Novel Triterpenoids from Cassia fistula Stem Bark Depreciates STZ-Induced Detrimental Changes in IRS-1/Akt-Mediated Insulin Signaling Mechanisms in Type-1 Diabetic Rats.

Here, we identified the mechanisms of action of antidiabetic activity of novel compounds isolated from Cassia fistula stem bark in STZ-diabetic animals. Novel triterpenoid compounds (C1, C2 and C3) were treated to STZ-administered diabetic animals at a concentration of 20mg/kg body weight orally for 60 days to assess their effects on plasma glucose, plasma insulin/C-peptide, serum lipid markers and the enzymes of carbohydrate metabolism, glucose oxidation and insulin signaling molecules. Oral administration of novel triterpenoid compounds to STZ-diabetic animals significantly decreased (p < 0.05) the plasma glucose concentration on the 7th, 15th, 30th, 45th and 60th daysin a duration-dependent manner (p < 0.05). Plasma insulin (p < 0.0001)/C-peptide (p < 0.0006), tissue glycogen (p < 0.0034), glycogen phosphorylase (p < 0.005), glucose 6-phosphatase (p < 0.0001) and lipid markers were significantly increased (p < 0.0001) in diabetic rats, whereas glucokinase (p < 0.0047), glycogen synthase (p < 0.003), glucose oxidation (p < 0.001), GLUT4 mRNA (p < 0.0463), GLUT4 protein (p < 0.0475) and the insulin-signaling molecules IR mRNA (p < 0.0195), IR protein (p < 0.0001), IRS-1 mRNA (p < 0.0478), p-IRS-1Tyr612 (p < 0.0185), Akt mRNA (p < 0.0394), p-AktSer473 (p < 0.0162), GLUT4 mRNA (p < 0.0463) and GLUT4 (p < 0.0475) were decreased in the gastrocnemius muscle. In silico analysis of C1-C3 with IRK and PPAR-γ protein coincided with in vivo findings. C1-C3 possessed promising antidiabetic activity by regulating insulin signaling mechanisms and carbohydrate metabolic enzymes.

Inhibition of cell survival and proliferation by nimbolide in human androgen-independent prostate cancer (PC-3) cells: involvement of the PI3K/Akt pathway.

Prostate cancer is most common malignancy among men in the world. PI3K-Akt signaling appears to be critical to prostate cancer cell proliferation and survival. Our earlier study reveals that nimbolide (2 µM) prevents cell survival via IGF signaling pathway through PI3K/Akt and induces apoptosis in PC-3 cell line. Akt mediates the phosphorylation and activation of mTOR that plays a critical role in the regulation of protein translation and synthesis, angiogenesis, and cell cycle progression. The present study was aimed to investigate the effect of nimbolide on tPI3K, tAkt, pAkt, tmTOR, GSK3ß, pGSK3ß, PCNA, c-Myc, Cyclin D1, and Survivin protein levels by western blot analysis. Apoptosis was visualized by Ao/EtBr dual staining (20×), and protein expression of PCNA by immunocytochemistry was performed. Molecular docking was performed to understand the possible interaction between nimbolide and Akt, PCNA, and Cyclin D1. Nimbolide altered the PI3K-Akt-mediated cell survival and proliferative molecules. Thus, nimbolide exerted anticancer effects in vitro by representing the PI3K-Akt-mTOR pathway in PC-3 cells. Thereby, it acts as a potent anticancer drug for prostate cancer.

Effects of dihydroxy gymnemic triacetate (DGT) on expression of apoptosis associated proteins in human prostate cancer cell lines (PC3).

Prostate cancer is the most common malignancies among men. The present study is aimed at the investigation of dihydroxy gymnemic triacetate (DGT) from Gymnema sylvestre on mitochondrial apoptotic pathway and cell cycle arrest. Treatment of DGT resulted in a dose-dependent inhibition of growth of PC-3 cells. The cell cycle arrest was observed at the G2/M phase and accumulation of apoptotic cells was observed in DGT-treated prostate cancer cell lines. The occurrence of apoptosis in these cells was observed by DNA fragmentation. These events were associated with increased levels of pro-apoptotic proteins Bax, Bad and reduced levels of the antiapoptotic proteins Bcl-2, Bcl-xL and Mcl-1. DGT also induces the activation of caspase-9 and caspase-3. The above results, clearly, suggest that DGT induces apoptosis by the intrinsic pathways which could be very useful for the treatment of prostate cancer.

Octyl Gallate and Gallic Acid Isolated from Terminalia bellirica Circumvent Breast Cancer Progression by Enhancing the Intrinsic Apoptotic Signaling Pathway and Elevating the Levels of Anti-oxidant Enzymes.

Exploration of new strategies and identification of less expensive novel chemoprevention agents against breast cancer progression have become the need of the hour. Thus, the present study aimed at evaluating the anti-cancer efficacies of octyl gallate (OG) and gallic acid (GA) isolated from Terminalia bellirica (T. bellirica) in breast cancer cell lines and DMBA-induced Sprague-Dawley animal model. The results of western blot analysis show significant (p < 0.05) downregulation of anti-apoptotic protein (Bcl-2 and Bcl-xL) expression and up-regulation of pro-apoptotic protein (Bak and Bax) expression in both MCF-7 and MDA-MB-231 cell lines. Our findings also show that DMBA-induced Sprague-Dawley rats (50-55 days old) orally administered with OG (20 mg/kg body wt.) and GA (20 mg/kg body wt.) for a treatment period of 14 weeks were observed for normalized body weight changes and hematological indices and significant reduction of tumor markers carcinoembryonic antigen (CEA), cancer antigen 15.3 (CA 15.3), and oxidative stress (TBARS) in serum, while the activity of anti-oxidant enzyme (SOD, CAT, and GPx) levels estimated in the mammary tissue was found restored back to normal. Computational molecular interaction study was also performed to substantiate the in vitro obtained results. The tissue histology reveals the therapeutic role of OG and GA. The study conducted brings to limelight of the molecular mechanisms of intrinsic apoptotic signaling pathway through which OG and GA exert their chemopreventive action. Both OG and GA can be explored further as chemotherapeutic natural drugs for their ability to prevent breast cancer progression.

Phytochemical screening and anti-obesity, anti- diabetic and anti-oxidant properties of Scoparia dulcis leaf decoction (crude).

Due to their rising prevalence, diabetes and obesity were both classified as epidemics by the World Health Organization.The natural leaf essence of Scoparia dulcis (S. dulcis), is used as herbal remedy for diabetes and obesity worldwide. However, the objective of the current research was to examine the effects of consuming commercially available S. dulcis porridge on both diabetes and obesity. The S. dulcis plants were collected and the essence was prepared in a traditional way. Phytochemical screening was carried out to identify the secondary metabolites present in the essence. The GC-MS analysis was carried out to identify the bioactive compounds present in the extracts. In order to understand the molecular interaction of identified compounds with selected target proteins from anti-diabetic, anti-oxidant, anti-obesity molecular docking studies was carried out. Results of this docking studies confirmed that identified compounds showed the strong interaction with all the selected target protein. Further experimental analysis is needed to confirm this activity.

DaiCee: A database for anti-cancer compounds with targets and side effect profiles.

Identification of the toxicity of compounds is more crucial before entering clinical trials. Awareness of physiochemical properties, possible targets and side effects has become a major public health issue to reduce risks. Experimental determination of analyzing the physiochemical properties of a drug, their interaction with specific receptors and identifying their side-effects remain challenging is time consuming and costly. We describe a manually compiled database named DaiCee database, which contains 2100 anticancer drugs with information on their physiochemical properties, targets of action and side effects. It includes both synthetic and herbal anti-cancer compounds. It allows the search for SMILES notation, Lipinski's and ADME/T properties, targets and side effect profiles of the drugs. This helps to identify drugs with effective anticancer properties, their toxic nature, drug-likeness for in-vitro and in-vivo experiments. It also used for comparative analysis and screening of effective anticancer drugs using available data for compounds in the database. The database will be updated regularly to provide the users with latest information. The database is available at the URL http://www.hccbif.org/usersearch.php.

Molecular docking analysis of alkaloid compounds with beta-catenin towards the treatment of colon cancer.

It is known that beta-catenin is associated with fibromatosis, sarcoma and mesenchymal tumor. Therefore, it is of interest to design an effective inhibtitor to the target protein beta-catenin. In this study, we report the molecular docking analysis of alkaloid compounds (aristolochicacid, cryptopleurine, demecolcine, fagaronine and thalicarpine) with beta-catenin for further consideration towards the design and development of potential inhintors for the treatmnet of colon cancer.

Molecular docking data of piperine with Bax, Caspase 3, Cox 2 and Caspase 9.

Several apoptotic signalling proteins such as Bax, Caspase 3, Cox 2 and Caspase 9 are known to be associated with colorectal cancer (CRC). It is of interest to study the interaction of these proteins with piperine a known drug candidate. We document the binding energy, hydrogen bond interaction and hydrophobic interaction between the piperine and apoptotic proteins for further consideration.

Molecular docking analysis of Bcl-2 with phyto-compounds.

The Bcl-2 protein is liked in several cancers and drug resistance to therapy is also known in this context. There are many Bcl-2 inhibitors under clinical trials. It is of further interest to design new Bcl2 inhibitors from phyto compounds such as artesunate, bruceantin, maytansin, Salvicine, indicine N-oxide, kamebanin and oxyacanthine. We report the optimal binding features of these compounds with Bcl-2 for further consideration towards in vitro and in vivo validation.

Metabolism-dependent cytotoxicity of citrinin and ochratoxin A alone and in combination as assessed adopting integrated discrete multiple organ co-culture (IdMOC).

Citrinin (CTN) and ochratoxin A (OTA) can be present as co-contaminants in cereals, foods and feed commodities, and can affect human health. Metabolism-dependent toxicity of these two mycotoxins, separately as well as in combination, is not yet understood. To fill this gap we adopted integrated discrete multiple organ co-culture (IdMOC) technique, which obviates animal experiments from the perspectives of species difference as well as animal welfare concerns. IdMOC facilitates co-culture of a metabolically competent cell (HepG2) and a metabolically incompetent cell (3T3) that are physically separated but provides for extracellular product(s) from one cell to interact with the other. After ascertaining that HepG2 is metabolically competent and 3T3 is not, adopting luciferin-IPA metabolism assay, CTN and OTA were tested separately and in combination in the co-culture set-up, when both proved to be metabolism-dependent cytotoxic agents. Hepatocytes metabolize CTN into a diffusible product that is cytotoxic to 3T3 cells but the cytotoxicity of OTA appears to be limited to the hepatocytes, i.e., local acting. As a combination at a concentration of 20% of IC50 of each, CTN forms a reactive metabolite that diffuses out of HepG2 to cause cytotoxicity to 3T3 cells synergistically with OTA parent molecule. The CYP isoenzymes involved in the metabolism OTA and CTN were identified adopting in silico methods which indicated that OTA and CTN can bind CYP proteins at specific sites.

ß-cell regenerative efficacy of a polysaccharide isolated from methanolic extract of Tinospora cordifolia stem on streptozotocin -induced diabetic Wistar rats.

The use of herbal supplements either as extracts or plant-derived individual molecules has significantly increased in the process of drug discovery and development for their potential efficacy or reduced risk in treating human disorders. Tinospora cordifolia (T. cordifolia) is a widely used herbal source to treat various human ailments, including diabetes mellitus. The present study was aimed on evaluating the antidiabetic property of a novel polysaccharide isolated from the methanolic extract of T. cordifolia stem. Bioassay guided fractionation was followed to isolate a compound from the methanol extract. The compound was administered orally at a dose of 20 mg/kg.b.wt for 60 days to control and STZ-induced diabetic male Wistar rats. It was found that plasma glucose was significantly (p < 0.05) reduced compared to normal. Oral administration of the compound significantly decreased HBA1c, triglycerides and total cholesterol and at the same time markedly increased hemoglobin, tissue glycogen and HDL cholesterol. Also the compounds restored the altered carbohydrate metabolizing enzymes, insulin, C-peptide, (14)C-glucose oxidation levels to near normal. In addition, the histological studies revealed that there was regeneration of ß-cells in the pancreatic sections. The expression of Glut-4 mRNA and protein in the gasrtocnemius muscle were significantly enhanced after the compound treatment. These results confirm that the novel polysaccharide possesses hypoglycemic, glucose oxidizing, hypolipidemic and ß-cell regenerative properties and hence it could be developed into potential oral hypoglycemic drug with lesser side effects.

Virtual screening of potential inhibitor against FtsZ protein from Staphylococcus aureus.

The gram-positive bacterium Staphylococcus aureus, responsible for a wide variety of diseases in human involve all organ systems ranging from localized skin infections to life-threatening systemic infections. FtsZ, the key protein of bacterial cell division was selected as a potent anti bacterial target. In order to identify the new compounds structure based screening process was carried out. An enrichment study was performed to select a suitable scoring function and to retrieve potential candidates against FtsZ from a large chemical database. The docking score and docking energy values were compared and their atomic interaction was also evaluated. Furthermore molecular dynamics simulation were also been performed to check the stability and the amino acids interacted towards the FtsZ. Finally we selected C ID 16284, 25916, 15894, 13403 as better lead compounds. From these results, we conclude that our insilico results will provide a framework for the detailed in vitro and in vivo studies about the FtsZ protein activity in drug development process.

Virtual screening of potential inhibitor against FtsZ protein from Staphylococcus aureus.

The gram-positive bacterium Staphylococcus aureus, responsible for a wide variety of diseases in human involve all organ systems ranging from localized skin infections to life-threatening systemic infections. FtsZ, the key protein of bacterial cell division was selected as a potent anti bacterial target. In order to identify the new compounds structure based screening process was carried out. An enrichment study was performed to select a suitable scoring function and to retrieve potential candidates against FtsZ from a large chemical database. The docking score and docking energy values were compared and their atomic interaction was also evaluated. Furthermore molecular dynamics simulation were also been performed to check the stability and the amino acids interacted towards the FtsZ. Finally we selected C ID 16284, 25916, 15894, 13403 as better lead compounds. From these results, we conclude that our insilico results will provide a framework for the detailed in vitro and in vivo studies about the FtsZ protein activity in drug development process.

A database for the predicted pharmacophoric features of medicinal compounds.

UNLABELLED: Pharmacophore feature is defined by a set of chemical structure patterns having the active site of drug like molecule. Pharmacophore can be used to assist in building hypothesis about desirable chemical properties in drug molecule and hence it can be used to refine and modify drug candidates. We predicted the pharmacophoric features of 150 medicinal compounds from plants for anti-cancer, anti-carcinogenic, anti-diabetic, anti-microbial, and anti-oxidant. Estimation of pharmacophoric feature is necessary to ensure the optimal supramolecular interaction with a biological target and to trigger or block its biological response. We subsequently make this data available to open access using a database at the URL: http://www.hccbif.info/index.htm AVAILABILITY: The database is available for free at http://www.hccbif.info/index.htm.