Application of Hayatine and analogue thereof in preparation of mTORC1 inhibitor
A technology of inhibitors and pharmaceutical preparations, applied in the directions of anti-inflammatory agents, drug combinations, non-central pain relievers, etc., to achieve the effect of preventing adverse effects
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Embodiment 1
[0036] Example 1 Screening of natural mTORC1 inhibitors
[0037] figure 2 In A, a virtual screening process is given to establish a cell size-dependent selection model to predict mTORC1 inhibitors, in which 141 natural compounds are combined with multiple classification methods to screen for mTORC1 inhibitors. To avoid other factors such as digestive enzymes that might affect drug selection, suspension B lymphocytes from multiple myeloma (H929) were used for compound identification. A total of 141 molecules were used in H929 cells, and to exclude false positives, 5 compounds (labeled #11, #17, #48, #153, #159) were selected ( figure 2 Middle B), according to an established criterion, the inhibition rate of mTORC1 activity-related functions is required to be greater than 20% [18] . In order to avoid acute cytotoxic effects affecting cell size, when performing the above compound selection, the cell death rate 48 hours after compound treatment was compared, and it was foun...
Embodiment 2
[0038] Example 2 Effect of Hayatine on mTORC1 Signal Transduction
[0039] In order to evaluate the effect of Hayatine on mTORC1 signal transduction, whether Hayatine can be used as a potential mTORC1 inhibitor was tested by detecting pT389-S6K protein expression and mTORC1-induced autophagy. The inhibitory activity of Hayatine on mTORC1 kinase in H929 and HCT116 cells was determined by the S6K phosphorylation activity using pT389-S6K protein as a specific mTORC1 substrate ( image 3 Middle A and image 3 Middle B). Such as image 3 As shown in C, treatment with Hayatine resulted in a dose-dependent inhibition of mTORC1 activity, suggesting that Hayatine acts as a potent mTORC1 inhibitor. In order to detect whether Hayatine affects mTORC1 activity-related functions, we measured the amount of autophagy in HCT116 cells using a fluorescently labeled LC3 indicator system, which is based on an obvious aggregation phenomenon of LC3 during autophagy formation, because when autopha...
Embodiment 3
[0040] Example 3 Hayatine inhibits the growth and cell migration of cancer cells.
[0041] Other mTORC1 activities such as controlling cell growth and migration are also closely related to tumor progression. To further confirm that Hayatine is an effective mTORC1 inhibitor, we performed the following experiments. First, we examined the effect of Hayatine on the growth of HCT116 cells. Our results showed that in the cell proliferation assay ( Figure 4 Middle A) and colony formation assay ( Figure 4 Medium B and Figure 4 Middle C), Hayatine significantly inhibited the cell growth of HCT116. Subsequently, we performed wound healing experiments and found that Hayatine hindered cell migration in a dose-dependent manner ( Figure 4 Medium D& Figure 4 Middle E), indicating that Hayatine plays a key role in downregulating mTORC1 activity in tumors.
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