Molecular Docking Studies of Botanical Beverage Mix Berries (LIFEGREENTM) against Breast Cancer Cells from Targeted Protein 1QQG, 7B5Q & 7B5O & Uterine Fibroid from Targeted Protein 2AYR, 6T41 & 3GRF ()
1. Introduction
1.1. LifeGreen™
LifeGreen™ Cactus Powder is known as a highly concentrated cactus extract which made using 22 different types of fruit and vegetable extracts, Italian mixed berries, and TRUEBROC® broccoli seed extract, a US-patented ingredient, Oxxynea® is a popular French health drink. According to studies, cactus polysaccharides having the ability to boost immunity and inhibit abnormal cell developments when consumed over time. In addition, Truebroc® broccoli seed extract promotes aberrant cell death, reduces abnormal cell blood supply, and prevents abnormal cell reproduction and spread. Figure 1 shows the packaging of the LifeGreen™ Beverage [1] .
1.2. Ingredients and Benefits
LifeGreen™ contains Italian Premium Mixed Berries (Blueberry, Blackcurrant, Raspberry, Elderberry, Red Grape, Strawberry, Cranberry), Cactus Powder, Oxxynea® (Green Tea Extract, Red Grape Extract, White Grape Extract, Bilberry, Carrot, Grapefruit, Papaya, Pineapple, Strawberry, Apple, Apricot, Cherry, Orange, Broccoli, Green Cabbage, Onion, Garlic, Olive, Cucumber, Blackcurrant, Tomato, Asparagus), TRUEBROC® broccoli seed extract, and immune deficiencies include fatigue and weakness, allergies, and the need to restore immunity. Moreover, LifeGreen™’s ingredients, particularly Cactus Powder, have been extensively researched and studied for their anticancer effects. Figure 2 shows the picture of LifeGreen™ Beverage drink [1] .
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Figure 1. Lifegreen™ Beverage sachet and packaging [1] .
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Figure 2. Lifegreen™ Beverage drink [1] .
1.3. Cancer Cell Growth
The body creates molecules are known as growth factors, which govern cell division. Growth factors occur in a number of types and each operates differently. Some growth factors advise cells on how to specialize and what type of cell they should become. Some cause cell division and proliferation to generate new cells. Cells can be told to cease growing or die. Growth factors operate by binding to cell surface receptors. This sends a signal to the cell’s inside, initiating a sequence
of complex chemical reactions. There are multiple different growth factors [2] . These include epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), platelet-derived endothelial growth factor (PDGF), and fibroblast growth factor (FGF), which all regulate cell growth. Each growth factor works by attaching to its corresponding cell surface receptor. For example, epidermal growth factor (EGF) interacts with the EGFR. Tyrosine Kinases are chemical messengers (enzymes) that control cells’ capacity to divide and grow. Similar to an “on-off” switch feature, Tyrosine kinase is activated when a growth factor attaches to a cell’s surface. This will prompt cell division as shown in Figure 3 [2] .
1.4. Uterine Fibroids
Fibroids, also known as leiomyoma or myoma, are frequent tumors that develop in the uterine wall or muscle. Fibroids in women of their reproductive years can present as single or multiple growths. Fibroids are uncommon in women who have not yet started menstruation, although they afflict around 20% of women of reproductive age. Usually, growth slows after menopause [3] . Fibroids can be classified into various types based on where they exist. The submucosal fibroid is a fibroid that usually develops inside the uterus while intramural fibroid is a fibroid that develops within the musculature of the uterine wall. Moreover, subserosa fibroid is known as a uterine fibroid that protrudes from the body. Lastly, a fibroid with a stalk that extent from the uterus into the pelvis or else discovered inside the inner uterine cavity and extends through cervix is known as pedunculated fibroid. Figure 4 shows the locations of the various types of fibroids [4] .
Despite their benign nature, they can grow rapidly and dramatically [5] . They produce heavy and irregular menstrual bleeding (HMB), which leads to severe anemia, dysmenorrhea, pelvic pressure and discomfort, urinary incontinence, dyspareunia, infertility, premature labor, and recurrent early and late pregnancy losses [6] . More than 70% of women have UFs, with only around 30% experiencing
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Figure 3. The cancer cells growth factor that affects the body [2] .
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Figure 4. The common locations for uterine fibroid to occur [4] .
symptoms, making UFs the most common clinical cause for hysterectomy, which removes a woman’s capacity to produce early [7] .
1.5. Breast Cancer Cells (IRS-1)
Breast cancer is the world’s second most frequent illness, affecting more women than any other malignancy. The two most common types of breast cancer are ductal and lobular. In situ (localized to a single location), ductal and lobular cancers account for 85% - 90% and 8% of all breast cancers, respectively. In addition, aggressive inflammatory breast cancers exist, as do invasive ductal and lobular tumors. Although chemotherapy has been demonstrated to improve breast cancer patients’ survival rates, a significant minority of individuals only have a brief response to the treatment before succumbing to metastatic disease. IRS1 has been shown to enhance breast cancer cell growth rather than preventing metastasis [8] . Figure 5 shows the process of IRS-1 occurring before metastasis happen in which responsible for the growth of breast cancer cell [9] .
1.6. BRCA 1 & BRCA 2
People protecting injurious variants in BRCA1 or BRCA2 genes aspect significantly higher risks of emerging various cancers, together with breast, ovarian, fallopian tube, and primary peritoneal cancers [10] . The incidence of these mutations significantly rises the lifetime risk of cancer onset, with pretentious people often facing earlier age of cancer diagnosis equated to the general population. While BRCA mutations are sturdily associated with increased cancer risk, the extent of risk variability amongst carriers is inclined by various factors [10] . These issues include environmental contacts, lifestyle choices, hormonal effects, and genetic modifiers that may interrelate with BRCA mutations to modulate cancer susceptibility. In spite of extensive research, some of these factors remain somewhat characterized, highlighting the need for further investigation into the complex interplay between genetic and environmental determinants of cancer risk [11] .
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Figure 5. The process of IRS-1 occurring before metastasis happens causes breast cancer cell growth.
1.7. MED12-a
Tumors did not exhibit more than one mutation, and previous research did not address the MED12 mutation status of multiple leiomyomas in one patient. Multiple uterine leiomyomas appeared to have a higher incidence of MED12 mutations compared to single uterine leiomyomas (72.73% versus 59.26%), although this difference was not statistically significant [12] . Moreover, patients with multiple leiomyomas exhibited smaller mean sizes of leiomyomas significantly, consistent with previous research. Nearly twofold difference in MED12 mutation frequency between multiple and single uterine leiomyomas from a cohort of 122 patients. However, they could not establish a significant association between MED12 mutation and tumor size [12] . Therefore, larger sample sizes are required to evaluate the relationship between MED12 mutation frequency and the number or size of uterine leiomyomas. MED12 mutation has also been detected in other uterine tumors such as leiomyosarcomas and smooth muscle tumors of uncertain malignant potential, but not in tumors of other organs. Interestingly, breast fibroadenoma also harbored highly frequent MED12 mutations. Whole exome sequencing revealed no genes other than MED12 mutation in MED12 mutation-positive and -negative leiomyomas, suggesting that MED12 mutation alone may be sufficient for leiomyoma tumorigenesis [12] .
1.8. Endometriosis
Endometriosis is a multifaceted gynecological state affecting 10% - 15% of reproductive-age females and around 70% of women facing persistent pelvic pain. While the ovaries and pelvic peritoneum are the primary sites of endometriotic lesions, they can also patent in various other locations inside the body [13] . The etiologic of endometriosis-associated pain remains poorly unspoken, with inflammation broadly believed to play a significant role. Despite fruitful surgical interventions, recurrence of symptoms is communal, underscoring the chronic nature of the condition. Endometriosis poses a substantial economic burden, particularly in countries like India, with costs estimated at approximately 1 to 2 lakhs per affected woman. The participation of structures such as the uterosacral ligaments, posterior vaginal wall, rectovaginal space, intestines, and urinary system is frequently experiential in endometriosis cases. Various studies have explored the molecular mechanisms underlying endometriosis-associated infertility, highlighting factors such as apoptosis, cell cycle alterations, and oxidative stress in granulosa cells. Moreover, recent research endeavors have sought to elucidate the role of MED12 mutations in endometriosis pathogenesis, revealing disruptions in cellular interactions and signaling pathways [13] .
2. Methodology
LifeGreen™ uses proteomic computing to identify compounds that can interact with IRS-1 (breast cancer cells) as well as BRCA 1 and 2 - 1QQG, 7B5Q & 7B5O and endometriosis and mutated Med-a (uterine fibroid)-2AYR, 3GRF & 6T41 proteins [14] . Furthermore, additional research and investigations on each effective molecule with the highest binding energy and its advantages to human health. Figure 6 shows the general methodology of this research while Figure 7
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Figure 6. Overview of the whole methodology
until Figure 10 shows the in-depth methodology.
2.1. Phase 1: Metabolomics Analysis
In Figure 7, in chromatography process, separation was performed using Thermo Scientific C18 column (AcclaimTM Polar Advantage II, 3 × 150 mm, 3 um particle size) on an UltiMate 3000 UHPLC system (Dionex) [15] . Gradient elution was performed at flow rate of 0.4 ml/min and 40˚C column temperature using H2O + 0.1% Formic Acid (A) and 100% ACN (B) with 22 minutes total run time. The injection volume of sample was 5 ul. The gradient started at 5% B (0 - 3 min); 80% B (3 - 10 min); 80% B (10 - 15 min) and 5% B (15 - 22 min). For mass-spectrophotometry, sample is analyzed with positive ionization parameter [15] , Table 1.
In data processing steps, the accurate mass data of the molecular ions, provided by the TOF analyzer, were processed by Compass Data Analysis software
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Figure 7. Phase 1: Metabolomics Analysis.
(Bruker Daltonik GmbH). Further process by using metfrag (In silico fragmentation for computer assisted identification of metabolite mass spectra) in order to pull out the list of compound present in each peak presented by LC-MS/TOF [16] gi-bin/portal.py#welcome.
2.2. Phase 2: Compound Analysis
Before proceeding with the analysis, each compound underwent meticulous quality control procedures employing ZoBio by NMR, ensuring the integrity and reliability of subsequent results. Following this initial step, the compounds were subjected to an array of sophisticated analyses aimed at elucidating their properties and functions. Significance testing was employed to discern meaningful patterns and deviations within the dataset, shedding light on potential biological implications. Quantitation-pattern recognition techniques were applied to discern quantitative relationships and trends within the data, facilitating a deeper understanding of compound behavior. Compound assignment methodologies were utilized to accurately identify and classify each compound, ensuring precise cataloguing and characterization. Finally, functional assessment protocols were implemented to assess the biological activities and potential applications of the compounds, providing valuable insights for further research, Figure 8.
2.3. Phase 3: Protein Analysis
During this third phase of the study Figure 9, an extensive protein analysis was undertaken utilizing resources available on the Protein Data Bank (PDB) https://www.rcsb.org/ website [17] . This involved a meticulous examination of protein structures and relevant data to glean insights into the molecular
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Figure 9. Phase 3: Protein analysis of 1QQG, 2AYR, 3GRF, 6T41, 7B5Q and 7B5O.
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Table 2. Breast cancer and uterine fibroid flow.
underpinnings of the investigated conditions. To augment our understanding, a thorough literature review was conducted. This involved delving into published studies and scientific literature to identify and elucidate the roles of specific proteins implicated in breast cancer and uterine fibroids. By synthesizing information from various sources, we aimed to pinpoint key proteins associated with these diseases, providing a comprehensive foundation for our research. Table 2 serves as a comprehensive repository of the gathered results pertaining to both breast cancer and uterine fibroids. This tabulated data offers a detailed overview of the proteins identified and their respective implications in the pathogenesis of these conditions. Through meticulous documentation and analysis, we aim to uncover potential biomarkers and therapeutic targets, contributing to the advancement of diagnostics and treatment strategies for breast cancer and uterine fibroids.
2.4. Phase 4 and Phase 5: Protein-Ligand Interactions and Molecular Docking
In Figure 10, proteomic molecular docking represents a sophisticated computational approach used in the field of proteomics to predict and analyses the
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Figure 10. Phase 4: Protein-Ligand Interaction (RPBS) and Phase 5: Molecular Docking.
interactions between proteins and other molecules, Achilles Blind Docking Server, https://bio-hpc.ucam.edu/achilles/. This method integrates principles from both proteomics and molecular docking, leveraging computational algorithms to simulate and predict the binding affinity and spatial orientation of proteins with various ligands, substrates, or inhibitors. The process typically begins with the identification of target proteins of interest through proteomic techniques such as mass spectrometry or protein microarrays. Once the target proteins are identified, molecular docking algorithms are employed to simulate the binding interactions between these proteins and small molecules, peptides, or other proteins. These docking algorithms use complex scoring functions and search algorithms to explore the conformational space and predict the most energetically favorable binding poses between the proteins and their ligands. By analyzing these predicted binding conformations, researchers can gain valuable insights into the molecular mechanisms underlying protein-ligand interactions, including the identification of key binding residues and structural determinants. Proteomic molecular docking holds significant promise for various applications in drug discovery, structural biology, and systems pharmacology. It enables the screening of large compound libraries to identify potential drug candidates or lead compounds that modulate the activity of target proteins implicated in diseases. Additionally, it facilitates the elucidation of protein-protein interaction networks and signaling pathways, providing valuable insights into complex biological processes and disease mechanisms.
3. Results
3.1. LC-TOF-MS
In Figure 11, the chemical composition of LifeGreenTM samples, LC-TOF MS analysis emerged as a pivotal investigative tool, yielding a comprehensive compound spectrum graph delineating the peaks indicative of molecular entities
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Figure 11. Shows the compound spectrum graph and its peaks.
present within the samples. Notably, the analysis revealed approximately 140 distinct compounds, with varying degrees of detectability attributable to the inherent limitations posed by sample composition and instrumental sensitivity. Within this array, certain compounds were readily discernible, while others remained undetected, a phenomenon attributed to the relatively sparse population of compounds within the samples.
In our examination of the compound spectrum graph, each peak was characterized by its corresponding retention time (RT) in minutes, intensity, signal-to-noise ratio, maximum mass-to-charge ratio (m/z), and area under the peak. These parameters collectively provided insights into the abundance, purity, and spectral characteristics of the identified compounds. Furthermore, to augment our understanding of the molecular identities associated with the observed peaks, collision energy (eV) values were assigned to selected peaks, facilitating subsequent tandem mass spectrometry (MS/MS) analyses.
To unveil the chemical identities corresponding to the observed peaks, we employed the computational tools afforded by MetFrag. Leveraging the spectral information encapsulated within the LC-TOF MS data, MetFrag facilitated the deconvolution of complex spectra, enabling the retrieval and annotation of putative compounds associated with each peak, Table 3.
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Table 3. Shows the compound spectrum list of LifeGreenTM.
3.2. MetFrag
In Table A1, through the utilization of MetFrag, a comprehensive compilation of compounds was extracted from each designated peak within the compound spectrum graph generated via LC-TOF MS analysis of the LifeGreenTM samples. These compounds were meticulously identified, characterized, and annotated, facilitating the elucidation of their chemical nature and potential functional attributes [18] .
Each identified compound was assigned a canonical SMILES (Simplified Molecular Input Line Entry System) representation, serving as a concise yet comprehensive descriptor of its molecular structure. Additionally, molecular formulas were determined, encapsulating the precise arrangement of atoms constituting each compound, thus providing crucial insights into their elemental composition and stoichiometry [18] . The benefits and potential applications associated with each identified compound were elucidated, leveraging existing knowledge and literature resources. These benefits encompassed a diverse array of domains, including pharmaceuticals, agriculture, food science, cosmetics, and environmental remediation, among others [18] .
By integrating the structural information derived from canonical SMILES notation and molecular formulas with the contextual understanding of their functional properties, a holistic perspective on the chemical constituents present within the LifeGreenTM samples was attained, Table 4.
3.3. LifeGreen™ Compounds
3.3.1. Swiss-ADME
The compiled list of 117 compounds extracted from the LC-TOF MS analysis of LifeGreenTM samples underwent comprehensive evaluation via SwissADME, a robust computational platform designed to assess various pharmacokinetic and pharmacodynamic parameters crucial for drug discovery and development [19] .
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Table 4. Shows list of compounds in LifeGreenTM.
This systematic scrutiny aimed to discern the compounds exhibiting favorable pharmacological properties conducive to combatting breast cancer and uterine fibroids, thus offering promising avenues for therapeutic intervention. Within this rigorous evaluation framework, several key metrics were scrutinized, including compliance with Lipinski’s Rule of Five, a pivotal criterion for predicting oral bioavailability and permeability of potential drug candidates. Additionally, lead-likeness, hydrogen bond donor and acceptor counts, and bioavailability scores were meticulously examined, offering valuable insights into the compounds’ drug-like properties and therapeutic potential [19] . By leveraging the insights gleaned from SwissADME analysis, compounds exhibiting optimal pharmacokinetic profiles and bioavailability were identified as prime candidates for further investigation and therapeutic development. These compounds, characterized by their propensity to permeate biological barriers, maintain favorable drug-like properties, and exhibit high bioavailability, hold promise in the targeted management and mitigation of breast cancer and uterine fibroids [19] . The systematic integration of LC-TOF MS analysis, compound identification, and SwissADME evaluation represents a pivotal step towards the rational design and discovery of novel therapeutics aimed at addressing the unmet clinical needs associated with breast cancer and uterine fibroids. Through this concerted effort, the identification of lead compounds with enhanced efficacy and safety profiles heralds a significant stride towards personalized and precision medicine approaches tailored to combat these debilitating diseases [19] .
3.3.2. RPBS
Following the initial filtration process employing SwissADME, which identified 51 compounds with potential efficacy against breast cancer and uterine fibroids, a subsequent analysis utilizing RPBS was conducted to evaluate rotatable bonds and energy profiles. This additional scrutiny yielded a refined subset of 34 compounds, each characterized by optimal structural flexibility and energetics conducive to molecular docking studies [20] . The RPBS assessment provided crucial insights into the molecular dynamics of the selected compounds, elucidating their ability to adopt diverse conformations and facilitating efficient interactions with target biomolecules implicated in disease pathogenesis [20] . By prioritizing compounds with favorable rotatable bond counts and energy profiles, this iterative screening process enhances the rational selection of lead candidates poised for further preclinical evaluation, Table 5 [20] .
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Table 5. Shows down streaming results from RPBS tools.
Out of the initial pool of 34 compounds, only 22 demonstrated the remarkable capability to bind with all six protein diseases associated with both breast cancer and uterine fibroid. This subset of compounds exhibits broad-spectrum activity, indicating their potential to target multiple pathological pathways implicated in the progression of these diseases. Their ability to interact with diverse protein targets underscores their versatility and promise as candidate therapeutics [20] . In contrast, the remaining 12 compounds displayed a more selective binding profile, interacting with a subset of two to four protein diseases. While these compounds may exhibit efficacy against specific disease subtypes or pathways, their narrower spectrum of activity suggests a more targeted mode of action. Despite this selectivity, these compounds still hold considerable therapeutic potential and merit further investigation for their specific applications in breast cancer and uterine fibroid management [20] .
3.3.3. Dogsitescorer
Dogsitescorer, a specialized computational tool, is employed to discern the highest binding energy exhibited by potential compounds against protein targets implicated in breast cancer and uterine fibroid pathogenesis. Subsequently, coordinates corresponding to the identified binding sites are extracted from the output generated by Dogsitescorer [21] . These coordinates play a pivotal role in elucidating the precise molecular interactions between the candidate compounds and their respective protein targets. By pinpointing the specific binding sites on the protein surfaces, these coordinates provide valuable insights into the molecular mechanisms underpinning the observed binding affinities [21] .
Furthermore, the coordinates obtained from Dogsitescorer serve as crucial input parameters for subsequent molecular docking simulations [21] . Leveraging advanced computational algorithms, molecular docking studies enable the prediction of the binding modes and affinities of the candidate compounds within the protein binding sites, offering valuable predictive insights into their therapeutic potential. Through the iterative integration of computational tools such as Dogsitescorer and molecular docking simulations, the identification of lead compounds with optimal binding energies and favorable interaction profiles against protein targets associated with breast cancer and uterine fibroids is facilitated. This systematic approach enhances the rational design and optimization of novel therapeutics aimed at mitigating the progression of these debilitating diseases [21] .
3.3.4. Molecular Docking - Achilles Blind Docking Server
The 34 identified compounds, targeting breast cancer and uterine fibroid-associated protein diseases, underwent comprehensive molecular docking simulations using the ACHILLES BLIND DOCKING SERVER [22] . This state-of-the-art computational tool facilitated the exploration of compound-protein interactions across multiple protein targets, providing valuable insights into their binding affinities and binding site preferences. Through the blind docking approach employed by ACHILLES, the compatibility between each compound and the diverse array of protein targets associated with breast cancer and uterine fibroids was systematically evaluated. By considering multiple protein structures representative of different disease states, this approach enabled a comprehensive assessment of compound efficacy across various pathological contexts [22] .
Furthermore, the molecular docking simulations facilitated the identification of potential binding sites within the protein coordinates for each compound. The number and distribution of these binding sites served as critical indicators of compound versatility and potential therapeutic efficacy. Compounds exhibiting a higher propensity to bind at multiple locations within the protein coordinates were deemed particularly promising, as they may exert broader therapeutic effects and target diverse disease mechanisms [22] . By integrating the insights gleaned from molecular docking simulations across multiple protein targets, a comprehensive understanding of compound-protein interactions and their potential implications for breast cancer and uterine fibroid management was attained, Table 6.
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Table 6. Shows the number of locations within protein coordinate set by dogsitescorer’s energy binding with a total residue > −6.
To visualize the binding profiles of the compounds across all six protein diseases, a graph can be constructed with the compounds on the x-axis and the number of binding locations within the protein coordinates on the y-axis. Each compound is represented by a bar, with the height of the bar indicating the number of binding locations within the protein coordinates for that compound. The graph provides an overview of the binding versatility of each compound across multiple protein targets associated with breast cancer and uterine fibroid, Graph 1.
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Graph 1. Show the graph for 22 compounds with the number of locations it’s able to bind.
The comprehensive analysis of compound-protein interactions reveals distinct binding profiles across multiple protein targets implicated in breast cancer and uterine fibroid pathogenesis. Specifically, against the 1QQG protein, 26 compounds exhibit diverse binding patterns, with each compound displaying a varying number of binding locations within the protein coordinates. Similarly, interactions with the 2AYR protein reveal comparable trends among 26 compounds, while interactions with the 3GRF, 6T41, 7B5Q, and 7B5O proteins demonstrate unique binding profiles for 29, 29, 33, and 28 compounds, respectively. These findings underscore the compound-specific nature of binding interactions and provide valuable insights for further exploration of therapeutic interventions targeting breast cancer and uterine fibroids in Graphs 2(a)-(f). Figures 12-17 show the one of the compounds of 10251 for each protein’s locations within set coordinates.
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Graph 2. (a) Shows protein 1QQG’s number of location binding for each compound; (b) Shows protein 2AYR’s number of location binding for each compound; (c) Shows protein 3GRF’s number of location binding for each compound; (d) Shows protein 6T41’s number of location binding for each compound; (e) Shows protein 7B5Q’s number of location binding for each compound; (f) Shows protein 7B5O’s number of location binding for each compound.
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Figure 12. Shows compound 10251 location within set coordinate in 1QQG protein.
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Figure 13. Shows compound 10251 location within set coordinate in 2AYR protein.
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Figure 14. Shows compound 10251 location within set coordinate in 3GRF protein.
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Figure 15. Shows compound 10251 location within set coordinate in 6T41 protein.
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Figure 16. Shows compound 10251 location within set coordinate in 7B5Q protein.
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Figure 17. Shows compound 10251 location within set coordinate in 7B5O protein.
The figure displays the distances between a specific compound and the amino acids of proteins associated with breast cancer and uterine fibroid pathogenesis. Specifically, it illustrates the distances to amino acids of proteins 1QQG, 7B5Q, and 7B5O, representing breast cancer-related proteins, as well as proteins 2AYR, 3GRF, and 6T41, which are associated with uterine fibroid disease. These distances serve as indicators of the successful rate of binding energy for the compound towards each protein disease. Variations in distance highlight the differing degrees of interaction between the compound and the amino acids within each protein structure. Shorter distances suggest stronger binding interactions, indicating a higher potential for therapeutic efficacy in reducing the propensity for breast cancer and uterine fibroid development. Conversely, longer distances may signify weaker binding interactions, suggesting a need for further investigation or optimization of the compound’s efficacy.
By analyzing the distances to amino acids in the proteins relevant to both breast cancer and uterine fibroids, this table and 4 figures for each proteins provides valuable insights into the compound’s potential effectiveness in targeting these diseases. This information aids in the identification and prioritization of compounds for further preclinical and clinical studies aimed at mitigating the progression of breast cancer and uterine fibroids, Figures 18-23.
4. Discussion
The compounds that have been found in the LifeGreenTM product provide various benefits towards human health, breast cancers and uterine fibroid. Isobergaptol is a compound found in certain plants, particularly in essential oils. It has been studied for its potential anti-inflammatory and antimicrobial properties, which could contribute to improved immune function and wound healing. Some research suggests that Isobergaptol may also have antioxidant properties, helping to neutralize harmful free radicals in the body and reduce oxidative stress, which is associated with various chronic diseases. 10-Hydroxycamptothecin is a naturally occurring compound found in the Camptotheca acuminata tree. It is a derivative of camptothecin, a well-known anticancer agent. Studies have shown that 10-Hydroxycamptothecin exhibits potent antitumor activity by inhibiting the enzyme topoisomerase I, which is involved in DNA replication. This action can prevent cancer cells from proliferating and induce apoptosis (programmed cell death) in cancer cells. Camptothecin and related analogs have shown promise as anticancer agents, which could lead to the death of tumor cells by targeting the nuclear enzyme, topoisomerase I, and inhibiting the relegation of the cleaved DNA strand. One of the camptothecin analogs, hydroxycamptothecin (HCPT), a plant alkaloid derived from Camptotheca acuminata, has demonstrated strong antitumor activity against gastric, lung, ovarian, breast, and pancreatic carcinomas [23] . 10-hydroxycamptothecin (10-HCPT) is an important class of antitumor agent, it inhibits the DNA topoisomerase I of tumors and suppresses the proliferation of cancer cells to elicit antitumor effect [24] . Studies in animal and human subjects have shown that 10-hydroxycamptothecin (HCPT) is more potent and less toxic than the parent compound CPT. Casticin is a polymethylflavone isolated from a traditional Chinese therapeutic plant named Vitex trifolia L. from the Verbenaceae family [25] . The plant contributes to improve many morbidities including premenstrual syndrome, mastalgia, inflammation and sexual dysfunction, and also helps to relieve pain, and possesses antinociceptive effects. This plant is useful in mild hyperprolactinemia and luteal phase defects. It is also helpful in alleviating menstruation, bleeding management uterine fibroids, polycystic ovarian syndrome, prostate disorders, migrainous women with premenstrual syndrome [26] . Casticin is a flavonoid compound found in several medicinal plants, including Vitex agnus-castus (chaste tree). It has been investigated for its various potential health benefits. Research suggests that casticin possesses anti-inflammatory and antioxidant properties, which may help reduce inflammation and oxidative stress in the body. This could potentially benefit conditions such as arthritis and cardiovascular disease. Some studies also indicate that casticin may have anticancer properties, inhibiting the growth and proliferation of cancer cells in certain types of cancer.
This flavonoid compound is found in various fruits and vegetables and has been studied for its potential health-promoting effects. Like other flavonoids, 3’,5’-dihydroxyflavanone exhibits antioxidant properties, which can help protect cells from oxidative damage and reduce the risk of chronic diseases such as heart disease and cancer. Additionally, some research suggests that this compound may have anti-inflammatory properties, which could help alleviate inflammation-related conditions such as arthritis and inflammatory bowel disease. This flavonoid compound is also found in various plants and has been investigated for its potential health benefits. Like other flavonoids, (2S)-2’,7-Dimethoxy-3’,5’-dihydroxy flavanone possesses antioxidant properties, which can help protect cells from oxidative damage and reduce the risk of chronic diseases. Some research suggests that this compound may also have anti-inflammatory effects, which could potentially benefit conditions such as arthritis and cardiovascular disease. Lupanine is a quinolizidine alkaloid found in various plant species, including lupin seeds. It has been studied for its potential health benefits. Research suggests that Lupanine may have hypotensive (blood pressure-lowering) effects, making it potentially beneficial for individuals with hypertension. Additionally, Lupanine has been investigated for its potential antidiabetic properties, with some studies indicating that it may help improve insulin sensitivity and glucose metabolism. Flavanones are a class of flavonoid compounds found in various fruits and vegetables, particularly citrus fruits. They have been studied for their numerous health benefits [27] .
Flavanones exhibit antioxidant properties, helping to neutralize harmful free radicals in the body and reduce oxidative stress, which is associated with various chronic diseases such as cardiovascular disease and cancer. Some flavanones, such as hesperidin and naringenin, have been shown to have anti-inflammatory effects, which may help reduce inflammation and alleviate symptoms of inflammatory conditions like arthritis. Flavanones have been found to have both antioxidant and anti-inflammatory properties. In particular, different studies have focused their attention on hesperidin and its aglycone form, hesperetin, which play an important role in the prevention of diseases associated with oxidative stress and inflammation, such as cancer and cardiovascular disease [28] . Glabranin is a compound found in licorice (Glycyrrhiza glabra) and has been studied for its potential health benefits. Research suggests that Glabranin may have anti-inflammatory properties, which could help reduce inflammation in the body and alleviate symptoms of inflammatory conditions such as arthritis. Additionally, Glabranin has been investigated for its potential antiviral and antimicrobial properties, which could contribute to its use in traditional medicine for treating infections. Citrinin is a mycotoxin produced by certain fungi, particularly species of Penicillium and Aspergillus. While it is primarily known for its toxic effects, some research has also explored potential health benefits. Limited studies suggest that Citrinin may have antioxidant properties. Glabridin is a flavonoid compound found in licorice root (Glycyrrhiza glabra) and has been studied for its various health benefits. Research suggests that Glabridin may have antioxidant properties, helping to protect cells from oxidative damage and reduce the risk of chronic diseases such as heart disease and cancer. Additionally, Glabridin has been investigated for its potential anti-inflammatory effects, which could help reduce inflammation in the body and alleviate symptoms of inflammatory conditions like arthritis. Glabridin is an isoflavan extracted from licorice (genus Glycyrrhiza) roots, which is also known as a phytoestrogen due to the similarity of its structure and lipophilicity to 17β-estradiol. Studies indicated that glabridin is able to bind to the ERs and induce estrogenic responses in cardiovascular and bone tissues, suggesting its possibility to be used in estrogen replacement therapy.
Mycocyclosin is a compound isolated from certain fungi, particularly marine-derived fungi. It has been studied for its potential pharmaceutical properties. Research suggests that Mycocyclosin may have antibacterial and antifungal properties, making it potentially useful in the development of new antibiotics or antifungal agents. Additionally, some studies indicate that Mycocyclosin may have cytotoxic effects on cancer cells, which could make it a candidate for further investigation as a potential anticancer agent. 2,3-Dehydro-UWM6 is a chemical compound with potential pharmacological applications. Prazepam is a benzodiazepine medication used to treat anxiety and panic disorders. It belongs to the class of psychoactive drugs known for their anxiolytic (anxiety-reducing), sedative, muscle relaxant, and anticonvulsant properties. Benefits of Prazepam include its ability to alleviate symptoms of anxiety and panic disorders, promote relaxation, and reduce muscle tension. Prazepam is often prescribed for short-term relief of anxiety symptoms and is considered effective when used as directed under medical supervision. (-)-Phaseollinisoflavan is a type of isoflavonoid compound found in certain plants, particularly in legumes like soybeans and chickpeas. Research suggests that isoflavonoids like (-)-Phaseollinisoflavan may have various health benefits, including potential anticancer, antioxidant, and anti-inflammatory properties. Some studies indicate that dietary intake of isoflavonoids may be associated with a reduced risk of certain cancers, such as breast and prostate cancer, as well as cardiovascular disease. Phaseollidin is a natural compound found in certain legumes, including beans and peas. Research suggests that Phaseollidin may have anticancer properties, as it has been shown to inhibit the growth of cancer cells in some studies. Additionally, Phaseollidin may possess anti-inflammatory and antioxidant properties, which could contribute to its potential health benefits.
Glycophymoline is a brand name for a topical solution containing various herbal extracts, including menthol, eucalyptol, and thymol. It is commonly used as a mouthwash and gargle for oral hygiene and minor throat irritations. Benefits may include its antiseptic and refreshing properties, which can help to kill bacteria in the mouth, reduce bad breath, and soothe sore throats. Abscisic acid (ABA) is a plant hormone involved in various physiological processes in plants, such as seed dormancy, bud dormancy, and response to environmental stress. While primarily studied for its role in plants, Abscisic acid (ABA) has also been investigated for its potential health benefits in humans. Research suggests that ABA may have anti-inflammatory, antioxidant, and immunomodulatory effects, which could potentially benefit human health. It has been studied for its potential therapeutic applications in conditions such as diabetes, obesity, inflammation, and autoimmune diseases. Dihydroresveratrol is a derivative of resveratrol, a polyphenolic compound found in various plants, including grapes, berries, and peanuts. Resveratrol and its derivatives have been extensively studied for their potential health benefits, including antioxidant, anti-inflammatory, cardioprotective, neuroprotective, and anticancer properties. While research on alpha-beta-dihydroresveratrol specifically may be limited, it likely shares some of the health-promoting properties of resveratrol due to its structural similarity. Bergaptol is a natural compound found in certain plants, particularly in the citrus family. It is structurally related to bergamottin and is primarily known for its photosensitizing effects. While Bergaptol itself may not have direct health benefits, it is used in combination with other compounds in phototherapy for the treatment of certain skin conditions, such as psoriasis and vitiligo.
Bergapten, also known as 5-methoxypsoralen, is a natural compound found in several plant species, including citrus fruits and certain herbs such as parsley and celery. Bergapten is primarily known for its photosensitizing effects, which have been utilized in the treatment of skin disorders such as psoriasis, vitiligo, and eczema through a process known as psoralen plus ultraviolet A (PUVA) therapy. Additionally, Bergapten has been studied for its potential anticancer properties, particularly in the treatment of cutaneous T-cell lymphoma (CTCL). Methylarbutin is a derivative of arbutin, a natural compound found in various plant species such as bearberry, cranberry, and blueberry. Arbutin is well-known for its skin-lightening and antioxidant properties. Methylarbutin is often used in cosmetic products for its potential to inhibit melanin production and reduce the appearance of hyperpigmentation, dark spots, and uneven skin tone. Additionally, arbutin and its derivatives like Methylarbutin have been studied for their potential antioxidant and anti-inflammatory effects, which could contribute to their skin-protective properties. Abscisic acid (ABA) is a plant hormone involved in various physiological processes, including seed dormancy, bud dormancy, and response to environmental stress. While primarily studied for its role in plants, Abscisic acid (ABA) has also been investigated for its potential health benefits in humans. Research suggests that Abscisic acid (ABA) may have anti-inflammatory, antioxidant, and immunomodulatory effects, which could potentially benefit human health. It has been studied for its potential therapeutic applications in conditions such as diabetes, obesity, inflammation, and autoimmune diseases.
(+)-8’-Hydroxyabscisic acid is a derivative of abscisic acid (ABA), a plant hormone involved in various physiological processes in plants, including seed dormancy, bud dormancy, and response to environmental stress. While primarily studied for its role in plants, some research suggests that abscisic acid and its derivatives may have potential health benefits in humans. Abscisic acid has been investigated for its potential anti-inflammatory, antioxidant, and immunomodulatory effects, which could potentially benefit human health. It has been studied for its potential therapeutic applications in conditions such as diabetes, obesity, inflammation, and autoimmune diseases. 4-hydroxycoumarin, also known as umbelliferon, is a natural compound found in various plants, including citrus fruits, and is also produced synthetically. Research suggests that 4-hydroxycoumarin may have antioxidant, anti-inflammatory, and antimicrobial properties. It has been studied for its potential use in the treatment of various conditions, including skin disorders, inflammatory diseases, and as a natural sunscreen agent. Scopoletin is a natural coumarin compound found in various plants, including members of the Apiaceae and Rutaceae families. Scopoletin has been studied for its potential pharmacological activities, including antioxidant, anti-inflammatory, antimicrobial, and antitumor properties. It has been investigated for its potential therapeutic applications in conditions such as diabetes, neurodegenerative diseases, cancer, and cardiovascular disorders.
5. Conclusion
In summary, using molecular docking studies, we effectively discovered the number of compounds present in the LifeGreenTM that have the potential efficacy against breast cancer and uterine fibroids. From the results, we identified that out of 117 compounds that have been extracted from the LC-TOF MS analysis of LifeGreenTM, only 34 compounds have the more binding profile against the protein diseases. A total of 22 compounds demonstrated the remarkable capability to bid with all the six protein diseases while the remaining 12 compounds displayed a more selective binding profile, where those only interact with a subset of two to four protein diseases. Finally, the identified 34 compounds, which are associated with breast cancer and uterine fibroids, studied further by conducting comprehensive molecular docking simulations to extract information on their binding affinities and binding site preferences. This study underscores the need for further analytical and experimental studies to establish the safety and efficacy of the identified compounds. In the future, this experiment will be conducted in animal studies for both breast cancer and uterine fibroids using the standard procedure dosage recommended by the WHO and also will proceed with the phytochemical studies in order to identify the similariton between the compounds that have been identified in this paper.
Authors’ Contribution
Conceptualization: Ummi Shahieda Lazaroo Binti Zurrein Shah Lazaroo, Chua Kia How. Methodology & Formal analysis: Ummi Shahieda Lazaroo Binti Zurrein Shah Lazaroo. Writing Original Draft: Ummi Shahieda Lazaroo Binti Zurrein Shah Lazaroo, Navanithan Sivanananthan. Discussion & Conclusion: Navanithan Sivanananthan. Writing Review & Editing: Ummi Shahieda Lazaroo Binti Zurrein Shah Lazaroo.
Appendix
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Table A1. Shows compound spectrum result list which consist of all the details from LC-TOF-MS analysis.