## 3 Results
### Identification of Phytochemicals and In Silico Molecular Docking Studies  
Gas chromatography coupled with mass spectrophotometry was used to identify phytochemicals present in crude extract of the shoots of _M. sinaica_, and the data are presented in Table 2. The major identified compound was 2-Tridecen-1-ol, which represents 35% of the crude extract, followed by 1,2-benzenedicarboxylic acid (19%); and hexadecanoic acid (17.4%).  
Figure 3: Cell cycle distribution profiles for the effects of the methanol extract of shoots of _M. sinaica_ on MDA-MB-231. Cells were incubated for 24 h (**A**) without (control) and (**B**) with the methanol extract of shoots of _M. sinaica_ and stained with PI. Following Fluorescence Activated Cell Sorting (FACS) analysis, DNA histograms were analyzed using CXP software V. 3.0. Data represent mean \(\pm\) S.D. and the difference was statistically significant at (\(p<0.05\)) compared to control.  
In order to investigate the possible mechanism through which the crude extract of shoots of _M. sinaica_ exert the anticancer activity, it is very important to determine the protein targets for each of these major compounds. The chemical structure for each compound was deduced from the PubChem database, and potential protein targets were obtained using an online protein target prediction tool [17]. The target prediction report for 2-Tridecen-1-ol is presented in Table S1. M-phase inducer phosphatases 1 (CDC25A; uniport ID P30304) and M-phase inducer phosphatases 2 (CDC25B; unipotent ID P30305) were identified as the proteins target for molecule \"2-Tridecen-1-ol\" with the highest probability. Table S2 represents the target prediction report for 1,2-benzenedicarboxylic acid. Tyrosyl-DNA phosphodiesterase 1 (TDP1; uniport port ID Q9 NUW8) was identified as a possible protein target for molecule 1,2-benzenedicarboxylic acid with the highest probability among other predicted protein targets. The protein target report for hexadecanoic acid is shown in Table S3. Various tissue-specific fatty acid-binding proteins have been predicted with the highest % probability to be the protein target for hexadecanoic acid.  
In silico molecular docking was used to find out that the binding of 2-Tridecen-1-ol might bind to the active domain of CDC25A and CDC25B. The three-dimensional structure of the catalytic domain of the human CDC25A (IC25) and CDC25 B (ICWR) was downloaded from RCSB protein data bank ([[https://www.rcsb.org/](https://www.rcsb.org/)]([https://www.rcsb.org/](https://www.rcsb.org/))). Molecular docking yielded nine different conformations in order of increasing binding energy. The docking result of the ligand with the lowest binding energy is analyzed here. The lowest binding energy for 1C25 and 1 CWR was found to be \(-\)4.2 and \(-\)4.4 kcal/mol, respectively. The docked structure of trans-2-tridecen-1-ol and 1C25 is shown in Figure 4A. Glu378 and Phe379 formed hydrogen bonds with 1C25 at a distance of 2.26 and 2.63 A, respectively. Other amino acids such as Val367, Ile376, and Ile381 are also involved in the binding through hydrophobic interactions. The docked trans-2-tridecen-1-ol in structure 1 CWR is shown in Figure 4B. The ligand interacted at the hydrophobic region of the protein,  
\begin{table}
\begin{tabular}{c c c c} \hline \hline
**Compound** & **Structure** & **Retention Time (s)** & **\% Area Covered** \\ \hline
2-Tridecen-1-ol & & 20.85 & 34.87 \\
7-Butyl-bicyclo [4.1.0]heptane & & 21.34 & 7.41 \\
1,2-Benzenedicarboxylic acid & & 22.32 & 18.62 \\
1,19-Eicosadiene & & 22.67 & 7.27 \\ Hexadecanoic acid & & 23.05 & 17.4 \\ Trans phytol & & 23.81 & 15.11 \\ \hline \hline \end{tabular}
\end{table}
Table 2: Photochemical present in MeOH extract of shoots of _M. sinaica._and Leu398, Cys484, Arg485, Arg488, and Met405 were involved in this protein-ligand complex formation.