# Molecular Targets of Minor Cannabinoids in Breast Cancer: In Silico and In Vitro Studies
## 2 Results
### Molecular Docking Results
\begin{tabular}{c} Possible hydrogen bond with His 524; close to Asp351 \\ with Trp383 \\ \end{tabular}  \\ \hline \({}^{7}\)-Eudesmyl-cannabigerolate & Cannabigerol & \(-7.7\) & \ \begin{tabular}{c} Close to Asp351; hydrophobic interactions \\ with Trp383 \\ \end{tabular}  \\ Cannabinerolic acid & Cannabigerol & \(-7.7\) & \begin{tabular}{c} Hydrogen bonds with His524 and Glu353; close to Asp351; hydrophobic interactions with Trp383 \\ \end{tabular}  \\ \hline \begin{table}
\begin{tabular}{c c c c} \hline \hline
**Cannabinoid Name** & **Cannabinoid Class** & \
\begin{tabular}{c} **Binding** \\ **Affinity (kcal/mol)** \\ \end{tabular} & **Relevant Interactions** \\ \hline Cannabichromenic acid & Cannabichromen & \(-\)7.7 &
2-Methyl-2-(4-methyl-2-pentyl)-7-propyl-2H-1-benzopyran-5-ol & Cannabichromene & \(-\)7.6 & \begin{tabular}{c} Close to Asp351; hydrophobic interactions \\ with Phe044 \\ \end{tabular} \\ \hline Cannabichromevarin & Cannabichromen & \(-\)7.6 & \begin{tabular}{c} Close to Asp351; hydrophobic interactions \\ with Phe044 \\ \end{tabular} \\ \hline Cannabichromevarin acid & Cannabichromene & \(-\)7.6 & \begin{tabular}{c} Hydrogen bond with Asp351; hydrophobic interactions \\ interactions with Trp383 \\ \end{tabular} \\ \hline (-)-3\({}^{\prime\prime}\)-Hydroxy-\(\Delta^{4^{\prime\prime}}\)-cannabichromene & Cannabichromen & \(-\)7.2 & \begin{tabular}{c} Close to Asp351; hydrophobic interactions \\ with Trp383 \\ \end{tabular} \\ \hline Cannabichromene & Cannabichromene & \(-\)7.0 & \begin{tabular}{c} Close to Asp351; hydrophobic interactions \\ with Trp383 \\ \end{tabular} \\ \hline Cannabinol & Cannabinol & \(-\)8.9 & \begin{tabular}{c} Hydrophobic interactions with Trp383 \\ and Phe044 \\ \end{tabular} \\ \hline Cannabinol methyl ether & Cannabinol & \(-\)8.4 & \begin{tabular}{c} Close to Asp351; hydrophobic interactions with \\ Trp383 and Phe404 \\ \end{tabular} \\ \hline Cannabinol-C\({}_{4}\) & Cannabinol & \(-\)6.6 & \
\begin{tabular}{c} Close to Asp351; hydrophobic interactions with \\ Trp383 and Phe404 \\ \end{tabular} \\ \hline (10S)-Hydroxycannabinol & Cannabinol & \(-\)6.4 &
\begin{tabular}{c} Hydrogen bond with Val534; close to Asp351; hydrophobic interactions with Trp383 \\ and Phe404 \\ \end{tabular} \\ \hline
8-Hydroxycannabinol & Cannabinol & \(-\)6.0 &
\begin{tabular}{c} Close to Asp351; hydrophobic interactions with \\ Trp383 and Phe404 \\ \end{tabular} \\ \hline
10\(\alpha\)-Hydroxyhexahydrocannabinol & Miscellaneous & \(-\)8.0 &
\begin{tabular}{c} Close to Asp351; hydrophobic interactions \\ with Trp383 \\ \end{tabular} \\ \hline
2-Geranyl-5-hydroxy-3-n-pentyl-1,4- & Miscellaneous & \(-\)7.9 &
\begin{tabular}{c} Close to Asp351; hydrophobic interactions \\ with Trp383 \\ \end{tabular} \\ \hline
9\(\alpha\)-Hydroxyhexahydrocannabinol & Miscellaneous & \(-\)7.9 &
\begin{tabular}{c} Close to Asp351; hydrophobic interactions \\ with Trp383 \\ \end{tabular} \\ \hline \hline \end{tabular}
\end{table} 
Table 2: Cont. Considering the overall results observed for these two targets, 36 out of the 129 phytocannabinoids were simultaneously considered as potential AIs and ER\(\alpha\) antagonists (Table 3, Supplementary Figure S3). However, only 7 out of those 36 selected cannabinoids, namely **CBDV**, **CBDB**, **CBDA**, **CBDM**, **CBCA**, **CBGVA**, and **CBC** (Figure 2), are commercially available and, thus, were selected for in vitro studies. Furthermore, two other cannabinoids, **CBG** and **CBN** (Figure 2), that did not show potential to act as AIs or ER\(\alpha\) antagonists, but have growing pharmacological interest and have been studied in other breast cancer subtypes [75,76], were also included in our study. Figure 2 highlights their structural and biosynthetic relationships. The docking positions of the nine selected minor cannabinoids in aromatase, as well as the most important residues for their binding, are represented in Figure 3.