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Exploring the Rational Designing of Protein Ligand Binding for Mouth Ulcer from Methanolic Extract of Olax Psittacorum

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Exploring the Rational Designing of Protein Ligand Binding for Mouth Ulcer from Methanolic Extract of Olax Psittacorum Original Article Exploring the Rational Designing of Protein Ligand Binding for Mouth Ulcer from Methanolic Extract of Olax psittacorum Choudhury Priyabati1,*, Adhikari Lopamudra2, Pattanaik Sovan2 1School of Pharmacy, ARKA JAIN University, Jharkhand, INDIA. 2Department of Pharmaceutical Analysis and Quality Assurance, School of Pharmaceutical Sciences, Siksha ‘O’ Anusandhan (Deemed to be University), Kalinga Nagar, Bhubaneswar, Odisha, INDIA. ABSTRACT Background: Olax psittacorum, a flowering plant grown in open forests, is consumed by the locals as dietary sources. Aim: This study was designed to check the phytochemical screening, antioxidant activity, antimicrobial studies and Chorioallantoic Membrane (CAM) study in the methanolic extract of Olax psittacorum. Further, in silico docking study was performed on the basis of GC-MS report of the methanolic extract. Methods: Antimicrobial activity was tested against five bacterial strains Escherichia coli, Salmonella typhi, Pseudomonas aeruginosa, Shigella flexneri, Straphylococcus aureus. In silico docking study was done by two software, Patchdock followed by Firedock and HEX 8.0.0. to find the best fit receptor ligand docked structure. Results: Methanolic extract of Olax psittacorum has exhibited appreciable antioxidant activity. The study showed that the methanolic extract of Olax psittacorum (10mg/ml) with DMSO shows very less activity against Escherichia coli, Salmonella typhi and did not show any activity in Pseudomonas aeruginosa, Shigella flexneri, Staphylococcus aureus. The result of the CAM study showed that the extract has good antiangiogenic activity. The methanolic extract containing phytoligand 1, 2 -Benzenedicarboxylic acid, butyl octyl ester against salivary peroxidase shows maximum Global energy of -69.40 kcal/mol. Conclusion: This study attempted a new dimension for establishing phytochemicals present over methanolic extract of Olax psittacorum which may be useful for mouth ulcer prevention. Science and technology are growing faster to make up things much better but is still lagging with some issues. In vitro and in vivo models help us in determining many facts about plant and its constituent but still it also has many drawbacks. In an intention to Submission Date: 02-06-2020; overcome the drawbacks an economic and moreover not harming any life creature CAM Revision Date: 02-12-2020; studies were performed. Accepted Date: 09-02-2021 Key words: Protein-ligand binding, Toxicity, Antioxidant study, Activity study, Methanolic DOI: 10.5530/ijper.55.2.83 extract. Correspondence: Dr. Lopamudra Adhikari School of Pharmaceutical Sciences, Siksha ‘O’ INTRODUCTION Anusandhan (Deemed to be University), Kalinga Nagar, An ulcer is an exposed sore in the lining ulcer. But sometimes the cause of a mouth Bhubaneswar-751003, of the epithelial cell or deep injury in a ulcer is unknown. Oral ulceration may occur Odisha, INDIA. Phone no: +91 9776354163 specific region of the body or organ causing due to viral infections or disruption in the Email id: lopamudraadhikari@ its degradation and disturbing the usual function of gastrointestinal, haematological, soa.ac.in physiology of the affected organ.1 Mouth immunological and dermatological systems.2 ulcers are development of small painful The oral mucosa is coated by saliva, shielding sores on the inside lining of the mouth. it leads to suffering during eating, swallowing They grow on the inside of the cheeks, lips and speaking. In people with reduced saliva, and underneath of the tongue. Destruction mouth soreness is very common. of the inner lining of the mouth can be The human salivary peroxidase system caused by various factors, leading to a mouth (SPS) is one of the non-immunoglobulin www.ijper.org Indian Journal of Pharmaceutical Education and Research | Vol 55 | Issue 2 | Apr-Jun, 2021 455 Priyabati, et al.: Protein Ligand Binding for Mouth Ulcer from Methanolic Extract of Olax psittacorum defence factors which regulate the quantity and species alkaloids, glycosides, carbohydrates, tannins, saponins, distribution of oral micro-organism that helps in the flavonoid, proteins steroids, oils, fats etc. maintenance of decent oral health. It prevents toxic In vitro antioxidant assay build-ups of hydrogen peroxide (H2O2) and deactivates many cancer-causing and mutagenic compounds.3 Under For determining the in vitro antioxidant capacity, two normal conditions a physiological balance exists, most acceptable method DPPH (2, 2´-diphenyl- but in case of mucosal injury the balance between 1picrylhydrazyl) and ABTS (2, 2´-azinobis the aggressive factor and the defensive mechanism is 3-ethylbenzthiozoline-6 sulphonic acid) were carried out disrupted thus leading to formation of mouth ulcer. separately.7 Extract solution of 1 mg/ml was prepared From the beginning of human history, the importance using methanol and from it further dilutions of 50, 100, of medicinal plant was known and it has been used in 200, 300, 400, 500 µg/ml was obtained. drug development and for curing different diseases. DPPH radical scavenging activity Researchers have always been fascinated by the traditional folk treatment consuming wild plants to 1 ml from each of the dilution was taken into different exploration for new medication to develop fit life for test tubes to which 3ml DPPH solution (methanolic) human.4 was added to each of the test tube. The mixture was Olax psittacorum a flowering plant grown in open forests, shaken and left in dark at room temperature. Absorbance belonging to the family Olacaceae, found throughout was measured at 517nm after 30 min against a blank. the topical region of the world. The plant contains Ascorbic acid was taken as the positive control. The saponin, which when given orally to mice shows percentage of inhibition of DPPH (I %) was calculated anti-inflammatory properties and decreases oedema. by means of the following equation: Olaxoside also shows laxative action. The leaf of Olax Inhibition (%) = [(Acontrol- Atest)/Acontrol] × 100 5 psittacorum also shows antioxidant activity. Where, Acontrol = the absorbance of ABTS solution without any sample MATERIALS AND METHODS Atest = the absorbance of ABTS solution with sample in different concentration Plant collection ABTS+ Scavenging Activity Fresh leaves of Olax psittacorum were collected from Regional Plant Research Centre (RPRC), Odisha. The A dark coloured greenish solution was obtained freshly collected leaves were washed, dried in shade for when the mixture of ABTS (7.0 mM) and Potassium about 7 days and then grinded to form coarsely grounded persulphate (140 mM) was kept in the dark for 16 h at powder. Authenticated field number RMOP-1 ambient temperature. The ABTS solution was diluted to obtain an absorbance of 0.700 ±0.05 at 754 nm Preparation of plant extract with methanol. Then 3ml of ABTS was added to 1ml The dry powder sample were placed in the glass Soxhlet of the sample concentration (50-500µg/ml). After 30 and successive extraction using Petroleum ether followed min the absorbance was taken at 754nm. The ABTS+ by methanol (600ml) for 2 days was carried out for total scavenging activity was calculated by means of the extraction process. At the end the plant methanolic following equation: extract was filtered using Whatman filter paper and was Inhibition (%) = [(Acontrol- Atest)/Acontrol] × 100 evaporated to obtain a concentrated leaf methanolic Where, Acontrol = the absorbance of ABTS solution extract (LME), which is a greenish black sticky mass. without any sample Percentage yield of the extract was calculated. Atest = the absorbance of ABTS solution with sample in Amount of grounded materials different concentration introduced for soxhlation % Yield = × 100 Antimicrobial Study Amount of concentrated extract obtained Bacterial strains Phytochemical screening The antibacterial activity of the methanolic extracts Using standard qualitative method screening for was evaluated using five bacterial strains. Four strains major phytochemical constituents was carried out.6 of gram-negative bacteria (Escherichia coli, Pseudomonas Phytochemical screening test was carried out from aeruginosa, Salmonella typhi, Shigellaf lexneri) and one gram- freshly prepared methanolic extract of the leaves for positive bacteria (Straphylococcus aureus). The bacterial 456 Indian Journal of Pharmaceutical Education and Research | Vol 55 | Issue 2 | Apr-Jun, 2021 Priyabati, et al.: Protein Ligand Binding for Mouth Ulcer from Methanolic Extract of Olax psittacorum strains were provided by RMRC (Regional Medical KMMTGELRNKLFQPTHRIHGFDLAAINTQRCR Research Centre), Bhubaneswar. DHGQPGYNSWRAFCDLSQPQTLEELNTVLKSK MLAKKLLGLYGTPDNIDIWIGAIAEPLVERGRV Inoculums preparation GPLLACLLGKQFQQIRDGDRFWWENPGVFTN Each bacterial strain was sub cultured overnight at 37°C EQKDSLQKMSFSRLVCDNTRITKVPRDPFWANS in Nutrient Agar media or Lysogeny Agar media. The YPYDFVDCSAIDKLDLSPWASVKN bacterial growth was harvested using 1ml of sterile saline water. Screening for best homologous templates Well diffusion method for the determination of Homology modelling approach was used for predicting zone of inhibition 3-Dstructure of salivary peroxidase which is based on pre-determined homologous template. In the retrieval Using well diffusion method the antibacterial activity of sequence of salivary peroxidase from NCBI database of the methanolic extract of Olax psittacorum was resulting query sequence with the accession AAC50717.1 performed. Mueller-Hilton agar (15ml) medium was were subjected by using PSI-BLAST. After analysis the
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