Chinese medicine lychee seed and pharmaceutical purpose of extractives of Chinese medicine lychee seed
A technology of lychee seed and extract, applied in the field of traditional Chinese medicine lychee seed and its extract, can solve the problem that HIV-1 inhibitor research has not been reported yet, etc.
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Embodiment 1
[0030] The preparation of embodiment 1 lychee core original drug medicine
[0031] The traditional Chinese medicine lychee core is taken as the crude drug, pulverized according to the conventional method, sieved, and made into litchi core fine powder with a particle size of 60-80 mesh, sterilized, and packaged.
Embodiment 2
[0032] The preparation of embodiment 2 litchi kernel organic solvent extract
[0033] Weigh 2.0 kg of lychee cores, add 8.0 liters of 95% ethanol to reflux and extract three times, each extraction takes 2 hours, combine the extracts obtained by reflux extraction, and concentrate to obtain an extract.
[0034] The screening protocol for HIV-1 protease in vitro inhibitors is described in detail below.
[0035] 1) Expression and purification of HIV-1 protease:
[0036] a) Amplification of HIV-1 protease fragment and construction of expression vector
[0037] Design primer PCR to amplify the HIV-1 protease coding fragment, recover the PCR product and pET-11a vector and digest it with Nde I and BamH I, recover the target fragment, connect the digested fragment with the linearized vector and transform E. coli.DH5α competent cells. The transformed clones were picked for PCR identification, and the positive clones were subjected to DNA sequencing.
[0038] Gene sequence of HIV-1 p...
Embodiment 3
[0053] Embodiment 3 measures the inhibitory rate of litchi core extract to HIV-1 protease
[0054] The extract obtained in Example 1 is used as a sample, and according to the method in Example 2, with HIV-1 protease as the target protein, the preliminary activity is carried out by in vitro enzyme activity high-throughput screening (HTS) as an activity evaluation method for preliminary screening. For evaluation, use a microplate reader to measure the fluorescence intensity value. Calculate the remaining activity of the sample according to the obtained enzyme activity value, repeat 3 times in parallel, and take the average value. The activity data is shown in Table 1. The logarithmic value of the diluted compound concentration is taken as the X-axis, and the corresponding remaining activity The value is the y-axis, and the corresponding IC is calculated by drawing a graph with GraphPad Prism 5 50 Value, IC of litchi core extract on HIV-1 protease inhibitory activity 50 The val...
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