Compounds inhibiting angiogenesis activity and their preparation methods and applications
A technology of angiogenesis and compounds, applied in biochemical equipment and methods, microorganism-based methods, microorganisms, etc.
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Embodiment 1
[0018] Example 1: Acquisition of bacterial strains: cut the epidermis of Apostichopus japonicus in Chengshantou sea area of Weihai into small pieces of 1cm×1cm, add streptomycin sulfate and ampicillin to PDA medium, cultivate at 25-30°C for 7-14 days, and select Pure strain, and using molecular biology methods combined with morphological identification to determine Aspergillus terreus ( Aspergillus terreus ).
[0019] Aspergillus terreus derived from the sea cucumber Aspergillus terreus , the initial stage of the colony culture is light khaki, and gradually becomes dark khaki, and the opposite side of the colony is dark yellow to brown; the conidia head is spherical at first, and then becomes a dense straight column.
[0020] The strain of Aspergillus terreus is preserved in the General Microorganism Center (CGMCC) of China Committee for Culture Collection of Microorganisms, and the preservation number is: CGMCC No.11545. Address: Institute of Microbiology, Chinese Academy...
Embodiment 2
[0021] Embodiment 2: a kind of preparation method of the compound of inhibiting angiogenesis activity is characterized in that it comprises the steps:
[0022] 1) Make seed culture solution: Aspergillus terreus Aspergillus terreus CGMCC No.11545 strain was inoculated on the slant of glucose, peptone and yeast extract medium: pH: 6.5-7.5, at a temperature of 25-30 ° C, a rotation speed of 150 rpm, and cultured for 120 hours to obtain a seed culture solution;
[0023] 2) Fermentation production in fermenter: transfer 500-1000 ml of seed culture solution into fermentation medium, and use a 30-liter fermenter to ferment for 72-96 hours at 25-30 °C; fermentation medium: glucose 1, yeast extract 0.1 , peptone 0.2, sodium chloride 4, water 100 (above is weight ratio). The ventilation rate is 0.4 (volume ratio, ventilation rate / fermentation volume), and the rotation speed is 120 rpm; the seed culture solution is inserted into the fermentation medium, and fermented at room temperatur...
Embodiment 3
[0025] Embodiment 3: use embodiment.
[0026]Inhibition of angiogenesis activity: Using normal zebrafish or transgenic zebrafish, the embryos were removed from the egg membrane by pronase E solution, and treated with this compound, and the number of intersegmental angiogenesis was calculated after 24-36 hours of growth, and the embryonic death was observed or deformities.
[0027] ♀♂The transgenic zebrafish were fed separately, alternately with 14h of light and 10h of darkness, and regularly fed with artificial granular bait and newly hatched Artemia nauplii. When collecting eggs, take healthy and sexually mature zebrafish and put them into the mating tank at a ratio of ♀♂1:1, and obtain fertilized eggs at 9-10 o'clock the next day. After the fertilized eggs were disinfected and washed, they were transferred to transgenic zebrafish embryo culture water (containing 5.0 mmol NaCl, 0.17 mmol KCl, 0.4 mmol CaCl 2 , 0.16 mmol MgSO 4 ), cultured at 28°C under light control.
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