Ester peptide compound as well as preparation method and application thereof
A compound and ester peptide technology, applied in the field of biopharmaceuticals, can solve problems such as insensitivity and drug resistance of cancer cells, achieve strong anti-tumor and anti-viral activities, and have good drug development prospects
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Embodiment 1
[0035] The preparation method of Jiangxi Peptidecin, the steps are as follows:
[0036] 1. Large-scale fermentation: Streak and revive Lentzea jiangxiensis CGMCC 4.6609 on YMG solid medium. After culturing for 10 days, pick a single colony and inoculate it into TSB liquid medium. After 3 days of cultivation, it was used as seed solution; then the seed solution (0.5mL / dish) was spread onto 200 dishes of YMG solid medium plates, and left to ferment at 30°C for 18 days.
[0037] 2. Obtaining of crude extract: cut the fermented solid medium plate into large bottles, then soak it in ethyl acetate for 12 hours and filter it with filter paper to obtain the ethyl acetate extract (repeat the operation 5 times); combine ethyl acetate The ester extract was then concentrated under reduced pressure to remove ethyl acetate to obtain a crude extract;
[0038] 3. Separation and purification of the compound: Dissolve the crude extract with an appropriate amount of methanol and put it into four
Embodiment 2
[0051] Determination of anti-tumor cell (human colon cancer cell HCT116) activity of Jiangxi Peptides:
[0052] 1. Preparation of the sample solution to be tested: the sample to be tested is the pure product of peptidin A, B and C prepared in the above-mentioned embodiment 1. The above three samples are accurately weighed and dissolved in dimethyl sulfoxide respectively (DMSO) to prepare ganxipeptide A, B and C solutions with a concentration of 5 mM.
[0053] 2. The activity of Peptides A, B and C on anti-tumor cells (human colon cancer cell line HCT116) was measured by cck8 assay. Experimental steps include:
[0054] (1) Cell culture. HCT116 cells were cultured with medium containing 1640+10% FBS+100x double antibody.
[0055] (2) Plank. Inoculate 28,000 / mL cell suspension in a 96-well plate, and add 200 μL / well of sterile PBS to the surrounding wells to prevent the evaporation of medium water. Incubate for 24 h in a humid 37°C incubator to allow the cells to adhere to
Embodiment 3
[0065] Determination of HIV Reverse Transcriptase Inhibitory Activity of Jiangxi Peptides:
[0066] 1. Preparation of the sample solution to be tested: the sample to be tested is the pure product of peptidin A, B and C prepared in the above-mentioned embodiment 1. The above three samples are accurately weighed and dissolved in dimethyl sulfoxide respectively (DMSO) to prepare ganxipeptide A, B and C solutions with a concentration of 5 mM. Then dilute with DMSO in turn to obtain jangsiopeptide A and B solutions with concentrations of 1000 μM, 300 μM, 120 μM, 60 μM, 24 μM, 8 μM, 3 μM, 1.2 μM, and 600 μM, 200 μM, 60 μM, 30 μM, 9 μM, 3 μM , 1.2μM, 0.6μM Jiangxipeptide C solution.
[0067] 2. Inhibition test of Jiangxi Peptides on HIV reverse transcriptase activity in vitro: The inhibitory activity of Jiangxi Peptides was measured using the HIV Reverse Transcriptase Activity Assay Kit purchased from Sigma-Aldrich Company (Sigma-Aldrich product number: 11468120910) conduct.
[0068]
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