Gas chromatographic-mass spectrometric (GC-MS) analysis and virtual screening of ripped fruit of Dialium guineense for potential inhibitors of integrase, cycloxygenase-1, and xanthine oxidase

Gas chromatographic-mass spectrometric (GC-MS) analysis and virtual screening of ripped fruit of Dialium guineense for potential inhibitors of integrase, cycloxygenase-1, and xanthine oxidase | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Gas chromatographic-mass spectrometric (GC-MS) analysis and virtual screening of ripped fruit of Dialium guineense for potential inhibitors of integrase, cycloxygenase-1, and xanthine oxidase Patrick Maduabuchi Aja, Peter Chinedu Agu, Boniface Anthony Ale, and 13 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-1727120/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Dialium guineense has a long history of medicinal use. This study evaluated the chemical constituents of the fruits using Gas chromatography-mass spectrometry (GC-MS) and virtual screening for potential inhibitors of three enzymes, integrase, cycloxygenase-1, and xanthine oxidase, to identify some of their therapeutic targets via bioinformatics tools. The results of the GC-MS revealed seventeen chemical constituents: 2,4-Dihydroxy-2,5-dimethyl-3(2H)-furanone (DG1), 3,5-Dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one (DG2), 5-Hydrxoymethylfurfural or 5-(hydroxymethyl)furan-2-carbaldehyde (DG3), 4-[( E )-(methoxyimino)methyl]phenol (DG4), 2,5-dihydroxy-6-methyl-2,3-dihydro-4 H -pyran-4-one (DG5), Lauric acid (DG6), Ethyl 5-methylnonanoate (DG7), Myristic acid, methyl ester (DG8), n-Tridecanoic acid methyl ester (DG9), Methyl 14-methylpentadecanoate (DG10), Palmitic acid, ethyl ester (DG11), Undecanoic acid (DG12), Stearic acid, ethyl ester (DG13), cis-11-Hexadecenal (DG14), 2-chloro- N -[(2-chlorophenyl)carbonyl]- N -(2methylpropyl)benzamide (DG15), Butyl-5-oxo-1-(trifluoroacetyl)pyrrolidine-2-carboxylate (DG16), and Glutaric acid, decylpentafluorobenzyl ester (DG17). Fifteen of these compounds, already deposited in the ChemSpider database, were retrieved and screened. Standard inhibitors of the enzymes validated the molecular docking. Only DG compounds that bind at the same site with the inhibitors were considered the potential enzyme inhibitors. DG1-17 demonstrated promising inhibition for at least one of the enzymes. The results also showed that DG1, 7, 11, 12, and 17 are integrase inhibitors, and DG1, 2, 4, 5, 6, 9, 10, and 12 are cycloxygenase-1 inhibitors, while DG1, 3, and 9 are xanthine oxidase inhibitors. The potential inhibitors' drug-likeness and ADMET properties revealed that DG11 and DG17 violated one of the Lipinski rules of five. All the Dialium guineense compounds showed good physiological and pharmacokinetic properties and were neither promiscuous to cytochrome p450 nor p-glycoprotein enzymes. Except for DG3, others are toxic to at least one of the organs, endocrines, and genomics. Our findings show that biologically active compounds may be responsible for the ethnomedical and nutritional uses of the fruits. Although some constituents have toxic effects, potential inhibitors can be optimized and synthesized as analogs of these standard inhibitors used in their chemotherapeutic targets. Dialium guineense fruit GC-MS analysis virtual screening potential inhibitors ADMET profile Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 Figure 13 Figure 14 Figure 15 Figure 16 Figure 17 Figure 18 Figure 19 Figure 20 Figure 21 1. Introduction Since time immemorial, the use of plants to maintain personal health and well-being has undoubtedly spurred the growth of scientific research. Some of these plants are widely used in non-industrialized societies and developing countries in Africa, Asia, and Southern America because they are considered less expensive than modern medicine and readily available (Awuchi, 2020 ). According to Aja et al. ( 2015 ), over 300 of these plants have been used in Nigeria to treat various diseases, including opportunistic infections such as HIV/AIDS, pneumonia, tuberculosis, diarrhea, typhoid fever, candidacies, and other ailments. More so, there has been growing interest in exploiting the biological activities of different ayurvedic medicinal herbs due to their natural origin, cost-effectiveness, and lesser side effects (Aja et al., 2014 ). However, the lack of nutrient compositional information on some of these plants has encouraged the population to ascribe some of their nutritional benefits to superstition (Achoba et al., 1992 ). Therefore, it has become imperative to evaluate the components of these plants to ascertain their safety applications and associated risks of toxicity in food and human nutrition. Studies have shown that solutions to food shortage and nutritional inadequacies in Africa lie in enhanced food production, processing, preservation, storage, and scientific evaluation of the nutrient content of several plants growing wild in bushes and forests (Achoba et al., 1992 ). Similarly, plants used as food ingredients, dietary supplements or nutraceuticals, pharmaceutical, and cosmetic products continue to be a point of reference in modern dietetics, chemotherapy, drug development, and cosmetology (Okeke et al., 2016 ; Besong et al., 2016 ). Surprisingly, some unanticipated plants or plant materials that can prove very potent in specific ailments have been neglected or abandoned for other preferred parts (Okeke et al., 2016 ). Dialium guineense ( D. guineense ), also known as Black Velvet Tamarind (BVT), is an indigenous tropical forest fruit tree of the family Leguminosae native to Southern Thailand and Malaysia (Osaigbovo and Nwaoguala, 2011 ). BVT is also found in West African countries such as Ghana, where it is known as Yoyi, Sierra Leone, Senegal (Asoiro et al., 2017 ), and Nigeria, where it is known as Tsamiyarkurmi in Hausa, Icheku in Igbo, Awin in the Yorubas, and Amugen in Edo language) (Osaigbovo and Nwaoguala, 2015). The plant is a non-timber multipurpose agroforestry crop with a high potential (Osaigbovo and Nwaoguala, 2011 ). According to Ewédjè and Tandjiékpon (2011), some older people consume non-alcoholic drinks made from fruit. In Nigeria, particularly in Abakaliki, the fruits are sold in local markets, and people of all ages eat them as a snack. Most women in South-eastern Nigeria chew the fruits to improve lactation and check genital infection (Asoiro et al., 2017 ). The minerals, sugars, and organic acids-rich fruits are used in the treatment and management of bacterial diseases and fever, diarrhea, and palpitations (Ewédjè and Tandjiékpon, 2011). Evidence suggests that apart from the traditional, medicinal, and nutritional uses of BVT, the pulp flour has desirable physicochemical and sensory properties, which may predispose it as a raw material for food industries, especially for the production of highly sought-after candy (Asoiro et al., 2017 ). The pulp is also a good source of simple sugars and vitamin C, although several factors could affect the fruit mposition (Arogba, et al., 1994 ). According to Nkanu et al. ( 2018 ), intraperitoneal administration of aqueous fruit pulp of D. guineense on streptozotocin-induced (55 mg/kg) diabetic rats significantly (p < 0.01) increased the serum angiotensin-converting enzyme (ACE) and blood glucose level and markedly decreased (p < 0.01) bilirubin, conjugated bilirubin, albumin, globulin, heme oxygenase-1 and insulin level in the diabetic control group. They further observed that D. guineense and Vitamin C treated groups showed no difference in ACE level, Bilirubin, Albumin, and globulin concentration, respectively, indicating the antibacterial and antioxidant potentials of the fruits. Interestingly, Ajiboye et al. ( 2018 ) discovered numerous phytochemicals with antimicrobial activity in the seeds, capable of exhibiting significant effects at various concentrations. This research corroborates previous findings by Utubaku et al. ( 2017 ) on secondary metabolites such as saponin, flavonoids, and phenolic compounds in fermented and unfermented D. guineense seeds with the potential to improve certain disease conditions. In recent years, Gas chromatography-mass spectrometry (GC-MS) analysis has become a gold standard for determining bioactive compounds of both plants and non-plant species (Kanthal et al., 2014 ). Several reviews on the D . guineense have shown that, despite its popularity and vast applications in traditional medicine for treating various disorders, there is still a scarcity of documented data available regarding GC-MS analysis of chemical constituents of D. guineense fruits. Structure-based drug design is becoming a valuable and integral part of the drug discovery process and has proven to be more effective than ligand-based drug design (Hirashima and Huang, 2008 ). Studies of interactions between protein domains and ligands are essential in virtual screening analysis (Powers, 2009 ). Virtual screening analysis can aid in the drug targets identification via bioinformatics tools. They are used to analyze target structures for potential binding sites, generate candidate molecules, test for drug-likeness, dock the molecules with the target, rank them based on binding affinities, and further optimize the molecules to improve binding characteristics (Breda et al., 2008 ). Autodock 4.2 is a suite of automated docking tools. It usually starts with the definition of a binding site, in general, at a restricted region of the protein. As an alternative to binding site prediction, a standard drug known to be an inhibitor of the target protein can be docked alongside other molecules being investigated to predict their chemotherapeutic target. Integrase (Int), Cycloxygenase1 (COX-1), and Xanthine oxidase (XO) are enzymes widely distributed among different species from bacteria to man and within the various tissues of mammals. Inhibition of these enzymes has been identified as a target for chemotherapy. For instance, integrase inhibitors rely on the fact that HIV needs integrase to replicate. These drugs stop HIV from being able to make integrase. Without the help of this enzyme, HIV cannot take over the T cells to copy itself. With a combination of other HIV medications, integrase inhibitors can help keep HIV under control ( https://www.healthline.com/health/hiv-aids/integrase-inhibitors ). Cyclooxygenase (COX) is the enzyme responsible for the conversion of arachidonic acid (AA) into prostanoids. It was identified over two decades ago, although aspirin, a cyclooxygenase inhibitor, has been commercially available (Bayer) since 1899 to treat inflammatory syndromes (Vane, 2000 ). In the last years, two isoforms of COX produced from different genes have been identified, COX-1 and COX-2 (Simmons et al., 2014 ). COX-1 is considered a “housekeeping gene” due to the constitutive low levels of expression in most cell types and tissues (Perrone et al., 2010 ). High levels of constitutive expression of COX-1 have, instead, been detected in the stomach and platelets. These differences in the regulation of COX-isozymes suggest that the significant action of COX-1 is to mediate gastrointestinal tract protection and modulate platelet function, whereas COX-2 is mainly involved in inflammation and pain. Traditional nonselective non-steroidal anti-inflammatory drugs [(t)NSAIDs], which inhibit both COX-1 and COX-2, exert their therapeutic effects through inhibition of COX-2-dependent prostanoid biosynthesis and cause gastrointestinal damage through the inhibition of COX-1 (Perrone et al., 2010 ). The unavailability of highly selective COX-1 inhibitors (Brenneis et al., 2006 ) led to the use of genetic mouse models of selective deletion of COX-1 as a strategy to enlighten the potential pathogenic contribution of prostanoids synthesized via COX-1, e.g. to inflammatory arthritis (Chen et al., 2008 ), atherogenesis (McClelland et al., 2004 ), intestinal polyposis (Chulada et al., 2000 ) and skin carcinogenesis (Tiano et al., 2002 ). It is now believed that COX-1 is responsible for the primary prostanoid response to inflammatory stimuli (particularly in cells and tissues, where it is constitutively and predominantly expressed). Besides, special attention will be given (a) to the assays used to determine the activity and selectivity (often discordant and assay dependent) of the COX inhibitors (selectivity values are used to classify the large number of clinically used NSAIDs), and (b) the primary structure-activity relationship investigations, so far reported, performed to identify the few known COX-1 selective inhibitors (Perrone et al., 2010 ). Xanthine oxidase is a member of a group of enzymes known as molybdenum iron-sulfur flavin hydroxylases (Symons et al., 1989 ). It catalyzes the oxidation of hypoxanthine to xanthine and then to uric acid, the final reaction in the metabolism of purine bases (Zarepour et al., 2010 ). The accumulation of uric acid in the body is responsible for several diseases, and thus it plays a vital role in hyperuricemia and gout (Egwim et al., 2005 ). Inherited xanthine oxidase reductase (XOR) deficiency leads to xanthinuria and multiple organ failure syndromes caused by the accumulation of xanthine in different tissues (Borges et al., 2002 ). The inhibitors XO, integrase, and cyclooxygenase-1 from plants are much more useful (Umamaheswar et al., 2013 ) since they possess lesser side effects compared to uricosuric and anti-inflammatory agents (Umamaheswari et al., 2009 ). As a result, the present study aimed to evaluate the GC-MS analysis of chemical constituents of the D. guineense fruits found in Abakaliki, South-eastern Nigeria. We further conducted virtual screening to predict the chemotherapeutic targets and drug-likeness of the chemical constituents. 2. Materials And Methods 2.1 Materials The equipment, chemicals, and reagents used in this research work were of analytical grade and quality. The fresh fruits of Dialium guineense were collected by hand plucking from the wild plants in March 2019 from the Abakaliki Area of Ebonyi State, Nigeria. The plant was classified and authenticated by a plant taxonomist Mr. O. E. Nwankwo of the Department of Applied Biology of Ebonyi State University, Abakaliki, Nigeria. Part of the identified plant was kept in the herbarium at the Applied Biology Department, Ebonyi State University, Abakaliki, Nigeria, for reference purposes. The molecular docking apparatus used was software (AutoDock Vina plugin Pyrx, UCSF Chimera, and Discovery studios 2020), database (PDB, PubChem, and ChemSpider), and web servers (SwissAdme, and AdmetSar). The standard inhibitors used are Raltegavir (Ralt), Ibuprofen (Ibu), and Caffeine (Caf) for Integrase, Cycloxygenase-1, and Xanthine oxidase, respectively. 2.2. Methods 2.2.1. Preparation and extraction of Dialium guineense Fruits The ripe fruits of Dialium guineense were destalked, deshelled and the fruits were carefully removed from the seeds. The fruits were shade dried at ambient temperature with constant turning to prevent fungal growth. The dried fruits were milled to obtain powdered fruit samples using an electric blender and were stored at 4 0 C temperature in a refrigerator in well-labelled airtight containers for analysis. Fifty grams of powdered fruit sample of D. guineense was extracted with 400 ml of methanol in an orbital shaker for 24 hrs at room temperature. The extract was filtered using Whatman No.1 filter paper to remove extractable substances, at every 4 hrs interval. The combined extracts were then evaporated with a rotary evaporator, and the dried extract was stored at 4°C in a sterile container. 2.2.3. GC-MS Analysis of the Dialium guineense Fruits GC-MS analysis of the methanol extract of D. guineense fruits was performed using Shimadzu Japan gas chromatography QP2010PLUS with a fused GC column (2010) coated with polymethyl silicon (0.25nm x 50m) and the conditions were as follows: Temperature programming from 80–200 0 C held at 80 0 C for 1 min, rate 5 0 C/min and at 200 0 C for 20 min. Field ionization detector (FID) temperature 300 0 C, injection temperature 220 0 C, carrier gas nitrogen at a flow rate of 1 ml/min, split ratio 1:75. Gas chromatography-mass spectrum was conducted using GC-MS –QP 2010 Plus Shimadzu Japan with injector temperature of 220 0 C and carrier gas pressure of 116.9 kpa. The column length is 30 m with a diameter of 0.25 mm and a flow rate of 50 ml/min. The elutes were automatically passed into a mass spectrometer with a dictator voltage set at 1.5 kV and a sampling rate of 0.2 sec. The mass spectrum was also equipped with a computer fed mass spectra data bank. Hermlez 233 M-Z centrifuge (Germany) was used. 2.2.3 Components Identification The method described by Aja et al. ( 2014 ) was used to identify the chemical constituents of the fruit extract by matching the peaks with Computer Wiley Micro-Soft libraries and confirmed by comparing mass spectra of the peaks and those from literature. 2.2.4 Virtual screening of Dialium guineense compounds The target proteins (integrase, cycloxygenase1, and xanthine oxidase), the standard inhibitors (Raltegavir, Ibuprofen, and Caffeine), and the identified compounds of Dialium guineese were retrieved from RCSB PDB, PubChem, and Chemspider databases respectively. The retrieved proteins and ligands were individually and in turns prepared for molecular docking using UCSF Chimera. The protein and ligand preparation were by removing non-standard ligand (where necessary) and structural editing/minimization in the default settings with adding hydrogen and charges Gastiger. All the proteins and ligands were saved in PDB file format after preparation. AutoDock vina plugin Pyrx was used for virtual screening (Trott and Olso, 2010) of the identified compounds for complementary binding with the standard inhibitor of each target protein. This program was run using a searching grid extended over the ligand molecules with box spacing and coordinates (x, y,z) respectively as follows: integrase (55x50x70 and 5.13, 31.40, and − 10.61); cycloxygenase-1 (87x84x73 and − 32.82, -49.17 and 2.92); xanthine oxidase (120x94x114 and 29.22, 14.25 and 168.34) with other parameters set as default. The identified compounds were docked into the crystal structure of ligand molecules, and the highest-scoring pose binding at the same site with the standard inhibitor was selected for each of the identified compounds. Those with the best docking poses are predicted to be inhibitors of the target enzymes. Further, the best poses were visualized using the UCSF Chimera and Discovery studio 2020 to ascertain the protein-ligand interactions. ADMET (Absorption, Distribution, Metabolism, Excretion, and Toxicity) prediction and significant descriptors of drug-likeness such as mutagenicity, toxicological dosage level, and pharmacologically relevant properties of the best pose compounds were predicted using Swissadme ( http://www.swissadme.ch ) and admetSAR (lmmd.ecust.edu.cn:8000) servers. 3. Results 3.1 GC-MS Constituents of Methanol Fruit-Extract of D. guineense The results of GC-MS constituents of methanol fruit-extract of D. guineense showed that seventeen compounds were present in the fruits of D. guineense (Table 1). These compounds comprise mainly saturated fatty acids, carboxylic acids, phenolic compounds, and esters. The composition of the extract comprises palmitic acid, ethyl ester, glutaric acid, decylpentafluorobenzyl ester, 4-[( E )-(methoxyimino) methyl] phenol, and5-Hydrxoymethylfurfural or 5-(hydroxymethyl) furan-2-carbaldehyde. The identified compounds and their corresponding molecular weight, retention time, percentage (%) base peaks, structures and their bioactivity are shown in Table 1. 3.3 Protein-Inhibitor Interaction visualization 3.3.1 Integrase vs DG best poses Figures 1-10 3.3.2 COX1 vs DG best poses Figures 11-18 3.3.3 Xanthine oxidase vs DG best poses Figures 19-21 4. Discussion Dialium guineense ( D. guineense ) is one of the emerging medicinal plants of interest (Besong et al ., 2016). Various parts of D. guineense have been reported in ethnomedicine to have medicinal uses. Studies by Okudu et al . (2017) on nutritional and sensory attributes of D. guineense revealed a reasonable number of antioxidants and numerous minerals such as magnesium, sodium, iron, potassium, beta-carotene (Vitamin A), copper, sugars and tartaric acid, citric acid, malic acid, ascorbic acid, and niacin. Similarly, Ogu et al . (2012) reported several phytochemicals on the methanolic stem bark extract of D . guineense : cardiac glycosides, tannins, phlobatannins, saponins, terpenoids, resins, steroids/triterpenes, alkaloids, flavonoids, reducing sugars and carbohydrates with anti-diarrheal effects. In the present study, the analysis of D . guineense fruits showed seventeen peaks from the GC-MS chromatogram. These peaks indicated the presence of seventeen compounds (DG1-17) depicted in Table 1. The composition comprises fatty acids, including Lauric acid (0.18%), Ethyl 5-methylnonanoate (2.30%), Myristic acid, methyl ester (2.39%), n-Tridecanoic acid methyl ester (2.32%), Methyl 14-methylpentadecanoate (5.21%), Palmitic acid, ethyl ester (34.23%), Undecanoic acid (4.52%) and Stearic acid, ethyl ester (1.37%). Thus, the fatty acid compositions of Dialium guineense were all saturated. Although eating foods containing saturated fats may raise low-density lipoprotein (LDL) cholesterol levels, thereby increasing the risk of cardiovascular diseases. Research has shown that saturated fats could undergo desaturation into their corresponding unsaturated forms. He also came to the conclusion that stearic acid is less likely to be incorporated into cholesterol esters. Also, a robust study by Emken (2018) on human isotope labeling showed that the fraction of dietary stearic acid that oxidatively desaturates to oleic acid is 2.4 times higher than the fraction of palmitic acid analogously converted to palmitoleic acid. He concluded further that stearic acid is less likely to be incorporated into cholesterol esters. Similarly, epidemiological and clinical studies by Hunter et al . (2010) showed that dietary stearic acid has favorable effects on plasma LDL cholesterol relative to cholesterol-raising saturated fatty acids, indicating its possible replacement for tans fat. This study also revealed cis-11-Hexadecenal (1.57%), 2-chloro- N -[(2 chlorophyll) carbonyl]- N -(2-methyl propyl)benzamide (1.72%), Butyl5-oxo-1-(trifluoroacetyl)pyrrolidine-2-carboxylate (1.24%), Glutaric acid, decylpentafluorobenzyl ester and 2,5-dihydroxy-6-methyl-2,3-dihydro-4 H -pyran-4-one (14.02%). Except cis-11-Hexadecenal, 2-chloro- N -[(2-chlorophenyl)carbonyl]- N -(2-methyl propyl)benzamide, Butyl 5-oxo-1-(trifluoroacetyl)pyrrolidine-2-carboxylate, Ethyl 5-methylnonanoate, 5-Hydrxoymethylfurfural or 5-(hydroxymethyl)furan-2-carbaldehyde and 4-[( E )-(methoxyimino)methyl]phenol which showed no bioactive, the other compounds identified displayed various bioactivities such as 11B-HSD-Inhibitor, 17-beta-hydroxysteroid dehydrogenase-Inhibitor, Catechol-O-Methyl-Transferase-Inhibitor, 5-HETE-Inhibitor, 5-HT-Inhibitor, Methyl-Guanidine-Inhibitor, Urine-Acidifier, uric acid production inhibitor and arachidonic acid inhibitor by 2,4-Dihydroxy-2,5-dimethyl-3(2H)-furanone, 3,5-Dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one, 5-dihydroxy-6-methyl-2,3-dihydro-4 H -pyran-4-one, lauric acid, Myristic acid, methyl ester, n-Tridecanoic acid methyl ester, Methyl 14-methylpentadecanoate, Palmitic acid, ethyl ester, Undecanoic acid, Stearic acid, ethyl ester and Glutaric acid, decylpentafluorobenzyl ester respectively. The endowment of DG with these constituents justifies its enormous folkloric uses. Notably, the wound-healing and antimicrobial potential of the dichloromethane fraction of D. guineense fruit coat have been documented by Okeke et al . (2016). Furthermore, the GC-MS analysis demonstrates that lauric acid (LA), a medium-chain fatty acid, had the lowest peak area (0.18%), indicating its low concentration in the D. guineense . On the other hand, the dominant class of the compounds was Palmitic acid, ethyl ester with a base peak area of 34.23%, and prominently urine acidifier. Lauric acid, on the other hand, has noticeable bioactivities as an acidifier, acidulant, and arachidonic acid inhibitor, which may modulate prostaglandin synthesis. Furthermore, studies by Lappano et al . (2017) established that LA induces apoptosis in cancer cells by increasing reactive oxygen species levels and stimulating the phosphorylation of epidermal growth factor receptor, extracellular-signal-regulated kinase, and induces the expression of proto-oncogenes. The presence of n-Tridecanoic acid methyl ester 2.32%), an Anaphylactic (antidote=Neostigmine), antitumor (Nasopharynx) also validates the application of D. guineense in traditional medicine for the treatment of allergy and cancer. Hence, supplementation with D. guineense fruits may be the potential natural therapy against many multidrug-resistant diseases. The virtual screening aimed to gain insight into the pharmacological effects of the bioactive compounds. Table 2 showed fifteen compounds were retrieved and used for virtual screening. DG1, DG2, DG3, DG4, DG5, DG6, DG7, DG9, DG10, DG11, DG12, and DG17 (Table 3) were identified as potential inhibitors of at least one of the enzymes used. The results also showed that DG1 to 7, 11, 12, and 17 are integrase inhibitors, DG1, 2, 4, 5, 6, 9, 10, and 12 are cycloxygenase-1 inhibitors, while DG1, 3, and 9 are xanthine oxidase inhibitors (Table 2). They have binding affinities of between 3.9-6.1Kcal/mol. Figures 1–21 show that potential inhibitors bind at the same binding sites as standard inhibitors for each enzyme. Furthermore, they interact with different amino acids while using similar bond types such as hydrogen bonds, van der Waals, unfavorable donors, pi-bonds, etc. The potential inhibitors have good drug-likeness (Table 4). Only DG11 and DG17 violate one each of the Lipinski Rules of five. As shown in Table 5, all potential inhibitors can be absorbed through the gastrointestinal tract and cross the blood-brain barrier. Others can achieve oral bioavailability. Others can attain oral bioavailability. They also do not interfere with the membrane transport proteins, the cytochrome p450, and p-glycoproteins. Thus, they have good physiological and pharmacokinetic properties. The toxicity profile showed that all the potential inhibitors except DG3 are toxic to the eyes. DG2 is mutagenic, and DG4, DG12, and DG17 are endocrine disruptors. 5. Conclusion In the present study, GC-MS analysis of D. guineense showed interesting classes of chemical compounds with remarkable biological activities, including 11B-HSD-Inhibitor, 17-beta-hydroxysteroid dehydrogenase-Inhibitor, Catechol-O-Methyl-Transferase-Inhibitor, 5-HETE-Inhibitor, 5-HT-Inhibitor, Methyl-Guanidine-Inhibitor, Urine-Acidifier, uric acid production inhibitor, and arachidonic acid inhibitor. The molecular docking showed varied classes of bioactive compounds that could be useful as integrase, cycloxygenase-1, and xanthine oxidase inhibitors. These effects could account for the potential use of D. guineense fruits in several disease conditions. However, virtual screening investigation of its pharmacological effects has given insight into its potential application in human nutrition and targets for chemotherapy. The optimization of the potential inhibitors to ward off the toxicity of the compounds can be a promising step toward drug discovery. Also, there is an urgent need to sensitize the public about the toxic bioactive compounds in the D. guineense fruit as an intervention in public health, especially in the rural area where it is consumed raw as food. Declarations Declaration of interests The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. References Achoba, I., Lori, J. A., Elegbede, J. A., and Kagbu, J. A. (1992). Nutritional Composition of Black (African) Velvet Tamarind ( Dialium guineense Wild) Seed and Pulp from Nigeria Journal of Food Biochemistry, 16 : 229-233 Aja, P. M., Nwachukwu, N., Ibiam, U. 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(2017).Traditional Medicine & Clinical Naturopathy Comparative Phytochemical Analysis of Fermented and Unfermented Seeds of Dialium giuneense . Journal of Traditional Medicine and Clinical Naturopathy , 6 (3): 2573-4555 Vane, J. R. (2000). The fight against rheumatism: from willow bark to COX-1 sparing drugs. Journal of Physiology & Pharmacology , 51 : 573-586. Simmons, D. L., Botting, R. M., and Hla, T. (2014). Cyclooxygenase isozymes: the biology of prostaglandin synthesis and inhibition. Pharmacology Rev iew, 56 : 387-437. Brenneis, C., Maier, T. J., Schmidt, R., Hofacker, A., Zulauf, L., Jakobsson, P. J., Scholich, K. and Geisslinger, G. (2006). Inhibition of prostaglandin E2 synthesis by SC-560 is independent of cyclooxygenase 1 inhibition. FASEB J ournal, 20 : 1352-1360. Chen, M., Boilard, E., Nigrovic, P. A., Clark, P., Xu, D., Fitzgerald, G. A., Audoly, L. P.and Lee, D. M. (2008). 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Deficiency of either cyclooxygenase (COX)-1 or COX-2 alters epidermal differentiation and reduces mouse skin tumorigenesis. Cancer Research , 62 : 3395-3401. Perrone, M. G., Scilimati, A., Simone, L., and Vitale, P. (2010). Selective COX-1 Inhibition: A Therapeutic Target to be Reconsidered. Current Medicinal Chemistry , 17: 3769-3805 3769 Umamaheswar, M., Madeswaran, A., and Asokkumar, K. (2013). Virtual Screening Analysis and In-vitro Xanthine Oxidase Inhibitory Activity of Some Commercially Available Flavonoids. Iranian Journal of Pharmaceutical Research , 12(3): 317-323 Symons, C. R. M., Taiwo, A. F., and Petersen, L. R. (1989). Electron addition to xanthine oxidase. An electron spins resonance study of the effects of ionizing radiation. Journal of Chemical Society, 85: 4063-74. Zarepour, M., Kaspari, K., Stagge, S., Rethmeier, R., Mendel, R. R., and Bittner, F. (2010). Xanthine dehydrogenase AtXDH1 from Arabidopsis thaliana is a potent producer of superoxide anions via its NADH oxidase activity. Plant Molecular Biol ogy, 72: 301-10. Egwim, E. C., Vanchi, M. A., and Egwin, P. O. (2005). Comparison of xanthine oxidase activities in cow and goat milk. Biokemistri, 17: 1-6. Borges, F., Fernandes, E., and Roleira, F. (2002). Progress towards the discovery of xanthine oxidase inhibitors. Current Medicinal Chemistry, 9: 195-217. Umamaheswari, M., Asokkumar, K., Sivashanmugam, A. T., Remyaraju, A., Subhadradevi, V., and Ravi, T. K. (2009). In-vitro xanthine oxidase inhibitory activity of the fractions of Erythrina stricta Roxb. Journal of Ethnopharmacol ogy , 124: 646-8. Breda, A., Basso, L. A., Santos, D. S., and Walter, F. (2008). Virtual screening of drugs: score functions, docking, and drug design. Current Computer-Aided Drug Design, 4: 265-72. Hirashima, A., and Huang, H. (2008). Homology modeling, agonist binding site identification, and docking in octopamine receptor of Periplaneta americana . Computational Biology & Chemistry, 32: 185-90. Powers, R. (2009). Advances in nuclear magnetic resonance for drug discovery. Expert Opinion in Drug Disc overy , 4: 1077-98 Tables 1-5 Tables 1-5 are available in the Supplementary Files section. Supplementary Files Tables.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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interaction\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"ScreenShot20220614at3.35.58PM.png","url":"https://assets-eu.researchsquare.com/files/rs-1727120/v1/9474ae81c1489b1b15b4e265.png"},{"id":22655964,"identity":"4fa69015-2fc7-4a55-8328-9c3ce71b67a5","added_by":"auto","created_at":"2022-06-14 20:24:23","extension":"png","order_by":18,"title":"Figure 18","display":"","copyAsset":false,"role":"figure","size":462936,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eCycloxygenase1 and Ibu/DG12 interaction\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"ScreenShot20220614at3.36.06PM.png","url":"https://assets-eu.researchsquare.com/files/rs-1727120/v1/34dcb9e2198562b76f55b546.png"},{"id":22655855,"identity":"d07d9420-c011-4aad-925b-36f9d42063c8","added_by":"auto","created_at":"2022-06-14 20:14:23","extension":"png","order_by":19,"title":"Figure 19","display":"","copyAsset":false,"role":"figure","size":539805,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eXanthine oxidase and Caf/DG1 interaction\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"ScreenShot20220614at3.36.13PM.png","url":"https://assets-eu.researchsquare.com/files/rs-1727120/v1/73498118417fb47f6c935e88.png"},{"id":22655856,"identity":"1b848b4c-7ea3-4b75-803a-44cef7954ec4","added_by":"auto","created_at":"2022-06-14 20:14:24","extension":"png","order_by":20,"title":"Figure 20","display":"","copyAsset":false,"role":"figure","size":506616,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eXanthine oxidase and Caf/DG3 interaction\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"ScreenShot20220614at3.36.21PM.png","url":"https://assets-eu.researchsquare.com/files/rs-1727120/v1/d1adeac560ccb1fb80c795fd.png"},{"id":22655841,"identity":"a0f414ab-9b9f-4f22-93c8-f86d4fe052b9","added_by":"auto","created_at":"2022-06-14 20:14:23","extension":"png","order_by":21,"title":"Figure 21","display":"","copyAsset":false,"role":"figure","size":435764,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eXanthine oxidase and Caf/DG9 interaction\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"ScreenShot20220614at3.36.28PM.png","url":"https://assets-eu.researchsquare.com/files/rs-1727120/v1/f4d00f61d5e8cb6281b01cc8.png"},{"id":22656088,"identity":"0509ac98-dda8-4f03-919e-59903421664f","added_by":"auto","created_at":"2022-06-14 20:29:33","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":10451087,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-1727120/v1/884592c8-0c61-424f-b20f-f9ede49881b3.pdf"},{"id":22655962,"identity":"090d2c95-7955-4bf1-97ee-b9fef2d34e67","added_by":"auto","created_at":"2022-06-14 20:24:23","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":2020753,"visible":true,"origin":"","legend":"","description":"","filename":"Tables.docx","url":"https://assets-eu.researchsquare.com/files/rs-1727120/v1/b33144bd840148f1225d8153.docx"}],"financialInterests":"","formattedTitle":"\u003cp\u003eGas chromatographic-mass spectrometric (GC-MS) analysis and virtual screening of ripped fruit of \u003cem\u003eDialium guineense \u003c/em\u003efor potential inhibitors of integrase, cycloxygenase-1, and xanthine oxidase\u003c/p\u003e","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eSince time immemorial, the use of plants to maintain personal health and well-being has undoubtedly spurred the growth of scientific research. Some of these plants are widely used in non-industrialized societies and developing countries in Africa, Asia, and Southern America because they are considered less expensive than modern medicine and readily available (Awuchi, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). According to Aja et al. (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), over 300 of these plants have been used in Nigeria to treat various diseases, including opportunistic infections such as HIV/AIDS, pneumonia, tuberculosis, diarrhea, typhoid fever, candidacies, and other ailments. More so, there has been growing interest in exploiting the biological activities of different ayurvedic medicinal herbs due to their natural origin, cost-effectiveness, and lesser side effects (Aja et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eHowever, the lack of nutrient compositional information on some of these plants has encouraged the population to ascribe some of their nutritional benefits to superstition (Achoba et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1992\u003c/span\u003e). Therefore, it has become imperative to evaluate the components of these plants to ascertain their safety applications and associated risks of toxicity in food and human nutrition. Studies have shown that solutions to food shortage and nutritional inadequacies in Africa lie in enhanced food production, processing, preservation, storage, and scientific evaluation of the nutrient content of several plants growing wild in bushes and forests (Achoba et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1992\u003c/span\u003e). Similarly, plants used as food ingredients, dietary supplements or nutraceuticals, pharmaceutical, and cosmetic products continue to be a point of reference in modern dietetics, chemotherapy, drug development, and cosmetology (Okeke et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Besong et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Surprisingly, some unanticipated plants or plant materials that can prove very potent in specific ailments have been neglected or abandoned for other preferred parts (Okeke et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cem\u003eDialium guineense\u003c/em\u003e (\u003cem\u003eD. guineense\u003c/em\u003e), also known as Black Velvet Tamarind (BVT), is an indigenous tropical forest fruit tree of the family Leguminosae native to Southern Thailand and Malaysia (Osaigbovo and Nwaoguala, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). BVT is also found in West African countries such as Ghana, where it is known as Yoyi, Sierra Leone, Senegal (Asoiro et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), and Nigeria, where it is known as Tsamiyarkurmi in Hausa, Icheku in Igbo, Awin in the Yorubas, and Amugen in Edo language) (Osaigbovo and Nwaoguala, 2015). The plant is a non-timber multipurpose agroforestry crop with a high potential (Osaigbovo and Nwaoguala, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). According to Ew\u0026eacute;dj\u0026egrave; and Tandji\u0026eacute;kpon (2011), some older people consume non-alcoholic drinks made from fruit. In Nigeria, particularly in Abakaliki, the fruits are sold in local markets, and people of all ages eat them as a snack. Most women in South-eastern Nigeria chew the fruits to improve lactation and check genital infection (Asoiro et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The minerals, sugars, and organic acids-rich fruits are used in the treatment and management of bacterial diseases and fever, diarrhea, and palpitations (Ew\u0026eacute;dj\u0026egrave; and Tandji\u0026eacute;kpon, 2011). Evidence suggests that apart from the traditional, medicinal, and nutritional uses of BVT, the pulp flour has desirable physicochemical and sensory properties, which may predispose it as a raw material for food industries, especially for the production of highly sought-after candy (Asoiro et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The pulp is also a good source of simple sugars and vitamin C, although several factors could affect the fruit mposition (Arogba, et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e1994\u003c/span\u003e). According to Nkanu et al. (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), intraperitoneal administration of aqueous fruit pulp of \u003cem\u003eD. guineense\u003c/em\u003e on streptozotocin-induced (55 mg/kg) diabetic rats significantly (p\u0026thinsp;\u0026lt;\u0026thinsp;0.01) increased the serum angiotensin-converting enzyme (ACE) and blood glucose level and markedly decreased (p\u0026thinsp;\u0026lt;\u0026thinsp;0.01) bilirubin, conjugated bilirubin, albumin, globulin, heme oxygenase-1 and insulin level in the diabetic control group. They further observed that \u003cem\u003eD. guineense\u003c/em\u003e and Vitamin C treated groups showed no difference in ACE level, Bilirubin, Albumin, and globulin concentration, respectively, indicating the antibacterial and antioxidant potentials of the fruits. Interestingly, Ajiboye et al. (\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) discovered numerous phytochemicals with antimicrobial activity in the seeds, capable of exhibiting significant effects at various concentrations. This research corroborates previous findings by Utubaku et al. (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) on secondary metabolites such as saponin, flavonoids, and phenolic compounds in fermented and unfermented \u003cem\u003eD. guineense\u003c/em\u003e seeds with the potential to improve certain disease conditions.\u003c/p\u003e \u003cp\u003eIn recent years, Gas chromatography-mass spectrometry (GC-MS) analysis has become a gold standard for determining bioactive compounds of both plants and non-plant species (Kanthal et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Several reviews on the \u003cem\u003eD\u003c/em\u003e. \u003cem\u003eguineense\u003c/em\u003e have shown that, despite its popularity and vast applications in traditional medicine for treating various disorders, there is still a scarcity of documented data available regarding GC-MS analysis of chemical constituents of \u003cem\u003eD. guineense\u003c/em\u003e fruits.\u003c/p\u003e \u003cp\u003eStructure-based drug design is becoming a valuable and integral part of the drug discovery process and has proven to be more effective than ligand-based drug design (Hirashima and Huang, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). Studies of interactions between protein domains and ligands are essential in virtual screening analysis (Powers, \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Virtual screening analysis can aid in the drug targets identification via bioinformatics tools. They are used to analyze target structures for potential binding sites, generate candidate molecules, test for drug-likeness, dock the molecules with the target, rank them based on binding affinities, and further optimize the molecules to improve binding characteristics (Breda et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). Autodock 4.2 is a suite of automated docking tools. It usually starts with the definition of a binding site, in general, at a restricted region of the protein. As an alternative to binding site prediction, a standard drug known to be an inhibitor of the target protein can be docked alongside other molecules being investigated to predict their chemotherapeutic target. Integrase (Int), Cycloxygenase1 (COX-1), and Xanthine oxidase (XO) are enzymes widely distributed among different species from bacteria to man and within the various tissues of mammals. Inhibition of these enzymes has been identified as a target for chemotherapy. For instance, integrase inhibitors rely on the fact that HIV needs integrase to replicate. These drugs stop HIV from being able to make integrase. Without the help of this enzyme, HIV cannot take over the T cells to copy itself. With a combination of other HIV medications, integrase inhibitors can help keep HIV under control (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.healthline.com/health/hiv-aids/integrase-inhibitors\u003c/span\u003e\u003cspan address=\"https://www.healthline.com/health/hiv-aids/integrase-inhibitors\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). Cyclooxygenase (COX) is the enzyme responsible for the conversion of arachidonic acid (AA) into prostanoids. It was identified over two decades ago, although aspirin, a cyclooxygenase inhibitor, has been commercially available (Bayer) since 1899 to treat inflammatory syndromes (Vane, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2000\u003c/span\u003e). In the last years, two isoforms of COX produced from different genes have been identified, COX-1 and COX-2 (Simmons et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). COX-1 is considered a \u0026ldquo;housekeeping gene\u0026rdquo; due to the constitutive low levels of expression in most cell types and tissues (Perrone et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). High levels of constitutive expression of COX-1 have, instead, been detected in the stomach and platelets. These differences in the regulation of COX-isozymes suggest that the significant action of COX-1 is to mediate gastrointestinal tract protection and modulate platelet function, whereas COX-2 is mainly involved in inflammation and pain. Traditional nonselective non-steroidal anti-inflammatory drugs [(t)NSAIDs], which inhibit both COX-1 and COX-2, exert their therapeutic effects through inhibition of COX-2-dependent prostanoid biosynthesis and cause gastrointestinal damage through the inhibition of COX-1 (Perrone et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). The unavailability of highly selective COX-1 inhibitors (Brenneis et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2006\u003c/span\u003e) led to the use of genetic mouse models of selective deletion of COX-1 as a strategy to enlighten the potential pathogenic contribution of prostanoids synthesized via COX-1, e.g. to inflammatory arthritis (Chen et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2008\u003c/span\u003e), atherogenesis (McClelland et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2004\u003c/span\u003e), intestinal polyposis (Chulada et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2000\u003c/span\u003e) and skin carcinogenesis (Tiano et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). It is now believed that COX-1 is responsible for the primary prostanoid response to inflammatory stimuli (particularly in cells and tissues, where it is constitutively and predominantly expressed). Besides, special attention will be given (a) to the assays used to determine the activity and selectivity (often discordant and assay dependent) of the COX inhibitors (selectivity values are used to classify the large number of clinically used NSAIDs), and (b) the primary structure-activity relationship investigations, so far reported, performed to identify the few known COX-1 selective inhibitors (Perrone et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Xanthine oxidase is a member of a group of enzymes known as molybdenum iron-sulfur flavin hydroxylases (Symons et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e1989\u003c/span\u003e). It catalyzes the oxidation of hypoxanthine to xanthine and then to uric acid, the final reaction in the metabolism of purine bases (Zarepour et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). The accumulation of uric acid in the body is responsible for several diseases, and thus it plays a vital role in hyperuricemia and gout (Egwim et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). Inherited xanthine oxidase reductase (XOR) deficiency leads to xanthinuria and multiple organ failure syndromes caused by the accumulation of xanthine in different tissues (Borges et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). The inhibitors XO, integrase, and cyclooxygenase-1 from plants are much more useful (Umamaheswar et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2013\u003c/span\u003e) since they possess lesser side effects compared to uricosuric and anti-inflammatory agents (Umamaheswari et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). As a result, the present study aimed to evaluate the GC-MS analysis of chemical constituents of the \u003cem\u003eD. guineense\u003c/em\u003e fruits found in Abakaliki, South-eastern Nigeria. We further conducted virtual screening to predict the chemotherapeutic targets and drug-likeness of the chemical constituents.\u003c/p\u003e"},{"header":"2. Materials And Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Materials\u003c/h2\u003e \u003cp\u003eThe equipment, chemicals, and reagents used in this research work were of analytical grade and quality. The fresh fruits of \u003cem\u003eDialium guineense\u003c/em\u003e were collected by hand plucking from the wild plants in March 2019 from the Abakaliki Area of Ebonyi State, Nigeria. The plant was classified and authenticated by a plant taxonomist Mr. O. E. Nwankwo of the Department of Applied Biology of Ebonyi State University, Abakaliki, Nigeria. Part of the identified plant was kept in the herbarium at the Applied Biology Department, Ebonyi State University, Abakaliki, Nigeria, for reference purposes. The molecular docking apparatus used was software (AutoDock Vina plugin Pyrx, UCSF Chimera, and Discovery studios 2020), database (PDB, PubChem, and ChemSpider), and web servers (SwissAdme, and AdmetSar). The standard inhibitors used are Raltegavir (Ralt), Ibuprofen (Ibu), and Caffeine (Caf) for Integrase, Cycloxygenase-1, and Xanthine oxidase, respectively.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Methods\u003c/h2\u003e \u003cdiv id=\"Sec5\" class=\"Section3\"\u003e \u003ch2\u003e2.2.1. Preparation and extraction of \u003cem\u003eDialium guineense\u003c/em\u003e Fruits\u003c/h2\u003e \u003cp\u003eThe ripe fruits of \u003cem\u003eDialium guineense\u003c/em\u003e were destalked, deshelled and the fruits were carefully removed from the seeds. The fruits were shade dried at ambient temperature with constant turning to prevent fungal growth. The dried fruits were milled to obtain powdered fruit samples using an electric blender and were stored at 4\u003csup\u003e0\u003c/sup\u003eC temperature in a refrigerator in well-labelled airtight containers for analysis. Fifty grams of powdered fruit sample of \u003cem\u003eD. guineense\u003c/em\u003e was extracted with 400 ml of methanol in an orbital shaker for 24 hrs at room temperature. The extract was filtered using Whatman No.1 filter paper to remove extractable substances, at every 4 hrs interval. The combined extracts were then evaporated with a rotary evaporator, and the dried extract was stored at 4\u0026deg;C in a sterile container.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section3\"\u003e \u003ch2\u003e2.2.3. GC-MS Analysis of the \u003cem\u003eDialium guineense\u003c/em\u003e Fruits\u003c/h2\u003e \u003cp\u003eGC-MS analysis of the methanol extract of \u003cem\u003eD. guineense\u003c/em\u003e fruits was performed using Shimadzu Japan gas chromatography QP2010PLUS with a fused GC column (2010) coated with polymethyl silicon (0.25nm x 50m) and the conditions were as follows: Temperature programming from 80\u0026ndash;200\u003csup\u003e0\u003c/sup\u003eC held at 80\u003csup\u003e0\u003c/sup\u003eC for 1 min, rate 5\u003csup\u003e0\u003c/sup\u003eC/min and at 200\u003csup\u003e0\u003c/sup\u003eC for 20 min. Field ionization detector (FID) temperature 300\u003csup\u003e0\u003c/sup\u003eC, injection temperature 220\u003csup\u003e0\u003c/sup\u003eC, carrier gas nitrogen at a flow rate of 1 ml/min, split ratio 1:75. Gas chromatography-mass spectrum was conducted using GC-MS \u0026ndash;QP 2010 Plus Shimadzu Japan with injector temperature of 220\u003csup\u003e0\u003c/sup\u003eC and carrier gas pressure of 116.9 kpa. The column length is 30 m with a diameter of 0.25 mm and a flow rate of 50 ml/min. The elutes were automatically passed into a mass spectrometer with a dictator voltage set at 1.5 kV and a sampling rate of 0.2 sec. The mass spectrum was also equipped with a computer fed mass spectra data bank. Hermlez 233 M-Z centrifuge (Germany) was used.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section3\"\u003e \u003ch2\u003e2.2.3 Components Identification\u003c/h2\u003e \u003cp\u003eThe method described by Aja et al. (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) was used to identify the chemical constituents of the fruit extract by matching the peaks with Computer Wiley Micro-Soft libraries and confirmed by comparing mass spectra of the peaks and those from literature.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section3\"\u003e \u003ch2\u003e2.2.4 Virtual screening of \u003cem\u003eDialium guineense\u003c/em\u003e compounds\u003c/h2\u003e \u003cp\u003eThe target proteins (integrase, cycloxygenase1, and xanthine oxidase), the standard inhibitors (Raltegavir, Ibuprofen, and Caffeine), and the identified compounds of \u003cem\u003eDialium guineese\u003c/em\u003e were retrieved from RCSB PDB, PubChem, and Chemspider databases respectively. The retrieved proteins and ligands were individually and in turns prepared for molecular docking using UCSF Chimera. The protein and ligand preparation were by removing non-standard ligand (where necessary) and structural editing/minimization in the default settings with adding hydrogen and charges Gastiger. All the proteins and ligands were saved in PDB file format after preparation. AutoDock vina plugin Pyrx was used for virtual screening (Trott and Olso, 2010) of the identified compounds for complementary binding with the standard inhibitor of each target protein. This program was run using a searching grid extended over the ligand molecules with box spacing and coordinates (x, y,z) respectively as follows: integrase (55x50x70 and 5.13, 31.40, and \u0026minus;\u0026thinsp;10.61); cycloxygenase-1 (87x84x73 and \u0026minus;\u0026thinsp;32.82, -49.17 and 2.92); xanthine oxidase (120x94x114 and 29.22, 14.25 and 168.34) with other parameters set as default. The identified compounds were docked into the crystal structure of ligand molecules, and the highest-scoring pose binding at the same site with the standard inhibitor was selected for each of the identified compounds. Those with the best docking poses are predicted to be inhibitors of the target enzymes. Further, the best poses were visualized using the UCSF Chimera and Discovery studio 2020 to ascertain the protein-ligand interactions. ADMET (Absorption, Distribution, Metabolism, Excretion, and Toxicity) prediction and significant descriptors of drug-likeness such as mutagenicity, toxicological dosage level, and pharmacologically relevant properties of the best pose compounds were predicted using Swissadme (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.swissadme.ch\u003c/span\u003e\u003cspan address=\"http://www.swissadme.ch\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) and admetSAR (lmmd.ecust.edu.cn:8000) servers.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"3. Results","content":"\u003cp\u003e\u003cstrong\u003e3.1\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eGC-MS Constituents of Methanol Fruit-Extract of \u003cem\u003eD. guineense\u0026nbsp;\u003c/em\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe results of GC-MS constituents of methanol fruit-extract of \u003cem\u003eD. guineense\u003c/em\u003e showed that seventeen compounds were present in the fruits of \u003cem\u003eD. guineense\u0026nbsp;\u003c/em\u003e(Table 1). These compounds comprise mainly\u0026nbsp;saturated fatty acids, carboxylic acids, phenolic compounds, and esters. The composition of the extract comprises palmitic acid, ethyl ester, glutaric acid, decylpentafluorobenzyl ester, 4-[(\u003cem\u003eE\u003c/em\u003e)-(methoxyimino) methyl] phenol, and5-Hydrxoymethylfurfural or 5-(hydroxymethyl) furan-2-carbaldehyde. The identified compounds and their corresponding molecular weight, retention time, percentage (%) base peaks, structures and their bioactivity are shown in Table 1.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.3 Protein-Inhibitor Interaction visualization\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.3.1 Integrase vs DG best poses\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFigures 1-10\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cstrong\u003e3.3.2 COX1 vs DG best poses\u003c/strong\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFigures 11-18\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cstrong\u003e\u003cstrong\u003e3.3.3 Xanthine oxidase vs DG best poses\u003c/strong\u003e\u003c/strong\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFigures 19-21\u003c/p\u003e"},{"header":"4. Discussion","content":"\u003cp\u003e\u003cem\u003eDialium guineense\u003c/em\u003e (\u003cem\u003eD.\u0026nbsp;\u003c/em\u003e\u003cem\u003eguineense\u003c/em\u003e)\u0026nbsp;is one of the emerging medicinal plants of interest (Besong \u003cem\u003eet al\u003c/em\u003e., 2016). Various parts of \u003cem\u003eD.\u003c/em\u003e \u003cem\u003eguineense\u003c/em\u003e have been reported in ethnomedicine to have medicinal uses.\u003cem\u003e\u0026nbsp;\u003c/em\u003eStudies by Okudu \u003cem\u003eet al\u003c/em\u003e. (2017) on nutritional and sensory attributes of \u003cem\u003eD.\u003c/em\u003e \u003cem\u003eguineense\u003c/em\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003erevealed a reasonable number of antioxidants and numerous minerals such as magnesium, sodium, iron, potassium, beta-carotene (Vitamin A), copper, sugars and tartaric acid, citric acid, malic acid, ascorbic acid, and niacin. Similarly, Ogu \u003cem\u003eet al\u003c/em\u003e. (2012) reported several phytochemicals on the methanolic stem bark extract of \u003cem\u003eD\u003c/em\u003e.\u0026nbsp;\u003cem\u003eguineense\u003c/em\u003e: cardiac glycosides, tannins, phlobatannins, saponins, terpenoids, resins, steroids/triterpenes, alkaloids, flavonoids, reducing sugars and carbohydrates with anti-diarrheal effects.\u003c/p\u003e\n\u003cp\u003eIn the present study, the analysis of \u003cem\u003eD\u003c/em\u003e.\u0026nbsp;\u003cem\u003eguineense\u003c/em\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003efruits showed seventeen peaks from the GC-MS chromatogram. These peaks indicated the presence of seventeen compounds (DG1-17) depicted in Table 1. The composition comprises fatty acids, including Lauric acid (0.18%), Ethyl 5-methylnonanoate (2.30%), Myristic acid, methyl ester (2.39%), n-Tridecanoic acid methyl ester (2.32%), Methyl 14-methylpentadecanoate (5.21%), Palmitic acid, ethyl ester (34.23%), Undecanoic acid (4.52%) and Stearic acid, ethyl ester (1.37%). Thus, the fatty acid compositions of \u003cem\u003eDialium guineense\u0026nbsp;\u003c/em\u003ewere all saturated. Although eating foods containing saturated fats may raise low-density lipoprotein (LDL) cholesterol levels, thereby increasing the risk of cardiovascular diseases. Research has shown that saturated fats could undergo desaturation into their corresponding unsaturated forms. He also came to the conclusion that stearic acid is less likely to be incorporated into cholesterol esters. Also, a robust study by Emken (2018) on human isotope labeling showed that the fraction of dietary stearic acid that oxidatively desaturates to oleic acid is 2.4 times higher than the fraction of palmitic acid analogously converted to palmitoleic acid. He concluded further that stearic acid is less likely to be incorporated into cholesterol esters. Similarly, epidemiological and clinical studies by Hunter \u003cem\u003eet al\u003c/em\u003e. (2010) showed that dietary stearic acid has favorable effects on plasma LDL cholesterol relative to cholesterol-raising saturated fatty acids, indicating its possible replacement for tans\u0026nbsp;fat.\u003c/p\u003e\n\u003cp\u003eThis study also revealed cis-11-Hexadecenal (1.57%), 2-chloro-\u003cem\u003eN\u003c/em\u003e-[(2 chlorophyll) carbonyl]-\u003cem\u003eN\u003c/em\u003e-(2-methyl propyl)benzamide (1.72%), Butyl5-oxo-1-(trifluoroacetyl)pyrrolidine-2-carboxylate (1.24%), Glutaric acid, decylpentafluorobenzyl ester and 2,5-dihydroxy-6-methyl-2,3-dihydro-4\u003cem\u003eH\u003c/em\u003e-pyran-4-one (14.02%). Except cis-11-Hexadecenal, 2-chloro-\u003cem\u003eN\u003c/em\u003e-[(2-chlorophenyl)carbonyl]-\u003cem\u003eN\u003c/em\u003e-(2-methyl propyl)benzamide, Butyl 5-oxo-1-(trifluoroacetyl)pyrrolidine-2-carboxylate, Ethyl 5-methylnonanoate, 5-Hydrxoymethylfurfural or 5-(hydroxymethyl)furan-2-carbaldehyde and 4-[(\u003cem\u003eE\u003c/em\u003e)-(methoxyimino)methyl]phenol which showed no bioactive, the other compounds identified displayed various bioactivities such as 11B-HSD-Inhibitor, 17-beta-hydroxysteroid dehydrogenase-Inhibitor, Catechol-O-Methyl-Transferase-Inhibitor, 5-HETE-Inhibitor, 5-HT-Inhibitor, Methyl-Guanidine-Inhibitor, Urine-Acidifier, uric acid production inhibitor and arachidonic acid inhibitor by 2,4-Dihydroxy-2,5-dimethyl-3(2H)-furanone, 3,5-Dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one, 5-dihydroxy-6-methyl-2,3-dihydro-4\u003cem\u003eH\u003c/em\u003e-pyran-4-one, lauric acid, Myristic acid, methyl ester, n-Tridecanoic acid methyl ester, Methyl 14-methylpentadecanoate, Palmitic acid, ethyl ester, Undecanoic acid, Stearic acid, ethyl ester and Glutaric acid, decylpentafluorobenzyl ester respectively. The endowment of DG\u003cem\u003e\u0026nbsp;\u003c/em\u003ewith these constituents justifies its enormous folkloric uses. Notably, the wound-healing and antimicrobial potential of the dichloromethane fraction of \u003cem\u003eD.\u003c/em\u003e \u003cem\u003eguineense\u003c/em\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003efruit coat have been documented by Okeke \u003cem\u003eet al\u003c/em\u003e. (2016).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eFurthermore, the GC-MS analysis demonstrates that lauric acid (LA), a medium-chain fatty acid, had the lowest peak area (0.18%), indicating its low concentration in the \u003cem\u003eD. guineense\u003c/em\u003e. On the other hand, the dominant class of the compounds was Palmitic acid, ethyl ester with a base peak area of 34.23%, and prominently urine acidifier. Lauric acid, on the other hand, has noticeable bioactivities as an acidifier, acidulant, and arachidonic acid inhibitor, which may modulate prostaglandin synthesis. Furthermore, studies by Lappano \u003cem\u003eet al\u003c/em\u003e. (2017) established that LA induces apoptosis in cancer cells by increasing reactive oxygen species levels and stimulating the phosphorylation of epidermal growth factor receptor, extracellular-signal-regulated kinase, and induces the expression of proto-oncogenes. The presence of n-Tridecanoic acid methyl ester 2.32%), an Anaphylactic (antidote=Neostigmine), antitumor (Nasopharynx) also validates the application of \u003cem\u003eD. guineense\u0026nbsp;\u003c/em\u003ein traditional medicine for the treatment of allergy and cancer. Hence, supplementation with \u003cem\u003eD. guineense\u0026nbsp;\u003c/em\u003efruits may be the potential natural therapy against many multidrug-resistant diseases.\u003c/p\u003e\n\u003cp\u003eThe virtual screening aimed to gain insight into the pharmacological effects of the bioactive compounds. Table 2 showed fifteen compounds were retrieved and used for virtual screening. DG1, DG2, DG3, DG4, DG5, DG6, DG7, DG9, DG10, DG11, DG12, and DG17 (Table 3) were identified as potential inhibitors of at least one of the enzymes used. The results also showed that DG1 to 7, 11, 12, and 17 are integrase inhibitors, DG1, 2, 4, 5, 6, 9, 10, and 12 are cycloxygenase-1 inhibitors, while DG1, 3, and 9 are xanthine oxidase inhibitors (Table 2). They have binding affinities of between 3.9-6.1Kcal/mol. Figures 1\u0026ndash;21 show that potential inhibitors bind at the same binding sites as standard inhibitors for each enzyme. Furthermore, they interact with different amino acids while using similar bond types such as hydrogen bonds, van der Waals, unfavorable donors, pi-bonds, etc. The potential inhibitors have good drug-likeness (Table 4). Only DG11 and DG17 violate one each of the Lipinski Rules of five. As shown in Table 5, all potential inhibitors can be absorbed through the gastrointestinal tract and cross the blood-brain barrier. Others can achieve oral bioavailability. Others can attain oral bioavailability. They also do not interfere with the membrane transport proteins, the cytochrome p450, and p-glycoproteins. Thus, they have good physiological and pharmacokinetic properties. The toxicity profile showed that all the potential inhibitors except DG3 are toxic to the eyes. DG2 is mutagenic, and DG4, DG12, and DG17 are endocrine disruptors.\u003c/p\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eIn the present study, GC-MS analysis of \u003cem\u003eD. guineense\u003c/em\u003e showed interesting classes of chemical compounds with remarkable biological activities, including 11B-HSD-Inhibitor, 17-beta-hydroxysteroid dehydrogenase-Inhibitor, Catechol-O-Methyl-Transferase-Inhibitor, 5-HETE-Inhibitor, 5-HT-Inhibitor, Methyl-Guanidine-Inhibitor, Urine-Acidifier, uric acid production inhibitor, and arachidonic acid inhibitor. The molecular docking showed varied classes of bioactive compounds that could be useful as integrase, cycloxygenase-1, and xanthine oxidase inhibitors. These effects could account for the potential use of \u003cem\u003eD. guineense\u003c/em\u003e fruits in several disease conditions. However, virtual screening investigation of its pharmacological effects has given insight into its potential application in human nutrition and targets for chemotherapy. The optimization of the potential inhibitors to ward off the toxicity of the compounds can be a promising step toward drug discovery. Also, there is an urgent need to sensitize the public about the toxic bioactive compounds in the \u003cem\u003eD. guineense\u003c/em\u003e fruit as an intervention in public health, especially in the rural area where it is consumed raw as food.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eDeclaration of interests\u003c/strong\u003e\u003cbr\u003e\u0026nbsp;The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e"},{"header":"References","content":"\u003cp\u003eAchoba, I., Lori, J. A., Elegbede, J. A., and Kagbu, J. A. 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The predominance of cyclooxygenase 1 over cyclooxygenase 2 in the generation of pro-inflammatory prostaglandins in autoantibody-driven K/BxN serum-transfer arthritis. \u003cem\u003eArthritis Rheumatoid\u003c/em\u003e, \u003cem\u003e58\u003c/em\u003e: 1354-1365.\u003c/p\u003e\n\u003cp\u003eMcClelland, S., Toomey, S., Hahren, B., Fitzgerald, D. J., and Belton, O. A. (2004). Cyclooxygenase-1 gene deletion inhibits atherosclerosis in the ApoE\u0026ndash;/\u0026ndash; mouse model. \u003cem\u003eArteriosclerosis Thrombosis Vascular Biology\u003c/em\u003e, \u003cem\u003e24\u003c/em\u003e: 73-79.\u003c/p\u003e\n\u003cp\u003eChulada, P. C., Thompson, M. B., Mahler, J. F., Doyle, C. M., Gaul, B. W., Lee, C., Tiano, H. F., Morham, S. G., Smithies, O. and Langenbach, R. (2000). Genetic disruption of Ptgs-1, as well as Ptgs-2, reduces intestinal tumorigenesis in Min mice. \u003cem\u003eCancer Research\u003c/em\u003e, \u003cem\u003e60\u003c/em\u003e: 4705-4708.\u003c/p\u003e\n\u003cp\u003eTiano, H. F., Loftin, C. D., Akunda, J., Lee, C. A., Spalding, J., Sessoms, A., Dunson, D. B., Rogan, E. G., Morham, G., Smart, R. C. and Langenbach, R. (2002). Deficiency of either cyclooxygenase (COX)-1 or COX-2 alters epidermal differentiation and reduces mouse skin tumorigenesis. \u003cem\u003eCancer Research\u003c/em\u003e, \u003cem\u003e62\u003c/em\u003e: 3395-3401.\u003c/p\u003e\n\u003cp\u003ePerrone, M. G., Scilimati, A., Simone, L., and Vitale, P. \u0026nbsp;(2010). Selective COX-1 Inhibition: A Therapeutic Target to be Reconsidered. \u003cem\u003eCurrent Medicinal Chemistry\u003c/em\u003e, \u003cem\u003e17:\u0026nbsp;\u003c/em\u003e3769-3805 3769\u003c/p\u003e\n\u003cp\u003eUmamaheswar, M., Madeswaran, A., and Asokkumar, K. (2013).\u0026nbsp;Virtual Screening Analysis and \u003cem\u003eIn-vitro\u0026nbsp;\u003c/em\u003eXanthine Oxidase Inhibitory Activity of Some Commercially Available Flavonoids. \u0026nbsp;\u0026nbsp;\u003cem\u003eIranian Journal of Pharmaceutical Research\u003c/em\u003e, 12(3): 317-323\u003c/p\u003e\n\u003cp\u003eSymons, C. R. M., Taiwo, A. F., and Petersen, L. R. (1989). Electron addition to xanthine oxidase. An electron spins resonance study of the effects of ionizing radiation. \u003cem\u003eJournal of Chemical Society,\u0026nbsp;\u003c/em\u003e85: 4063-74.\u003c/p\u003e\n\u003cp\u003eZarepour, M., Kaspari, K., Stagge, S., Rethmeier, R., Mendel, R. R., and Bittner, F. (2010). Xanthine dehydrogenase AtXDH1 from \u003cem\u003eArabidopsis thaliana\u0026nbsp;\u003c/em\u003eis a potent producer of superoxide anions via its NADH oxidase activity. \u003cem\u003ePlant Molecular Biol\u003c/em\u003eogy, 72: 301-10.\u003c/p\u003e\n\u003cp\u003eEgwim, E. C., Vanchi, M. A., and Egwin, P. O. (2005). Comparison of xanthine oxidase activities in cow and goat milk. \u003cem\u003eBiokemistri,\u0026nbsp;\u003c/em\u003e17: 1-6.\u003c/p\u003e\n\u003cp\u003eBorges, F., Fernandes, E., and Roleira, F. (2002). Progress towards the discovery of xanthine oxidase inhibitors. \u003cem\u003eCurrent Medicinal Chemistry,\u003c/em\u003e 9: 195-217.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eUmamaheswari, M., Asokkumar, K., Sivashanmugam, A. T., Remyaraju, A., Subhadradevi, V., and Ravi, T. K. (2009). \u003cem\u003eIn-vitro\u0026nbsp;\u003c/em\u003exanthine oxidase inhibitory activity of the fractions of \u003cem\u003eErythrina stricta\u0026nbsp;\u003c/em\u003eRoxb. \u003cem\u003eJournal of Ethnopharmacol\u003c/em\u003e\u003cem\u003eogy\u003c/em\u003e, 124: 646-8.\u003c/p\u003e\n\u003cp\u003eBreda, A., Basso, L. A., Santos, D. S., and Walter, F. (2008). Virtual screening of drugs: score functions, docking, and drug design. \u003cem\u003eCurrent Computer-Aided Drug Design,\u003c/em\u003e 4: 265-72.\u003c/p\u003e\n\u003cp\u003eHirashima, A., and Huang, H. (2008). Homology modeling, agonist binding site identification, and docking in octopamine receptor of \u003cem\u003ePeriplaneta americana\u003c/em\u003e. \u003cem\u003eComputational Biology \u0026amp; Chemistry,\u003c/em\u003e 32: 185-90.\u003c/p\u003e\n\u003cp\u003ePowers, R. (2009). Advances in nuclear magnetic resonance for drug discovery. \u003cem\u003eExpert Opinion in Drug Disc\u003c/em\u003e\u003cem\u003eovery\u003c/em\u003e, 4: 1077-98\u003c/p\u003e"},{"header":"Tables 1-5","content":"\u003cp\u003eTables 1-5 are available in the Supplementary Files section.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Dialium guineense fruit, GC-MS analysis, virtual screening, potential inhibitors, ADMET profile","lastPublishedDoi":"10.21203/rs.3.rs-1727120/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-1727120/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e \u003cem\u003eDialium guineense\u003c/em\u003e has a long history of medicinal use. This study evaluated the chemical constituents of the fruits using Gas chromatography-mass spectrometry (GC-MS) and virtual screening for potential inhibitors of three enzymes, integrase, cycloxygenase-1, and xanthine oxidase, to identify some of their therapeutic targets via bioinformatics tools. The results of the GC-MS revealed seventeen chemical constituents: 2,4-Dihydroxy-2,5-dimethyl-3(2H)-furanone (DG1), 3,5-Dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one (DG2), 5-Hydrxoymethylfurfural or 5-(hydroxymethyl)furan-2-carbaldehyde (DG3), 4-[(\u003cem\u003eE\u003c/em\u003e)-(methoxyimino)methyl]phenol (DG4), 2,5-dihydroxy-6-methyl-2,3-dihydro-4\u003cem\u003eH\u003c/em\u003e-pyran-4-one (DG5), Lauric acid (DG6), Ethyl 5-methylnonanoate (DG7), Myristic acid, methyl ester (DG8), n-Tridecanoic acid methyl ester (DG9), Methyl 14-methylpentadecanoate (DG10), Palmitic acid, ethyl ester (DG11), Undecanoic acid (DG12), Stearic acid, ethyl ester (DG13), cis-11-Hexadecenal (DG14), 2-chloro-\u003cem\u003eN\u003c/em\u003e-[(2-chlorophenyl)carbonyl]-\u003cem\u003eN\u003c/em\u003e-(2methylpropyl)benzamide (DG15), Butyl-5-oxo-1-(trifluoroacetyl)pyrrolidine-2-carboxylate (DG16), and Glutaric acid, decylpentafluorobenzyl ester (DG17). Fifteen of these compounds, already deposited in the ChemSpider database, were retrieved and screened. Standard inhibitors of the enzymes validated the molecular docking. Only DG compounds that bind at the same site with the inhibitors were considered the potential enzyme inhibitors. DG1-17 demonstrated promising inhibition for at least one of the enzymes. The results also showed that DG1, 7, 11, 12, and 17 are integrase inhibitors, and DG1, 2, 4, 5, 6, 9, 10, and 12 are cycloxygenase-1 inhibitors, while DG1, 3, and 9 are xanthine oxidase inhibitors. The potential inhibitors' drug-likeness and ADMET properties revealed that DG11 and DG17 violated one of the Lipinski rules of five. All the \u003cem\u003eDialium guineense\u003c/em\u003e compounds showed good physiological and pharmacokinetic properties and were neither promiscuous to cytochrome p450 nor p-glycoprotein enzymes. Except for DG3, others are toxic to at least one of the organs, endocrines, and genomics. Our findings show that biologically active compounds may be responsible for the ethnomedical and nutritional uses of the fruits. Although some constituents have toxic effects, potential inhibitors can be optimized and synthesized as analogs of these standard inhibitors used in their chemotherapeutic targets.\u003c/p\u003e","manuscriptTitle":"Gas chromatographic-mass spectrometric (GC-MS) analysis and virtual screening of ripped fruit of Dialium guineense for potential inhibitors of integrase, cycloxygenase-1, and xanthine oxidase","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2022-06-14 20:14:20","doi":"10.21203/rs.3.rs-1727120/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"67ef89fa-7170-4a2a-b371-42006b0e409e","owner":[],"postedDate":"June 14th, 2022","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2023-03-06T03:29:17+00:00","versionOfRecord":[],"versionCreatedAt":"2022-06-14 20:14:20","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-1727120","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-1727120","identity":"rs-1727120","version":["v1"]},"buildId":"_2-kVJe1T_tPrBINL-cwx","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}