Application of dendrobium officinale polysaccharide in preparation of drug for treating tumors by targeting tumor-associated macrophages
A technology related to Dendrobium officinale polysaccharide and tumor, which is applied in the directions of antitumor drugs, drug combinations, and pharmaceutical formulations to achieve the effects of promoting tumor killing activity, inhibiting immune escape, and inhibiting tumor growth.
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Embodiment 1
[0028] Embodiment 1: the preparation of dendrobium officinale polysaccharide
[0029] Dendrobium officinale Kimura et Migo was collected from Huoshan, and its fresh strips were dried at 70°C to constant weight and crushed for later use. Accurately weigh 100g of dried Dendrobium officinale powder, add 80% ethanol according to the ratio of material to liquid 1:60, stir and extract at room temperature for 24 hours, centrifuge at 5000g for 10 minutes, collect the precipitate, dry the precipitate to remove ethanol, press 1: Add distilled water at a liquid ratio of 60, stir and extract at 80°C for 2 hours, collect the supernatant by gauze filtration, repeat the water extraction of the residue twice, and finally combine the filtrate three times, and concentrate the extracted filtrate under reduced pressure (65°C), and precipitate with 80% ethanol , deproteinized by Sevag method, dialyzed (3500Da, 24h), and freeze-dried to obtain the crude polysaccharide of Dendrobium officinale. The...
Embodiment 2
[0033] Example 2: DOP anti-tumor activity evaluation
[0034] After the mice were adaptively fed for one week, 150 male C57BL / 6J mice were selected. Except for the normal group (Normal, 10-15 mice), the rest of the mice were inoculated with Hepa 1-6 liver cancer cells, and the growing Hepa1-6 cell suspension in logarithmic phase (2×10 6 each / mL), inoculated in the right armpit of 100 C57BL / 6J mice after disinfection, and each mouse was inoculated with 0.2mL (2×10 6 cells / mL), and press the injection point to prevent cell leakage. During the modeling period, the tumor formation in mice was closely observed. After 3-7 days, when the tumor grew to the touch (the tumor grew to about 0.5cm×0.5cm), use The long diameter (a) and short diameter (b) of the tumor were measured with a vernier caliper, and those with too large or small tumor mass were excluded. Finally, the experiment finally divided the tumor-bearing mice that met the criteria into 5 groups randomly: the model group (Mo...
Embodiment 3
[0038] Example 3: DOP anti-tumor immunomodulatory effect
[0039] After the tumor tissues of each group were taken out and weighed, they were respectively placed in sterile plates containing PBS (FBS, 0.01M, pH7.4) buffer solution containing an appropriate amount of 5% fetal bovine serum, and the tumor tissues were cut with ophthalmic surgical scissors on ice. Minced (1mm), add 2mL of digestion solution containing type IV collagenase (1.5mg / mL, 500U / mg) and DNase-I (0.5mg / mL, 3000U / mg) to every 100mg of tumor tissue, at 37°C, 120rpm After digestion for 60 min under the condition of / mim, transfer to ice and add PBS buffer solution containing 20% fetal bovine serum to stop the digestion, filter through 200 mesh gauze, collect the cell filtrate, centrifuge at 400g for 5 min at 4°C, discard the supernatant to collect the precipitate, After resuspending the pellet in FBS buffer, add red blood cell lysate at a ratio of 1:3, lyse on ice for 15 minutes, centrifuge at 400g for 5 min...
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