Pharmaceutical composition for synergistically inhibiting liver cancer sorafenib drug resistance and application thereof
A technology of synergistic inhibition and composition, applied in the field of pharmaceutical composition for synergistic inhibition of sorafenib drug resistance in liver cancer
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0027] Example 1: The killing effect of melittin and bufalin single drug on SMMC-7721-R
[0028] cell line
[0029] Sorafenib-acquired drug-resistant human liver cancer cell line: SMMC-7721-R, which was constructed and preserved in the Institute of Tumor Research of Traditional Chinese Medicine of the People's Liberation Army (Liu Dong, Fan Yifu, Li Jun, Cheng Binbin, Lin Wanfu, Li Xiaoyan, DuJuan* , Ling Changquan*. Inhibition of cFLIP overcomes acquired resistance to Sorafenib via reducing ER stress-related autophagy in hepatocellular carcinoma. Oncol Rep 2018; 40(4):2206-14.).
[0030] 1. Plating: take normal growing SMMC-7721-R cells, digest with trypsin, centrifuge (1000rpm, 5min), count under a microscope with a cell counting plate; dilute the cell concentration to 5.0×10 4 cells / mL of cell suspension, which was added to a 96-well plate (100 μL / well) and placed in a cell culture incubator (37°C, 5% CO 2 ), incubated for 12h.
[0031] 2. Dosing: 12 hours after the cell...
Embodiment 2
[0037]Embodiment 2: The killing effect of melittin and bufalin combined on SMMC-7721-R
[0038] The difference between melittin and bufalin at 48h and 72h is relatively small (the IC of melittin and bufalin at 48h 50 0.38±0.13μmol / L, 1.64±0.54μmol / L, respectively). Therefore, in the next step, we choose the action time of the two drugs to be 48h. We set 5 concentrations of the two drugs respectively (the set concentration is IC 50 and two concentrations above and below). The concentration gradient of bufalin is: 0.1μmol / L, 0.2μmol / L, 0.4μmol / L, 0.8μmol / L, 1.6μmol / L; the concentration gradient of melittin is: 0.5μmol / L, 1μmol / L , 2 μmol / L, 4 μmol / L, 8 μmol / L; paired in pairs to form 25 groups, which were tested for CCK-8 activity.
[0039] 1. As shown in Table 1, two groups were combined, a total of 25 groups; a negative control group (only containing cells, no drug treatment) and a blank group (only containing complete medium) were set at the same time, with 6 replicate we...
Embodiment 3
[0045] Example 3: Effects of melittin and bufalin combined on the apoptosis of SMMC-7721-R
[0046] The comprehensive index analysis has verified that 1.0 μmol / L melittin + 0.2 μmol / L bufolin is the optimal synergistic drug ratio. In order to verify this result, we further used flow cytometry to detect the combined use of melittin / bufalin and the two drugs alone (the single drug concentration of melittin was 2.0 μmol / L, and the concentration of bufalin single drug was 0.4 μmol / L). ) on the apoptosis of liver cancer cells. Melittin and bufalin were combined to act on SMMC-7721-R cells for 48 hours, and cell apoptosis was detected after Annexin V-APC / PI double staining.
[0047] 1. Plating and dosing: Plate SMMC-7721 and SMMC-7721-R on a 12-well plate; after 12 hours of wall attachment, add DMEM containing 0 μmol / L, 5 μmol / L, 10 μmol / L and 20 μmol / L Sorafenib respectively Above 12-well plate culture medium.
[0048] 2. Aspirate the cell culture solution into the centrifuge tu...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com