Potential activity of Madhuca longifolia leaf extract: through In vitro, Pharmacological and In silico studies

Jerine Peter S¹, Ram Kumar K¹, Manisha P¹, Sangeetha N¹, Arun Raj N¹, Usha Kumari², Evan Prince Sabina¹*

¹School of Biosciences and Technology, VIT, Vellore, India.

²Faculty of Medicine, AIMST University, Malayasia

*Corresponding Author E-mail: eps674@gmail.com

ABSTRACT:

Madhuca longifolia is a universal panacea in the treatment as ayurvedic medicine which is traditionally valued for timber, flowers, fruits, leaves and seeds. The leaf of M.longifolia is known to have anti-diabetic, anti-oxidant, anti-ulcer and anti-inflammatory activities. The aim of our research is to know the beneficial activity of the leaf of M.longifolia through in vitro, pharmacological and in silico studies. The in vitro assays like Total phenolic content, DPPH assay, catalase activity and peroxidase activity was performed in ethanolic, methanolic and aqueous leaf extract of M.longifolia on serial dilution. The pharmacological activities were observed through the hot-plate method, analgesic method, antipyretic method and ulcerogenic method in female Wistar albino rats. The in silico analysis was performed through patch dock online server against the nuclear receptor and the active compounds of M.longifolia leaf. The 3-dimensional structures of the nuclear receptors were retrieved from RCBS protein data bank and the ligands were obtained from Corina molecular network. The in vitro assay demonstrated the antioxidant activity of all the leaf extract in which aqueous extract shown a potential antioxidant activity in 1:2 serial dilutions. The aqueous leaf extract on rats at 500 mg/kg. b.w. p.o. dosage has M.longifolia beneficial pharmacological activities than other extract. The ligands like myricetin 3-O-L-rhamnoside, quercetin 3-galactoside, a-D-Mannopyranoside, D-Allose, myricetin, Quercetin, myricetin 3- O-arabinoside and 3'-Hydroxy-4'-methoxydiclofenac has M.longifolia potential binding affinity with the receptors. Thus, the antioxidant, pharmacological and potential binding affinity of M.longifolia leaf was demonstrated in our research.

KEYWORDS: Madhuca longifolia leaf extract, in vitro analysis, antioxidant, pharmacological activities, in silico docking.

1. INTRODUCTION:

Madhuca longifolia is wild shady and cultivated large deciduous tree which found predominately in central India. It is commonly termed as mahua and buttercup. The tree is traditionally valued for timber, flowers, fruits, leaves and seeds¹. This tree is worshipped as the Goddess because it is known to be used in the maintaining the human health for more than thousand years. M.longifolia is considered as universal panacea in the treatment as ayurvedic medicine².

Leaves of M.longifolia are used for the preparation of ghee, in the treatment of eczema, respiratory infections, emaciation, debility and intestinal worms³. The bark is used in the treatment of diabetes, rheumatism and teeth problems. The flower of M.longifolia is used for the enhancing the breast milk, treatment of chest problems like bronchitis and it is known for its nutritional value. The flower is also used in the production of liquor and for the manufacture of vinegar⁴. The fruit and seed of M.longifolia are rich in fat content that is used in the treatment of piles, skin disease, headache and rheumatism. The fat-rich part of the plant is used in the manufacture of soap and the seed cake is used for fishing and as pesticide and insecticide⁵. The phytochemical analysis on M.longifolia has M.longifolia activity as steroids, flavonoids, saponins, carbohydrates, glycosides and triterpenoids⁶. During the chemical reaction, some free radicals are produced inside the body which will lead to the accumulation of toxic metabolites. The accumulation of these toxic metabolites is termed as oxidative stress that ultimately results in organ damage. Natural enzymes like superoxide dismutase, Catalase, reduced glutathione, glutathione peroxidase are present in the body which degrades the accumulation of free radical that prevent the system from the formation of oxidative stress⁷. 2, 2‑diphenyl‑1‑picrylhydrazyl (DPPH) is commonly used to estimate the antioxidant activity of the plant extract. Recent research has discovered an instrument to identify the amount of antioxidant found in food and beverage which involve the electrode with redox probe and potentiometric⁸. DPPH assay is reliable and rapid assay to determine the free radical scavenging level of the antioxidants⁹. Toxicology methods through in silico techniques are much useful in the manufacturing of potential drug and optimizing the lead compound that were predominant at the designing stage in pharmaceutical industry¹⁰. These methods will help in identifying the active ingredients of the compound that are low liable to toxicity. The methods known for the in silico toxicology analysis are QSAR (Predictive Quantitative Structure Activity Relationship), e-tox, predictive ADMET, i-drug discovery, etc¹¹. The aim of our research is to know the beneficial activity of the leaf of M.longifolia through in vitro, pharmacological and in silico studies.

2 MATERIALS AND METHODS:

2.1 In vitro studies:

The required chemicals were obtained from Sigma Adrich, India. The Madhuca longifolia leaves were obtained and were authenticated by Prof. Jayaraman, Director of Institute of Herbal Botany, Plant Anatomy Research Centre (PARC), Chennai, India. The taxonomical identification number for the specimen is PARC/2016/3322. The waste particles in leaves were removed by rinsing it in running water then it was dried for a month in room temperature. The fine powered leaves were used for the preparation of Ethanolic, Methonolic and Aqueous extract.

The Ethanolic, Methonolic and aqueous extract were prepared by dissolving 10g of the fine grinded leaves of Madhuca longifolia in 100ml of ethanol, methanol and distilled water respectively in different conical flash which is then incubated in 25°C for 48 hours and 24 hours respectively at constant shaking. Later it was filtered by Whatman filter paper-1, then concentrated and stored in 4°C for future use. All the three extracts were diluted by serial dilution at the concentration of A (1:2), B (1:4), C (1:8), D (1:16) and E (1:32). The serial diluted samples were analysed to observe the level of total phenolic content¹², DPPH assay¹³, catalase activity¹⁴, and peroxidase activity^15(p) for each extract respectively.

2.2 Pharmacological studies:

The studies to observe the pharmacological activity was carried out on female Wistar albino rats of 180 to 220 grams weight according to the CPCSEA guidelines which was obtained and approved (Reg no: VIT/IAEC/13/Feb13/21) by institutional animal ethical committee, VIT, Vellore, India. The rats are provided with the suitable environment and food according to the guidelines. Indomethacin tablet was purchased from Sun Pharma Ltd, Tamil Nadu, India.

The rats were divided into five group and six rats in each group. Group-I is normal control, group-II is treated with indomethacin (10mg/kg/b.wt., i.p), group-III is treated with aqueous leaf extract of Madhuca longifolia (ALMEL) at the dosage of 500mg/kg/b.wt., p.o., group-IV is treated with ethanolic leaf extract of Madhuca longifolia (ELMEL) at the dosage of 500mg/kg/b.wt., p.o. and group-V is treated with methanolic leaf extract of Madhuca longifolia (MLEML) at the dosage of 500 mg/kg/b.wt., p.o. The pharmacological activity was analysed through Hot- Plate test, Analgesic Test¹⁶, Antipyretic Test¹⁷, Ulcerogenic Test¹⁸, Stomach and intestine Mucosa.

2.3 Computational analysis:

2.3.1 Receptor designing:

The receptors that are responsible for the hepto, renal and gastro toxicity were selected from the literature. The PDB structures of the receptors were retrieved from RCBS (Research Collaborator for Structural Bioinformatics) protein data bank. The PDB Id for each receptor is Apo Human Pregnane X Receptor (PXR): 1ILG, Constitutive androstane receptor (CAR): 1XNX, Nuclear bile acid receptor (FXR): 1OSH, Liver X receptors alpha (LXR): 5AVI and Nuclear factor kappa-light-chain-enhancer of activated B cells (Nf-kB): 1NFK. Each receptor was prepared for docking by removing the hydrogen atom and water molecule.

2.3.2 Ligand designing:

The active compounds of the leaf of Madhuca longifolia is selected as the ligands for the docking procedure. PubChem database is used to obtain the canonical smile of the ligands which is submitted to Corina Molecular Network to retrieve their PDB structure. The ligand representation and its PubChem Id are: Propanoic acid (1032), N-Methoxy-N-methylacetamide (537505), Furfural (7362), Butanoic acid (2462), 2-Furancarboxaldehyde, 5-methyl (12097), phenol (996), 2-Hydroxy-gamma-butyrolactone (545831), 1-Amino-2, 6-dimethylpiperidine (123482), 4-Hydroxy-2, 5-dimethyl-3(2H)-furanone (19309), 3H-Pyrazol-3-one, 2, 4-dihydro-2, 4, 5-trimethyl (580964), 3-Hexanone, 2, 5-dimethy (15901), 4H-Pyran-4-one, 2, 3-dihydro-3, 5-dihydroxy-6-methyl (119838), Benzenecarboxylic acid (243), 2-Furancarboxaldehyde, 5-(hydroxymethyl) (237332), Hydroquinone (785), Cyclohexanecarboxylic acid 2-methyl, (85811), D-Allose (439507), 1-Methyl-2-pyrrolidone-4-carboxamide (541476), a-D-Mannopyranoside, methyl (101798), β-carotene (5280489), xanthophylls (24728610), erthrodiol (101761), palmitic acid (985), myricetin (5281672), myricetin 3- O-arabinoside (21672568), myricetin 3-O-L-rhamnoside (5352000), Quercetin (3, 3’, 4’, 5, 7-pentahydroxyflavone) (5280343), quercetin 3-galactoside (5281643), oleanolic acid (10494), β-sitosterol (222284), stigmasterol (5280794), β-sitosterol- β-Dglucoside (β-Dglucoside of β-sitosterol) (12309055), n-hexacosanol (68171), n-octacosanol (68406), 3'-Hydroxy-4'-methoxydiclofenac (129615), 3'-hydroxydiclofenac (112230), (3Z)-Phycocyanobilin (5280816).

2.3.3 In silico docking and analysis of docked complex:

The molecular docking procedure was carried out in PatchDock online server which is based on the principle of shape complementary algorithm. The receptors and ligands were submitted in the server on PDB format and the results were received through mail id. The best interactions were selected for the experiment based on Score, Area, and ACE. The docked complex were visualised and analysed in PyMOL software. The docked complex was analysed of interacting residues and atoms along with bond length which was labelled and recorded using the software.

2.4 Statistical analysis:

The statistical analysis was carried out using Analysis of Variance (ANOVA) which represents the significance difference between each groups using Students Newmam-Keul’s test using Graph pad software InStat version 3.10. For significance, p<0.05 is considered. The results were expressed in mean ± SD.

3 RESULTS:

3.1 In vitro studies on the leaf extract of M.longifolia:

3.1.1 Outcome of M.longifolia leaf extract on Total Phenolic content:

The activity of M.longifolia leaf extract on Total Phenolic content is represented in Figure 1a. It is observed that the dilution of A (1:2) has M.longifolia better result on compared to all other dilution in all the extract. Activity of total phenolic content reduces respective to the ratio of serial dilution. Compared to all extract, aqueous extract shown a better effect in total phenolic content. On compared to ethanolic and methanolic, there was better activity found in the methanolic extract of M.longifolia.

3.1.2 Outcome of M.longifolia leaf extract on DPPH assay:

Aqueous extract possessed better result in DPPH assay than all other extract of M.longifolia (Figure 1b). In all the extract the dilution of 1:2 shown good activity on compared to all other dilution. The ratio of serial dilution is directly proportional to the activity of the DPPH assay. Methanolic extract of M.longifolia shown a better activity than the ethanolic extract.

3.1.3 Outcome of M.longifolia leaf extract on Catalase activity:

Figure 1c represent the activity of M.longifolia leaf extract on Catalase assay. The sample with the dilution of 1:2 has show potential activity in catalase assay in all the extract on compared to the other dilution like 1:4, 1:8, 1:16 and 1:32. Aqueous extract shown a potential effect on compared to all other extracts.

3.1.4 Outcome of M.longifolia leaf extract on Peroxidase activity:

Peroxidase activity of the ethanolic, methanolic and aqueous extract of M.longifolia leaf extract is represented in Figure 1d. A gradual reduction in the level of peroxidase activity was observed in all the extract with respect to the decrease in dilution. Compared all other dilutions, the dilution of 1:2 shown potential activity. The aqueous extract possessed better activity on compared to all other extract.

3.2 Pharmacological studies:

3.2.1 Response of M.longifolia leaf extract on Hot- Plate test:

Figure 2a represents the effect of the leaf extract of M.longifolia on hot plate. The ability of rats in withstanding the heat in normal control group is 5 second. The reaction time of Indomethacin treated rats to jump out of the hot plate is significantly (p<0.05) high on compared to normal control rats. Whereas the reaction time of ALEML and MLEML is significantly decrease on compared to group-II, but it is similar to normal control group. The reaction time of rats treated with ELEML was also decrease on compared to group-II. Compared to all other extract, the rats treated with ALEML shown the better tolerance to hot plate in withstanding the heat which is similar to normal control group. ALEML shown better result on compared to group-II.

3.2.2 Response of M.longifolia leaf extract on Analgesic Test:

The number of writhining in 30 minutes of control group induced with acetic acid is 67 (Figure 2b). The number of writhining of ALEML treated rats is 50 which significantly decrease on compared to group-I that was similar to indomethacin-treated rats. The writhining of ELEML and MLEML is also similar to group-II.

3.2.3 Response of M.longifolia leaf extract on Antipyretic Test:

Figure 2c represents the rectal temperature of the leaf extract of M.longifolia on exposed to baker’s yeast. At 0^th hour the rectal temperature of the entire group is in normal level. After the administration of yeast there is a slight increase in the temperature at the 18^th hour. The rectal temperature of the rats of control group has shown a significant increase on the 22^nd hour. Indomethacin and the leaf extract treated rats were able to maintain the temperature in the normal range on every time interval.

3.2.4 Response of M.longifolia leaf extract on Ulcerogenic Test:

The ulcerogenic test on the leaf extract of M.longifolia was represented in Figure 2d. After the fasting period of 16 hours, there is much elevation in the ulcer score of the control group.

3.3 Computational analysis:

3.3.1 Docked complex of 1ILG receptor with ligands:

Among the docked complex with 1ILG and ligands, the interaction were found in the ligands like Butanoic acid, 2-Hydroxy-gamma-butyrolactone, 2-Furancarboxaldehyde, 5-(hydroxymethyl), D-Allose, a-D-Mannopyranoside, methyl, myricetin, myricetin 3- O-arabinoside, myricetin 3-O-L-rhamnoside, Quercetin, quercetin 3-galactoside, β-sitosterol- β-Dglucoside, 3'-Hydroxy-4'-methoxydiclofenac and 3'-hydroxydiclofenac. Other ligands have not shown any bonding. Among the interacting ligands with 1ILG, myricetin 3-O-L-rhamnoside and quercetin 3-galactoside were found to have potential interactive residues. Both myricetin 3-O-L-rhamnoside and quercetin 3-galactoside were observed to have five interaction which is M.longifolia in figure 4a and figure 4b. The residues name and bond length of the docked complex of myricetin 3-O-L-rhamnoside with 1ILG is MET-243:-2.2, SER-247:-3.4, GNL-285:-3.5, HIS-407:-3.6 and SER-208:-2.8. The residues name and bond length of the docked complex of quercetin 3-galactoside and 1ILG are SER-247:-2.3, SER-247:-2.2, ARG-410:-2.3, SER-208:-2.4 and GLU-321:-2.8

3.3.2 Docked complex of 1XNX receptor with ligands:

The ligands like Propanoic acid, Butanoic acid, 2-Hydroxy-gamma-butyrolactone, 4-Hydroxy-2, 5-dimethyl-3(2H)-furanone, 3H-Pyrazol-3-one, 2, 4-dihydro-2, 4, 5-trimethyl, 4H-Pyran-4-one, 2, 3-dihydro-3, 5-dihydroxy-6-methyl, Benzenecarboxylic acid, 2-Furancarboxaldehyde, 5-(hydroxymethyl), Hydroquinone, Cyclohexanecarboxylic acid 2-methyl,, D-Allose, a-D-Mannopyranoside, methyl, erthrodiol, myricetin, myricetin 3- O-arabinoside, myricetin 3-O-L-rhamnoside, Quercetin, quercetin 3-galactoside, oleanolic acid, β-sitosterol- β-Dglucoside, 3'-hydroxydiclofenac and (3Z)-Phycocyanobilin has shown bonding and others have not shown any bonding. The docked complex of a-D-Mannopyranoside, methyl with 1XNX and D-Allose with 1XNX was observed to have potential interaction. The docked complex of a-D-Mannopyranoside, methyl with 1XNX has shown six interacting residues (Figure 4c), the residue and bond of this interacting complex are SER-315:-3.1, ARG-312:-2.5, HIS-263:-1.7, GLN-314:-2.9, SER-315:-2.8 and SER-315:-3.5. Five interacting residues was found in the docked complex of D-Allose with 1XNX (Figure 4d), the residues name and bond length are HIS-263:-2.7, ARG-312:-2.7, SER-315:-3.3, SER-315:-2.8 and SER-315:-3.2.

3.3.3 Docked complex of 1OSH receptor with ligands:

The score, area and ACE of the docked complex were mentioned in table 3. The interacting residues were observed in 4-Hydroxy-2, 5-dimethyl-3(2H)-furanone, 4H-Pyran-4-one, 2, 3-dihydro-3, 5-dihydroxy-6-methyl, Benzenecarboxylic acid, Hydroquinone, D-Allose, 1-Methyl-2-pyrrolidone-4-carboxamide, a-D-Mannopyranoside, methyl, erthrodiol, myricetin, myricetin 3- O-arabinoside, myricetin 3-O-L-rhamnoside, Quercetin, quercetin 3-galactoside, oleanolic acid, β-sitosterol, n-octacosanol, 3'-Hydroxy-4'-methoxydiclofenac and (3Z)-Phycocyanobilin. Other ligands have not shown any bonding. Among the other docked complex with 1OSH, myricetin and Quercetin has possessed a potential interaction with the receptor. Five interaction was found between myricetin and 1OSH, their residues and bond length are SER-336:- 3.4, SER-336:-1.9, TYR-373:-2.8, TYR-365:-2.4 and LEU-291:-2 (Figure 4e).

3.3.4 Docked complex of 5AVI receptor with ligands:

Interacting residues were found in the docked of 5AVI with the ligands like Butanoic acid, phenol, 2-Hydroxy-gamma-butyrolactone, 4-Hydroxy-2, 5-dimethyl-3(2H)-furanone, 3H-Pyrazol-3-one, 2, 4-dihydro-2, 4, 5-trimethyl, 3-Hexanone, 2, 5-dimethy, 4H-Pyran-4-one, 2, 3-dihydro-3, 5-dihydroxy-6-methyl, 2-Furancarboxaldehyde, 5-(hydroxymethyl), Hydroquinone, Cyclohexanecarboxylic acid 2-methyl, D-Allose, 1-Methyl-2-pyrrolidone-4-carboxamide, a-D-Mannopyranoside, methyl, erthrodiol, palmitic acid, myricetin, myricetin 3- O-arabinoside, myricetin 3-O-L-rhamnoside, Quercetin, quercetin 3-galactoside, β-sitosterol- β-Dglucoside, 3'-Hydroxy-4'-methoxydiclofenac and (3Z)-Phycocyanobilin (table 4). Others have not shown any bonding. Quercetin has shown eight potential interactions with 5AVI than the other docked ligands with 5AVI (Figure 4f). The residues and bond length of the docked complex of Quercetin and 5AVI are GLU-348:-2.4, ARG-344:-2.4, ARG-344:-2.4, GLN-375:-2.1, GLN-375:-3.0, ASP-368:-3.1 and ALA-367:-3.3.

Others have not shown any bonding. The potential interaction was found between the docked complex with the ligands like D-Allose, a-D-Mannopyranoside, myricetin 3- O-arabinoside, myricetin 3-O-L-rhamnoside, quercetin 3-galactoside and 3'-Hydroxy-4'-methoxydiclofenac methyl. The interacting residue and bond length of the 1NFK with D-allose are DA-5:-2.1, LYS-272:-2.2, ARG-305:-2.3, ARG-305:-2.7, DA-6:-2.6, DA-6:-2.4, DT-7:-2.3 and DA-6:-2.1(Figure 5a). The interacting residue and bond length of the 1NFK with a-D-Mannopyranoside are DA-5:-2.1, DA-6:-2.0, DA-6:-1.9, DA-6:-2.0, DA-6:-2.4 and DA-6:-1.4 (Figure 5b). The interacting residue and bond length of the 1NFK with myricetin 3- O-arabinoside are DG-2:-1.4, LYS-249:-2.2, LYS-249:-1.7, ARG-305:-2.1, DG-4:-2.5, SER-240:-3.1, LYS-241:-2.5, LYS-272:-1.4, ASN-247:-1.6 and DA-5:-2.6 (Figure 5c). The interacting residue and bond length of the 1NFK with myricetin 3-O-L-rhamnoside are DA-5:-1.8, LYS-241:-2.5, SER-246:-2.8, SER-240:-2.8, ASN-247:-1.9, DG-3:-2.6, LYS-249:-2.4, ASP-271:-2.8 and ARG-305:-1.5 (Figure 5d). The interacting residue and bond length of the 1NFK with Quercetin 3-Galactoside are LYS-249:- 1.2, ASN-247: 2.7, DG-4: 1.1, ARG-305: 2.6, ASN-247:2.2, DG-4: 3.3 and ARG-305: 1.5 (Figure 5e). The interacting residue and bond length of the 1NFK with 3'-Hydroxy-4'-methoxydiclofenac methyl are DA-5:-2.8, LYS-249:-2.6, LYS-249:-2.1, ASP-271:-2.6, DG-4:-2.8, LYS-272:-2.7, LYS-241:-2.5 and LYS-241:-2.8 (Figure 5f).

4. DISCUSSION:

In vitro assays like Total phenolic content, DPPH assay, Catlase activity and Peroxidase activity were observed in the extract of M.longifolia which shown potential activity. Compared to all other extracts, aqueous leaf extract of M.longifolia has shown better activity in which the dilution of 1:2 shown good activity among other dilution. The studies on Onobrychis argyrea extract has also demonstrated its antioxidant activity through DPPH assay which is similar to our studies¹⁹. DPPH is a stable free radical that helps in evaluating the antioxidant properties²⁰. Catalase is one of the antioxidant enzymes which inhibit the process of oxidative stress by forming water and oxygen through degrading hydrogen peroxidase that were formed during the metabolic reactions²¹. Peroxidase is an enzyme that plays an important role in the process of detoxifying the hydrogen peroxidase and biosynthesis. It is found in plants, animals, bacteria and fungi that have the property of degrading rich content of hydrogen peroxidase and it is a rich scavenger of reactive oxygen species²². The beneficial activities of the extract on in vitro assay prove its potential activity against the formation of oxidative stress by scavenging the free radicals that exhibit its antioxidant activity. Its potential activity on catalase activity represents its ability in inhibiting the oxidative stress by converting the toxic metabolites into water and oxygen. Its peroxidase activity shows its ability in detoxifying hydrogen peroxidase. The extract of M.longifolia has possessed potential activity Our research was observed to a show potential effect in analgesic activity which is due to the effect extract in suppressing the synthesis of arachidonic acid²³. Similar to our result on the analgesic test, the research on the analgesic activity of Aconitum carmichaelii has also shown potential activity²⁴. The ulcerogenic activity of the plant extract is directly related to the anti-inflammatory activity of the extract. The plant extract of M.longifolia has shown potential anti-ulcerogenic activity which is also similar in other plant extracts like Castilleja tenuiflora¹⁸, Haplophyllum tuberculatum leaves²⁵, Achillea fragrantissima²⁶, Styrax liquidus²⁷ and Quassia amara²⁸. The potential activity of the extract on antipyretic assay demonstrates the activity of M. longifolia in controlling the elevation of the hypothalamus. Its potential activity on ulcerogenic test shows its activity in controlling augmentation of prostaglandins. Our docking experiment, Nf-kB have shown potential binding affinity with the compound like D-Allose, a-D-Mannopyranoside, methyl, myricetin 3- O-arabinoside, myricetin 3-O-L-rhamnoside, quercetin 3-galactoside, 3'-Hydroxy-4'-methoxydiclofenac. The potential binding affinity of these ligands in the docked complex with receptors shows its benefits in maintaining the body haemostasis by activating or inhibiting the nuclear receptors.

5. CONCLUSION:

We have examined the potential activity of the leaf extract of M.longifolia through in vitro, pharmacological and in silico studies. The in vitro assay has demonstrated the antioxidant activity of the ethanolic, methanolic and aqueous leaf extract of M.longifolia in which aqueous leaf extract at 1:2 ratio was observed to exhibit more antioxidant activity on compared to all other extract and serial dilution. The various pharmacological activities were observed in the aqueous, ethanolic and methanolic extract at 500mg/kg. b.w. p.o. in which aqueous leaf extract has shown better pharmacological properties on compared to other extracts. The in silico docking was performed with the orphan nuclear receptor and the active compound of the leaf of M.longifolia in which the ligands like myricetin 3-O-L-rhamnoside, quercetin 3-galactoside, a-D-Mannopyranoside, D-Allose, myricetin, Quercetin, myricetin 3- O-arabinoside and 3'-Hydroxy-4'-methoxydiclofenac has shown the potential binding interaction. Further, this ligand can be studied through in vivo and gene expression studies to prove are beneficial mechanism against toxicity.

6. ACKNOWLEDGEMENT:

The authors thank VIT for providing ‘VIT SEED GRANT’ for carrying out this research work.

7. CONFLICTS OF INTEREST:

The authors declare that they have no conflicts of interest.

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Received on 09.05.2020 Modified on 16.06.2019

Research J. Pharm. and Tech 2020; 13(3): 1083-1091.

DOI: 10.5958/0974-360X.2020.00199.7