Immune activation type antibody and application thereof
An immune activation and antibody technology, applied in the direction of antibodies, immunoglobulins, antibody medical components, etc., can solve the problems of normal tissue damage and side effects, and achieve the effects of avoiding damage and side effects, good immune regulation and therapeutic effect
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preparation example Construction
[0160] The synthetic method of compound 1, comprises the steps:
[0161] Dissolve 2.37 grams of compound Pro-1 and 1.2 grams of bromopropionitrile in 50 mL of acetonitrile, and add 1.5 grams of K 2 CO 3 , and stirred at room temperature for 12 hours. After filtration, 2 mL of iodotrimethylsilane was added, and stirred at room temperature for 10 hours. Add 5 mL of saturated NaHCO 3 Solution, the solvent was distilled off under reduced pressure, separated by silica gel column chromatography (methanol:dichloromethane=1:10 volume ratio), the product solution was collected, concentrated under reduced pressure to remove the dissolved solution, and the solid product was obtained as compound 1.
[0162]
[0163] The synthesis method of compound 2 and compound 3 is basically the same as that of compound 1, except that when compound 2 is synthesized, compound Pro-1 is replaced by compound 9; when compound 2 is synthesized, compound Pro-1 is replaced by compound 10 . Among them, ...
Embodiment 1
[0357] TLR7 Activation Detection of the Compound of Example 1 (HEK-BlueTM Detection)
[0358]Take HEK-Blue TMh TLR7 cells in the logarithmic growth phase (purchased from InvivoGen), discard the growth medium (Gibco, C11995500BT, Invivo Gen, ant-nr), and take an appropriate amount of 37°C PBS (Hyclone, SH30256.01) Wash the cells twice by lightly rinsing, and discard the PBS. Add 2-5mL 37°C PBS, incubate for 1-2min, scrape the cells with a cell scraper and blow gently to disperse them into a single-cell suspension. The cells were counted and the cell concentration was calculated using a hemocytometer, and the cell suspension was adjusted to 2.5×104 / 180 μL per well using HEK-Blue™ Dectetion solution (purchased from Invivo Gen) for cell plating on a 96-well cell culture plate. According to the compound or drug concentration (for example, 0.01 μM, 0.1 μM, 1 μM, 5 μM, 15 μM, 30 μM, 40 μM), HEK-BlueTM hTLR7 cells were stimulated, and three replicate wells were set for each concentra...
Embodiment 2
[0359] Example 2 Detection of the TLR7 agonist release effect of each immune-activating antibody
[0360] Take each immune-activating antibody and add it to HER2-positive SKBR3 (10 5 1) cells in RPMI1640 medium. After mixing and culturing for 12 hours, the supernatants were taken out, and the TLR7 activation effect was tested according to the method in Example 1. The result is as figure 2 and image 3 shown.
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