°C for 1.5h, remove the virus solution, wash with PBS three times, add 50μg / mL L 4 , L 5 cell maintenance solution. Cells and supernatant culture fluid were collected at 12h and 36h, and after three freeze-thaw lysis at -20°C and 37°C, TCID 50 Methods To determine the titer of CVB3 virus.
[0050] The result is as image 3 As shown, the CVB3 virus control group showed obvious virus titers at 12 hours after infection, and the virus titers rose rapidly by about 3.0log until 36 hours after infection. while 40μg / mL L 4 , L 5 The virus titer of the treatment group was lower than that of the virus control group under the same time conditions, the increase was small from 12h to 36h after virus i...
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