Preparation of kakkatin derivatives and their anti-tumor activity

Figure 1 Molecular structure and preparation process of 6-(hept-6-yn-1-yloxy)-3-(4-hydroxyphenyl)-7-methoxy-4H-chromen-4-one. A: Molecular structure of the kakkatin derivative; B: Process preparation diagram of 6-(hept-6-yn-1-yloxy)-3-(4-hydroxyphenyl)-7-methoxy-4H-chromen-4-one.

Figure 2 Characterization of 6-(hept-6-yn-1-yloxy)-3-(4-hydroxyphenyl)-7-methoxy-4H-chromen-4-one. A: Liquid chromatography-mass spectrometry of 6-(hept-6-yn-1-yloxy)-3-(4-hydroxyphenyl)-7-methoxy-4H-chromen-4-one (HK); B: Nuclear magnetic hydrogen spectroscopy of HK.

Figure 3 The 6-(hept-6-yn-1-yloxy)-3-(4-hydroxyphenyl)-7-methoxy-4H-chromen-4-one inhibited the proliferation, invasion and metastasis of hepatocellular carcinoma SMMC-7721 cells. A: Cloning assay evaluated the effect of 6-(hept-6-yn-1-yloxy)-3-(4-hydroxyphenyl)-7-methoxy-4H-chromen-4-one (HK) on the proliferation of SMMC-7721 cells; B: Colony formation assay to quantify cell proliferation; C: Transwell assay to detect the inhibitory effect of HK on the invasion of SMMC-7721 cells; D: Transwell assay to quantify SMMC-7721 cells; E: Scratch assay to detect the inhibitory effect of HK on the lateral migration of SMMC-7721 cells; F: Scratch assay to quantify SMMC-7721 cells. ^aP < 0.05; ^bP < 0.01; ^cP < 0.05; ^dP < 0.01; ^eP < 0.05; ^fP < 0.01; ^gP < 0.05; ^hP < 0.01. CDDP: Cisplatin; HK: 6-(hept-6-yn-1-yloxy)-3-(4-hydroxyphenyl)-7-methoxy-4H-chromen-4-one.

Figure 4 The 6-(hept-6-yn-1-yloxy)-3-(4-hydroxyphenyl)-7-methoxy-4H-chromen-4-one induces apoptosis and cycle arrest in hepatocellular carcinoma SMMC-7721 cells. A: Effect of 6-(hept-6-yn-1-yloxy)-3-(4-hydroxyphenyl)-7-methoxy-4H-chromen-4-one (HK) on apoptosis of SMMC-7721 cells at 24 hours and 48 hours; B: Quantification of cell apoptosis; C: Effect of HK on SMMC-7721 cell cycle arrest at 24 hours; D: Quantification of cell cycle arrest. ^aP < 0.05; ^bP < 0.01; ^cP < 0.05; ^dP < 0.01; ^eP < 0.05; ^fP < 0.01; ^gP < 0.05; ^hP < 0.01.

Figure 5 Network pharmacological effects of 6-(hept-6-yn-1-yloxy)-3-(4-hydroxyphenyl)-7-methoxy-4H-chromen-4-one and hepatocellular carcinoma SMMC-7721 cells. A: Cross-Venn diagram of 6-(hept-6-yn-1-yloxy)-3-(4-hydroxyphenyl)-7-methoxy-4H-chromen-4-one (HK)-related targets and SMMC-7721 cells; B: Gene Ontology enrichment analysis of 49 potential targets of HK in the treatment of SMMC-7721 cells; C: Kyoto Encyclopedia of Genes and Genomes enrichment analysis of 49 potential targets of HK in the treatment of SMMC-7721 cells; D: Correlation network analysis of 49 potential targets of HK in the treatment of SMMC-7721 cells; E: Correlation of 49 targets with HK. GO: Gene Ontology; KEGG: Kyoto Encyclopedia of Genes and Genomes.

Figure 6 Molecular docking diagram of 6-(hept-6-yn-1-yloxy)-3-(4-hydroxyphenyl)-7-methoxy-4H-chromen-4-one and genes. A: Molecular docking between PDE3B and 6-(hept-6-yn-1-yloxy)-3-(4-hydroxyphenyl)-7-methoxy-4H-chromen-4-one (HK); B: Molecular docking between ARBD1 and HK; C: Molecular docking between NFKB1 and HK; D: Molecular docking between PRKACB and HK.

Figure 7 Real-time PCR assays validation. ^aP < 0.05; ^bP < 0.01.