**4. Conclusions**

**References**

**4. Conclusions** The title compound **5**, C19H16O6, was crystallized in the monoclinic space group P21/c and consists of two independent molecules **5I** and **5II**. Both molecules exhibit the disorder of each methylene groups present in the 1,4-dioxane ring with occupancies 0.6289 (17) and 0.3711 (17), respectively. Based on the Hirshfeld analysis, two independent crystals (**5I** and **5II**) showed differences in the dnorm, shape index (SI), curvedness and the overall contribution to the total Hirshfeld surface. In the crystal, the molecule **5II** forms inversion dimers which are linked into chains along the a axis direction by intermolecular C–H···O interactions. Western blot analysis demonstrated that compound **5** inhibited the TNFα-induced phosphorylation of IKKα/β. A molecular docking study predicted the possible binding of compound **5** to the active site of IKKβ. We found that isoflavone derivative **5** inhibited the TNFα-induced phosphorylation of IKKα/β and its downstream IκB and p65/RelA NF-κB, which suggest that compound **5** can be used as a platform for the The title compound **5**, C19H16O6, was crystallized in the monoclinic space group P21/c and consists of two independent molecules **5I** and **5II**. Both molecules exhibit the disorder of each methylene groups present in the 1,4-dioxane ring with occupancies 0.6289 (17) and 0.3711 (17), respectively. Based on the Hirshfeld analysis, two independent crystals (**5I** and **5II**) showed differences in the dnorm, shape index (SI), curvedness and the overall contribution to the total Hirshfeld surface. In the crystal, the molecule **5II** forms inversion dimers which are linked into chains along the a axis direction by intermolecular C–H···O interactions. Western blot analysis demonstrated that compound **5** inhibited the TNFα-induced phosphorylation of IKKα/β. A molecular docking study predicted the possible binding of compound **5** to the active site of IKKβ. We found that isoflavone derivative **5** inhibited the TNFα-induced phosphorylation of IKKα/β and its downstream IκB and p65/RelA NF-κB, which suggest that compound **5** can be used as a platform for the development of anti-cancer agent targeting IKKα/β.

development of anti-cancer agent targeting IKKα/β. **Supplementary Materials:** The following are available online at www.mdpi.com/xxx/s1, CIF file and NMR spectra for reaction intermediates compounds **2**, **3**, and final product compound **5**: Figure S1: 1H NMR spectrum of compound **2**; Figure S2: 13C-NMR spectrum of compound **2**; Figure S3: 1H-NMR spectrum of compound **3**.; Figure S4: 13C -NMR spectrum of compound **3**; Figure S5: 1H-NMR spectrum of compound **5**; Figure S6: 13C-NMR spectrum of compound **5**; Figure S7: 2D-HMBC spectrum of compound **5**; Figure S8: 2D-HMQC spectrum **Supplementary Materials:** The following are available online at http://www.mdpi.com/2073-4352/10/10/911/s1, CIF file and NMR spectra for reaction intermediates compounds **2**, **3**, and final product compound **5**: Figure S1: 1H NMR spectrum of compound **2**; Figure S2: 13C-NMR spectrum of compound **2**; Figure S3: 1H-NMR spectrum of compound **3**.; Figure S4: 13C-NMR spectrum of compound **3**; Figure S5: 1H-NMR spectrum of compound **5**; Figure S6: 13C-NMR spectrum of compound **5**; Figure S7: 2D-HMBC spectrum of compound **5**; Figure S8: 2D-HMQC spectrum of compound **5**; Figure S9: 2D-TOSCY spectrum of compound **5**.

of compound **5**; Figure S9: 2D-TOSCY spectrum of compound **5**. **Author Contributions:** Conceptualization, D.K. and S.Y.S; investigation, S.A., J.H.L. and M.Y.; validation, **Author Contributions:** Conceptualization, D.K. and S.Y.S.; investigation, S.A., J.H.L. and M.Y.; validation, Y.H.L., H.J.L. and Y.L.; writing—original draft, S.A., D.K. writing—review and editing, S.Y.S. All authors have read and agreed to the published version of the manuscript.

Y.H.L., H.J.L. and Y.L.; writing—original draft, S.A., D.K. writing—review and editing, S.Y.S. All authors have read and agreed to the published version of the manuscript. **Funding:** The authors acknowledge financial support from the Basic Science Research Program (award No. NRF-2019R1F1A1058747). S.Y. Shin was supported by the KU Research Professor Program of Konkuk University.

2019R1F1A1058747). S.Y. Shin was supported by the KU Research Professor Program of Konkuk University.

**Funding:** The authors acknowledge financial support from the Basic Science Research Program (award No. NRF-**Conflicts of Interest:** The authors declare no conflict of interest.

**Conflicts of Interest:** The authors declare no conflict of interest.

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