In Silico Study on the Interactions, Molecular Docking, Dynamics and Simulation of Potential Compounds from Withania somnifera (L.) Dunal Root against Cancer by Targeting KAT6A.

Cancer is characterized by the abnormal development of cells that divide in an uncontrolled manner and further take over the body and destroy the normal cells of the body. Although several therapies are practiced, the demand and need for new therapeutic agents are ever-increasing because of issues with the safety, efficacy and efficiency of old drugs. Several plant-based therapeutics are being used for treatment, either as conjugates with existing drugs or as standalone formulations. Withania somnifera (L.) Dunal is a highly studied medicinal plant which is known to possess immunomodulatory activity as well as anticancer properties. The pivotal role of KAT6A in major cellular pathways and its oncogenic nature make it an important target in cancer treatment. Based on the literature and curated datasets, twenty-six compounds from the root of W. somnifera and a standard inhibitor were docked with the target KAT6A using Autodock vina. The compounds and the inhibitor complexes were subjected to molecular dynamics simulation (50 ns) using Desmond to understand the stability and interactions. The top compounds (based on the docking score of less than -8.5 kcal/mol) were evaluated in comparison to the inhibitor. Based on interactions at ARG655, LEU686, GLN760, ARG660, LEU689 and LYS763 amino acids with the inhibitor WM-8014, the compounds from W. somnifera were evaluated. Withanolide D, Withasomniferol C, Withanolide E, 27-Hydroxywithanone, Withanolide G, Withasomniferol B and Sitoindoside IX showed high stability with the residues of interest. The cell viability of human breast cancer MCF-7 cells was evaluated by treating them with W. Somnifera root extract using an MTT assay, which showed inhibitory activity with an IC50 value of 45 µg/mL. The data from the study support the traditional practice of W. somnifera as an anticancer herb.

Evaluation of Vrikshayurveda treatments on physiological attributes and production of diterpenoids in Andrographis paniculata (Burm.f.) Nees.

Background: An ancient text on plant life Vrikshayurveda mentions the use of horticulture treatments, Kunapa Jala (KJ) and Panchagavya (PG) (cow milk, cow ghee, cow curd, cow dung, and cow urine) to enhance the efficiency of plants. Aim: An experiment was conducted to evaluate the effect of KJ and PG application on total leaf area, leaf area index (LAI) leaf area duration (LAD), crop growth rate (CGR), relative growth rate (RGR), and net assimilation rate (NAR); production of andrographolide (A1), neoandrographolide (A2), and 14-deoxy-11,12-didehydroandrographolide (A3) of the Andrographis paniculata (Burm.f.) Nees. Materials and methods: The experiment was conducted in randomized block design with six treatments, namely control, KJ, PG, farmyard manure, inorganic fertilizer, and humic acid. Simultaneous detection of contents was carried out using reversed-phase-ultra-flow liquid chromatography (RP-UFLC). The parameters were studied at 30, 60, 90, and 120 days after sowing (DAS). Results: KJ produced higher leaf area and LAI at 90 DAS, LAD between 90 and 120 DAS. PG produced higher CGR, RGR, and NAR between 60 and 90 DAS. RP-UFLC analysis revealed the maximum amount of ingredients at 120 DAS. At this stage, PG treatment recorded the maximum amount of A1, A2, and A3. Conclusion: Kunapa Jala and Panchagavya treatments were better or at par on physiological parameters and production of diterpenoids of A. paniculata.

Traditional Horticulture Practices Increase the Production of Selected Withanolides in Withania Somnifera (L.) Dunal-A RP-UFLC Analysis.

The study evaluates the effect of two traditional horticulture treatments mentioned in Vrikshayurveda, a text from ancient India on the science of plant life, namely Kunapa jala (KJ) and Pancha gavya (PG) on the production of Withaferin A (WFA), withanolide A (WIA) and Withanolide B (WIB) in Withania somnifera (L) Dunal. Leaves and roots of W. somnifera were collected from different treated groups viz. control, KJ, PG, farmyard manure (FYM) and inorganic fertilizer (NPK). Reverse phase ultra-flow liquid chromatography (RP-UFLC) method was developed, validated for simultaneous detection of WFA, WIA and WIB. Statistical analysis of data was performed by ANOVA and tested for significance by the Dunnett multiple comparison test and data were expressed as mean ± standard deviation (SD). Results revealed, leaves possessed highest WFA content and roots possessed highest content of WIA and WIB. PG treated leaves were observed highest WFA (18.29 mg/g) and roots were observed highest WIA (19.63 mg/g) and WIB (1.36 mg/g). Conclusively, RP-UFLC method for simultaneous detection of withanolides has been developed and validated to evaluate the effect of traditional horticulture treatments. It is concluded that the enhanced production of withanolides can be achieved by the application of PG when compared to NPK application.

Nutrient analysis of Kunapa jala and Pancha gavya and their evaluation on germination of Ashwagandha and Kalamegha seeds: A comparative study.

BACKGROUND: Vrikshayurveda, an ancient science of plant life described by Surapala, deals with healthy growth and productivity of plants, which has clearly outlined a systematized agricultural practice that insisted the use of Kunapa jala (KJ) and Pancha gavya (PG). OBJECTIVE: An experiment was conducted to validate KJ and PG by nutrient analysis and their effect on the germination parameters of Ashwagandha and Kalamegha seeds in comparison with other treatment groups. MATERIALS AND METHODS: KJ and PG were prepared according to the classical references. The nutrient contents and germination parameters of KJ and PG were compared with other groups namely control (Contr), farmyard manure (FYM), humic acid (HA) and NPK. RESULTS: The pH and EC were 5.793, 2.653 dS/m and 5.584, 2.216 dS/m for KJ and PG respectively. KJ possess highest nutrient contents followed by PG. The germination parameters revealed the better activity of NPK followed by KJ, PG, HA, FYM and Contr. CONCLUSION: KJ and PG were found to be good in nutrient contents and were found to be effective on studied germination parameters of Ashwagandha and Kalamegha seeds.

Evaluation of Kunapa jala and Pancha gavya on plant quality, physiology, biochemistry, yield and growth attributes - A case study of Withania somnifera Dun.

BACKGROUND: 'Vrikshayurveda', an ancient science of plant life described by Surapala, has clearly outlined a systematized agricultural practice that insisted on the use of Kunapa jala (KJ) and Pancha gavya (PG) to enhance the yield and quality of plants. OBJECTIVE: An experiment was conducted to evaluate the effect of KJ and PG on growth, physiological, biochemical, quality attributes and yield of Withania somnifera Dun. MATERIALS AND METHODS: The effect of KJ and PG was evaluated in comparison with control, organic (farmyard manure and humic acid) and inorganic (NPK) fertilizer at 60, 90, 120 and 150 days after sowing (DAS). The study was conducted in randomized complete block design method. RESULTS: KJ group were higher in, total leaf area (1707.89 cm2) at 120 DAS, leaf area index (3.795) at 120 DAS, crop growth rate (0.256 g m-2 day-1) at 60-90 DAS, leaf area duration (101.909) at 120-150 DAS, relative growth rate (0.0170 g g-1 day-1) at 60-90 DAS, net assimilation rate (0.0537 g m-2 day-1) at 60-90 DAS. Leaf area ratio was higher in PG group (37.937 m2 g-1) at 60 DAS. The higher levels of chlorophyll a, b and carotenoids were in KJ group (1.877, 0.745 and 1318.14 mg g-1 respectively) at 90 DAS. Yield and quality attributes at harvest (150 DAS) indicated, higher dry root yield (5.93 quintal hectare-1), root length (15.66 cm) were higher in KJ group and root diameter was higher in PG group (1.36 cm). CONCLUSION: Vrikshayurveda practices viz. Kunapa jala and Pancha gavya were effective on studied parameters of W. somnifera. Economical and eco-friendly Vrikhayurveda practices can be initiated for sustainable agriculture.

Pharmacognostic Screening of Piper trichostachyon Fruits and its Comparative Analysis with Piper nigrum Using Chromatographic Techniques.

BACKGROUND: Piper trichostachyon is a wild, endemic Piper species from Western Ghats of India. The folklore healers of Belagavi region use this plant, similar to Piper nigrum. AIMS: The present study investigates the comparison between P. nigrum and P. trichostachyon using pharmacognostic parameters. MATERIALS AND METHODS: Pharmacognostic evaluation was carried out in terms of morphological, microscopic characters, and phytochemical analysis using standard methods. Comparative physicochemical analysis between P. trichostachyon and P. nigrum was also carried out through estimation of micro-macro nutrients, high-performance thin layer chromatography (HPTLC) investigation and using piperine as a marker compound for reversed phase-ultra flow liquid chromatographic (RP-UFLC) technique. RESULTS: P. trichostachyon grows in the forests, and the fruits are morphologically similar to P. nigrum fruits, so the name in Kannada "Kaadu Kalu menasu" (wild/forest black pepper). The microscopy revealed the presence of stone cells, starch grains, oil cells and globules, beaker cells, and yellowish brown pigment layer, parenchymatous cells. The presence of alkaloids, oil, and tannins were observed in P. trichostachyon fruits. The HPTLC studies visibly indicated differences among two species with 12 peaks and varied banding pattern. RP-UFLC results showed less amount of piperine in P. trichostachyon (0.05 ± 0.002 mg/g) than in P. nigrum (16.14 ± 0.807 mg/g). CONCLUSION: The study reports on pharmacognostic parameters of P. trichostachyon for the 1(st) time and will be useful for the identification and authentication. The comparative HPTLC and RP-UFLC studies resolve the differentiation impasse among two species. However, further biological efficacy studies are required to establish its use in traditional medicine. SUMMARY: Piper trichostachyon grows in the forests, and the fruits are morphologically similar to Piper nigrum fruitsThe microscopy of P. trichostachyon revealed the presence of stone cells, starch grains, oil cells and globules, beaker cells and yellowish brown pigment layer, parenchymatous cellsThe high-performance thin layer chromatography studies visibly indicated differences among two species with varied banding patternReversed phase-ultra flow liquid chromatographic results showed less amount of piperine in P. trichostachyon than in P. nigrum. Abbreviation used: HPTLC: High Performance Thin Layer Chromatography, RP-UFLC: Reversed phase-ultra flow liquid chromatographic analysis, DST: Length of line, Maj: Length of large half axis for ellipse RDS - radius for circle, Rf: Retention Factor, TS: Transverse Section, TLC: Thin Layer Chromatography.

Simultaneous determination of vasicine and vasicinone by High-performance liquid chromatography in roots of eight Sida species.

INTRODUCTION: Sida L. is a medicinally important genus widely used in conventional systems of medicine in India. AIM: The present study aims toward simultaneous determination of two bioactive compounds vasicine and vasicinone in root extracts of eight Sida spp. from Western Ghats, India. MATERIALS AND METHODS: Determination of vasicine and vasicinone was undertaken in methanolic root extracts (10% w/v) of Sida acuta, Sida cordata, Sida cordifolia, Sida rhombifolia, Sida spinosa, Sida indica, Sida retusa and Sida mysorensis by high performance liquid chromatography (HPLC) method. The standards were prepared with the concentration of mg/mL. Data were expressed as mean values of three reading and relative standard deviations. The separation was achieved on a Waters, Nova-Pack, C18 (250 mm × 4.6 mm, 5 µ) column, with acetonitrile - 0.1 M phosphate buffer-glacial acetic acid (15: 85: 1, v/v/v) as solvent system at a flow-rate of 1.0 mL/min. The effluent was monitored using ultraviolet detection at a wavelength of 300 nm. RESULTS: Both calibration curves of standard showed good linear regression (R2 > 0.994). The limit of detection and the limit of quantification for vasicine was 0.110 and 0.333 µg/mL and for vasicinone was 0.059 and 0.179 µg/mL respectively. The vasicine content was highest in S. cordifolia (9.891 ± 0.495 µg/100 mg) and vasicinone content was rich in S. cordata (33.013 ± 1.651 µg/100 mg.) The content of vasicinone was higher than vasicine. CONCLUSION: HPLC method provides simple, accurate, and reproducible quantitative analysis for simultaneous determination of vasicine and vasicinone. Among the selected Sida species, S. cordifolia and S. cordata were found to be rich in the vasicine and vasicinone contents, respectively.

Preliminary pharmacognostic screening of Achyranthes coynei stem.

Achyranthes coynei is a rare, endemic perennial shrub reported from Karnataka and Maharashtra states of India. The plant is used to treat various disorders by folk healers and was proven to have antimicrobial and antioxidant properties. The present study was undertaken to evaluate microscopic and macroscopic characters of A. coynei stem, along with its physicochemical parameters. ProgRes(®) CapturePro and Microsoft Excel were used for statistical analysis. Perennial, shrubby nature and woody stem were the distinguishing morphological characters observed. Transverse section (TS) illustrated quadrangular outline of the stem and showed the presence of two types of trichomes on the thick-walled epidermis. TS also showed number of rosette calcium oxalates crystals; prismatic and microsphenoid crystals; conjoint, collateral open secondary vascular bundles; and two amphixylic medullary bundles in the pith. Ash and extractive values, micro and macro elements and nutritive factors were estimated in the present study. The presence of alkaloids, saponins and triterpenoids were observed in preliminary phytochemical screening. High-performance thin layer chromatographic analysis yielded different bands and also indicated the presence of oleanolic acid. The studied parameters for A. coynei stem will be useful for identification and authentication of the plant material.

Total polyphenolic contents and in vitro antioxidant properties of eight Sida species from Western Ghats, India.

BACKGROUND: Sida L., is a medicinally important genus, the species of which are widely used in traditional systems of medicine in India. Pharmacologically, roots are known for anti-tumor, anti-HIV, hepatoprotective, and many other properties. Phenolic antioxidants help in reducing oxidative stress occurring during treatment of such diseases. OBJECTIVE: The study aimed to evaluate and compare polyphenol contents and antioxidant properties of eight selected species of Sida from Western Ghats, India. MATERIALS AND METHODS: Methanolic root extracts (10% w/v) of Sida species, viz., S. acuta, S. cordata, S. cordifolia, S. indica, S. mysorensis, S. retusa, S. rhombifolia, and S. spinosa were analyzed. RESULTS: Sida cordifolia possessed highest total phenolic content (TPC: 1.92 ± 0.10 mg Caffeic Acid Equivalent/g and 2.13 ± 0.11 mg Tannic Acid Equivalant/g), total flavonoid content (TF: 2.60 ± 0.13 mg Quercetin Equivalent/g) and also possessed highest antioxidant activities in 2,2-diphenylpicrylhydrazyl (DPPH) radical scavenging (51.31 ± 2.57% Radical Scavenging Activity, (RSA); Trolox Equivalent Antioxidant Capacity: 566.25 ± 28.31µM; Ascorbic acid Equivalent Antioxidant Capacity: 477.80 ± 23.89 µM) and Ferric Reducing Antioxidant Power assays (TEAC: 590.67 ± 29.53 µM; AEAC: 600.67 ± 30.03 µM). Unlike DPPH and Ferric Reducing Antioxidant Power (FRAP) activity, 2, 2'-Azinobis (3-ethyl Benzo Thiazoline-6-Sulfonic acid) ABTS(+) antioxidant activity was highest in S. indica (TEAC: 878.44 ± 43.92 µM; AEAC 968.44 ± 48.42 µM). It was significant to note that values of AEAC (µM) for all the antioxidant activities analyzed were higher than that of TEAC. CONCLUSION: The high contents of phenolic compounds in the root extracts of selected Sida species have direct correlation with their antioxidant properties. Conclusively, roots of S. cordifolia can be considered as the potential source of polyphenols and antioxidants.

Accumulation and trends in distribution of three triterpenoids in various parts of Achyranthes coynei determined using RP-UFLC analysis.

BACKGROUND: Achyranthes coynei Sant. (Family: Amaranthaceae) is a rare endemic medicinal plant used by local traditional practitioners to treat various diseases. The plant has been reported for promising antibacterial and antioxidant activities. However, the plant is not explored for its phytocompounds, especially triterpenoids. OBJECTIVE: TO STUDY THE ACCUMULATION AND TRENDS IN DISTRIBUTION OF TRITERPENOIDS: betulinic, oleanolic and ursolic acids (BA, OA and UA) in leaf, stem, root and inflorescence of A. coynei. MATERIALS AND METHODS: Extraction was achieved using refluxing and reversed phase-ultra flow liquid chromatographic (RP-UFLC) technique was employed for determination. Separation of triterpenoids was achieved on a Hibar 250-4.6 mm, 5 µ, Lichrospher 100, C18e column using methanol and water (90:10) as mobile phase (pH adjusted to 5.0 using GAA) in an isocratic mode. RESULTS: Oleanolic acid was higher in leaf (0.172 ± 0.009%) followed by stem (0.035 ± 0.002%) and root (0.028 ± 0.001%). Ursolic acid was accounted to be highest in the inflorescence (0.099 ± 0.005%). The contents of BA and UA were lower than OA in leaf and stem while it was remained undetected in roots of A. coynei. CONCLUSION: THE TRITERPENOIDS: BA, OA and UA were detected, quantified and reported for the first time from A. coynei. In the present study leaves were found to be the major source of BA and OA, whereas inflorescence was for UA.