Synthesis of compounds based on the active domain of cabotegravir and their application in inhibiting tumor cells activity

Abstract

Structural modification based on existing drugs, which ensures the safety of marketed drugs, is an essential approach in developing new drugs. In this study, we modified the structure of cabotegravir by introducing the front alkyne on the core structure through chemical reaction, resulting in the synthesis of 9 compounds resembling 1,2,3-triazoles. The potential of these new cabotegravir derivatives as tumor suppressors in gastrointestinal tumors was investigated. Based on the MTT experiment, most compounds showed a reduction in the viability of KYSE30 and HCT116 cells. Notably, derivatives 5b and 5h exhibited the most significant inhibitory effects. To further explore the effects of derivatives 5b and 5h on gastrointestinal tumors, KYSE30 cells were chosen as a representative cell line. Both derivatives can effectively curtail the migration and invasion capabilities of KYSE30 cells and induce apoptosis in a dose-dependent manner. We further demonstrated these derivatives induce cell apoptosis in KYSE30 cells by inhibiting the expression of Stat3 protein and Smad2/3 protein. Based on the above results, we suggest they show promise in developing drugs for esophageal squamous cell carcinoma.

Keywords: Cabotegravir; ESCC; KYSE30; apoptosis; derivatives.

Scheme 1

The reaction routes to compounds 5a–5i.

Figure 1

Compounds 5b and 5 h inhibited the migration ability of KYSE30 cells. Photographs of cells at 0 h, 24 h and 48 h after treatment with different concentrations of 5b (a) and 5 h (b). (Scale bar=200 μm). (c): Effect of 5b on the migration rate of KYSE30 cells. (d): Effect of 5 h on the migration rate of YSE30 cells (n=3, *P<0.05). Data are shown as mean±SD and analyzed by one‐way ANOVA. (*P<0.05, **P<0.01, ****P<0.0001 compared to control group).

Figure 2

Effect of 5b and 5 h on the invasive ability of KYSE30 cells. (a)(b): Using microscopy to record cell invasion after cells were treated with different concentrations of compounds for 24 h. (Scale bar=200 μm). Effects of different concentrations of 5b (c) and 5 h (d) on invasion rate of KYSE30 cells (n=3). Data are shown as mean±SD and analyzed by one‐way ANOVA. (*P<0.05, **P<0.01, ****P<0.0001 compared to control group).

Figure 3

Effects of 5b and 5 h on apoptosis of KYSE30 cells. (a): Apoptosis flow results. (b): Effect of different concentrations of 5b on apoptosis of KYSE30 cells (n=3). (c): Effect of different concentrations of 5 h on apoptosis of KYSE30 cell(n=3). Data are shown as the mean±SD and were analyzed by one‐way ANOVA. (*P<0.05, **P<0.01 and ****P<0.0001, compared with the control group).

Figure 4

TUNEL staining to observe the effect of different drugs on DNA breakage in KYSE30 cells. (Scale bar =100 μm).

Figure 5

(a): Western blot was used to detect the protein expression of KYSE30 cells treated with different concentrations of compounds. (b): Western blot results showed that the expression of Stat3 protein was significantly decreased after treatment with different concentrations of the compounds. (c): The expression of Smad2/3 protein was significantly decreased after treatment with different concentrations of drugs. (d): The expression of cleaved caspase3 protein was significantly increased after treatment with different drug concentrations. Data are shown as the mean±SD and were analyzed with one‐way ANOVA. (*P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001, compared with the control group).