Fixed dose combination of cannabinoids and medical mushrooms for prevention and treatment of cancer, inflammatory or immune-mediated inflammatory diseases
An inflammatory disease, immune-mediated technology, applied in the direction of resistance to vector-borne diseases, drug combinations, allergic diseases, etc., can solve problems such as destroying tumors, reducing cancer immunity, anti-tumor effects, and cancer progression damage
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example 1
[0243] To assess the anti-inflammatory properties of the composition of the invention, assays of nitric oxide (NO) and cytokine secretion from RAW cells following incubation with mushroom extracts and cannabinoids, as well as viability assays were performed.
[0244] Table 1: *Suggested Assay Design
[0245]
[0246]
[0247] *Rows marked in red are plate set up
[0248] **Only in CBD / THC board
[0249] Table 2: Plate setup for mushroom extracts (4 mushrooms / plate)
[0250]
[0251]
[0252] Table 3: Plate setup for cannabinoid extracts
[0253]
[0254] Cell proliferation and viability (XTT assay)
[0255] a. Preparation of XTT reaction solution.
[0256] b. Add 50 μL of XTT reaction solution to each well.
[0257] c. Place the plate in a humid atmosphere (e.g. 37°C, 5% CO 2 ) incubate.
[0258] d. Shake the plate gently to distribute the dye evenly in the wells.
[0259] e. Measure the absorbance of the samples with a spectrophotometer (ELISA reader) a...
example 2
[0359] Viability assay of panc-1 and gl-1 cells after treatment with various combinations of mushroom extracts and cannabinoids
[0360] test system
[0361] a. PANC-1 cells (human pancreatic cancer cells) (American Type Culture Collection, Rockville, MD, USA).
[0362] b. EGL-1 cells (human extrahepatic cholangiocarcinoma)
[0363] Cell growth:
[0364] Growth medium suitable for PANC-1 / EGL-1 cells: Dulbecco's (S. ) Modified Eagle's medium (DMEM) high glucose.
[0365] Suitable growth medium for EGL-1 cells: RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine and 1% penicillin-streptomycin solution (PS).
[0366] Assay medium suitable for PANC-1: Assay medium contains only 1% FBS.
[0367] Assay medium suitable for EGL-1: Assay medium contains only 1% FBS in DMEM low glucose (1 g / L)
[0368] Experimental Design: Incubate cells with test item:
[0369] a. Cells were harvested at 80% confluency, counted and resuspended in growth medium to a ...
example 3
[0393] Cancer Study Results: Mushroom Extracts
[0394]
[0395] a. Seven mushroom extracts were incubated with 0.5-10 mg / ml of PANC-1 or EGL-1 for 72 hours.
[0396] b. Tables 12 and 13 describe the doses of mushrooms that resulted in the greatest kill (lowest viability).
[0397] c. Test the viability of the cells (XTT assay).
[0398] d. EGL-1 cells are much more resistant to cell death than PANC-1 cells. Cordyceps M (RM) was the most effective mushroom, followed by Ganoderma lucidum.
[0399] e. PANC-1 cells: Chaga was the most effective mushroom, followed by Ganoderma lucidum and Turkey Tail (MS).
[0400] f. The results are shown in Figures 7 and 8.
[0401] Table 12: Summary of PANC-1 Cell Viability*
[0402]
[0403]
[0404]
[0405] Table 13: Summary of viability in EGL-1 cells
[0406]
[0407]
[0408]
[0409] The results showed that both EGL-1 and PANC-1 cells incubated with Agaricus and Cordyseps Cs-4(MS) exhibited a % viability higher...
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