Reactive oxygen species (ROS) are cellular byproducts that comprise both free and non-free radicals, such as hydrogen peroxide. Reactive oxygen species (ROS) are overproduced when the body's inherent defences against free radicals are compromised. These ROS, which result from the body's oxidative stress process, have an effect on macromolecules like protein, DNA, and lipid[25]. Many phyto-constituents, including carotenoids, flavonoids, and anthocyanins, contain phenolic moieties that cooperate to inhibit the ROS-induced cascade of events.[26].Antioxidants are able to perform their function through altering DNA's binding mode[27]. Based on its DPPH and iron chelating scavenging activity, this study found that the crude chloroform extract possesses potent antioxidant properties. The DPPH quenching IC50 value is recorded at 38.691g/mL, which is the lowest of all the extracts and is nearly equivalent to the standard. The iron reduction IC50 for the crude chloroform extract was 59.81 g/mL. Our research indicates that the extract has a high total phenolic content, as well as high levels of flavonoids and antioxidant phytochemicals. We tested the extracts' anticancer effects on a human lung cancer cell line. Our research showed that the chloroform extract of the same source presented remarkable anti-mutagenic response against lung cancer cell line as well, even though methanol extract of the areal part of F. indica was shown to inhibit proliferation of human colon cancer cell via inhibition of ROS generation [9]. Cancer cells were inhibited by 22.83% in a crude chloroform extract. The demise of cells was clearly seen under an inverted microscope. Changes in the cell's morphology, such as the fragmentation of the nucleus, the destruction of the mitochondria, or the clustering of the chromosomes, can all lead to cell death. The GCMS analysis was performed to identify the active chemicals in various biological processes. Although GCMS analysis of plant root extract has been previously reported[13], the examination of the plant's aerial component was new to this investigation. Phytol was the most abundant component in both the chloroform and methanol extracts, as determined by our research. Phytol, an essential oil extracted from plants, has been shown to have beneficial effects as an antioxidant, anti-inflammatory, anti-hyperlipidemic, and antibacterial [28]. Inducing apoptosis of human gastric cancer cells has been demonstrated for this compound[29], albeit the mechanism of action is poorly explained. Antioxidant, antibacterial, and antifungal properties have all been attributed to cyclopentapropanoic acid,2-undecyl,methyl ester, another significant component in crude chloroform extract [30].There are also significant amounts of chemicals such deacane, benzyl alcohol, pentadecanoic acid, 14-methyl, undecane, and 9,12-octadecadienoic acid. Hepatoprotective, antihistaminic, and hypocholesterolemic effects of 9,12-octadecadienoic acid have been described [31].
Our investigation in computer-aided in-silico methods was supported by a molecular docking analysis. Methods like these also show how ligands interact with receptor proteins to boost the biological effects of phyto-constituents. The proteins topoisomerase II and calvasculin were chosen because they are unique to human lung cancer cell lines, and porin was chosen because it has been shown to have greater expression in the A549 cell line[32]. The binding affinity was best expressed by topoisomerase II, which was chosen along with two other proteins. The highest binding affinities (-6.6kcal/mol and − 6.7kcal/mol, respectively) with topoisomerase II were demonstrated by phytol and pentadecanoic acid,14-methyl. These substances in the extracts may be responsible for inhibiting the growth of human lung cancer cells because the protein is specific to this cell line. Binding affinities more than − 5kcl/mol were also observed for benzyl alcohol and neophytadiene. Therefore, with the necessary structural alterations, these phyto components can be considered lead compounds to create medicines for human lung cancer. The reaction can be transmitted via contact between the chemical and the target protein. Because of its ability to reduce cellular stress, aid in DNA damage repair, and limit oncogene expression, TP53 is considered a key tumor suppressor protein[36]. Breast cancer cells can be stimulated to express TP53 by a wide variety of phytochemicals, including curcumin, coumarin, and vanillin[33]. The expression of TP53 in cancer cells is one mechanism by which the active ingredients can confer anticancer effects. In our PASS study, we have carried out prediction study against numerous cell line of human lung cancer correlated to our in-vitro investigation. In that case, pentadecanoic acid,14 methyl seemed to exhibit prominent anticancer activity with Pa value 0.543 against non-small cell lung cancer. Our molecular docking score of the phyto-constituents with TP53 protein reveals that phytol can be a promising lead compound to exhibit anticancer property. Phytol has been proved to interact with active sites of both topoisomerase and TP53 protein. In case of topoisomerase, hydrophobic interaction was carried out by pi alkyl and alkyl bond formation with Ile A: 217, Ile:125, Ile:121l and Phe A:142. Hydrogen bond was formed with Asn A: 121 while topoisomerase was interacted with phytol. Phytol formed bonds by both hydrophobic-interaction with alkyl and pi-alkyl (Phe A:1553; Ile A:1587; Leu A:1547) interlinkage as well as polar interaction with conventional hydrogen bond (Asp A:1521). Pentadecanoic acid, 14 methyl was identified which showed maximum docking affinity (-6.7 kcal/mol) against topoisomerase contrast to the other compounds in the experiment. Moreover, the compound showed promising interaction to this particular protein rather than other proteins. As topoisomeraseII is a specific protein involved in expression of lung cancer cell line, it can be hypothesised that pentadecanoic acid,14 methyl can be a lead compound to be anti-carcinogenic via a particular pathway to inhibit expression of that specific protein.