Molecular simulations probing Kushecarpin A as a new lipoxygenase inhibitor

Zizyphus oxyphylla Edgew is used in Pakistan as a folk medicinal remedy for inflammatory conditions, pains especially rheumatic pain, fevers, allergy and diabetes. The aim of the current study was to scientifically validate the folk use of Z. oxyphylla Edgew by using the isolated compound in vitro and in vivo levels. Kushecarpin A was isolated from ethyl acetate fraction of the plant crude extract. Molecular docking simulations predicted Kushecarpin A as a potential new lipoxygenase (LOX) inhibitor. Kushecarpin A showed significant lipoxygenase inhibition (IC50: 7.2 ± 0.02 μM) thus validated computational prediction. It also exhibited significant and highly significant inhibition (p < 0.05 and p < 0.01) of carrageenan-induced hind paw oedema at the doses of 5, 10 and 20 mg/kg. Kushecarpin A seems to be a potentially new anti-inflammatory compound responsible for anti-inflammatory activities of Z. oxyphylla Edgew. In vitro and in vivo anti-inflammatory inflammatory activities were found in good agreement with the folk medicinal use of Z. oxyphylla Edgew in inflammatory disorders.     

Molecular simulations of Taxawallin I inside classical taxol binding site of β-tubulin

A new taxoid Taxawallin I (1) along with two known taxoids (2-3) were isolated from methanolic bark extract of Taxus wallichiana Zucc. Structural characterization was confirmed by mass and NMR spectral techniques. Taxawallin I exhibited significant in-vitro anticancer activity against HepG2, A498, NCI-H226 and MDR 2780AD cancer lines. Tubulin binding assay was performed to assess its tubulin binding activity. Molecular docking analysis was performed to study the potential binding mode inside the taxol binding site of β-tubulin.     

Pistagremic acid,a glucosidase inhibitor from Pistacia integerrima

Pistacia integerrima Stewart in traditionally used as folk remedy for various pathological conditions including diabetes. In order to identify the bioactive compound responsible for its folk use in diabetes, a phytochemical and biological study was conducted. Pistagremic acid (PA) was isolated from the dried galls extract of P. integerrima. Strong α-glucosidase inhibitory potential of PA was predicted using its molecular docking simulations against yeast α-glucosidase as a therapeutic target. Significant experimental α-glucosidase inhibitory activity of PA confirmed the computational predictions. PA showed potent enzyme inhibitory activity both against yeast (IC₅₀: 89.12 ± 0.12 μM) and rat intestinal (IC₅₀: 62.47 ± 0.09 μM) α-glucosidases. Interestingly, acarbose was found to be more than 12 times more potent an inhibitor against mammalian (rat intestinal) enzyme (having IC₅₀ value 62.47 ± 0.09 μM), as compared to the microbial (yeast) enzyme (with IC₅₀ value 780.21 μM). Molecular binding mode was explored via molecular docking simulations, which revealed hydrogen bonding interactions between PA and important amino acid residues (Asp60, Arg69 and Asp 70 (3.11 Å)), surrounding the catalytic site of the α-glucosidase. These interactions could be mainly responsible for their role in potent inhibitory activity of PA. PA has a strong potential to be further investigated as a new lead compound for better management of diabetes.     

Anti-inflammatory activities of Taxusabietane A isolated from Taxus wallichiana Zucc

Current study was conducted to identify constituents of Taxus wallichiana Zucc. that might be responsible for its folk use in anti-inflammatory conditions. Taxusabietane A was isolated from the bark extract of Taxus wallichiana Zucc. Taxusabietane A was analyzed for in-vitro and in-vivo anti-inflammatory activities using Lipoxygenase (LOX) inhibition assay and carrageenan-induced paw edema model. Taxusabietane A revealed considerable LOX inhibitory activity with the IC₅₀ value being 57 ± 0.31. Standard compound Baicalein showed the IC₅₀ value being 22.1 ± 0.03 μM. Taxusabietane A also showed significant (5 and 10 mg/kg) anti-inflammatory activity induced by carrageenan. However, this study highlighted the potential of Taxusabietane A to be further explored as a new lead compound for management of conditions associated with inflammation.     

Dry Matter Partitioning and Harvest Index Differ in Rice Genotypes with Variable Rates of Phosphorus and Zinc Nutrition

Phosphorus(P) and zinc(Zn) deficiencies are the major problems that decrease crop productivity under rice-wheat cropping system. Field experiments were conducted to investigate impacts of P(0, 40, 80 and 120 kg/hm~2) and Zn levels(0, 5, 10 and 15 kg/hm~2) on dry matter(DM) accumulation and partitioning, and harvest index of three rice genotypes ‘fine(Bamati-385) vs. coarse(F-Malakand and Pukhraj)' at various growth stages(tillering, heading and physiological maturity). The experiments were conducted at farmers' field at Batkhela in Northwestern Pakistan for two years in summer 2011 and 2012. The two year pooled data reveled that there were no differences in percent of DM partitioning into leaves and culms with application of different P and Zn levels, and genotypes at tillering. The highest P level(120 kg/hm~2) partitioned more DM into panicles than leaves and culms at heading and physiological maturity stages. The highest Zn level(15 kg/hm~2) accumulated more DM and partitioned more DM into panicles than leaves and culms at heading and physiological maturity stages. The hybrid rice(Pukhraj) produced and partitioned more DM into panicles than F-Malakand and Bamati-385 at heading and physiological maturity stages. Higher DM accumulation and greater amounts of partitioning into panicles at heading and physiological maturity stages was noticed with increase in P and Zn levels, and the increase was significantly higher in the coarse rice genotypes than fine. We concluded that the growing hybrid rice with application of 120 kg/hm~2 P + 15 kg/hm~2 Zn not only increases total DM accumulation and partitioned greater amounts into the reproductive plant parts(panicles) but also results in higher harvest index.     

Structural insights to investigate Conypododiol as a dual cholinesterase inhibitor from Asparagus adscendens

The main aim of the current study was to explore molecular insights for potentially new dual cholinesterase inhibitor(s) from Asparagus adscendens via molecular docking. This medicinal plant is traditionally used as a nerve tonic and remedy for memory impairments. Conypododiol was isolated from the chloroform fraction of methanolic extract of A. adscendens, based on bioactivity guided isolation. Conypododiol exhibited significant inhibition of both acetylcholinesterase and butyrylcholinesterase, having the IC₅₀ values 2.17 ± 0.1 μM and 11.21 ± 0.1 μM, respectively. IC₅₀ values of the standard compound Galanthamine for both the enzymes were 0.537 ± 0.018 μM and 8.6 ± 0.27 μM, respectively. Based on MTT cytotoxicity assay, Conypododiol was found safe against LCMK-2 monkey kidney epithelial cells and mice hepatocytes. Molecular docking studies revealed the hydrogen bonding interactions of Conypododiol with His440 and Ser200 at esteratic site (ES), and also with Tyr334 at peripheral anionic site (PAS) of the aromatic gorge of acetylcholinesterase. Simultaneous contacts of Conypododiol with PAS and ES shows its significance as a bivalent ligand. This preliminary study highlighted the potential of Conypododiol to be further developed and modified as new lead compound identified by its folk use.