# Simple Thalidomide Analogs in Melanoma: Synthesis and Biological Activity
## 4 Conclusions  
Thalidomide, a drug well-known in the pharmaceutical history for its teratogenicity has been re-evaluated due to its antiangiogenic and immunomodulatory actions, which have made it effective against several malignancies. As a result, in the last decade, many researchers have concentrated their efforts on the design and development of new thalidomide analogs with improved efficacy and no or reduced toxicity. The focal point of our work was the synthesis and the biological evaluation of a series of thalidomide analogs (**2a-I**) for their potential anticancer activity toward a series of cell lines, including melanoma cells. Interestingly, compound **2k**, bearing a methyl group at the ortho position of the phenoxy moiety, exhibited a noteworthy cytotoxic effect against A2058 cells, thus becoming the most efficient compound of the series. A weak or no activity was detected for the other compounds. All the compounds under investigation have proven to be safe on the non-tumoral human embryonic renal cell line Hek-293. Furthermore, several studies agree on the importance of microtubule dynamics in governing the main cell functions, such as mitosis, maintaining of cell shape, cell division, intracellular transport, and chromosomes  
Figure 6: A schematic representation of the tetrameric 2\(\alpha\)2\(\beta\) tubulin structure. \(\alpha\) chain is reported as cyan ribbons, \(\beta\) as Golden ribbons (**A**). Compounds **2d**, **2k** and **2i** superpose to vinblastine binding site (**B**). (**C–E**) report the compounds **2d**, **2k** and **2i**, respectively, drawn as white, green and dark magenta sticks, bound to the vinblastine binding task of the protein.  
segregation. Following this, we tested the ability of compound **2k** to target tubulin polymerization performing both an immunofluorescence assay in A2058 cells and an in vitro tubulin-polymerization assay. Data extrapolated from both the experiments evidenced that the compound **2k** interferes with the microtubule dynamics because of its capability of preventing tubulin polymerization. These interesting findings were also supported by docking studies that shown the ability of **2k** to superpose to vinblastine binding site in tubulin structure. Next, we performed a TUNEL assay proving that compound **2k** can induce the apoptotic process in A2058 melanoma cell line. To conclude, although most of the examined compounds did not display an encouraging activity, we aimed to focus our attention on a single compound (**2k**) from which we obtained auspicious results that make it a promising ally in the fight against cancer. However, further studies must be carried out to optimize this candidate and validate its efficacy in vivo.  
Conceptualization, M.S.S.; methodology, D.I. and A.C. ([PERSON]); software, C.R.; validation, C.F., formal analysis, A.B. and A.C. ([PERSON]); investigation, A.B. and A.C. ([PERSON]); data curation, D.I. and J.C.; writing--original draft preparation, A.C. ([PERSON]) and A.B.; supervision, M.S.S. and C.F. All authors have read and agreed to the published version of the manuscript.  
The work of [PERSON] has been partially supported by a grant from the Italian Ministry of Health (Ricerca Corrente).  
Not applicable.  
The authors declare no conflict of interest.