Applications of ursodeoxycholic acid or pharmaceutical salts of ursodeoxycholic acid in preparation of anti-tumor medicines, and anti-tumor medicines
An anti-tumor drug, ursodeoxycholic acid technology, applied in the field of biomedicine, can solve the problems of distant metastasis, hidden tumor onset, difficulty in early diagnosis, etc., and achieve the effects of improving therapeutic effect, good inhibitory effect, and good application prospect.
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Embodiment 1
[0027] The surface of Treg cells can express CD4 and CD25 molecules. In addition, its characteristic marker is its high expression of the transcription factor Foxp3. Using the above characteristics, flow cytometry was used to explore the effect of ursodeoxycholic acid on the differentiation of Treg cells in vitro, specifically Proceed as follows:
[0028] T cell sorting: Mouse T cells were sorted by immunomagnetic bead method. The mice were sacrificed by cervical dislocation, and the spleen and lymph nodes were aseptically removed, ground and filtered, centrifuged at 400 g for 5 min, and then resuspended cells with sorting buffer. After counting the total number of cells with 3% glacial acetic acid, follow-up operations were performed strictly according to the instructions of the sorting kit.
[0029] sorting CD4+ T cells (CD4 positive naive T cells) were selected using CD4 negative selection kit (EasySep TM MouseCD4+T Cell Isolation Kit, #19852A) combined with Biotin sort...
Embodiment 2
[0039] Because Treg cells not only have the ability to suppress a wide range of anti-tumor immune responses, but also promote angiogenesis in the tumor microenvironment. In order to explore the effect of ursodeoxycholic acid on Treg cells in vivo, female SPF grade C57BL / 6 was used as the experimental object, and tumor-bearing mice were constructed by using mouse colon cancer cell line MC38, lung cancer cell line LLC-luci, and melanoma cell line B16 Model. In order to better promote the absorption of ursodeoxycholic acid in the body, the ursodeoxycholic acid sodium salt URSO (SANTA CRUZBIOTECHNOLOGY, #2898-95-5) was used instead of the above ursodeoxycholic acid.
[0040] The control group (Control) and the experimental group were injected with ddH every day 2 O and URSO (30mg / kg), the tumor size was observed and recorded. The result is as figure 2 A, image 3 A, Figure 4 As shown in A, compared with the control group, the volume of the tumor-bearing mice in the treatmen...
Embodiment 3
[0050] Ursodeoxycholic acid inhibits tumor growth mainly by reducing the proportion of Treg cells in the tumor microenvironment, while SHR-1210, as a humanized anti-PD-1 antibody (Hengrui Medicine), can specifically block PD- 1 binds to PD-L1 and terminates the PD-1 immunosuppressive signal caused by the interaction between PD-1 and PD-L1 in T cells.
[0051] In order to verify whether the combination therapy of ursodeoxycholic acid and anti-PD-1 antibody can improve the anti-tumor efficacy, colon cancer cells (MC38), melanoma cell line (B16), lung cancer cell line (LLC) were subcutaneously inoculated into human PD-1 1 On transgenic mice, when the tumor volume reaches 100mm 3 , were randomly divided into four groups, the treatment group was treated with SHR-1210, URSO or the combination of SHR-1210 and URSO, water for injection and human IgG4 (SHRR-1210 isotype control antibody) were used as the control group, and SHR-1210 was given once every two days , the dosage is 10mg / kg...
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