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Application of ursolic acid to antitumor immunity

A technology of anti-tumor immunity and ursolic acid

Inactive Publication Date: 2013-06-12
XINJIANG TECHN INST OF PHYSICS & CHEM CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a kind of purposes of ursolic acid in anti-tumor immunity for the low selectivity of current anti-tumor drugs, obvious toxic and side effects and suppression of immune system. Experiments in vivo in animals and mice show that: Ursolic acid can inhibit the proliferation of mouse liver cancer cells and induce apoptosis; ursolic acid can effectively inhibit the proliferation of liver cancer transplanted tumors, and has no obvious side effects on mice; ursolic acid can inhibit the abnormal growth caused by liver tumors. Large spleen index, which can significantly promote the proliferation of T lymphocytes and B lymphocytes, and can significantly increase CD4 + Content of T lymphocyte subsets and CD4 + / CD8 + Proportion of T cell subsets, promote the expression of serum cytokines IL-2 and TNF-α, and reduce the expression of IL-4

Method used

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  • Application of ursolic acid to antitumor immunity
  • Application of ursolic acid to antitumor immunity
  • Application of ursolic acid to antitumor immunity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The influence of embodiment 1 ursolic acid on H22 cell proliferation inhibition:

[0023] (1) H22 cells were grown to the logarithmic phase, and 2.0×10 4 Individual / mL single cell suspension, 180 μL / well was inserted into a 96-well culture plate and cultured overnight;

[0024] (2) Add 20 μL of different concentrations of UA to each well, the final concentrations are 0 (control group), 5, 10, 20, 40 μmol / L (medication group), and each concentration is replicated in 3 wells, cultured for 48 hours;

[0025] (3) Add 20 μL / well of MTT with a concentration of 5 mg / mL and place in CO 2 After continuing to culture in the incubator for 4 hours, discard the supernatant in the wells of the culture plate, add 150 μL of DMSO to each well, shake well to fully dissolve the blue-purple precipitate;

[0026] (4) Measure the absorbance A value at 570nm with a microplate reader, calculate the cell growth inhibition rate, and use the Bliss method to obtain the half inhibitory concentrat...

Embodiment 2

[0029] Example 2 The effect of ursolic acid on inducing the apoptosis of mouse hepatoma cell H22 cells:

[0030] (1) Cells in logarithmic growth phase 0.8×10 6 inoculated in 100mm cell culture dish overnight;

[0031] (2) Add 20 μmol / L ursolic acid to act for 24 hours and 48 hours respectively. After trypsinization, collect the cells, wash twice with pre-cooled PBS, and wash with 1.0×10 6 Cells per mL were resuspended in 500 μL of 1× binding buffer, and 5 μL of Annexin V-fluorescein isothiocyanate (Annexin V-FITC) and 10 μL of propidium iodide (PI) dye were added , mix gently, and react at room temperature for 15 minutes in the dark;

[0032] (3) Cell apoptosis rate was measured by flow cytometry, and the experiment was repeated three times;

[0033] (4) The results showed that ursolic acid could induce H22 tumor cell apoptosis in a time-dependent manner ( figure 2 ).

Embodiment 3

[0034] Embodiment 3 ursolic acid is to the growth inhibitory action of H22 mouse xenograft tumor:

[0035] (1) Centrifuge H22 cells in the logarithmic growth phase at 1000r / min for 5min, wash the cell pellet with saline three times, discard the supernatant, resuspend in PBS, and count the viable cell rate by staining with 0.2% trypan blue. When the viable cell rate is >95% , adjust the number of cells to 1×10 7 Each / ml, 0.2ml was inoculated in the abdominal cavity of mice;

[0036] (2) Randomly select 50 healthy Kunming female mice, absorb H22 cells that have been passaged in the mice for 7-10 days under sterile conditions, and adjust the cell density to 1×10 with normal saline. 7 Each / ml, inoculate 0.2ml subcutaneously in the armpit of the left forelimb of the mouse;

[0037](3) After modeling, according to the weight of the mice, the mice were randomly grouped into blocks and divided into 5 groups, with 10 mice in each group, which were high (40mg / kg.d), medium (20mg / kg.d)...

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Abstract

The invention relates to an application of ursolic acid to antitumor immunity. The ursolic acid has the functions of inhibiting the proliferation and inducing the apoptosis of the hepatoma carcinoma cell of a mouse. The in vivo experiments on the mouse show that the ursolic acid has the function of effectively inhibiting the proliferation of the transplanted hepatocellular carcinoma and has no significant side and toxic effects on the mouse. The ursolic acid has the functions of inhibiting the spleen index which increases abornormally due to hepatic tumor, promoting the proliferation of the T lymphocyte and the B lymphocyte significantly, increasing the proportion of the content of the CD4<+>T lymphocyte subset to that of the content of the CD4<+> / CD8<+T> lymphocyte subset, promoting the expression of IL-2 (interleukin-2) which is a serum cell factor, the TNF-alpha (tumor necrosis factor alpha) and reducing the expression of the IL-4 (interleukin-4). The ursolic acid is similar to the cyclophosphamide which is a low-dose antitumor medicament in the antitumor effect and the immunoregulation function, has low toxicity and can be applied to antitumor immunity to provide a theoretical basis for the development of an antitumor immunity medicament.

Description

technical field [0001] The invention belongs to the technical field of medicinal chemistry, and relates to the use of ursolic acid, in particular to the use of ursolic acid in anti-liver tumor immune drugs. Background technique [0002] Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in humans. The incidence of HCC in my country ranks first in the world, and its mortality rate ranks second in the mortality rate of malignant tumors. Surgery is the most effective treatment for HCC. However, due to the concealment before the onset of the disease, patients have lost the opportunity for surgery when they see a doctor. For most postoperative patients, because HCC has a high tendency of vascular metastasis, it is easy to invade the portal vein or other parts of the liver, causing liver cancer recurrence and intrahepatic metastasis, and the long-term survival is not optimistic. Although the current HCC treatment method has developed from a single surgical...

Claims

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Application Information

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IPC IPC(8): A61K31/56A61P35/00A61P37/04
Inventor 李晓波李艳红肖向文陆雪莹
Owner XINJIANG TECHN INST OF PHYSICS & CHEM CHINESE ACAD OF SCI
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