The roots of Dechaschistia crotonifolia Wight & Arn. belonging to the family to Ebaenaceae were collected from surroundings of Tirumala, Andhra Pradesh, India in the month of June and it was authenticated by Dr. K. Madhava Chetty, Head of Department, Department of Botany, SV University, Tirupati. Voucher Specimen (PHCOG/VVIPS/056) were preserved. The roots of Decaschistia crotonifolia were shade dried, powdered and stored in well closed container.
Preparation of Extracts
About 300gm of dried root powdered drug of Dechaschistia crotonifolia Wight & Arn. was extracted by successive solvent extraction using chloroform, ethylacetate and ethanol by Soxhlet extraction for 72 hours. The extract was made concentrated by rotary evaporator and placed in desiccator for further use.
Evaluation of Anthelmintic Property
Anthelmintic property of chloroform, ethylacetate and ethanol root extracts of Dechaschistia crotonifolia Wight & Arn. was examined by using an Indian earthworm Pheretima posthuma [5,6]. Choosing of Pheretima posthuma is made as it resembles identical towards anatomy and physiology of round worm parasite which occurs in alimentary tract of Homosapiens.
Adult earth worms measures an average size 4-7cm in length and 0.3 – 0.7 cm of width was collected from medicinal garden of V. V. Institute of Pharmaceutical Sciences and proper washings are carried out to remove extraneous matter. The extract at concentration of 30mg/ml, 60mg/ml & 80mg/ml was used to examine the time of paralysis (Pt) and Death (Dt). The selected earthworms are categorized into 11 groups of 6 each viz., control group treated with 2% Tween 80 in distilled water, 9 Test groups treated with concentrations of 30mg/ml, 60mg/ml & 80mg/ml of each Chloroform, Ethylacetate and Ethanol extract of Dechaschistia crotonifolia Wight & Arn. and standard group treated with 10mg/ml concentration of Albendazole. Earthworms are treated with volume of 10ml of each concentration of standard, control and test solutions respectively. The time taken for Paralysis (Pt) and Death (Dt) was noted.
Pharmacokinetic Evaluation of phytoconstituents is necessary as it effects binding of compounds in specific active target site [7,8]. Prior docking studies of Phytochemicals, it is very much needed to qualify drug-likeness test, i.e., they have to obey Lipinski rule. The canonical smiles of phytocompounds Parvifloral A (PubChem CID: 90470346), Syriacusin A (PubChem CID: 9991528), Syriacusin B (PubChem CID: 10015552), Syriacusin C (PubChem CID: 10105245), Scopoletin (PubChem CID: 5280460), Stigmasterol (PubChem CID: 5280794) and Standard drug Albendazole (PubChem CID: 2082)was obtained from Pubchem (pubchem.ncbi.nlm.nih.gov) predicted their drug likeness test using SwissADME (SwissADME) and their physico chemical parameters.
For molecular docking study [10,11,12] Autodock vina is used for prediction of potent phytocompounds of Dechaschistia viz., Parvifloral, Syriacusin A, Syriacusin B, Syriacusin C, Scopoletin and Stigmasterol against active site of β-tubulin. The chemical structures of phytoconstituents Parvifloral, Syriacusin A, Syriacusin B, Syriacusin C, Scopoletin and Stigmasterol were obtained from Pubchem Project Database shown in Fig. 3. They were structurally plotted in Discovery Studio Biovia. The 3D structure of protein β-tubulin (PDB ID: 1oj0) is collected from Protein Data Bank (www.rcsb.org/pdb) shown in Fig. 2. The x, y & z attributes along with radius is noted. Further the structure is prepared by removing water, adds up polar hydrogen bond and made torsion free.
The values were represented as mean ± S.D; via one-way ANOVA. The analysis was carried out by using Graph pad Prism (Version 3, U.S.A.) software program. P < 0.05 was taken into statistically significant.