## 4 Discussion
Furthermore, as they are not P-gp substrates, their efficacy in therapies against resistance mediated by this protein is likely to be increased. Only the sterol derivatives (ergosta-4,6,8(14),22-tetra-3-one; (22E,245)-ergosta-4,22-dien-3-one, and (22E)-19-norergosta-5,7,9,22-tetra-3\(\beta\)-ol) could cross the BBB. Some P-gp modulatorsare competitive inhibitors of the ability of anticancer drugs to bind to P-gp. However, they are also P-gp substrates and are expelled from cells [52]. Only the compounds (22\(E\),24\(S\))-ergosta-4,22-dien-3-one and (22\(E\))-19-noregosta-5,7,9,22-tetra-3\(\beta\)-ol violated [PERSON]'s Rule of Five--specifically, the criterion MLOGP > 4.15--indicating high lipophilicity. Additionally, the control compound tariquidar violated the rule regarding the molecular weight (MW > 500 Da). Although they pose a challenge, these violations are common in natural products, which often have high biological potential. Optimization strategies can overcome these limitations. Tariquidar is an example of this; despite its violations and toxicity, it has progressed to clinical trials. Therefore, such cases should be evaluated flexibly in the early stages of development. This research's findings align with previous studies that have shown that cancer cells with microRNA-378 are more aggressive and resistant to drugs, requiring higher doses of chemotherapy to be eradicated. However, therapy with ergosterol peroxide, obtained from _Ganoderma lucidum_, overcomes this resistance, inducing the cell's death more efficiently, even at lower doses [53]. In a more recent study, an ethanolic extract of _Ganoderma lucidum_ showed an ability to reverse MDR in cells and mice with resistant hepatocellular carcinoma, inhibiting the activity of P-gp without modifying its expression levels [54]. Although no studies have been found on the modulation of P-gp by the compound ergosta-4,6,8(14),22-tetraen-3-one, some studies have highlighted the antitumor potential of this compound and its derivatives. One study demonstrated antiproliferative activity in hepatocellular carcinoma cells (HepG2), induced by caspase-mediated apoptosis, with the interruption of the cell cycle in G2/M and changes in the regulation of Bax and Bcl-2 [55]. In another study, the same compound extracted from _Antrodia cinnamonea_ reduced cancer-initiating cells in head and neck carcinoma, inhibiting STAT3 and Src, promoting cell differentiation, and restoring chemosensitivity in combination with chemotherapy [56]. We found no specific studies evaluating lucidumol A in the modulation of P-gp. However, research has shown that lucidumol D, extracted from _Ganoderma lingzhi_, exhibits selective cytotoxicity towards the MCF-7, HepG2, HeLa, Caco-2, and HCT-116 cancer cell lines [57]. Additionally, in another study, lucidumol A extracted from _Ganoderma lucidum_ was shown to have an effective anticancer and anti-inflammatory effects against colorectal cancer and RAW264.7 cells. In vitro studies, this compound reduced the metastatic potential at low concentrations and reduced inflammatory markers and cytokine levels in RAW264.7 cells subjected to treatment [20]. It is important to note that these cell lines were not described by the authors as cells with the overexpression of P-gp. The results of this study and previous research show that certain compounds from mushrooms may overcome MDR. These compounds are predicted to have a high affinity for P-gp, exhibit low toxicity, and have interesting pharmacokinetic properties, suggesting that they may render cancer cells more sensitive to chemotherapy treatments, according to molecular docking studies. These findings imply that these substances could be used as possible aids in cancer therapy, increasing the effectiveness of medication and decreasing the resistance to traditional treatments. This in silico work provides a predictive model, but more research is needed to elucidate and validate the therapeutic safety of these drugs.